Single-shot quantitative phase microscopy with color-multiplexed differential phase contrast (cDPC)

PubMed Central

2017-01-01

We present a new technique for quantitative phase and amplitude microscopy from a single color image with coded illumination. Our system consists of a commercial brightfield microscope with one hardware modification—an inexpensive 3D printed condenser insert. The method, color-multiplexed Differential Phase Contrast (cDPC), is a single-shot variant of Differential Phase Contrast (DPC), which recovers the phase of a sample from images with asymmetric illumination. We employ partially coherent illumination to achieve resolution corresponding to 2× the objective NA. Quantitative phase can then be used to synthesize DIC and phase contrast images or extract shape and density. We demonstrate amplitude and phase recovery at camera-limited frame rates (50 fps) for various in vitro cell samples and c. elegans in a micro-fluidic channel. PMID:28152023

Quantitative phase imaging of arthropods

PubMed Central

Sridharan, Shamira; Katz, Aron; Soto-Adames, Felipe; Popescu, Gabriel

2015-01-01

Abstract. Classification of arthropods is performed by characterization of fine features such as setae and cuticles. An unstained whole arthropod specimen mounted on a slide can be preserved for many decades, but is difficult to study since current methods require sample manipulation or tedious image processing. Spatial light interference microscopy (SLIM) is a quantitative phase imaging (QPI) technique that is an add-on module to a commercial phase contrast microscope. We use SLIM to image a whole organism springtail Ceratophysella denticulata mounted on a slide. This is the first time, to our knowledge, that an entire organism has been imaged using QPI. We also demonstrate the ability of SLIM to image fine structures in addition to providing quantitative data that cannot be obtained by traditional bright field microscopy. PMID:26334858

Isotropic differential phase contrast microscopy for quantitative phase bio-imaging.

PubMed

Chen, Hsi-Hsun; Lin, Yu-Zi; Luo, Yuan

2018-05-16

Quantitative phase imaging (QPI) has been investigated to retrieve optical phase information of an object and applied to biological microscopy and related medical studies. In recent examples, differential phase contrast (DPC) microscopy can recover phase image of thin sample under multi-axis intensity measurements in wide-field scheme. Unlike conventional DPC, based on theoretical approach under partially coherent condition, we propose a new method to achieve isotropic differential phase contrast (iDPC) with high accuracy and stability for phase recovery in simple and high-speed fashion. The iDPC is simply implemented with a partially coherent microscopy and a programmable thin-film transistor (TFT) shield to digitally modulate structured illumination patterns for QPI. In this article, simulation results show consistency of our theoretical approach for iDPC under partial coherence. In addition, we further demonstrate experiments of quantitative phase images of a standard micro-lens array, as well as label-free live human cell samples. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Quantitative phase-digital holographic microscopy: a new imaging modality to identify original cellular biomarkers of diseases

NASA Astrophysics Data System (ADS)

Marquet, P.; Rothenfusser, K.; Rappaz, B.; Depeursinge, C.; Jourdain, P.; Magistretti, P. J.

2016-03-01

Quantitative phase microscopy (QPM) has recently emerged as a powerful label-free technique in the field of living cell imaging allowing to non-invasively measure with a nanometric axial sensitivity cell structure and dynamics. Since the phase retardation of a light wave when transmitted through the observed cells, namely the quantitative phase signal (QPS), is sensitive to both cellular thickness and intracellular refractive index related to the cellular content, its accurate analysis allows to derive various cell parameters and monitor specific cell processes, which are very likely to identify new cell biomarkers. Specifically, quantitative phase-digital holographic microscopy (QP-DHM), thanks to its numerical flexibility facilitating parallelization and automation processes, represents an appealing imaging modality to both identify original cellular biomarkers of diseases as well to explore the underlying pathophysiological processes.

Quantitative label-free sperm imaging by means of transport of intensity

NASA Astrophysics Data System (ADS)

Poola, Praveen Kumar; Pandiyan, Vimal Prabhu; Jayaraman, Varshini; John, Renu

2016-03-01

Most living cells are optically transparent which makes it difficult to visualize them under bright field microscopy. Use of contrast agents or markers and staining procedures are often followed to observe these cells. However, most of these staining agents are toxic and not applicable for live cell imaging. In the last decade, quantitative phase imaging has become an indispensable tool for morphological characterization of the phase objects without any markers. In this paper, we report noninterferometric quantitative phase imaging of live sperm cells by solving transport of intensity equations with recorded intensity measurements along optical axis on a commercial bright field microscope.

Quantitative x-ray phase-contrast imaging using a single grating of comparable pitch to sample feature size.

PubMed

Morgan, Kaye S; Paganin, David M; Siu, Karen K W

2011-01-01

The ability to quantitatively retrieve transverse phase maps during imaging by using coherent x rays often requires a precise grating or analyzer-crystal-based setup. Imaging of live animals presents further challenges when these methods require multiple exposures for image reconstruction. We present a simple method of single-exposure, single-grating quantitative phase contrast for a regime in which the grating period is much greater than the effective pixel size. A grating is used to create a high-visibility reference pattern incident on the sample, which is distorted according to the complex refractive index and thickness of the sample. The resolution, along a line parallel to the grating, is not restricted by the grating spacing, and the detector resolution becomes the primary determinant of the spatial resolution. We present a method of analysis that maps the displacement of interrogation windows in order to retrieve a quantitative phase map. Application of this analysis to the imaging of known phantoms shows excellent correspondence.

A comparison of phase imaging and quantitative susceptibility mapping in the imaging of multiple sclerosis lesions at ultrahigh field.

PubMed

Cronin, Matthew John; Wharton, Samuel; Al-Radaideh, Ali; Constantinescu, Cris; Evangelou, Nikos; Bowtell, Richard; Gowland, Penny Anne

2016-06-01

The aim of this study was to compare the use of high-resolution phase and QSM images acquired at ultra-high field in the investigation of multiple sclerosis (MS) lesions with peripheral rings, and to discuss their usefulness for drawing inferences about underlying tissue composition. Thirty-nine Subjects were scanned at 7 T, using 3D T 2*-weighted and T 1-weighted sequences. Phase images were then unwrapped and filtered, and quantitative susceptibility maps were generated using a thresholded k-space division method. Lesions were compared visually and using a 1D profiling algorithm. Lesions displaying peripheral rings in the phase images were identified in 10 of the 39 subjects. Dipolar projections were apparent in the phase images outside of the extent of several of these lesions; however, QSM images showed peripheral rings without such projections. These projections appeared ring-like in a small number of phase images where no ring was observed in QSM. 1D profiles of six well-isolated example lesions showed that QSM contrast corresponds more closely to the magnitude images than phase contrast. Phase images contain dipolar projections, which confounds their use in the investigation of tissue composition in MS lesions. Quantitative susceptibility maps correct these projections, providing insight into the composition of MS lesions showing peripheral rings.

Quantitative phase imaging using four interferograms with special phase shifts by dual-wavelength in-line phase-shifting interferometry

NASA Astrophysics Data System (ADS)

Xu, Xiaoqing; Wang, Yawei; Ji, Ying; Xu, Yuanyuan; Xie, Ming; Han, Hao

2018-05-01

A new approach of quantitative phase imaging using four interferograms with special phase shifts in dual-wavelength in-line phase-shifting interferometry is presented. In this method, positive negative 2π phase shifts are employed to easily separate the incoherent addition of two single-wavelength interferograms by combining the phase-shifting technique with the subtraction procedure, then the quantitative phase at one of both wavelengths can be achieved based on two intensities without the corresponding dc terms by the use of the character of the trigonometric function. The quantitative phase of the other wavelength can be retrieved from two dc-term suppressed intensities obtained by employing the two-step phase-shifting technique or the filtering technique in the frequency domain. The proposed method is illustrated with theory, and its effectiveness is demonstrated by simulation experiments of the spherical cap and the HeLa cell, respectively.

Quantitative dispersion microscopy

PubMed Central

Fu, Dan; Choi, Wonshik; Sung, Yongjin; Yaqoob, Zahid; Dasari, Ramachandra R.; Feld, Michael

2010-01-01

Refractive index dispersion is an intrinsic optical property and a useful source of contrast in biological imaging studies. In this report, we present the first dispersion phase imaging of living eukaryotic cells. We have developed quantitative dispersion microscopy based on the principle of quantitative phase microscopy. The dual-wavelength quantitative phase microscope makes phase measurements at 310 nm and 400 nm wavelengths to quantify dispersion (refractive index increment ratio) of live cells. The measured dispersion of living HeLa cells is found to be around 1.088, which agrees well with that measured directly for protein solutions using total internal reflection. This technique, together with the dry mass and morphology measurements provided by quantitative phase microscopy, could prove to be a useful tool for distinguishing different types of biomaterials and studying spatial inhomogeneities of biological samples. PMID:21113234

Ptychography: use of quantitative phase information for high-contrast label free time-lapse imaging of living cells

NASA Astrophysics Data System (ADS)

Suman, Rakesh; O'Toole, Peter

2014-03-01

Here we report a novel label free, high contrast and quantitative method for imaging live cells. The technique reconstructs an image from overlapping diffraction patterns using a ptychographical algorithm. The algorithm utilises both amplitude and phase data from the sample to report on quantitative changes related to the refractive index (RI) and thickness of the specimen. We report the ability of this technique to generate high contrast images, to visualise neurite elongation in neuronal cells, and to provide measure of cell proliferation.

Phase noise optimization in temporal phase-shifting digital holography with partial coherence light sources and its application in quantitative cell imaging.

PubMed

Remmersmann, Christian; Stürwald, Stephan; Kemper, Björn; Langehanenberg, Patrik; von Bally, Gert

2009-03-10

In temporal phase-shifting-based digital holographic microscopy, high-resolution phase contrast imaging requires optimized conditions for hologram recording and phase retrieval. To optimize the phase resolution, for the example of a variable three-step algorithm, a theoretical analysis on statistical errors, digitalization errors, uncorrelated errors, and errors due to a misaligned temporal phase shift is carried out. In a second step the theoretically predicted results are compared to the measured phase noise obtained from comparative experimental investigations with several coherent and partially coherent light sources. Finally, the applicability for noise reduction is demonstrated by quantitative phase contrast imaging of pancreas tumor cells.

Diffraction enhance x-ray imaging for quantitative phase contrast studies

DOE Office of Scientific and Technical Information (OSTI.GOV)

Agrawal, A. K.; Singh, B., E-mail: balwants@rrcat.gov.in; Kashyap, Y. S.

2016-05-23

Conventional X-ray imaging based on absorption contrast permits limited visibility of feature having small density and thickness variations. For imaging of weakly absorbing material or materials possessing similar densities, a novel phase contrast imaging techniques called diffraction enhanced imaging has been designed and developed at imaging beamline Indus-2 RRCAT Indore. The technique provides improved visibility of the interfaces and show high contrast in the image forsmall density or thickness gradients in the bulk. This paper presents basic principle, instrumentation and analysis methods for this technique. Initial results of quantitative phase retrieval carried out on various samples have also been presented.

Identification of ginseng root using quantitative X-ray microtomography.

PubMed

Ye, Linlin; Xue, Yanling; Wang, Yudan; Qi, Juncheng; Xiao, Tiqiao

2017-07-01

The use of X-ray phase-contrast microtomography for the investigation of Chinese medicinal materials is advantageous for its nondestructive, in situ , and three-dimensional quantitative imaging properties. The X-ray phase-contrast microtomography quantitative imaging method was used to investigate the microstructure of ginseng, and the phase-retrieval method is also employed to process the experimental data. Four different ginseng samples were collected and investigated; these were classified according to their species, production area, and sample growth pattern. The quantitative internal characteristic microstructures of ginseng were extracted successfully. The size and position distributions of the calcium oxalate cluster crystals (COCCs), important secondary metabolites that accumulate in ginseng, are revealed by the three-dimensional quantitative imaging method. The volume and amount of the COCCs in different species of the ginseng are obtained by a quantitative analysis of the three-dimensional microstructures, which shows obvious difference among the four species of ginseng. This study is the first to provide evidence of the distribution characteristics of COCCs to identify four types of ginseng, with regard to species authentication and age identification, by X-ray phase-contrast microtomography quantitative imaging. This method is also expected to reveal important relationships between COCCs and the occurrence of the effective medicinal components of ginseng.

Dual function microscope for quantitative DIC and birefringence imaging

NASA Astrophysics Data System (ADS)

Li, Chengshuai; Zhu, Yizheng

2016-03-01

A spectral multiplexing interferometry (SXI) method is presented for integrated birefringence and phase gradient measurement on label-free biological specimens. With SXI, the retardation and orientation of sample birefringence are simultaneously encoded onto two separate spectral carrier waves, generated by a crystal retarder oriented at a specific angle. Thus sufficient information for birefringence determination can be obtained from a single interference spectrum, eliminating the need for multiple acquisitions with mechanical rotation or electrical modulation. In addition, with the insertion of a Nomarski prism, the setup can then acquire quantitative differential interference contrast images. Red blood cells infected by malaria parasites are imaged for birefringence retardation as well as phase gradient. The results demonstrate that the SXI approach can achieve both quantitative phase imaging and birefringence imaging with a single, high-sensitivity system.

Photothermal quantitative phase imaging of living cells with nanoparticles utilizing a cost-efficient setup

NASA Astrophysics Data System (ADS)

Turko, Nir A.; Isbach, Michael; Ketelhut, Steffi; Greve, Burkhard; Schnekenburger, Jürgen; Shaked, Natan T.; Kemper, Björn

2017-02-01

We explored photothermal quantitative phase imaging (PTQPI) of living cells with functionalized nanoparticles (NPs) utilizing a cost-efficient setup based on a cell culture microscope. The excitation light was modulated by a mechanical chopper wheel with low frequencies. Quantitative phase imaging (QPI) was performed with Michelson interferometer-based off-axis digital holographic microscopy and a standard industrial camera. We present results from PTQPI observations on breast cancer cells that were incubated with functionalized gold NPs binding to the epidermal growth factor receptor. Moreover, QPI was used to quantify the impact of the NPs and the low frequency light excitation on cell morphology and viability.

Dynamic phase differences based on quantitative phase imaging for the objective evaluation of cell behavior.

PubMed

Krizova, Aneta; Collakova, Jana; Dostal, Zbynek; Kvasnica, Lukas; Uhlirova, Hana; Zikmund, Tomas; Vesely, Pavel; Chmelik, Radim

2015-01-01

Quantitative phase imaging (QPI) brought innovation to noninvasive observation of live cell dynamics seen as cell behavior. Unlike the Zernike phase contrast or differential interference contrast, QPI provides quantitative information about cell dry mass distribution. We used such data for objective evaluation of live cell behavioral dynamics by the advanced method of dynamic phase differences (DPDs). The DPDs method is considered a rational instrument offered by QPI. By subtracting the antecedent from the subsequent image in a time-lapse series, only the changes in mass distribution in the cell are detected. The result is either visualized as a two dimensional color-coded projection of these two states of the cell or as a time dependence of changes quantified in picograms. Then in a series of time-lapse recordings, the chain of cell mass distribution changes that would otherwise escape attention is revealed. Consequently, new salient features of live cell behavior should emerge. Construction of the DPDs method and results exhibiting the approach are presented. Advantage of the DPDs application is demonstrated on cells exposed to an osmotic challenge. For time-lapse acquisition of quantitative phase images, the recently developed coherence-controlled holographic microscope was employed.

Dynamic phase differences based on quantitative phase imaging for the objective evaluation of cell behavior

NASA Astrophysics Data System (ADS)

Krizova, Aneta; Collakova, Jana; Dostal, Zbynek; Kvasnica, Lukas; Uhlirova, Hana; Zikmund, Tomas; Vesely, Pavel; Chmelik, Radim

2015-11-01

Quantitative phase imaging (QPI) brought innovation to noninvasive observation of live cell dynamics seen as cell behavior. Unlike the Zernike phase contrast or differential interference contrast, QPI provides quantitative information about cell dry mass distribution. We used such data for objective evaluation of live cell behavioral dynamics by the advanced method of dynamic phase differences (DPDs). The DPDs method is considered a rational instrument offered by QPI. By subtracting the antecedent from the subsequent image in a time-lapse series, only the changes in mass distribution in the cell are detected. The result is either visualized as a two-dimensional color-coded projection of these two states of the cell or as a time dependence of changes quantified in picograms. Then in a series of time-lapse recordings, the chain of cell mass distribution changes that would otherwise escape attention is revealed. Consequently, new salient features of live cell behavior should emerge. Construction of the DPDs method and results exhibiting the approach are presented. Advantage of the DPDs application is demonstrated on cells exposed to an osmotic challenge. For time-lapse acquisition of quantitative phase images, the recently developed coherence-controlled holographic microscope was employed.

Accurate single-shot quantitative phase imaging of biological specimens with telecentric digital holographic microscopy.

PubMed

Doblas, Ana; Sánchez-Ortiga, Emilio; Martínez-Corral, Manuel; Saavedra, Genaro; Garcia-Sucerquia, Jorge

2014-04-01

The advantages of using a telecentric imaging system in digital holographic microscopy (DHM) to study biological specimens are highlighted. To this end, the performances of nontelecentric DHM and telecentric DHM are evaluated from the quantitative phase imaging (QPI) point of view. The evaluated stability of the microscope allows single-shot QPI in DHM by using telecentric imaging systems. Quantitative phase maps of a section of the head of the drosophila melanogaster fly and of red blood cells are obtained via single-shot DHM with no numerical postprocessing. With these maps we show that the use of telecentric DHM provides larger field of view for a given magnification and permits more accurate QPI measurements with less number of computational operations.

Review of quantitative phase-digital holographic microscopy: promising novel imaging technique to resolve neuronal network activity and identify cellular biomarkers of psychiatric disorders

PubMed Central

Marquet, Pierre; Depeursinge, Christian; Magistretti, Pierre J.

2014-01-01

Abstract. Quantitative phase microscopy (QPM) has recently emerged as a new powerful quantitative imaging technique well suited to noninvasively explore a transparent specimen with a nanometric axial sensitivity. In this review, we expose the recent developments of quantitative phase-digital holographic microscopy (QP-DHM). Quantitative phase-digital holographic microscopy (QP-DHM) represents an important and efficient quantitative phase method to explore cell structure and dynamics. In a second part, the most relevant QPM applications in the field of cell biology are summarized. A particular emphasis is placed on the original biological information, which can be derived from the quantitative phase signal. In a third part, recent applications obtained, with QP-DHM in the field of cellular neuroscience, namely the possibility to optically resolve neuronal network activity and spine dynamics, are presented. Furthermore, potential applications of QPM related to psychiatry through the identification of new and original cell biomarkers that, when combined with a range of other biomarkers, could significantly contribute to the determination of high risk developmental trajectories for psychiatric disorders, are discussed. PMID:26157976

Review of quantitative phase-digital holographic microscopy: promising novel imaging technique to resolve neuronal network activity and identify cellular biomarkers of psychiatric disorders.

PubMed

Marquet, Pierre; Depeursinge, Christian; Magistretti, Pierre J

2014-10-01

Quantitative phase microscopy (QPM) has recently emerged as a new powerful quantitative imaging technique well suited to noninvasively explore a transparent specimen with a nanometric axial sensitivity. In this review, we expose the recent developments of quantitative phase-digital holographic microscopy (QP-DHM). Quantitative phase-digital holographic microscopy (QP-DHM) represents an important and efficient quantitative phase method to explore cell structure and dynamics. In a second part, the most relevant QPM applications in the field of cell biology are summarized. A particular emphasis is placed on the original biological information, which can be derived from the quantitative phase signal. In a third part, recent applications obtained, with QP-DHM in the field of cellular neuroscience, namely the possibility to optically resolve neuronal network activity and spine dynamics, are presented. Furthermore, potential applications of QPM related to psychiatry through the identification of new and original cell biomarkers that, when combined with a range of other biomarkers, could significantly contribute to the determination of high risk developmental trajectories for psychiatric disorders, are discussed.

Portable smartphone based quantitative phase microscope

NASA Astrophysics Data System (ADS)

Meng, Xin; Tian, Xiaolin; Yu, Wei; Kong, Yan; Jiang, Zhilong; Liu, Fei; Xue, Liang; Liu, Cheng; Wang, Shouyu

2018-01-01

To realize portable device with high contrast imaging capability, we designed a quantitative phase microscope using transport of intensity equation method based on a smartphone. The whole system employs an objective and an eyepiece as imaging system and a cost-effective LED as illumination source. A 3-D printed cradle is used to align these components. Images of different focal planes are captured by manual focusing, followed by calculation of sample phase via a self-developed Android application. To validate its accuracy, we first tested the device by measuring a random phase plate with known phases, and then red blood cell smear, Pap smear, broad bean epidermis sections and monocot root were also measured to show its performance. Owing to its advantages as accuracy, high-contrast, cost-effective and portability, the portable smartphone based quantitative phase microscope is a promising tool which can be future adopted in remote healthcare and medical diagnosis.

Quantitative phase imaging method based on an analytical nonparaxial partially coherent phase optical transfer function.

PubMed

Bao, Yijun; Gaylord, Thomas K

2016-11-01

Multifilter phase imaging with partially coherent light (MFPI-PC) is a promising new quantitative phase imaging method. However, the existing MFPI-PC method is based on the paraxial approximation. In the present work, an analytical nonparaxial partially coherent phase optical transfer function is derived. This enables the MFPI-PC to be extended to the realistic nonparaxial case. Simulations over a wide range of test phase objects as well as experimental measurements on a microlens array verify higher levels of imaging accuracy compared to the paraxial method. Unlike the paraxial version, the nonparaxial MFPI-PC with obliquity factor correction exhibits no systematic error. In addition, due to its analytical expression, the increase in computation time compared to the paraxial version is negligible.

Computational method for multi-modal microscopy based on transport of intensity equation

NASA Astrophysics Data System (ADS)

Li, Jiaji; Chen, Qian; Sun, Jiasong; Zhang, Jialin; Zuo, Chao

2017-02-01

In this paper, we develop the requisite theory to describe a hybrid virtual-physical multi-modal imaging system which yields quantitative phase, Zernike phase contrast, differential interference contrast (DIC), and light field moment imaging simultaneously based on transport of intensity equation(TIE). We then give the experimental demonstration of these ideas by time-lapse imaging of live HeLa cell mitosis. Experimental results verify that a tunable lens based TIE system, combined with the appropriate post-processing algorithm, can achieve a variety of promising imaging modalities in parallel with the quantitative phase images for the dynamic study of cellular processes.

Influence of sample preparation and reliability of automated numerical refocusing in stain-free analysis of dissected tissues with quantitative phase digital holographic microscopy

NASA Astrophysics Data System (ADS)

Kemper, Björn; Lenz, Philipp; Bettenworth, Dominik; Krausewitz, Philipp; Domagk, Dirk; Ketelhut, Steffi

2015-05-01

Digital holographic microscopy (DHM) has been demonstrated to be a versatile tool for high resolution non-destructive quantitative phase imaging of surfaces and multi-modal minimally-invasive monitoring of living cell cultures in-vitro. DHM provides quantitative monitoring of physiological processes through functional imaging and structural analysis which, for example, gives new insight into signalling of cellular water permeability and cell morphology changes due to toxins and infections. Also the analysis of dissected tissues quantitative DHM phase contrast prospects application fields by stain-free imaging and the quantification of tissue density changes. We show that DHM allows imaging of different tissue layers with high contrast in unstained tissue sections. As the investigation of fixed samples represents a very important application field in pathology, we also analyzed the influence of the sample preparation. The retrieved data demonstrate that the quality of quantitative DHM phase images of dissected tissues depends strongly on the fixing method and common staining agents. As in DHM the reconstruction is performed numerically, multi-focus imaging is achieved from a single digital hologram. Thus, we evaluated the automated refocussing feature of DHM for application on different types of dissected tissues and revealed that on moderately stained samples highly reproducible holographic autofocussing can be achieved. Finally, it is demonstrated that alterations of the spatial refractive index distribution in murine and human tissue samples represent a reliable absolute parameter that is related of different degrees of inflammation in experimental colitis and Crohn's disease. This paves the way towards the usage of DHM in digital pathology for automated histological examinations and further studies to elucidate the translational potential of quantitative phase microscopy for the clinical management of patients, e.g., with inflammatory bowel disease.

Refractive index variance of cells and tissues measured by quantitative phase imaging.

PubMed

Shan, Mingguang; Kandel, Mikhail E; Popescu, Gabriel

2017-01-23

The refractive index distribution of cells and tissues governs their interaction with light and can report on morphological modifications associated with disease. Through intensity-based measurements, refractive index information can be extracted only via scattering models that approximate light propagation. As a result, current knowledge of refractive index distributions across various tissues and cell types remains limited. Here we use quantitative phase imaging and the statistical dispersion relation (SDR) to extract information about the refractive index variance in a variety of specimens. Due to the phase-resolved measurement in three-dimensions, our approach yields refractive index results without prior knowledge about the tissue thickness. With the recent progress in quantitative phase imaging systems, we anticipate that using SDR will become routine in assessing tissue optical properties.

High resolution quantitative phase imaging of live cells with constrained optimization approach

NASA Astrophysics Data System (ADS)

Pandiyan, Vimal Prabhu; Khare, Kedar; John, Renu

2016-03-01

Quantitative phase imaging (QPI) aims at studying weakly scattering and absorbing biological specimens with subwavelength accuracy without any external staining mechanisms. Use of a reference beam at an angle is one of the necessary criteria for recording of high resolution holograms in most of the interferometric methods used for quantitative phase imaging. The spatial separation of the dc and twin images is decided by the reference beam angle and Fourier-filtered reconstructed image will have a very poor resolution if hologram is recorded below a minimum reference angle condition. However, it is always inconvenient to have a large reference beam angle while performing high resolution microscopy of live cells and biological specimens with nanometric features. In this paper, we treat reconstruction of digital holographic microscopy images as a constrained optimization problem with smoothness constraint in order to recover only complex object field in hologram plane even with overlapping dc and twin image terms. We solve this optimization problem by gradient descent approach iteratively and the smoothness constraint is implemented by spatial averaging with appropriate size. This approach will give excellent high resolution image recovery compared to Fourier filtering while keeping a very small reference angle. We demonstrate this approach on digital holographic microscopy of live cells by recovering the quantitative phase of live cells from a hologram recorded with nearly zero reference angle.

Smartphone based hand-held quantitative phase microscope using the transport of intensity equation method.

PubMed

Meng, Xin; Huang, Huachuan; Yan, Keding; Tian, Xiaolin; Yu, Wei; Cui, Haoyang; Kong, Yan; Xue, Liang; Liu, Cheng; Wang, Shouyu

2016-12-20

In order to realize high contrast imaging with portable devices for potential mobile healthcare, we demonstrate a hand-held smartphone based quantitative phase microscope using the transport of intensity equation method. With a cost-effective illumination source and compact microscope system, multi-focal images of samples can be captured by the smartphone's camera via manual focusing. Phase retrieval is performed using a self-developed Android application, which calculates sample phases from multi-plane intensities via solving the Poisson equation. We test the portable microscope using a random phase plate with known phases, and to further demonstrate its performance, a red blood cell smear, a Pap smear and monocot root and broad bean epidermis sections are also successfully imaged. Considering its advantages as an accurate, high-contrast, cost-effective and field-portable device, the smartphone based hand-held quantitative phase microscope is a promising tool which can be adopted in the future in remote healthcare and medical diagnosis.

Diagnosis of breast cancer biopsies using quantitative phase imaging

NASA Astrophysics Data System (ADS)

Majeed, Hassaan; Kandel, Mikhail E.; Han, Kevin; Luo, Zelun; Macias, Virgilia; Tangella, Krishnarao; Balla, Andre; Popescu, Gabriel

2015-03-01

The standard practice in the histopathology of breast cancers is to examine a hematoxylin and eosin (H&E) stained tissue biopsy under a microscope. The pathologist looks at certain morphological features, visible under the stain, to diagnose whether a tumor is benign or malignant. This determination is made based on qualitative inspection making it subject to investigator bias. Furthermore, since this method requires a microscopic examination by the pathologist it suffers from low throughput. A quantitative, label-free and high throughput method for detection of these morphological features from images of tissue biopsies is, hence, highly desirable as it would assist the pathologist in making a quicker and more accurate diagnosis of cancers. We present here preliminary results showing the potential of using quantitative phase imaging for breast cancer screening and help with differential diagnosis. We generated optical path length maps of unstained breast tissue biopsies using Spatial Light Interference Microscopy (SLIM). As a first step towards diagnosis based on quantitative phase imaging, we carried out a qualitative evaluation of the imaging resolution and contrast of our label-free phase images. These images were shown to two pathologists who marked the tumors present in tissue as either benign or malignant. This diagnosis was then compared against the diagnosis of the two pathologists on H&E stained tissue images and the number of agreements were counted. In our experiment, the agreement between SLIM and H&E based diagnosis was measured to be 88%. Our preliminary results demonstrate the potential and promise of SLIM for a push in the future towards quantitative, label-free and high throughput diagnosis.

Quantitative X-ray Differential Interference Contrast Microscopy

NASA Astrophysics Data System (ADS)

Nakamura, Takashi

Full-field soft x-ray microscopes are widely used in many fields of sciences. Advances in nanofabrication technology enabled short wavelength focusing elements with significantly improved spatial resolution. In the soft x-ray spectral region, samples as small as 12 nm can be resolved using micro zone-plates as the objective lens. In addition to conventional x-ray microscopy in which x-ray absorption difference provides the image contrast, phase contrast mechanisms such as differential phase contrast (DIC) and Zernike phase contrast have also been demonstrated These phase contrast imaging mechanisms are especially attractive at the x-ray wavelengths where phase contrast of most materials is typically 10 times stronger than the absorption contrast. With recent progresses in plasma-based x- ray sources and increasing accessibility to synchrotron user facilities, x-ray microscopes are quickly becoming standard measurement equipment in the laboratory. To further the usefulness of x-ray DIC microscopy this thesis explicitly addresses three known issues with this imaging modality by introducing new techniques and devices First, as opposed to its visible-light counterpart, no quantitative phase imaging technique exists for x-ray DIC microscopy. To address this issue, two nanoscale x-ray quantitative phase imaging techniques, using exclusive OR (XOR) patterns and zone-plate doublets, respectively, are proposed. Unlike existing x-ray quantitative phase imaging techniques such as Talbot interferometry and ptychography, no dedicated experimental setups or stringent illumination coherence are needed for quantitative phase retrieval. Second, to the best of our knowledge, no quantitative performance characterization of DIC microscopy exists to date. Therefore the imaging system's response to sample's spatial frequency is not known In order to gain in-depth understanding of this imaging modality, performance of x-ray DIC microscopy is quantified using modulation transfer function. A new illumination apparatus required for the transfer function analysis under partially coherent illumination is also proposed. Such a characterization is essential for a proper selection of DIC optics for various transparent samples under study. Finally, optical elements used for x-ray DIC microscopy are highly absorptive and high brilliance x-ray sources such as synchrotrons are generally needed for image contrast. To extend the use of x-ray DIC microscopy to a wider variety of applications, a high efficiency large numerical aperture optical element consisting of high reflective Bragg reflectors is proposed. Using Bragg reflectors, which have 70% ˜99% reflectivity at extreme ultraviolet and soft x-rays for all angles of glancing incidence, the first order focusing efficiency is expected to increase by ˜ 8 times compared to that of a typical Fresnel zone-plate. This thesis contributes to current nanoscale x-ray phase contrast imaging research and provides new insights for biological, material, and magnetic sciences

Measurements of morphology and refractive indexes on human downy hairs using three-dimensional quantitative phase imaging.

PubMed

Lee, SangYun; Kim, Kyoohyun; Lee, Yuhyun; Park, Sungjin; Shin, Heejae; Yang, Jongwon; Ko, Kwanhong; Park, HyunJoo; Park, YongKeun

2015-01-01

We present optical measurements of morphology and refractive indexes (RIs) of human downy arm hairs using three-dimensional (3-D) quantitative phase imaging techniques. 3-D RI tomograms and high-resolution two-dimensional synthetic aperture images of individual downy arm hairs were measured using a Mach–Zehnder laser interferometric microscopy equipped with a two-axis galvanometer mirror. From the measured quantitative images, the RIs and morphological parameters of downy hairs were noninvasively quantified including the mean RI, volume, cylinder, and effective radius of individual hairs. In addition, the effects of hydrogen peroxide on individual downy hairs were investigated.

Quantitative Visualization of Salt Concentration Distributions in Lithium-Ion Battery Electrolytes during Battery Operation Using X-ray Phase Imaging.

PubMed

Takamatsu, Daiko; Yoneyama, Akio; Asari, Yusuke; Hirano, Tatsumi

2018-02-07

A fundamental understanding of concentrations of salts in lithium-ion battery electrolytes during battery operation is important for optimal operation and design of lithium-ion batteries. However, there are few techniques that can be used to quantitatively characterize salt concentration distributions in the electrolytes during battery operation. In this paper, we demonstrate that in operando X-ray phase imaging can quantitatively visualize the salt concentration distributions that arise in electrolytes during battery operation. From quantitative evaluation of the concentration distributions at steady states, we obtained the salt diffusivities in electrolytes with different initial salt concentrations. Because of no restriction on samples and high temporal and spatial resolutions, X-ray phase imaging will be a versatile technique for evaluating electrolytes, both aqueous and nonaqueous, of many electrochemical systems.

Automated classification of cell morphology by coherence-controlled holographic microscopy

NASA Astrophysics Data System (ADS)

Strbkova, Lenka; Zicha, Daniel; Vesely, Pavel; Chmelik, Radim

2017-08-01

In the last few years, classification of cells by machine learning has become frequently used in biology. However, most of the approaches are based on morphometric (MO) features, which are not quantitative in terms of cell mass. This may result in poor classification accuracy. Here, we study the potential contribution of coherence-controlled holographic microscopy enabling quantitative phase imaging for the classification of cell morphologies. We compare our approach with the commonly used method based on MO features. We tested both classification approaches in an experiment with nutritionally deprived cancer tissue cells, while employing several supervised machine learning algorithms. Most of the classifiers provided higher performance when quantitative phase features were employed. Based on the results, it can be concluded that the quantitative phase features played an important role in improving the performance of the classification. The methodology could be valuable help in refining the monitoring of live cells in an automated fashion. We believe that coherence-controlled holographic microscopy, as a tool for quantitative phase imaging, offers all preconditions for the accurate automated analysis of live cell behavior while enabling noninvasive label-free imaging with sufficient contrast and high-spatiotemporal phase sensitivity.

Automated classification of cell morphology by coherence-controlled holographic microscopy.

PubMed

Strbkova, Lenka; Zicha, Daniel; Vesely, Pavel; Chmelik, Radim

2017-08-01

In the last few years, classification of cells by machine learning has become frequently used in biology. However, most of the approaches are based on morphometric (MO) features, which are not quantitative in terms of cell mass. This may result in poor classification accuracy. Here, we study the potential contribution of coherence-controlled holographic microscopy enabling quantitative phase imaging for the classification of cell morphologies. We compare our approach with the commonly used method based on MO features. We tested both classification approaches in an experiment with nutritionally deprived cancer tissue cells, while employing several supervised machine learning algorithms. Most of the classifiers provided higher performance when quantitative phase features were employed. Based on the results, it can be concluded that the quantitative phase features played an important role in improving the performance of the classification. The methodology could be valuable help in refining the monitoring of live cells in an automated fashion. We believe that coherence-controlled holographic microscopy, as a tool for quantitative phase imaging, offers all preconditions for the accurate automated analysis of live cell behavior while enabling noninvasive label-free imaging with sufficient contrast and high-spatiotemporal phase sensitivity. (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Simultaneous off-axis multiplexed holography and regular fluorescence microscopy of biological cells.

PubMed

Nygate, Yoav N; Singh, Gyanendra; Barnea, Itay; Shaked, Natan T

2018-06-01

We present a new technique for obtaining simultaneous multimodal quantitative phase and fluorescence microscopy of biological cells, providing both quantitative phase imaging and molecular specificity using a single camera. Our system is based on an interferometric multiplexing module, externally positioned at the exit of an optical microscope. In contrast to previous approaches, the presented technique allows conventional fluorescence imaging, rather than interferometric off-axis fluorescence imaging. We demonstrate the presented technique for imaging fluorescent beads and live biological cells.

Wavefront image sensor chip

PubMed Central

Cui, Xiquan; Ren, Jian; Tearney, Guillermo J.; Yang, Changhuei

2010-01-01

We report the implementation of an image sensor chip, termed wavefront image sensor chip (WIS), that can measure both intensity/amplitude and phase front variations of a light wave separately and quantitatively. By monitoring the tightly confined transmitted light spots through a circular aperture grid in a high Fresnel number regime, we can measure both intensity and phase front variations with a high sampling density (11 µm) and high sensitivity (the sensitivity of normalized phase gradient measurement is 0.1 mrad under the typical working condition). By using WIS in a standard microscope, we can collect both bright-field (transmitted light intensity) and normalized phase gradient images. Our experiments further demonstrate that the normalized phase gradient images of polystyrene microspheres, unstained and stained starfish embryos, and strongly birefringent potato starch granules are improved versions of their corresponding differential interference contrast (DIC) microscope images in that they are artifact-free and quantitative. Besides phase microscopy, WIS can benefit machine recognition, object ranging, and texture assessment for a variety of applications. PMID:20721059

Using digital inline holographic microscopy and quantitative phase contrast imaging to assess viability of cultured mammalian cells

NASA Astrophysics Data System (ADS)

Missan, Sergey; Hrytsenko, Olga

2015-03-01

Digital inline holographic microscopy was used to record holograms of mammalian cells (HEK293, B16, and E0771) in culture. The holograms have been reconstructed using Octopus software (4Deep inwater imaging) and phase shift maps were unwrapped using the FFT-based phase unwrapping algorithm. The unwrapped phase shifts were used to determine the maximum phase shifts in individual cells. Addition of 0.5 mM H2O2 to cell media produced rapid rounding of cultured cells, followed by cell membrane rupture. The cell morphology changes and cell membrane ruptures were detected in real time and were apparent in the unwrapped phase shift images. The results indicate that quantitative phase contrast imaging produced by the digital inline holographic microscope can be used for the label-free real time automated determination of cell viability and confluence in mammalian cell cultures.

Quantitative breast tissue characterization using grating-based x-ray phase-contrast imaging

NASA Astrophysics Data System (ADS)

Willner, M.; Herzen, J.; Grandl, S.; Auweter, S.; Mayr, D.; Hipp, A.; Chabior, M.; Sarapata, A.; Achterhold, K.; Zanette, I.; Weitkamp, T.; Sztrókay, A.; Hellerhoff, K.; Reiser, M.; Pfeiffer, F.

2014-04-01

X-ray phase-contrast imaging has received growing interest in recent years due to its high capability in visualizing soft tissue. Breast imaging became the focus of particular attention as it is considered the most promising candidate for a first clinical application of this contrast modality. In this study, we investigate quantitative breast tissue characterization using grating-based phase-contrast computed tomography (CT) at conventional polychromatic x-ray sources. Different breast specimens have been scanned at a laboratory phase-contrast imaging setup and were correlated to histopathology. Ascertained tumor types include phylloides tumor, fibroadenoma and infiltrating lobular carcinoma. Identified tissue types comprising adipose, fibroglandular and tumor tissue have been analyzed in terms of phase-contrast Hounsfield units and are compared to high-quality, high-resolution data obtained with monochromatic synchrotron radiation, as well as calculated values based on tabulated tissue properties. The results give a good impression of the method’s prospects and limitations for potential tumor detection and the associated demands on such a phase-contrast breast CT system. Furthermore, the evaluated quantitative tissue values serve as a reference for simulations and the design of dedicated phantoms for phase-contrast mammography.

Single-exposure quantitative phase imaging in color-coded LED microscopy.

PubMed

Lee, Wonchan; Jung, Daeseong; Ryu, Suho; Joo, Chulmin

2017-04-03

We demonstrate single-shot quantitative phase imaging (QPI) in a platform of color-coded LED microscopy (cLEDscope). The light source in a conventional microscope is replaced by a circular LED pattern that is trisected into subregions with equal area, assigned to red, green, and blue colors. Image acquisition with a color image sensor and subsequent computation based on weak object transfer functions allow for the QPI of a transparent specimen. We also provide a correction method for color-leakage, which may be encountered in implementing our method with consumer-grade LEDs and image sensors. Most commercially available LEDs and image sensors do not provide spectrally isolated emissions and pixel responses, generating significant error in phase estimation in our method. We describe the correction scheme for this color-leakage issue, and demonstrate improved phase measurement accuracy. The computational model and single-exposure QPI capability of our method are presented by showing images of calibrated phase samples and cellular specimens.

New approaches for the analysis of confluent cell layers with quantitative phase digital holographic microscopy

NASA Astrophysics Data System (ADS)

Pohl, L.; Kaiser, M.; Ketelhut, S.; Pereira, S.; Goycoolea, F.; Kemper, Björn

2016-03-01

Digital holographic microscopy (DHM) enables high resolution non-destructive inspection of technical surfaces and minimally-invasive label-free live cell imaging. However, the analysis of confluent cell layers represents a challenge as quantitative DHM phase images in this case do not provide sufficient information for image segmentation, determination of the cellular dry mass or calculation of the cell thickness. We present novel strategies for the analysis of confluent cell layers with quantitative DHM phase contrast utilizing a histogram based-evaluation procedure. The applicability of our approach is illustrated by quantification of drug induced cell morphology changes and it is shown that the method is capable to quantify reliable global morphology changes of confluent cell layers.

Quantitative Phase Fraction Detection in Organic Photovoltaic Materials through EELS Imaging

DOE PAGES

Dyck, Ondrej; Hu, Sheng; Das, Sanjib; ...

2015-11-24

Organic photovoltaic materials have recently seen intense interest from the research community. Improvements in device performance are occurring at an impressive rate; however, visualization of the active layer phase separation still remains a challenge. Our paper outlines the application of two electron energy-loss spectroscopic (EELS) imaging techniques that can complement and enhance current phase detection techniques. Specifically, the bulk plasmon peak position, often used to produce contrast between phases in energy filtered transmission electron microscopy (EFTEM), is quantitatively mapped across a sample cross section. One complementary spectrum image capturing the carbon and sulfur core loss edges is compared with themore » plasmon peak map and found to agree quite well, indicating that carbon and sulfur density differences between the two phases also allows phase discrimination. Additionally, an analytical technique for determining absolute atomic areal density is used to produce an absolute carbon and sulfur areal density map. We also show how these maps may be re-interpreted as a phase ratio map, giving quantitative information about the purity of the phases within the junction.« less

Quantitative investigation of the edge enhancement in in-line phase contrast projections and tomosynthesis provided by distributing microbubbles on the interface between two tissues: a phantom study

NASA Astrophysics Data System (ADS)

Wu, Di; Donovan Wong, Molly; Li, Yuhua; Fajardo, Laurie; Zheng, Bin; Wu, Xizeng; Liu, Hong

2017-12-01

The objective of this study was to quantitatively investigate the ability to distribute microbubbles along the interface between two tissues, in an effort to improve the edge and/or boundary features in phase contrast imaging. The experiments were conducted by employing a custom designed tissue simulating phantom, which also simulated a clinical condition where the ligand-targeted microbubbles are self-aggregated on the endothelium of blood vessels surrounding malignant cells. Four different concentrations of microbubble suspensions were injected into the phantom: 0%, 0.1%, 0.2%, and 0.4%. A time delay of 5 min was implemented before image acquisition to allow the microbubbles to become distributed at the interface between the acrylic and the cavity simulating a blood vessel segment. For comparison purposes, images were acquired using three system configurations for both projection and tomosynthesis imaging with a fixed radiation dose delivery: conventional low-energy contact mode, low-energy in-line phase contrast and high-energy in-line phase contrast. The resultant images illustrate the edge feature enhancements in the in-line phase contrast imaging mode when the microbubble concentration is extremely low. The quantitative edge-enhancement-to-noise ratio calculations not only agree with the direct image observations, but also indicate that the edge feature enhancement can be improved by increasing the microbubble concentration. In addition, high-energy in-line phase contrast imaging provided better performance in detecting low-concentration microbubble distributions.

Comparison of qualitative and quantitative evaluation of diffusion-weighted MRI and chemical-shift imaging in the differentiation of benign and malignant vertebral body fractures.

PubMed

Geith, Tobias; Schmidt, Gerwin; Biffar, Andreas; Dietrich, Olaf; Dürr, Hans Roland; Reiser, Maximilian; Baur-Melnyk, Andrea

2012-11-01

The objective of our study was to compare the diagnostic value of qualitative diffusion-weighted imaging (DWI), quantitative DWI, and chemical-shift imaging in a single prospective cohort of patients with acute osteoporotic and malignant vertebral fractures. The study group was composed of patients with 26 osteoporotic vertebral fractures (18 women, eight men; mean age, 69 years; age range, 31 years 6 months to 86 years 2 months) and 20 malignant vertebral fractures (nine women, 11 men; mean age, 63.4 years; age range, 24 years 8 months to 86 years 4 months). T1-weighted, STIR, and T2-weighted sequences were acquired at 1.5 T. A DW reverse fast imaging with steady-state free precession (PSIF) sequence at different delta values was evaluated qualitatively. A DW echo-planar imaging (EPI) sequence and a DW single-shot turbo spin-echo (TSE) sequence at different b values were evaluated qualitatively and quantitatively using the apparent diffusion coefficient. Opposed-phase sequences were used to assess signal intensity qualitatively. The signal loss between in- and opposed-phase images was determined quantitatively. Two-tailed Fisher exact test, Mann-Whitney test, and receiver operating characteristic analysis were performed. Sensitivities, specificities, and accuracies were determined. Qualitative DW-PSIF imaging (delta = 3 ms) showed the best performance for distinguishing between benign and malignant fractures (sensitivity, 100%; specificity, 88.5%; accuracy, 93.5%). Qualitative DW-EPI (b = 50 s/mm(2) [p = 1.00]; b = 250 s/mm(2) [p = 0.50]) and DW single-shot TSE imaging (b = 100 s/mm(2) [p = 1.00]; b = 250 s/mm(2) [p = 0.18]; b = 400 s/mm(2) [p = 0.18]; b = 600 s/mm(2) [p = 0.39]) did not indicate significant differences between benign and malignant fractures. DW-EPI using a b value of 500 s/mm(2) (p = 0.01) indicated significant differences between benign and malignant vertebral fractures. Quantitative DW-EPI (p = 0.09) and qualitative opposed-phase imaging (p = 0.06) did not exhibit significant differences, quantitative DW single-shot TSE imaging (p = 0.002) and quantitative chemical-shift imaging (p = 0.01) showed significant differences between benign and malignant fractures. The DW-PSIF sequence (delta = 3 ms) had the highest accuracy in differentiating benign from malignant vertebral fractures. Quantitative chemical-shift imaging and quantitative DW single-shot TSE imaging had a lower accuracy than DW-PSIF imaging because of a large overlap. Qualitative assessment of opposed-phase, DW-EPI, and DW single-shot TSE sequences and quantitative assessment of the DW-EPI sequence were not suitable for distinguishing between benign and malignant vertebral fractures.

Theory and preliminary experimental verification of quantitative edge illumination x-ray phase contrast tomography.

PubMed

Hagen, C K; Diemoz, P C; Endrizzi, M; Rigon, L; Dreossi, D; Arfelli, F; Lopez, F C M; Longo, R; Olivo, A

2014-04-07

X-ray phase contrast imaging (XPCi) methods are sensitive to phase in addition to attenuation effects and, therefore, can achieve improved image contrast for weakly attenuating materials, such as often encountered in biomedical applications. Several XPCi methods exist, most of which have already been implemented in computed tomographic (CT) modality, thus allowing volumetric imaging. The Edge Illumination (EI) XPCi method had, until now, not been implemented as a CT modality. This article provides indications that quantitative 3D maps of an object's phase and attenuation can be reconstructed from EI XPCi measurements. Moreover, a theory for the reconstruction of combined phase and attenuation maps is presented. Both reconstruction strategies find applications in tissue characterisation and the identification of faint, weakly attenuating details. Experimental results for wires of known materials and for a biological object validate the theory and confirm the superiority of the phase over conventional, attenuation-based image contrast.

Total internal reflection holographic microscopy (TIRHM) for quantitative phase characterization of cell-substrate adhesion

NASA Astrophysics Data System (ADS)

Ash, William Mason, III

Total Internal Reflection Holographic Microscopy (TIRHM) combines near-field microscopy with digital holography to produce a new form of near-field phase microscopy. Using a prism in TIR as a near-field imager, the presence of microscopic organisms, cell-substrate interfaces, and adhesions, causes relative refractive index (RRI) and frustrated TIR (f-TIR) to modulate the object beam's evanescent wave phase front. Quantitative phase images of test specimens such as Amoeba proteus, Dictyostelium Discoideum and cells such as SKOV-3 ovarian cancer and 3T3 fibroblasts are produced without the need to introduce stains or fluorophores. The angular spectrum method of digital holography to compensate for tilt anamorphism due to the inclined TIR plane is also discussed. The results of this work conclusively demonstrate, for the first time, the integration of near-field microscopy with digital holography. The cellular images presented show a correlation between the physical extent of the Amoeba proteus plasma membrane and the adhesions that are quantitatively profiled by phase cross-sectioning of the holographic images obtained by digital holography. With its ability to quantitatively characterise cellular adhesion and motility, it is anticipated that TIRHM can be a tool for characterizing and combating cancer metastasis, as well as improving our understanding of morphogenesis and embryogenesis itself.

Integrated quantitative phase and birefringence microscopy for imaging malaria-infected red blood cells.

PubMed

Li, Chengshuai; Chen, Shichao; Klemba, Michael; Zhu, Yizheng

2016-09-01

A dual-modality birefringence/phase imaging system is presented. The system features a crystal retarder that provides polarization mixing and generates two interferometric carrier waves in a single signal spectrum. The retardation and orientation of sample birefringence can then be measured simultaneously based on spectral multiplexing interferometry. Further, with the addition of a Nomarski prism, the same setup can be used for quantitative differential interference contrast (DIC) imaging. Sample phase can then be obtained with two-dimensional integration. In addition, birefringence-induced phase error can be corrected using the birefringence data. This dual-modality approach is analyzed theoretically with Jones calculus and validated experimentally with malaria-infected red blood cells. The system generates not only corrected DIC and phase images, but a birefringence map that highlights the distribution of hemozoin crystals.

Integrated quantitative phase and birefringence microscopy for imaging malaria-infected red blood cells

NASA Astrophysics Data System (ADS)

Li, Chengshuai; Chen, Shichao; Klemba, Michael; Zhu, Yizheng

2016-09-01

A dual-modality birefringence/phase imaging system is presented. The system features a crystal retarder that provides polarization mixing and generates two interferometric carrier waves in a single signal spectrum. The retardation and orientation of sample birefringence can then be measured simultaneously based on spectral multiplexing interferometry. Further, with the addition of a Nomarski prism, the same setup can be used for quantitative differential interference contrast (DIC) imaging. Sample phase can then be obtained with two-dimensional integration. In addition, birefringence-induced phase error can be corrected using the birefringence data. This dual-modality approach is analyzed theoretically with Jones calculus and validated experimentally with malaria-infected red blood cells. The system generates not only corrected DIC and phase images, but a birefringence map that highlights the distribution of hemozoin crystals.

Intracellular subsurface imaging using a hybrid shear-force feedback/scanning quantitative phase microscopy technique

NASA Astrophysics Data System (ADS)

Edward, Kert

Quantitative phase microscopy (QPM) allows for the imaging of translucent or transparent biological specimens without the need for exogenous contrast agents. This technique is usually applied towards the investigation of simple cells such as red blood cells which are typically enucleated and can be considered to be homogenous. However, most biological cells are nucleated and contain other interesting intracellular organelles. It has been established that the physical characteristics of certain subsurface structures such as the shape and roughness of the nucleus is well correlated with onset and progress of pathological conditions such as cancer. Although the acquired quantitative phase information of biological cells contains surface information as well as coupled subsurface information, the latter has been ignored up until now. A novel scanning quantitative phase imaging system unencumbered by 2pi ambiguities is hereby presented. This system is incorporated into a shear-force feedback scheme which allows for simultaneous phase and topography determination. It will be shown how subsequent image processing of these two data sets allows for the extraction of the subsurface component in the phase data and in vivo cell refractometry studies. Both fabricated samples and biological cells ranging from rat fibroblast cells to malaria infected human erythrocytes were investigated as part of this research. The results correlate quite well with that obtained via other microscopy techniques.

Quantitative evaluation of phase processing approaches in susceptibility weighted imaging

NASA Astrophysics Data System (ADS)

Li, Ningzhi; Wang, Wen-Tung; Sati, Pascal; Pham, Dzung L.; Butman, John A.

2012-03-01

Susceptibility weighted imaging (SWI) takes advantage of the local variation in susceptibility between different tissues to enable highly detailed visualization of the cerebral venous system and sensitive detection of intracranial hemorrhages. Thus, it has been increasingly used in magnetic resonance imaging studies of traumatic brain injury as well as other intracranial pathologies. In SWI, magnitude information is combined with phase information to enhance the susceptibility induced image contrast. Because of global susceptibility variations across the image, the rate of phase accumulation varies widely across the image resulting in phase wrapping artifacts that interfere with the local assessment of phase variation. Homodyne filtering is a common approach to eliminate this global phase variation. However, filter size requires careful selection in order to preserve image contrast and avoid errors resulting from residual phase wraps. An alternative approach is to apply phase unwrapping prior to high pass filtering. A suitable phase unwrapping algorithm guarantees no residual phase wraps but additional computational steps are required. In this work, we quantitatively evaluate these two phase processing approaches on both simulated and real data using different filters and cutoff frequencies. Our analysis leads to an improved understanding of the relationship between phase wraps, susceptibility effects, and acquisition parameters. Although homodyne filtering approaches are faster and more straightforward, phase unwrapping approaches perform more accurately in a wider variety of acquisition scenarios.

High-speed transport-of-intensity phase microscopy with an electrically tunable lens.

PubMed

Zuo, Chao; Chen, Qian; Qu, Weijuan; Asundi, Anand

2013-10-07

We present a high-speed transport-of-intensity equation (TIE) quantitative phase microscopy technique, named TL-TIE, by combining an electrically tunable lens with a conventional transmission microscope. This permits the specimen at different focus position to be imaged in rapid succession, with constant magnification and no physically moving parts. The simplified image stack collection significantly reduces the acquisition time, allows for the diffraction-limited through-focus intensity stack collection at 15 frames per second, making dynamic TIE phase imaging possible. The technique is demonstrated by profiling of microlens array using optimal frequency selection scheme, and time-lapse imaging of live breast cancer cells by inversion the defocused phase optical transfer function to correct the phase blurring in traditional TIE. Experimental results illustrate its outstanding capability of the technique for quantitative phase imaging, through a simple, non-interferometric, high-speed, high-resolution, and unwrapping-free approach with prosperous applications in micro-optics, life sciences and bio-photonics.

Quantitative phase imaging of biological cells and tissues using singleshot white light interference microscopy and phase subtraction method for extended range of measurement

NASA Astrophysics Data System (ADS)

Mehta, Dalip Singh; Sharma, Anuradha; Dubey, Vishesh; Singh, Veena; Ahmad, Azeem

2016-03-01

We present a single-shot white light interference microscopy for the quantitative phase imaging (QPI) of biological cells and tissues. A common path white light interference microscope is developed and colorful white light interferogram is recorded by three-chip color CCD camera. The recorded white light interferogram is decomposed into the red, green and blue color wavelength component interferograms and processed it to find out the RI for different color wavelengths. The decomposed interferograms are analyzed using local model fitting (LMF)" algorithm developed for reconstructing the phase map from single interferogram. LMF is slightly off-axis interferometric QPI method which is a single-shot method that employs only a single image, so it is fast and accurate. The present method is very useful for dynamic process where path-length changes at millisecond level. From the single interferogram a wavelength-dependent quantitative phase imaging of human red blood cells (RBCs) are reconstructed and refractive index is determined. The LMF algorithm is simple to implement and is efficient in computation. The results are compared with the conventional phase shifting interferometry and Hilbert transform techniques.

Quantitative phase imaging of human red blood cells using phase-shifting white light interference microscopy with colour fringe analysis

NASA Astrophysics Data System (ADS)

Singh Mehta, Dalip; Srivastava, Vishal

2012-11-01

We report quantitative phase imaging of human red blood cells (RBCs) using phase-shifting interference microscopy. Five phase-shifted white light interferograms are recorded using colour charge coupled device camera. White light interferograms were decomposed into red, green, and blue colour components. The phase-shifted interferograms of each colour were then processed by phase-shifting analysis and phase maps for red, green, and blue colours were reconstructed. Wavelength dependent refractive index profiles of RBCs were computed from the single set of white light interferogram. The present technique has great potential for non-invasive determination of refractive index variation and morphological features of cells and tissues.

Quantitative phase tomography by using x-ray microscope with Foucault knife-edge scanning filter

DOE Office of Scientific and Technical Information (OSTI.GOV)

Watanabe, Norio; Tsuburaya, Yuji; Shimada, Akihiro

2016-01-28

Quantitative phase tomography was evaluated by using a differential phase microscope with a Foucault knife-edge scanning filter. A 3D x-ray phase image of polystyrene beads was obtained at 5.4 keV. The reconstructed refractive index was fairly good agreement with the Henke’s tabulated data.

White-light diffraction phase microscopy at doubled space-bandwidth product.

PubMed

Shan, Mingguang; Kandel, Mikhail E; Majeed, Hassaan; Nastasa, Viorel; Popescu, Gabriel

2016-12-12

White light diffraction microscopy (wDPM) is a quantitative phase imaging method that benefits from both temporal and spatial phase sensitivity, granted, respectively, by the common-path geometry and white light illumination. However, like all off-axis quantitative phase imaging methods, wDPM is characterized by a reduced space-bandwidth product compared to phase shifting approaches. This happens essentially because the ultimate resolution of the image is governed by the period of the interferogram and not just the diffraction limit. As a result, off-axis techniques generates single-shot, i.e., high time-bandwidth, phase measurements, at the expense of either spatial resolution or field of view. Here, we show that combining phase-shifting and off-axis, the original space-bandwidth is preserved. Specifically, we developed phase-shifting diffraction phase microscopy with white light, in which we measure and combine two phase shifted interferograms. Due to the white light illumination, the phase images are characterized by low spatial noise, i.e., <1nm pathlength. We illustrate the operation of the instrument with test samples, blood cells, and unlabeled prostate tissue biopsy.

Quantitative phase imaging of living cells with a swept laser source

NASA Astrophysics Data System (ADS)

Chen, Shichao; Zhu, Yizheng

2016-03-01

Digital holographic phase microscopy is a well-established quantitative phase imaging technique. However, interference artifacts from inside the system, typically induced by elements whose optical thickness are within the source coherence length, limit the imaging quality as well as sensitivity. In this paper, a swept laser source based technique is presented. Spectra acquired at a number of wavelengths, after Fourier Transform, can be used to identify the sources of the interference artifacts. With proper tuning of the optical pathlength difference between sample and reference arms, it is possible to avoid these artifacts and achieve sensitivity below 0.3nm. Performance of the proposed technique is examined in live cell imaging.

Quantitative phase-filtered wavelength-modulated differential photoacoustic radar tumor hypoxia imaging toward early cancer detection.

PubMed

Dovlo, Edem; Lashkari, Bahman; Soo Sean Choi, Sung; Mandelis, Andreas; Shi, Wei; Liu, Fei-Fei

2017-09-01

Overcoming the limitations of conventional linear spectroscopy used in multispectral photoacoustic imaging, wherein a linear relationship is assumed between the absorbed optical energy and the absorption spectra of the chromophore at a specific location, is crucial for obtaining accurate spatially-resolved quantitative functional information by exploiting known chromophore-specific spectral characteristics. This study introduces a non-invasive phase-filtered differential photoacoustic technique, wavelength-modulated differential photoacoustic radar (WM-DPAR) imaging that addresses this issue by eliminating the effect of the unknown wavelength-dependent fluence. It employs two laser wavelengths modulated out-of-phase to significantly suppress background absorption while amplifying the difference between the two photoacoustic signals. This facilitates pre-malignant tumor identification and hypoxia monitoring, as minute changes in total hemoglobin concentration and hemoglobin oxygenation are detectable. The system can be tuned for specific applications such as cancer screening and SO 2 quantification by regulating the amplitude ratio and phase shift of the signal. The WM-DPAR imaging of a head and neck carcinoma tumor grown in the thigh of a nude rat demonstrates the functional PA imaging of small animals in vivo. The PA appearance of the tumor in relation to tumor vascularity is investigated by immunohistochemistry. Phase-filtered WM-DPAR imaging is also illustrated, maximizing quantitative SO 2 imaging fidelity of tissues. Oxygenation levels within a tumor grown in the thigh of a nude rat using the two-wavelength phase-filtered differential PAR method. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Versatile quantitative phase imaging system applied to high-speed, low noise and multimodal imaging (Conference Presentation)

NASA Astrophysics Data System (ADS)

Federici, Antoine; Aknoun, Sherazade; Savatier, Julien; Wattellier, Benoit F.

2017-02-01

Quadriwave lateral shearing interferometry (QWLSI) is a well-established quantitative phase imaging (QPI) technique based on the analysis of interference patterns of four diffraction orders by an optical grating set in front of an array detector [1]. As a QPI modality, this is a non-invasive imaging technique which allow to measure the optical path difference (OPD) of semi-transparent samples. We present a system enabling QWLSI with high-performance sCMOS cameras [2] and apply it to perform high-speed imaging, low noise as well as multimodal imaging. This modified QWLSI system contains a versatile optomechanical device which images the optical grating near the detector plane. Such a device is coupled with any kind of camera by varying its magnification. In this paper, we study the use of a sCMOS Zyla5.5 camera from Andor along with our modified QWLSI system. We will present high-speed live cell imaging, up to 200Hz frame rate, in order to follow intracellular fast motions while measuring the quantitative phase information. The structural and density information extracted from the OPD signal is complementary to the specific and localized fluorescence signal [2]. In addition, QPI detects cells even when the fluorophore is not expressed. This is very useful to follow a protein expression with time. The 10 µm spatial pixel resolution of our modified QWLSI associated to the high sensitivity of the Zyla5.5 enabling to perform high quality fluorescence imaging, we have carried out multimodal imaging revealing fine structures cells, like actin filaments, merged with the morphological information of the phase. References [1]. P. Bon, G. Maucort, B. Wattellier, and S. Monneret, "Quadriwave lateral shearing interferometry for quantitative phase microscopy of living cells," Opt. Express, vol. 17, pp. 13080-13094, 2009. [2] P. Bon, S. Lécart, E. Fort and S. Lévêque-Fort, "Fast label-free cytoskeletal network imaging in living mammalian cells," Biophysical journal, 106(8), pp. 1588-1595, 2014

Microscopic optical path length difference and polarization measurement system for cell analysis

NASA Astrophysics Data System (ADS)

Satake, H.; Ikeda, K.; Kowa, H.; Hoshiba, T.; Watanabe, E.

2018-03-01

In recent years, noninvasive, nonstaining, and nondestructive quantitative cell measurement techniques have become increasingly important in the medical field. These cell measurement techniques enable the quantitative analysis of living cells, and are therefore applied to various cell identification processes, such as those determining the passage number limit during cell culturing in regenerative medicine. To enable cell measurement, we developed a quantitative microscopic phase imaging system based on a Mach-Zehnder interferometer that measures the optical path length difference distribution without phase unwrapping using optical phase locking. The applicability of our phase imaging system was demonstrated by successful identification of breast cancer cells amongst normal cells. However, the cell identification method using this phase imaging system exhibited a false identification rate of approximately 7%. In this study, we implemented a polarimetric imaging system by introducing a polarimetric module to one arm of the Mach-Zehnder interferometer of our conventional phase imaging system. This module was comprised of a quarter wave plate and a rotational polarizer on the illumination side of the sample, and a linear polarizer on the optical detector side. In addition, we developed correction methods for the measurement errors of the optical path length and birefringence phase differences that arose through the influence of elements other than cells, such as the Petri dish. As the Petri dish holding the fluid specimens was transparent, it did not affect the amplitude information; however, the optical path length and birefringence phase differences were affected. Therefore, we proposed correction of the optical path length and birefringence phase for the influence of elements other than cells, as a prerequisite for obtaining highly precise phase and polarimetric images.

Quantitative phase imaging of retinal cells (Conference Presentation)

NASA Astrophysics Data System (ADS)

LaForest, Timothé; Carpentras, Dino; Kowalczuk, Laura; Behar-Cohen, Francine; Moser, Christophe

2017-02-01

Vision process is ruled by several cells layers of the retina. Before reaching the photoreceptors, light entering the eye has to pass through a few hundreds of micrometers thick layer of ganglion and neurons cells. Macular degeneration is a non-curable disease of themacula occurring with age. This disease can be diagnosed at an early stage by imaging neuronal cells in the retina and observing their death chronically. These cells are phase objects locatedon a background that presents an absorption pattern and so difficult to see with standard imagingtechniques in vivo. Phase imaging methods usually need the illumination system to be on the opposite side of the sample with respect to theimaging system. This is a constraintand a challenge for phase imaging in-vivo. Recently, the possibility of performing phase contrast imaging from one side using properties of scattering media has been shown. This phase contrast imaging is based on the back illumination generated by the sample itself. Here, we present a reflection phase imaging technique based on oblique back-illumination. The oblique back-illumination creates a dark field image of the sample. Generating asymmetric oblique illumination allows obtaining differential phase contrast image, which in turn can be processed to recover a quantitative phase image. In the case of the eye, a transcleral illumination can generate oblique incident light on the retina and the choroidal layer.The back reflected light is then collected by the eye lens to produce dark field image. We show experimental results of retinal phase imagesin ex vivo samples of human and pig retina.

Large field of view quantitative phase imaging of induced pluripotent stem cells and optical pathlength reference materials

NASA Astrophysics Data System (ADS)

Kwee, Edward; Peterson, Alexander; Stinson, Jeffrey; Halter, Michael; Yu, Liya; Majurski, Michael; Chalfoun, Joe; Bajcsy, Peter; Elliott, John

2018-02-01

Induced pluripotent stem cells (iPSCs) are reprogrammed cells that can have heterogeneous biological potential. Quality assurance metrics of reprogrammed iPSCs will be critical to ensure reliable use in cell therapies and personalized diagnostic tests. We present a quantitative phase imaging (QPI) workflow which includes acquisition, processing, and stitching multiple adjacent image tiles across a large field of view (LFOV) of a culture vessel. Low magnification image tiles (10x) were acquired with a Phasics SID4BIO camera on a Zeiss microscope. iPSC cultures were maintained using a custom stage incubator on an automated stage. We implement an image acquisition strategy that compensates for non-flat illumination wavefronts to enable imaging of an entire well plate, including the meniscus region normally obscured in Zernike phase contrast imaging. Polynomial fitting and background mode correction was implemented to enable comparability and stitching between multiple tiles. LFOV imaging of reference materials indicated that image acquisition and processing strategies did not affect quantitative phase measurements across the LFOV. Analysis of iPSC colony images demonstrated mass doubling time was significantly different than area doubling time. These measurements were benchmarked with prototype microsphere beads and etched-glass gratings with specified spatial dimensions designed to be QPI reference materials with optical pathlength shifts suitable for cell microscopy. This QPI workflow and the use of reference materials can provide non-destructive traceable imaging method for novel iPSC heterogeneity characterization.

Quantitative x-ray phase imaging at the nanoscale by multilayer Laue lenses

PubMed Central

Yan, Hanfei; Chu, Yong S.; Maser, Jörg; Nazaretski, Evgeny; Kim, Jungdae; Kang, Hyon Chol; Lombardo, Jeffrey J.; Chiu, Wilson K. S.

2013-01-01

For scanning x-ray microscopy, many attempts have been made to image the phase contrast based on a concept of the beam being deflected by a specimen, the so-called differential phase contrast imaging (DPC). Despite the successful demonstration in a number of representative cases at moderate spatial resolutions, these methods suffer from various limitations that preclude applications of DPC for ultra-high spatial resolution imaging, where the emerging wave field from the focusing optic tends to be significantly more complicated. In this work, we propose a highly robust and generic approach based on a Fourier-shift fitting process and demonstrate quantitative phase imaging of a solid oxide fuel cell (SOFC) anode by multilayer Laue lenses (MLLs). The high sensitivity of the phase to structural and compositional variations makes our technique extremely powerful in correlating the electrode performance with its buried nanoscale interfacial structures that may be invisible to the absorption and fluorescence contrasts. PMID:23419650

Time-resolved imaging refractometry of microbicidal films using quantitative phase microscopy.

PubMed

Rinehart, Matthew T; Drake, Tyler K; Robles, Francisco E; Rohan, Lisa C; Katz, David; Wax, Adam

2011-12-01

Quantitative phase microscopy is applied to image temporal changes in the refractive index (RI) distributions of solutions created by microbicidal films undergoing hydration. We present a novel method of using an engineered polydimethylsiloxane structure as a static phase reference to facilitate calibration of the absolute RI across the entire field. We present a study of dynamic structural changes in microbicidal films during hydration and subsequent dissolution. With assumptions about the smoothness of the phase changes induced by these films, we calculate absolute changes in the percentage of film in regions across the field of view.

Time-resolved imaging refractometry of microbicidal films using quantitative phase microscopy

PubMed Central

Rinehart, Matthew T.; Drake, Tyler K.; Robles, Francisco E.; Rohan, Lisa C.; Katz, David; Wax, Adam

2011-01-01

Quantitative phase microscopy is applied to image temporal changes in the refractive index (RI) distributions of solutions created by microbicidal films undergoing hydration. We present a novel method of using an engineered polydimethylsiloxane structure as a static phase reference to facilitate calibration of the absolute RI across the entire field. We present a study of dynamic structural changes in microbicidal films during hydration and subsequent dissolution. With assumptions about the smoothness of the phase changes induced by these films, we calculate absolute changes in the percentage of film in regions across the field of view. PMID:22191912

Quantitative phase and amplitude imaging using Differential-Interference Contrast (DIC) microscopy

NASA Astrophysics Data System (ADS)

Preza, Chrysanthe; O'Sullivan, Joseph A.

2009-02-01

We present an extension of the development of an alternating minimization (AM) method for the computation of a specimen's complex transmittance function (magnitude and phase) from DIC images. The ability to extract both quantitative phase and amplitude information from two rotationally-diverse DIC images (i.e., acquired by rotating the sample) extends previous efforts in computational DIC microscopy that have focused on quantitative phase imaging only. Simulation results show that the inverse problem at hand is sensitive to noise as well as to the choice of the AM algorithm parameters. The AM framework allows constraints and penalties on the magnitude and phase estimates to be incorporated in a principled manner. Towards this end, Green and De Pierro's "log-cosh" regularization penalty is applied to the magnitude of differences of neighboring values of the complex-valued function of the specimen during the AM iterations. The penalty is shown to be convex in the complex space. A procedure to approximate the penalty within the iterations is presented. In addition, a methodology to pre-compute AM parameters that are optimal with respect to the convergence rate of the AM algorithm is also presented. Both extensions of the AM method are investigated with simulations.

Measuring the Nonuniform Evaporation Dynamics of Sprayed Sessile Microdroplets with Quantitative Phase Imaging.

PubMed

Edwards, Chris; Arbabi, Amir; Bhaduri, Basanta; Wang, Xiaozhen; Ganti, Raman; Yunker, Peter J; Yodh, Arjun G; Popescu, Gabriel; Goddard, Lynford L

2015-10-13

We demonstrate real-time quantitative phase imaging as a new optical approach for measuring the evaporation dynamics of sessile microdroplets. Quantitative phase images of various droplets were captured during evaporation. The images enabled us to generate time-resolved three-dimensional topographic profiles of droplet shape with nanometer accuracy and, without any assumptions about droplet geometry, to directly measure important physical parameters that characterize surface wetting processes. Specifically, the time-dependent variation of the droplet height, volume, contact radius, contact angle distribution along the droplet's perimeter, and mass flux density for two different surface preparations are reported. The studies clearly demonstrate three phases of evaporation reported previously: pinned, depinned, and drying modes; the studies also reveal instances of partial pinning. Finally, the apparatus is employed to investigate the cooperative evaporation of the sprayed droplets. We observe and explain the neighbor-induced reduction in evaporation rate, that is, as compared to predictions for isolated droplets. In the future, the new experimental methods should stimulate the exploration of colloidal particle dynamics on the gas-liquid-solid interface.

Phase-space evolution of x-ray coherence in phase-sensitive imaging.

PubMed

Wu, Xizeng; Liu, Hong

2008-08-01

X-ray coherence evolution in the imaging process plays a key role for x-ray phase-sensitive imaging. In this work we present a phase-space formulation for the phase-sensitive imaging. The theory is reformulated in terms of the cross-spectral density and associated Wigner distribution. The phase-space formulation enables an explicit and quantitative account of partial coherence effects on phase-sensitive imaging. The presented formulas for x-ray spectral density at the detector can be used for performing accurate phase retrieval and optimizing the phase-contrast visibility. The concept of phase-space shearing length derived from this phase-space formulation clarifies the spatial coherence requirement for phase-sensitive imaging with incoherent sources. The theory has been applied to x-ray Talbot interferometric imaging as well. The peak coherence condition derived reveals new insights into three-grating-based Talbot-interferometric imaging and gratings-based x-ray dark-field imaging.

Flipping interferometry and its application for quantitative phase microscopy in a micro-channel.

PubMed

Roitshtain, Darina; Turko, Nir A; Javidi, Bahram; Shaked, Natan T

2016-05-15

We present a portable, off-axis interferometric module for quantitative phase microscopy of live cells, positioned at the exit port of a coherently illuminated inverted microscope. The module creates on the digital camera an interference pattern between the image of the sample and its flipped version. The proposed simplified module is based on a retro-reflector modification in an external Michelson interferometer. The module does not contain any lenses, pinholes, or gratings and its alignment is straightforward. Still, it allows full control of the off-axis angle and does not suffer from ghost images. As experimentally demonstrated, the module is useful for quantitative phase microscopy of live cells rapidly flowing in a micro-channel.

High-speed quantitative phase imaging using time-stretch spectral shearing contrast (Conference Presentation)

NASA Astrophysics Data System (ADS)

Bosworth, Bryan; Foster, Mark A.

2017-02-01

Photonic time-stretch microscopy (TSM) provides an ideal platform for high-throughput imaging flow cytometry, affording extremely high shutter speeds and frame rates with high sensitivity. In order to resolve weakly scattering cells in biofluid and solve the issue of signal-to-noise in cell labeling specificity of biomarkers in imaging flow cytometry, several quantitative phase (QP) techniques have recently been adapted to TSM. However, these techniques have relied primarily on sensitive free-space optical configurations to generate full electric field measurements. The present work draws from the field of ultrashort pulse characterization to leverage the coherence of the ultrashort optical pulses integral to all TSM systems in order to do self-referenced single-shot quantitative phase imaging in a TSM system. Self-referencing is achieved via spectral shearing interferometry in an exceptionally stable and straightforward Sagnac loop incorporating an electro-optic phase modulator and polarization-maintaining fiber that produce sheared and unsheared copies of the pulse train with an inter-pulse delay determined by polarization mode dispersion. The spectral interferogram then yields a squared amplitude and a phase derivative image that can be integrated for conventional phase. We apply this spectral shearing contrast microscope to acquire QP images on a high-speed flow microscope at 90-MHz line rates with <400 pixels per line. We also consider the extension of this technique to compressed sensing (CS) acquisition by intensity modulating the interference spectra with pseudorandom binary waveforms to reconstruct the images from a highly sub-Nyquist number of random inner products, providing a path to even higher operating rates and reduced data storage requirements.

Quantitative phase microscopy for cellular dynamics based on transport of intensity equation.

PubMed

Li, Ying; Di, Jianglei; Ma, Chaojie; Zhang, Jiwei; Zhong, Jinzhan; Wang, Kaiqiang; Xi, Teli; Zhao, Jianlin

2018-01-08

We demonstrate a simple method for quantitative phase imaging of tiny transparent objects such as living cells based on the transport of intensity equation. The experiments are performed using an inverted bright field microscope upgraded with a flipping imaging module, which enables to simultaneously create two laterally separated images with unequal defocus distances. This add-on module does not include any lenses or gratings and is cost-effective and easy-to-alignment. The validity of this method is confirmed by the measurement of microlens array and human osteoblastic cells in culture, indicating its potential in the applications of dynamically measuring living cells and other transparent specimens in a quantitative, non-invasive and label-free manner.

3D/4D multiscale imaging in acute lymphoblastic leukemia cells: visualizing dynamics of cell death

NASA Astrophysics Data System (ADS)

Sarangapani, Sreelatha; Mohan, Rosmin Elsa; Patil, Ajeetkumar; Lang, Matthew J.; Asundi, Anand

2017-06-01

Quantitative phase detection is a new methodology that provides quantitative information on cellular morphology to monitor the cell status, drug response and toxicity. In this paper the morphological changes in acute leukemia cells treated with chitosan were detected using d'Bioimager a robust imaging system. Quantitative phase image of the cells was obtained with numerical analysis. Results show that the average area and optical volume of the chitosan treated cells is significantly reduced when compared with the control cells, which reveals the effect of chitosan on the cancer cells. From the results it can be attributed that d'Bioimager can be used as a non-invasive imaging alternative to measure the morphological changes of the living cells in real time.

Quantitative studies on inner interfaces in conical metal joints using hard x-ray inline phase contrast radiography

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zabler, S.; Rack, T.; Nelson, K.

2010-10-15

Quantitative investigation of micrometer and submicrometer gaps between joining metal surfaces is applied to conical plug-socket connections in dental titanium implants. Microgaps of widths well beyond the resolving power of industrial x-ray systems are imaged by synchrotron phase contrast radiography. Furthermore, by using an analytical model for the relatively simple sample geometry and applying it to numerical forward simulations of the optical Fresnel propagation, we show that quantitative measurements of the microgap width down to 0.1 {mu}m are possible. Image data recorded at the BAMline (BESSY-II light source, Germany) are presented, with the resolving power of the imaging system beingmore » 4 {mu}m in absorption mode and {approx}14 {mu}m in phase contrast mode (z{sub 2}=0.74 m). Thus, phase contrast radiography, combined with numerical forward simulations, is capable of measuring the widths of gaps that are two orders of magnitude thinner than the conventional detection limit.« less

Hard x-ray phase contrastmicroscopy - techniques and applications

NASA Astrophysics Data System (ADS)

Holzner, Christian

In 1918, Einstein provided the first description of the nature of the refractive index for X-rays, showing that phase contrast effects are significant. A century later, most x-ray microscopy and nearly all medical imaging remains based on absorption contrast, even though phase contrast offers orders of magnitude improvements in contrast and reduced radiation exposure at multi-keV x-ray energies. The work presented is concerned with developing practical and quantitative methods of phase contrast for x-ray microscopy. A theoretical framework for imaging in phase contrast is put forward; this is used to obtain quantitative images in a scanning microscope using a segmented detector, and to correct for artifacts in a commercial phase contrast x-ray nano-tomography system. The principle of reciprocity between scanning and full-field microscopes is then used to arrive at a novel solution: Zernike contrast in a scanning microscope. These approaches are compared on a theoretical and experimental basis in direct connection with applications using multi-keV x-ray microscopes at the Advanced Photon Source at Argonne National Laboratory. Phase contrast provides the best means to image mass and ultrastructure of light elements that mainly constitute biological matter, while stimulated x-ray fluorescence provides high sensitivity for studies of the distribution of heavier trace elements, such as metals. These approaches are combined in a complementary way to yield quantitative maps of elemental concentration from 2D images, with elements placed in their ultrastructural context. The combination of x-ray fluorescence and phase contrast poses an ideal match for routine, high resolution tomographic imaging of biological samples in the future. The presented techniques and demonstration experiments will help pave the way for this development.

Automated Detection of P. falciparum Using Machine Learning Algorithms with Quantitative Phase Images of Unstained Cells.

PubMed

Park, Han Sang; Rinehart, Matthew T; Walzer, Katelyn A; Chi, Jen-Tsan Ashley; Wax, Adam

2016-01-01

Malaria detection through microscopic examination of stained blood smears is a diagnostic challenge that heavily relies on the expertise of trained microscopists. This paper presents an automated analysis method for detection and staging of red blood cells infected by the malaria parasite Plasmodium falciparum at trophozoite or schizont stage. Unlike previous efforts in this area, this study uses quantitative phase images of unstained cells. Erythrocytes are automatically segmented using thresholds of optical phase and refocused to enable quantitative comparison of phase images. Refocused images are analyzed to extract 23 morphological descriptors based on the phase information. While all individual descriptors are highly statistically different between infected and uninfected cells, each descriptor does not enable separation of populations at a level satisfactory for clinical utility. To improve the diagnostic capacity, we applied various machine learning techniques, including linear discriminant classification (LDC), logistic regression (LR), and k-nearest neighbor classification (NNC), to formulate algorithms that combine all of the calculated physical parameters to distinguish cells more effectively. Results show that LDC provides the highest accuracy of up to 99.7% in detecting schizont stage infected cells compared to uninfected RBCs. NNC showed slightly better accuracy (99.5%) than either LDC (99.0%) or LR (99.1%) for discriminating late trophozoites from uninfected RBCs. However, for early trophozoites, LDC produced the best accuracy of 98%. Discrimination of infection stage was less accurate, producing high specificity (99.8%) but only 45.0%-66.8% sensitivity with early trophozoites most often mistaken for late trophozoite or schizont stage and late trophozoite and schizont stage most often confused for each other. Overall, this methodology points to a significant clinical potential of using quantitative phase imaging to detect and stage malaria infection without staining or expert analysis.

Automated Detection of P. falciparum Using Machine Learning Algorithms with Quantitative Phase Images of Unstained Cells

PubMed Central

Park, Han Sang; Rinehart, Matthew T.; Walzer, Katelyn A.; Chi, Jen-Tsan Ashley; Wax, Adam

2016-01-01

Malaria detection through microscopic examination of stained blood smears is a diagnostic challenge that heavily relies on the expertise of trained microscopists. This paper presents an automated analysis method for detection and staging of red blood cells infected by the malaria parasite Plasmodium falciparum at trophozoite or schizont stage. Unlike previous efforts in this area, this study uses quantitative phase images of unstained cells. Erythrocytes are automatically segmented using thresholds of optical phase and refocused to enable quantitative comparison of phase images. Refocused images are analyzed to extract 23 morphological descriptors based on the phase information. While all individual descriptors are highly statistically different between infected and uninfected cells, each descriptor does not enable separation of populations at a level satisfactory for clinical utility. To improve the diagnostic capacity, we applied various machine learning techniques, including linear discriminant classification (LDC), logistic regression (LR), and k-nearest neighbor classification (NNC), to formulate algorithms that combine all of the calculated physical parameters to distinguish cells more effectively. Results show that LDC provides the highest accuracy of up to 99.7% in detecting schizont stage infected cells compared to uninfected RBCs. NNC showed slightly better accuracy (99.5%) than either LDC (99.0%) or LR (99.1%) for discriminating late trophozoites from uninfected RBCs. However, for early trophozoites, LDC produced the best accuracy of 98%. Discrimination of infection stage was less accurate, producing high specificity (99.8%) but only 45.0%-66.8% sensitivity with early trophozoites most often mistaken for late trophozoite or schizont stage and late trophozoite and schizont stage most often confused for each other. Overall, this methodology points to a significant clinical potential of using quantitative phase imaging to detect and stage malaria infection without staining or expert analysis. PMID:27636719

Label-free imaging of intracellular motility by low-coherent quantitative phase microscope in reflection geometry

NASA Astrophysics Data System (ADS)

Yamauchi, Toyohiko; Iwai, Hidenao; Yamashita, Yutaka

2011-11-01

We demonstrate tomographic imaging of intracellular activity of living cells by a low-coherent quantitative phase microscope. The intracellular organelles, such as the nucleus, nucleolus, and mitochondria, are moving around inside living cells, driven by the cellular physiological activity. In order to visualize the intracellular motility in a label-free manner we have developed a reflection-type quantitative phase microscope which employs the phase shifting interferometric technique with a low-coherent light source. The phase shifting interferometry enables us to quantitatively measure the intensity and phase of the optical field, and the low-coherence interferometry makes it possible to selectively probe a specific sectioning plane in the cell volume. The results quantitatively revealed the depth-resolved fluctuations of intracellular surfaces so that the plasma membrane and the membranes of intracellular organelles were independently measured. The transversal and the vertical spatial resolutions were 0.56 μm and 0.93 μm, respectively, and the mechanical sensitivity of the phase measurement was 1.2 nanometers. The mean-squared displacement was applied as a statistical tool to analyze the temporal fluctuation of the intracellular organelles. To the best of our knowledge, our system visualized depth-resolved intracellular organelles motion for the first time in sub-micrometer resolution without contrast agents.

Dual-modality wide-field photothermal quantitative phase microscopy and depletion of cell populations

NASA Astrophysics Data System (ADS)

Turko, Nir A.; Barnea, Itay; Blum, Omry; Korenstein, Rafi; Shaked, Natan T.

2015-03-01

We review our dual-modality technique for quantitative imaging and selective depletion of populations of cells based on wide-field photothermal (PT) quantitative phase imaging and simultaneous PT cell extermination. The cells are first labeled by plasmonic gold nanoparticles, which evoke local plasmonic resonance when illuminated by light in a wavelength corresponding to their specific plasmonic resonance peak. This reaction creates changes of temperature, resulting in changes of phase. This phase changes are recorded by a quantitative phase microscope (QPM), producing specific imaging contrast, and enabling bio-labeling in phase microscopy. Using this technique, we have shown discrimination of EGFR over-expressing (EGFR+) cancer cells from EGFR under-expressing (EGFR-) cancer cells. Then, we have increased the excitation power in order to evoke greater temperatures, which caused specific cell death, all under real-time phase acquisition using QPM. Close to 100% of all EGFR+ cells were immediately exterminated when illuminated with the strong excitation beam, while all EGFR- cells survived. For the second experiment, in order to simulate a condition where circulating tumor cells (CTCs) are present in blood, we have mixed the EGFR+ cancer cells with white blood cells (WBCs) from a healthy donor. Here too, we have used QPM to observe and record the phase of the cells as they were excited for selective visualization and then exterminated. The WBCs survival rate was over 95%, while the EGFR+ survival rate was under 5%. The technique may be the basis for real-time detection and controlled treatment of CTCs.

Analyser-based phase contrast image reconstruction using geometrical optics.

PubMed

Kitchen, M J; Pavlov, K M; Siu, K K W; Menk, R H; Tromba, G; Lewis, R A

2007-07-21

Analyser-based phase contrast imaging can provide radiographs of exceptional contrast at high resolution (<100 microm), whilst quantitative phase and attenuation information can be extracted using just two images when the approximations of geometrical optics are satisfied. Analytical phase retrieval can be performed by fitting the analyser rocking curve with a symmetric Pearson type VII function. The Pearson VII function provided at least a 10% better fit to experimentally measured rocking curves than linear or Gaussian functions. A test phantom, a hollow nylon cylinder, was imaged at 20 keV using a Si(1 1 1) analyser at the ELETTRA synchrotron radiation facility. Our phase retrieval method yielded a more accurate object reconstruction than methods based on a linear fit to the rocking curve. Where reconstructions failed to map expected values, calculations of the Takagi number permitted distinction between the violation of the geometrical optics conditions and the failure of curve fitting procedures. The need for synchronized object/detector translation stages was removed by using a large, divergent beam and imaging the object in segments. Our image acquisition and reconstruction procedure enables quantitative phase retrieval for systems with a divergent source and accounts for imperfections in the analyser.

Speckle-free and halo-free low coherent Mach-Zehnder quantitative-phase-imaging module as a replacement of objective lens in conventional inverted microscopes

NASA Astrophysics Data System (ADS)

Yamauchi, Toyohiko; Yamada, Hidenao; Matsui, Hisayuki; Yasuhiko, Osamu; Ueda, Yukio

2018-02-01

We developed a compact Mach-Zehnder interferometer module to be used as a replacement of the objective lens in a conventional inverted microscope (Nikon, TS100-F) in order to make them quantitative phase microscopes. The module has a 90-degree-flipped U-shape; the dimensions of the module are 160 mm by 120 mm by 40 mm and the weight is 380 grams. The Mach-Zehnder interferometer equipped with the separate reference and sample arms was implemented in this U-shaped housing and the path-length difference between the two arms was manually adjustable. The sample under test was put on the stage of the microscope and a sample light went through it. Both arms had identical achromatic lenses for image formation and the lateral positions of them were also manually adjustable. Therefore, temporally and spatially low coherent illumination was applicable because the users were able to balance precisely the path length of the two arms and to overlap the two wavefronts. In the experiment, spectrally filtered LED light for illumination (wavelength = 633 nm and bandwidth = 3 nm) was input to the interferometer module via a 50 micrometer core optical fiber. We have successfully captured full-field interference images by a camera put on the trinocular tube of the microscope and constructed quantitative phase images of the cultured cells by means of the quarter-wavelength phase shifting algorithm. The resultant quantitative phase images were speckle-free and halo-free due to spectrally and spatially low coherent illumination.

Photon-counting-based diffraction phase microscopy combined with single-pixel imaging

NASA Astrophysics Data System (ADS)

Shibuya, Kyuki; Araki, Hiroyuki; Iwata, Tetsuo

2018-04-01

We propose a photon-counting (PC)-based quantitative-phase imaging (QPI) method for use in diffraction phase microscopy (DPM) that is combined with a single-pixel imaging (SPI) scheme (PC-SPI-DPM). This combination of DPM with the SPI scheme overcomes a low optical throughput problem that has occasionally prevented us from obtaining quantitative-phase images in DPM through use of a high-sensitivity single-channel photodetector such as a photomultiplier tube (PMT). The introduction of a PMT allowed us to perform PC with ease and thus solved a dynamic range problem that was inherent to SPI. As a proof-of-principle experiment, we performed a comparison study of analogue-based SPI-DPM and PC-SPI-DPM for a 125-nm-thick indium tin oxide (ITO) layer coated on a silica glass substrate. We discuss the basic performance of the method and potential future modifications of the proposed system.

High-speed and high-resolution quantitative phase imaging with digital-micromirror device-based illumination (Conference Presentation)

NASA Astrophysics Data System (ADS)

Zhou, Renjie; Jin, Di; Yaqoob, Zahid; So, Peter T. C.

2017-02-01

Due to the large number of available mirrors, the patterning speed, low-cost, and compactness, digital-micromirror devices (DMDs) have been extensively used in biomedical imaging system. Recently, DMDs have been brought to the quantitative phase microscopy (QPM) field to achieve synthetic-aperture imaging and tomographic imaging. Last year, our group demonstrated using DMD for QPM, where the phase-retrieval is based on a recently developed Fourier ptychography algorithm. In our previous system, the illumination angle was varied through coding the aperture plane of the illumination system, which has a low efficiency on utilizing the laser power. In our new DMD-based QPM system, we use the Lee-holograms, which is conjugated to the sample plane, to change the illumination angles for much higher power efficiency. Multiple-angle illumination can also be achieved with this method. With this versatile system, we can achieve FPM-based high-resolution phase imaging with 250 nm lateral resolution using the Rayleigh criteria. Due to the use of a powerful laser, the imaging speed would only be limited by the camera acquisition speed. With a fast camera, we expect to achieve close to 100 fps phase imaging speed that has not been achieved in current FPM imaging systems. By adding reference beam, we also expect to achieve synthetic-aperture imaging while directly measuring the phase of the sample fields. This would reduce the phase-retrieval processing time to allow for real-time imaging applications in the future.

Collaborative Initiative in Biomedical Imaging to Study Complex Diseases

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lin, Weili; Fiddy, Michael A.

2012-03-31

The work reported addressed these topics: Fluorescence imaging; Optical coherence tomography; X-ray interferometer/phase imaging system; Quantitative imaging from scattered fields, Terahertz imaging and spectroscopy; and Multiphoton and Raman microscopy.

Noninvasive characterization of the fission yeast cell cycle by monitoring dry mass with digital holographic microscopy.

PubMed

Rappaz, Benjamin; Cano, Elena; Colomb, Tristan; Kühn, Jonas; Depeursinge, Christian; Simanis, Viesturs; Magistretti, Pierre J; Marquet, Pierre

2009-01-01

Digital holography microscopy (DHM) is an optical technique which provides phase images yielding quantitative information about cell structure and cellular dynamics. Furthermore, the quantitative phase images allow the derivation of other parameters, including dry mass production, density, and spatial distribution. We have applied DHM to study the dry mass production rate and the dry mass surface density in wild-type and mutant fission yeast cells. Our study demonstrates the applicability of DHM as a tool for label-free quantitative analysis of the cell cycle and opens the possibility for its use in high-throughput screening.

Spatiotemporal Characterization of a Fibrin Clot Using Quantitative Phase Imaging

PubMed Central

Gannavarpu, Rajshekhar; Bhaduri, Basanta; Tangella, Krishnarao; Popescu, Gabriel

2014-01-01

Studying the dynamics of fibrin clot formation and its morphology is an important problem in biology and has significant impact for several scientific and clinical applications. We present a label-free technique based on quantitative phase imaging to address this problem. Using quantitative phase information, we characterized fibrin polymerization in real-time and present a mathematical model describing the transition from liquid to gel state. By exploiting the inherent optical sectioning capability of our instrument, we measured the three-dimensional structure of the fibrin clot. From this data, we evaluated the fractal nature of the fibrin network and extracted the fractal dimension. Our non-invasive and speckle-free approach analyzes the clotting process without the need for external contrast agents. PMID:25386701

Influence of sample preparation and identification of subcelluar structures in quantitative holographic phase contrast microscopy

NASA Astrophysics Data System (ADS)

Kemper, Björn; Schmidt, Lisa; Przibilla, Sabine; Rommel, Christina; Vollmer, Angelika; Ketelhut, Steffi; Schnekenburger, Jürgen; von Bally, Gert

2010-04-01

Digital holographic microscopy (DHM) provides label-free quantitative phase contrast with low demands on sample preparation. Nevertheless, for DHM measurements on fixed cells the mounting medium has to be considered while the phase contrast of living cells may be influenced by the used buffer solution. To quantify these effects, the maximum cell caused phase contrast and the visibility of the nucleoli were analyzed. A second aim of the study was to identify subcellular components in DHM phase contrast images. Therefore, comparative investigations using bright field imaging, DHM and fluorescence microscopy with 4',6- Diamidino-2-phenylindol (DAPI) staining were performed. DAPI-staining visualizes cell components containing DNA. The obtained results demonstrate exemplarily for two tumor cell lines that from DHM phase contrast images of fixed cells in phosphate buffer saline (PBS) cell thickness values are obtained which are comparable to living cells. Furthermore, it is shown that in many cases nucleus components can be identified only by DHM phase contrast.

Fourier phase in Fourier-domain optical coherence tomography.

PubMed

Uttam, Shikhar; Liu, Yang

2015-12-01

Phase of an electromagnetic wave propagating through a sample-of-interest is well understood in the context of quantitative phase imaging in transmission-mode microscopy. In the past decade, Fourier-domain optical coherence tomography has been used to extend quantitative phase imaging to the reflection-mode. Unlike transmission-mode electromagnetic phase, however, the origin and characteristics of reflection-mode Fourier phase are poorly understood, especially in samples with a slowly varying refractive index. In this paper, the general theory of Fourier phase from first principles is presented, and it is shown that Fourier phase is a joint estimate of subresolution offset and mean spatial frequency of the coherence-gated sample refractive index. It is also shown that both spectral-domain phase microscopy and depth-resolved spatial-domain low-coherence quantitative phase microscopy are special cases of this general theory. Analytical expressions are provided for both, and simulations are presented to explain and support the theoretical results. These results are further used to show how Fourier phase allows the estimation of an axial mean spatial frequency profile of the sample, along with depth-resolved characterization of localized optical density change and sample heterogeneity. Finally, a Fourier phase-based explanation of Doppler optical coherence tomography is also provided.

Quantitative refractive index distribution of single cell by combining phase-shifting interferometry and AFM imaging.

PubMed

Zhang, Qinnan; Zhong, Liyun; Tang, Ping; Yuan, Yingjie; Liu, Shengde; Tian, Jindong; Lu, Xiaoxu

2017-05-31

Cell refractive index, an intrinsic optical parameter, is closely correlated with the intracellular mass and concentration. By combining optical phase-shifting interferometry (PSI) and atomic force microscope (AFM) imaging, we constructed a label free, non-invasive and quantitative refractive index of single cell measurement system, in which the accurate phase map of single cell was retrieved with PSI technique and the cell morphology with nanoscale resolution was achieved with AFM imaging. Based on the proposed AFM/PSI system, we achieved quantitative refractive index distributions of single red blood cell and Jurkat cell, respectively. Further, the quantitative change of refractive index distribution during Daunorubicin (DNR)-induced Jurkat cell apoptosis was presented, and then the content changes of intracellular biochemical components were achieved. Importantly, these results were consistent with Raman spectral analysis, indicating that the proposed PSI/AFM based refractive index system is likely to become a useful tool for intracellular biochemical components analysis measurement, and this will facilitate its application for revealing cell structure and pathological state from a new perspective.

Simultaneous fluorescence and quantitative phase microscopy with single-pixel detectors

NASA Astrophysics Data System (ADS)

Liu, Yang; Suo, Jinli; Zhang, Yuanlong; Dai, Qionghai

2018-02-01

Multimodal microscopy offers high flexibilities for biomedical observation and diagnosis. Conventional multimodal approaches either use multiple cameras or a single camera spatially multiplexing different modes. The former needs expertise demanding alignment and the latter suffers from limited spatial resolution. Here, we report an alignment-free full-resolution simultaneous fluorescence and quantitative phase imaging approach using single-pixel detectors. By combining reference-free interferometry with single-pixel detection, we encode the phase and fluorescence of the sample in two detection arms at the same time. Then we employ structured illumination and the correlated measurements between the sample and the illuminations for reconstruction. The recovered fluorescence and phase images are inherently aligned thanks to single-pixel detection. To validate the proposed method, we built a proof-of-concept setup for first imaging the phase of etched glass with the depth of a few hundred nanometers and then imaging the fluorescence and phase of the quantum dot drop. This method holds great potential for multispectral fluorescence microscopy with additional single-pixel detectors or a spectrometer. Besides, this cost-efficient multimodal system might find broad applications in biomedical science and neuroscience.

Label-free cell-cycle analysis by high-throughput quantitative phase time-stretch imaging flow cytometry

NASA Astrophysics Data System (ADS)

Mok, Aaron T. Y.; Lee, Kelvin C. M.; Wong, Kenneth K. Y.; Tsia, Kevin K.

2018-02-01

Biophysical properties of cells could complement and correlate biochemical markers to characterize a multitude of cellular states. Changes in cell size, dry mass and subcellular morphology, for instance, are relevant to cell-cycle progression which is prevalently evaluated by DNA-targeted fluorescence measurements. Quantitative-phase microscopy (QPM) is among the effective biophysical phenotyping tools that can quantify cell sizes and sub-cellular dry mass density distribution of single cells at high spatial resolution. However, limited camera frame rate and thus imaging throughput makes QPM incompatible with high-throughput flow cytometry - a gold standard in multiparametric cell-based assay. Here we present a high-throughput approach for label-free analysis of cell cycle based on quantitative-phase time-stretch imaging flow cytometry at a throughput of > 10,000 cells/s. Our time-stretch QPM system enables sub-cellular resolution even at high speed, allowing us to extract a multitude (at least 24) of single-cell biophysical phenotypes (from both amplitude and phase images). Those phenotypes can be combined to track cell-cycle progression based on a t-distributed stochastic neighbor embedding (t-SNE) algorithm. Using multivariate analysis of variance (MANOVA) discriminant analysis, cell-cycle phases can also be predicted label-free with high accuracy at >90% in G1 and G2 phase, and >80% in S phase. We anticipate that high throughput label-free cell cycle characterization could open new approaches for large-scale single-cell analysis, bringing new mechanistic insights into complex biological processes including diseases pathogenesis.

Time-resolved quantitative-phase microscopy of laser-material interactions using a wavefront sensor.

PubMed

Gallais, Laurent; Monneret, Serge

2016-07-15

We report on a simple and efficient technique based on a wavefront sensor to obtain time-resolved amplitude and phase images of laser-material interactions. The main interest of the technique is to obtain quantitative self-calibrated phase measurements in one shot at the femtosecond time-scale, with high spatial resolution. The technique is used for direct observation and quantitative measurement of the Kerr effect in a fused silica substrate and free electron generation by photo-ionization processes in an optical coating.

Quantitative Imaging of Single Unstained Magnetotactic Bacteria by Coherent X-ray Diffraction Microscopy.

PubMed

Fan, Jiadong; Sun, Zhibin; Zhang, Jian; Huang, Qingjie; Yao, Shengkun; Zong, Yunbing; Kohmura, Yoshiki; Ishikawa, Tetsuya; Liu, Hong; Jiang, Huaidong

2015-06-16

Novel coherent diffraction microscopy provides a powerful lensless imaging method to obtain a better understanding of the microorganism at the nanoscale. Here we demonstrated quantitative imaging of intact unstained magnetotactic bacteria using coherent X-ray diffraction microscopy combined with an iterative phase retrieval algorithm. Although the signal-to-noise ratio of the X-ray diffraction pattern from single magnetotactic bacterium is weak due to low-scattering ability of biomaterials, an 18.6 nm half-period resolution of reconstructed image was achieved by using a hybrid input-output phase retrieval algorithm. On the basis of the quantitative reconstructed images, the morphology and some intracellular structures, such as nucleoid, polyβ-hydroxybutyrate granules, and magnetosomes, were identified, which were also confirmed by scanning electron microscopy and energy dispersive spectroscopy. With the benefit from the quantifiability of coherent diffraction imaging, for the first time to our knowledge, an average density of magnetotactic bacteria was calculated to be ∼1.19 g/cm(3). This technique has a wide range of applications, especially in quantitative imaging of low-scattering biomaterials and multicomponent materials at nanoscale resolution. Combined with the cryogenic technique or X-ray free electron lasers, the method could image cells in a hydrated condition, which helps to maintain their natural structure.

Active illumination using a digital micromirror device for quantitative phase imaging.

PubMed

Shin, Seungwoo; Kim, Kyoohyun; Yoon, Jonghee; Park, YongKeun

2015-11-15

We present a powerful and cost-effective method for active illumination using a digital micromirror device (DMD) for quantitative phase-imaging techniques. Displaying binary illumination patterns on a DMD with appropriate spatial filtering, plane waves with various illumination angles are generated and impinged onto a sample. Complex optical fields of the sample obtained with various incident angles are then measured via Mach-Zehnder interferometry, from which a high-resolution 2D synthetic aperture phase image and a 3D refractive index tomogram of the sample are reconstructed. We demonstrate the fast and stable illumination-control capability of the proposed method by imaging colloidal spheres and biological cells. The capability of high-speed optical diffraction tomography is also demonstrated by measuring 3D Brownian motion of colloidal particles with the tomogram acquisition rate of 100 Hz.

Quantitative phase-contrast digital holographic microscopy for cell dynamic evaluation

NASA Astrophysics Data System (ADS)

Yu, Lingfeng; Mohanty, Samarendra; Berns, Michael W.; Chen, Zhongping

2009-02-01

The laser microbeam uses lasers to alter and/or to ablate intracellular organelles and cellular and tissue samples, and, today, has become an important tool for cell biologists to study the molecular mechanism of complex biological systems by removing individual cells or sub-cellular organelles. However, absolute quantitation of the localized alteration/damage to transparent phase objects, such as the cell membrane or chromosomes, was not possible using conventional phase-contrast or differential interference contrast microscopy. We report the development of phase-contrast digital holographic microscopy for quantitative evaluation of cell dynamic changes in real time during laser microsurgery. Quantitative phase images are recorded during the process of laser microsurgery and thus, the dynamic change in phase can be continuously evaluated. Out-of-focus organelles are re-focused by numerical reconstruction algorithms.

Transportable and vibration-free full-field low-coherent quantitative phase microscope

NASA Astrophysics Data System (ADS)

Yamauchi, Toyohiko; Yamada, Hidenao; Goto, Kentaro; Matsui, Hisayuki; Yasuhiko, Osamu; Ueda, Yukio

2018-02-01

We developed a transportable Linnik-type full-field low-coherent quantitative phase microscope that is able to compensate for optical path length (OPL) disturbance due to environmental mechanical noises. Though two-beam interferometers such as Linnik ones suffer from unstable OPL difference, we overcame this problem with a mechanical feedback system based on digital signal-processing that controls the OPL difference in sub-nanometer resolution precisely with a feedback bandwidth of 4 kHz. The developed setup has a footprint of 200 mm by 200 mm, a height of 500 mm, and a weight of 4.5 kilograms. In the transmission imaging mode, cells were cultured on a reflection-enhanced glass-bottom dish, and we obtained interference images sequentially while performing stepwise quarter-wavelength phase-shifting. Real-time image processing, including retrieval of the unwrapped phase from interference images and its background correction, along with the acquisition of interference images, was performed on a laptop computer. Emulation of the phase contrast (PhC) images and the differential interference contrast (DIC) images was also performed in real time. Moreover, our setup was applied for full-field cell membrane imaging in the reflection mode, where the cells were cultured on an anti-reflection (AR)-coated glass-bottom dish. The phase and intensity of the light reflected by the membrane revealed the outer shape of the cells independent of the refractive index. In this paper, we show imaging results on cultured cells in both transmission and reflection modes.

Comparative study of quantitative phase imaging techniques for refractometry of optical fibers

NASA Astrophysics Data System (ADS)

de Dorlodot, Bertrand; Bélanger, Erik; Bérubé, Jean-Philippe; Vallée, Réal; Marquet, Pierre

2018-02-01

The refractive index difference profile of optical fibers is the key design parameter because it determines, among other properties, the insertion losses and propagating modes. Therefore, an accurate refractive index profiling method is of paramount importance to their development and optimization. Quantitative phase imaging (QPI) is one of the available tools to retrieve structural characteristics of optical fibers, including the refractive index difference profile. Having the advantage of being non-destructive, several different QPI methods have been developed over the last decades. Here, we present a comparative study of three different available QPI techniques, namely the transport-of-intensity equation, quadriwave lateral shearing interferometry and digital holographic microscopy. To assess the accuracy and precision of those QPI techniques, quantitative phase images of the core of a well-characterized optical fiber have been retrieved for each of them and a robust image processing procedure has been applied in order to retrieve their refractive index difference profiles. As a result, even if the raw images for all the three QPI methods were suffering from different shortcomings, our robust automated image-processing pipeline successfully corrected these. After this treatment, all three QPI techniques yielded accurate, reliable and mutually consistent refractive index difference profiles in agreement with the accuracy and precision of the refracted near-field benchmark measurement.

Shot noise-limited Cramér-Rao bound and algorithmic sensitivity for wavelength shifting interferometry

NASA Astrophysics Data System (ADS)

Chen, Shichao; Zhu, Yizheng

2017-02-01

Sensitivity is a critical index to measure the temporal fluctuation of the retrieved optical pathlength in quantitative phase imaging system. However, an accurate and comprehensive analysis for sensitivity evaluation is still lacking in current literature. In particular, previous theoretical studies for fundamental sensitivity based on Gaussian noise models are not applicable to modern cameras and detectors, which are dominated by shot noise. In this paper, we derive two shot noiselimited theoretical sensitivities, Cramér-Rao bound and algorithmic sensitivity for wavelength shifting interferometry, which is a major category of on-axis interferometry techniques in quantitative phase imaging. Based on the derivations, we show that the shot noise-limited model permits accurate estimation of theoretical sensitivities directly from measured data. These results can provide important insights into fundamental constraints in system performance and can be used to guide system design and optimization. The same concepts can be generalized to other quantitative phase imaging techniques as well.

Quantitative 3D imaging of yeast by hard X-ray tomography.

PubMed

Zheng, Ting; Li, Wenjie; Guan, Yong; Song, Xiangxia; Xiong, Ying; Liu, Gang; Tian, Yangchao

2012-05-01

Full-field hard X-ray tomography could be used to obtain three-dimensional (3D) nanoscale structures of biological samples. The image of the fission yeast, Schizosaccharomyces pombe, was clearly visualized based on Zernike phase contrast imaging technique and heavy metal staining method at a spatial resolution better than 50 nm at the energy of 8 keV. The distributions and shapes of the organelles during the cell cycle were clearly visualized and two types of organelle were distinguished. The results for cells during various phases were compared and the ratios of organelle volume to cell volume can be analyzed quantitatively. It showed that the ratios remained constant between growth and division phase and increased strongly in stationary phase, following the shape and size of two types of organelles changes. Our results demonstrated that hard X-ray microscopy was a complementary method for imaging and revealing structural information for biological samples. Copyright © 2011 Wiley Periodicals, Inc.

Fourier phase in Fourier-domain optical coherence tomography

PubMed Central

Uttam, Shikhar; Liu, Yang

2015-01-01

Phase of an electromagnetic wave propagating through a sample-of-interest is well understood in the context of quantitative phase imaging in transmission-mode microscopy. In the past decade, Fourier-domain optical coherence tomography has been used to extend quantitative phase imaging to the reflection-mode. Unlike transmission-mode electromagnetic phase, however, the origin and characteristics of reflection-mode Fourier phase are poorly understood, especially in samples with a slowly varying refractive index. In this paper, the general theory of Fourier phase from first principles is presented, and it is shown that Fourier phase is a joint estimate of subresolution offset and mean spatial frequency of the coherence-gated sample refractive index. It is also shown that both spectral-domain phase microscopy and depth-resolved spatial-domain low-coherence quantitative phase microscopy are special cases of this general theory. Analytical expressions are provided for both, and simulations are presented to explain and support the theoretical results. These results are further used to show how Fourier phase allows the estimation of an axial mean spatial frequency profile of the sample, along with depth-resolved characterization of localized optical density change and sample heterogeneity. Finally, a Fourier phase-based explanation of Doppler optical coherence tomography is also provided. PMID:26831383

Holographic 3D imaging through diffuse media by compressive sampling of the mutual intensity

NASA Astrophysics Data System (ADS)

Falldorf, Claas; Klein, Thorsten; Agour, Mostafa; Bergmann, Ralf B.

2017-05-01

We present a method for holographic imaging through a volume scattering material, which is based on selfreference and light with good spatial but limited temporal coherence. In contrast to existing techniques, we do not require a separate reference wave, thus our approach provides great advantages towards the flexibility of the measurement system. The main applications are remote sensing and investigation of moving objects through gaseous streams, bubbles or foggy water for example. Furthermore, due to the common path nature, the system is also insensitive to mechanical disturbances. The measurement result is a complex amplitude which is comparable to a phase shifted digital hologramm and therefore allows 3D imaging, numerical refocusing and quantitative phase contrast imaging. As an example of application, we present measurements of the quantitative phase contrast of the epidermis of an onion through a volume scattering material.

Optical properties of acute kidney injury measured by quantitative phase imaging

PubMed Central

Ban, Sungbea; Min, Eunjung; Baek, Songyee; Kwon, Hyug Moo; Popescu, Gabriel

2018-01-01

The diagnosis of acute kidney disease (AKI) has been examined mainly by histology, immunohistochemistry and western blot. Though these approaches are widely accepted in the field, it has an inherent limitation due to the lack of high-throughput and quantitative information. For a better understanding of prognosis in AKI, we present a new approach using quantitative phase imaging combined with a wide-field scanning platform. Through the phase-delay information from the tissue, we were able to predict a stage of AKI based on various optical properties such as light scattering coefficient and anisotropy. These optical parameters quantify the deterioration process of the AKI model of tissue. Our device would be a very useful tool when it is required to deliver fast feedback of tissue pathology or when diseases are related to mechanical properties such as fibrosis. PMID:29541494

Cerebral Metabolic Rate of Oxygen (CMRO2 ) Mapping by Combining Quantitative Susceptibility Mapping (QSM) and Quantitative Blood Oxygenation Level-Dependent Imaging (qBOLD).

PubMed

Cho, Junghun; Kee, Youngwook; Spincemaille, Pascal; Nguyen, Thanh D; Zhang, Jingwei; Gupta, Ajay; Zhang, Shun; Wang, Yi

2018-03-07

To map the cerebral metabolic rate of oxygen (CMRO 2 ) by estimating the oxygen extraction fraction (OEF) from gradient echo imaging (GRE) using phase and magnitude of the GRE data. 3D multi-echo gradient echo imaging and perfusion imaging with arterial spin labeling were performed in 11 healthy subjects. CMRO 2 and OEF maps were reconstructed by joint quantitative susceptibility mapping (QSM) to process GRE phases and quantitative blood oxygen level-dependent (qBOLD) modeling to process GRE magnitudes. Comparisons with QSM and qBOLD alone were performed using ROI analysis, paired t-tests, and Bland-Altman plot. The average CMRO 2 value in cortical gray matter across subjects were 140.4 ± 14.9, 134.1 ± 12.5, and 184.6 ± 17.9 μmol/100 g/min, with corresponding OEFs of 30.9 ± 3.4%, 30.0 ± 1.8%, and 40.9 ± 2.4% for methods based on QSM, qBOLD, and QSM+qBOLD, respectively. QSM+qBOLD provided the highest CMRO 2 contrast between gray and white matter, more uniform OEF than QSM, and less noisy OEF than qBOLD. Quantitative CMRO 2 mapping that fits the entire complex GRE data is feasible by combining QSM analysis of phase and qBOLD analysis of magnitude. © 2018 International Society for Magnetic Resonance in Medicine.

Technique to quantitatively measure magnetic properties of thin structures at <10 NM spatial resolution

DOEpatents

Bajt, Sasa

2003-07-08

A highly sensitive and high resolution magnetic microscope images magnetic properties quantitatively. Imaging is done with a modified transmission electron microscope that allows imaging of the sample in a zero magnetic field. Two images from closely spaced planes, one in focus and one slightly out of focus, are sufficient to calculate the absolute values of the phase change imparted to the electrons, and hence obtain the magnetization vector field distribution.

Susceptibility-Weighted Imaging and Quantitative Susceptibility Mapping in the Brain

PubMed Central

Liu, Chunlei; Li, Wei; Tong, Karen A.; Yeom, Kristen W.; Kuzminski, Samuel

2015-01-01

Susceptibility-weighted imaging (SWI) is a magnetic resonance imaging (MRI) technique that enhances image contrast by using the susceptibility differences between tissues. It is created by combining both magnitude and phase in the gradient echo data. SWI is sensitive to both paramagnetic and diamagnetic substances which generate different phase shift in MRI data. SWI images can be displayed as a minimum intensity projection that provides high resolution delineation of the cerebral venous architecture, a feature that is not available in other MRI techniques. As such, SWI has been widely applied to diagnose various venous abnormalities. SWI is especially sensitive to deoxygenated blood and intracranial mineral deposition and, for that reason, has been applied to image various pathologies including intracranial hemorrhage, traumatic brain injury, stroke, neoplasm, and multiple sclerosis. SWI, however, does not provide quantitative measures of magnetic susceptibility. This limitation is currently being addressed with the development of quantitative susceptibility mapping (QSM) and susceptibility tensor imaging (STI). While QSM treats susceptibility as isotropic, STI treats susceptibility as generally anisotropic characterized by a tensor quantity. This article reviews the basic principles of SWI, its clinical and research applications, the mechanisms governing brain susceptibility properties, and its practical implementation, with a focus on brain imaging. PMID:25270052

Multi-spectral digital holographic microscopy for enhanced quantitative phase imaging of living cells

NASA Astrophysics Data System (ADS)

Kemper, Björn; Kastl, Lena; Schnekenburger, Jürgen; Ketelhut, Steffi

2018-02-01

Main restrictions of using laser light in digital holographic microscopy (DHM) are coherence induced noise and parasitic reflections in the experimental setup which limit resolution and measurement accuracy. We explored, if coherence properties of partial coherent light sources can be generated synthetically utilizing spectrally tunable lasers. The concept of the method is demonstrated by label-free quantitative phase imaging of living pancreatic tumor cells and utilizing an experimental configuration including a commercial microscope and a laser source with a broad tunable spectral range of more than 200 nm.

Assessment of imaging quality in magnified phase CT of human bone tissue at the nanoscale

NASA Astrophysics Data System (ADS)

Yu, Boliang; Langer, Max; Pacureanu, Alexandra; Gauthier, Remy; Follet, Helene; Mitton, David; Olivier, Cecile; Cloetens, Peter; Peyrin, Francoise

2017-10-01

Bone properties at all length scales have a major impact on the fracture risk in disease such as osteoporosis. However, quantitative 3D data on bone tissue at the cellular scale are still rare. Here we propose to use magnified X-ray phase nano-CT to quantify bone ultra-structure in human bone, on the new setup developed on the beamline ID16A at the ESRF, Grenoble. Obtaining 3D images requires the application of phase retrieval prior to tomographic reconstruction. Phase retrieval is an ill-posed problem for which various approaches have been developed. Since image quality has a strong impact on the further quantification of bone tissue, our aim here is to evaluate different phase retrieval methods for imaging bone samples at the cellular scale. Samples from femurs of female donors were scanned using magnified phase nano-CT at voxel sizes of 120 and 30 nm with an energy of 33 keV. Four CT scans at varying sample-to-detector distances were acquired for each sample. We evaluated three phase retrieval methods adapted to these conditions: Paganin's method at single distance, Paganin's method extended to multiple distances, and the contrast transfer function (CTF) approach for pure phase objects. These methods were used as initialization to an iterative refinement step. Our results based on visual and quantitative assessment show that the use of several distances (as opposed to single one) clearly improves image quality and the two multi-distance phase retrieval methods give similar results. First results on the segmentation of osteocyte lacunae and canaliculi from such images are presented.

Quantitative phase imaging of cell division in yeast cells and E.coli using digital holographic microscopy

NASA Astrophysics Data System (ADS)

Pandiyan, Vimal Prabhu; John, Renu

2015-12-01

Digital holographic microscope (DHM) is an emerging quantitative phase imaging technique with unique imaging scales and resolutions leading to multitude of applications. DHM is promising as a novel investigational and applied tool for cell imaging, studying the morphology and real time dynamics of cells and a number of related applications. The use of numerical propagation and computational digital optics offer unique flexibility to tune the depth of focus, and compensate for image aberrations. In this work, we report imaging the dynamics of cell division in E.coli and yeast cells using a DHM platform. We demonstrate 3-D and depth imaging as well as reconstruction of phase profiles of E.coli and yeast cells using the system. We record a digital hologram of E.coli and yeast cells and reconstruct the image using Fresnel propagation algorithm. We also use aberration compensation algorithms for correcting the aberrations that are introduced by the microscope objective in the object path using linear least square fitting techniques. This work demonstrates the strong potential of a DHM platform in 3-D live cell imaging, fast clinical quantifications and pathological applications.

Isotropic scalar image visualization of vector differential image data using the inverse Riesz transform.

PubMed

Larkin, Kieran G; Fletcher, Peter A

2014-03-01

X-ray Talbot moiré interferometers can now simultaneously generate two differential phase images of a specimen. The conventional approach to integrating differential phase is unstable and often leads to images with loss of visible detail. We propose a new reconstruction method based on the inverse Riesz transform. The Riesz approach is stable and the final image retains visibility of high resolution detail without directional bias. The outline Riesz theory is developed and an experimentally acquired X-ray differential phase data set is presented for qualitative visual appraisal. The inverse Riesz phase image is compared with two alternatives: the integrated (quantitative) phase and the modulus of the gradient of the phase. The inverse Riesz transform has the computational advantages of a unitary linear operator, and is implemented directly as a complex multiplication in the Fourier domain also known as the spiral phase transform.

Isotropic scalar image visualization of vector differential image data using the inverse Riesz transform

PubMed Central

Larkin, Kieran G.; Fletcher, Peter A.

2014-01-01

X-ray Talbot moiré interferometers can now simultaneously generate two differential phase images of a specimen. The conventional approach to integrating differential phase is unstable and often leads to images with loss of visible detail. We propose a new reconstruction method based on the inverse Riesz transform. The Riesz approach is stable and the final image retains visibility of high resolution detail without directional bias. The outline Riesz theory is developed and an experimentally acquired X-ray differential phase data set is presented for qualitative visual appraisal. The inverse Riesz phase image is compared with two alternatives: the integrated (quantitative) phase and the modulus of the gradient of the phase. The inverse Riesz transform has the computational advantages of a unitary linear operator, and is implemented directly as a complex multiplication in the Fourier domain also known as the spiral phase transform. PMID:24688823

Quantitative energy-filtered TEM imaging of interfaces

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bentley, J.; Kenik, E.A.; Siangchaew, K.

Quantitative elemental mapping by inner shell core-loss energy-filtered transmission electron microscopy (TEM) with a Gatan Imaging Filter (GIF) interfaced to a Philips CM30 TEM operated with a LaB{sub 6} filament at 300 kV has been applied to interfaces in a range of materials. In sensitized type 304L stainless steel aged 15 h at 600{degrees}C, grain-boundary Cr depletion occurs between Cr-rich intergranular M{sub 23}C{sub 6} particles. Images of net Cr L{sub 23} intensity show segregation profiles that agree quantitatively with focused-probe spectrum-line measurements recorded with a Gatan PEELS on a Philips EM400T/FEG (0.8 nA in 2-nm-diam probe) of the same regions.more » Rare-earth oxide additives that are used for the liquid-phase sintering of Si{sub 3}N{sub 4} generate second phases of complex composition at grain boundaries and edges. These grain boundary phases often control corrosion, crack growth and creep damage behavior. High resolution imaging has been widely and with focused probes can be compromised by beam damage, but elemental mapping by EFTEM appears not to cause appreciable beam damage.« less

Common-path digital holographic microscopy based on a beam displacer unit

NASA Astrophysics Data System (ADS)

Di, Jianglei; Zhang, Jiwei; Song, Yu; Wang, Kaiqiang; Wei, Kun; Zhao, Jianlin

2018-02-01

Digital holographic microscopy (DHM) has become a novel tool with advantages of full field, non-destructive, high-resolution and 3D imaging, which captures the quantitative amplitude and phase information of microscopic specimens. It's a well-established method for digital recording and numerical reconstructing the full complex field of wavefront of the samples with a diffraction-limited lateral resolution down to 0.3 μm depending on the numerical aperture of microscope objective. Meanwhile, its axial resolution through axial direction is less than 10 nm due to the interferometric nature in phase imaging. Compared with the typical optical configurations such as Mach-Zehnder interferometer and Michelson interferometer, the common-path DHM has the advantages of simple and compact configuration, high stability, and so on. Here, a simple, compact, and low-cost common-path DHM based on a beam displacer unit is proposed for quantitative phase imaging of biological cells. The beam displacer unit is completely compatible with commercial microscope and can be easily set up in the output port of the microscope as a compact independent device. This technique can be used to achieve the quantitative phase measurement of biological cells with an excellent temporal stability of 0.51 nm, which makes it having a good prospect in the fields of biological and medical science. Living mouse osteoblastic cells are quantitatively measured with the system to demonstrate its capability and applicability.

The importance of the boundary condition in the transport of intensity equation based phase measurement

NASA Astrophysics Data System (ADS)

Zhang, Jialin; Chen, Qian; Li, Jiaji; Zuo, Chao

2017-02-01

The transport of intensity equation (TIE) is a powerful tool for direct quantitative phase retrieval in microscopy imaging. However, there may be some problems when dealing with the boundary condition of the TIE. The previous work introduces a hard-edged aperture to the camera port of the traditional bright field microscope to generate the boundary signal for the TIE solver. Under this Neumann boundary condition, we can obtain the quantitative phase without any assumption or prior knowledge about the test object and the setup. In this paper, we will demonstrate the effectiveness of this method based on some experiments in practice. The micro lens array will be used for the comparison of two TIE solvers results based on introducing the aperture or not and this accurate quantitative phase imaging technique allows measuring cell dry mass which is used in biology to follow cell cycle, to investigate cell metabolism, or to address effects of drugs.

Accurate and quantitative polarization-sensitive OCT by unbiased birefringence estimator with noise-stochastic correction

NASA Astrophysics Data System (ADS)

Kasaragod, Deepa; Sugiyama, Satoshi; Ikuno, Yasushi; Alonso-Caneiro, David; Yamanari, Masahiro; Fukuda, Shinichi; Oshika, Tetsuro; Hong, Young-Joo; Li, En; Makita, Shuichi; Miura, Masahiro; Yasuno, Yoshiaki

2016-03-01

Polarization sensitive optical coherence tomography (PS-OCT) is a functional extension of OCT that contrasts the polarization properties of tissues. It has been applied to ophthalmology, cardiology, etc. Proper quantitative imaging is required for a widespread clinical utility. However, the conventional method of averaging to improve the signal to noise ratio (SNR) and the contrast of the phase retardation (or birefringence) images introduce a noise bias offset from the true value. This bias reduces the effectiveness of birefringence contrast for a quantitative study. Although coherent averaging of Jones matrix tomography has been widely utilized and has improved the image quality, the fundamental limitation of nonlinear dependency of phase retardation and birefringence to the SNR was not overcome. So the birefringence obtained by PS-OCT was still not accurate for a quantitative imaging. The nonlinear effect of SNR to phase retardation and birefringence measurement was previously formulated in detail for a Jones matrix OCT (JM-OCT) [1]. Based on this, we had developed a maximum a-posteriori (MAP) estimator and quantitative birefringence imaging was demonstrated [2]. However, this first version of estimator had a theoretical shortcoming. It did not take into account the stochastic nature of SNR of OCT signal. In this paper, we present an improved version of the MAP estimator which takes into account the stochastic property of SNR. This estimator uses a probability distribution function (PDF) of true local retardation, which is proportional to birefringence, under a specific set of measurements of the birefringence and SNR. The PDF was pre-computed by a Monte-Carlo (MC) simulation based on the mathematical model of JM-OCT before the measurement. A comparison between this new MAP estimator, our previous MAP estimator [2], and the standard mean estimator is presented. The comparisons are performed both by numerical simulation and in vivo measurements of anterior and posterior eye segment as well as in skin imaging. The new estimator shows superior performance and also shows clearer image contrast.

An iterative method for near-field Fresnel region polychromatic phase contrast imaging

NASA Astrophysics Data System (ADS)

Carroll, Aidan J.; van Riessen, Grant A.; Balaur, Eugeniu; Dolbnya, Igor P.; Tran, Giang N.; Peele, Andrew G.

2017-07-01

We present an iterative method for polychromatic phase contrast imaging that is suitable for broadband illumination and which allows for the quantitative determination of the thickness of an object given the refractive index of the sample material. Experimental and simulation results suggest the iterative method provides comparable image quality and quantitative object thickness determination when compared to the analytical polychromatic transport of intensity and contrast transfer function methods. The ability of the iterative method to work over a wider range of experimental conditions means the iterative method is a suitable candidate for use with polychromatic illumination and may deliver more utility for laboratory-based x-ray sources, which typically have a broad spectrum.

Understanding the optics to aid microscopy image segmentation.

PubMed

Yin, Zhaozheng; Li, Kang; Kanade, Takeo; Chen, Mei

2010-01-01

Image segmentation is essential for many automated microscopy image analysis systems. Rather than treating microscopy images as general natural images and rushing into the image processing warehouse for solutions, we propose to study a microscope's optical properties to model its image formation process first using phase contrast microscopy as an exemplar. It turns out that the phase contrast imaging system can be relatively well explained by a linear imaging model. Using this model, we formulate a quadratic optimization function with sparseness and smoothness regularizations to restore the "authentic" phase contrast images that directly correspond to specimen's optical path length without phase contrast artifacts such as halo and shade-off. With artifacts removed, high quality segmentation can be achieved by simply thresholding the restored images. The imaging model and restoration method are quantitatively evaluated on two sequences with thousands of cells captured over several days.

Identification of malaria infected red blood samples by digital holographic quantitative phase microscope

NASA Astrophysics Data System (ADS)

Patel, Nimit R.; Chhaniwal, Vani K.; Javidi, Bahram; Anand, Arun

2015-07-01

Development of devices for automatic identification of diseases is desired especially in developing countries. In the case of malaria, even today the gold standard is the inspection of chemically treated blood smears through a microscope. This requires a trained technician/microscopist to identify the cells in the field of view, with which the labeling chemicals gets attached. Bright field microscopes provide only low contrast 2D images of red blood cells and cell thickness distribution cannot be obtained. Quantitative phase contrast microscopes can provide both intensity and phase profiles of the cells under study. The phase information can be used to determine thickness profile of the cell. Since cell morphology is available, many parameters pertaining to the 3D shape of the cell can be computed. These parameters in turn could be used to decide about the state of health of the cell leading to disease diagnosis. Here the investigations done on digital holographic microscope, which provides quantitative phase images, for comparison of parameters obtained from the 3D shape profile of objects leading to identification of diseased samples is described.

Non-interferometric phase retrieval using refractive index manipulation.

PubMed

Chen, Chyong-Hua; Hsu, Hsin-Feng; Chen, Hou-Ren; Hsieh, Wen-Feng

2017-04-07

We present a novel, inexpensive and non-interferometric technique to retrieve phase images by using a liquid crystal phase shifter without including any physically moving parts. First, we derive a new equation of the intensity-phase relation with respect to the change of refractive index, which is similar to the transport of the intensity equation. The equation indicates that this technique is unneeded to consider the variation of magnifications between optical images. For proof of the concept, we use a liquid crystal mixture MLC 2144 to manufacture a phase shifter and to capture the optical images in a rapid succession by electrically tuning the applied voltage of the phase shifter. Experimental results demonstrate that this technique is capable of reconstructing high-resolution phase images and to realize the thickness profile of a microlens array quantitatively.

Ultra-fast quantitative imaging using ptychographic iterative engine based digital micro-mirror device

NASA Astrophysics Data System (ADS)

Sun, Aihui; Tian, Xiaolin; Kong, Yan; Jiang, Zhilong; Liu, Fei; Xue, Liang; Wang, Shouyu; Liu, Cheng

2018-01-01

As a lensfree imaging technique, ptychographic iterative engine (PIE) method can provide both quantitative sample amplitude and phase distributions avoiding aberration. However, it requires field of view (FoV) scanning often relying on mechanical translation, which not only slows down measuring speed, but also introduces mechanical errors decreasing both resolution and accuracy in retrieved information. In order to achieve high-accurate quantitative imaging with fast speed, digital micromirror device (DMD) is adopted in PIE for large FoV scanning controlled by on/off state coding by DMD. Measurements were implemented using biological samples as well as USAF resolution target, proving high resolution in quantitative imaging using the proposed system. Considering its fast and accurate imaging capability, it is believed the DMD based PIE technique provides a potential solution for medical observation and measurements.

Automatic phase aberration compensation for digital holographic microscopy based on deep learning background detection.

PubMed

Nguyen, Thanh; Bui, Vy; Lam, Van; Raub, Christopher B; Chang, Lin-Ching; Nehmetallah, George

2017-06-26

We propose a fully automatic technique to obtain aberration free quantitative phase imaging in digital holographic microscopy (DHM) based on deep learning. The traditional DHM solves the phase aberration compensation problem by manually detecting the background for quantitative measurement. This would be a drawback in real time implementation and for dynamic processes such as cell migration phenomena. A recent automatic aberration compensation approach using principle component analysis (PCA) in DHM avoids human intervention regardless of the cells' motion. However, it corrects spherical/elliptical aberration only and disregards the higher order aberrations. Traditional image segmentation techniques can be employed to spatially detect cell locations. Ideally, automatic image segmentation techniques make real time measurement possible. However, existing automatic unsupervised segmentation techniques have poor performance when applied to DHM phase images because of aberrations and speckle noise. In this paper, we propose a novel method that combines a supervised deep learning technique with convolutional neural network (CNN) and Zernike polynomial fitting (ZPF). The deep learning CNN is implemented to perform automatic background region detection that allows for ZPF to compute the self-conjugated phase to compensate for most aberrations.

Brain Injury Lesion Imaging Using Preconditioned Quantitative Susceptibility Mapping without Skull Stripping.

PubMed

Soman, S; Liu, Z; Kim, G; Nemec, U; Holdsworth, S J; Main, K; Lee, B; Kolakowsky-Hayner, S; Selim, M; Furst, A J; Massaband, P; Yesavage, J; Adamson, M M; Spincemallie, P; Moseley, M; Wang, Y

2018-04-01

Identifying cerebral microhemorrhage burden can aid in the diagnosis and management of traumatic brain injury, stroke, hypertension, and cerebral amyloid angiopathy. MR imaging susceptibility-based methods are more sensitive than CT for detecting cerebral microhemorrhage, but methods other than quantitative susceptibility mapping provide results that vary with field strength and TE, require additional phase maps to distinguish blood from calcification, and depict cerebral microhemorrhages as bloom artifacts. Quantitative susceptibility mapping provides universal quantification of tissue magnetic property without these constraints but traditionally requires a mask generated by skull-stripping, which can pose challenges at tissue interphases. We evaluated the preconditioned quantitative susceptibility mapping MR imaging method, which does not require skull-stripping, for improved depiction of brain parenchyma and pathology. Fifty-six subjects underwent brain MR imaging with a 3D multiecho gradient recalled echo acquisition. Mask-based quantitative susceptibility mapping images were created using a commonly used mask-based quantitative susceptibility mapping method, and preconditioned quantitative susceptibility images were made using precondition-based total field inversion. All images were reviewed by a neuroradiologist and a radiology resident. Ten subjects (18%), all with traumatic brain injury, demonstrated blood products on 3D gradient recalled echo imaging. All lesions were visible on preconditioned quantitative susceptibility mapping, while 6 were not visible on mask-based quantitative susceptibility mapping. Thirty-one subjects (55%) demonstrated brain parenchyma and/or lesions that were visible on preconditioned quantitative susceptibility mapping but not on mask-based quantitative susceptibility mapping. Six subjects (11%) demonstrated pons artifacts on preconditioned quantitative susceptibility mapping and mask-based quantitative susceptibility mapping; they were worse on preconditioned quantitative susceptibility mapping. Preconditioned quantitative susceptibility mapping MR imaging can bring the benefits of quantitative susceptibility mapping imaging to clinical practice without the limitations of mask-based quantitative susceptibility mapping, especially for evaluating cerebral microhemorrhage-associated pathologies, such as traumatic brain injury. © 2018 by American Journal of Neuroradiology.

Breast cancer diagnosis using spatial light interference microscopy

NASA Astrophysics Data System (ADS)

Majeed, Hassaan; Kandel, Mikhail E.; Han, Kevin; Luo, Zelun; Macias, Virgilia; Tangella, Krishnarao; Balla, Andre; Popescu, Gabriel

2015-11-01

The standard practice in histopathology of breast cancers is to examine a hematoxylin and eosin (H&E) stained tissue biopsy under a microscope to diagnose whether a lesion is benign or malignant. This determination is made based on a manual, qualitative inspection, making it subject to investigator bias and resulting in low throughput. Hence, a quantitative, label-free, and high-throughput diagnosis method is highly desirable. We present here preliminary results showing the potential of quantitative phase imaging for breast cancer screening and help with differential diagnosis. We generated phase maps of unstained breast tissue biopsies using spatial light interference microscopy (SLIM). As a first step toward quantitative diagnosis based on SLIM, we carried out a qualitative evaluation of our label-free images. These images were shown to two pathologists who classified each case as either benign or malignant. This diagnosis was then compared against the diagnosis of the two pathologists on corresponding H&E stained tissue images and the number of agreements were counted. The agreement between SLIM and H&E based diagnosis was 88% for the first pathologist and 87% for the second. Our results demonstrate the potential and promise of SLIM for quantitative, label-free, and high-throughput diagnosis.

Automatic Gleason grading of prostate cancer using quantitative phase imaging and machine learning

NASA Astrophysics Data System (ADS)

Nguyen, Tan H.; Sridharan, Shamira; Macias, Virgilia; Kajdacsy-Balla, Andre; Melamed, Jonathan; Do, Minh N.; Popescu, Gabriel

2017-03-01

We present an approach for automatic diagnosis of tissue biopsies. Our methodology consists of a quantitative phase imaging tissue scanner and machine learning algorithms to process these data. We illustrate the performance by automatic Gleason grading of prostate specimens. The imaging system operates on the principle of interferometry and, as a result, reports on the nanoscale architecture of the unlabeled specimen. We use these data to train a random forest classifier to learn textural behaviors of prostate samples and classify each pixel in the image into different classes. Automatic diagnosis results were computed from the segmented regions. By combining morphological features with quantitative information from the glands and stroma, logistic regression was used to discriminate regions with Gleason grade 3 versus grade 4 cancer in prostatectomy tissue. The overall accuracy of this classification derived from a receiver operating curve was 82%, which is in the range of human error when interobserver variability is considered. We anticipate that our approach will provide a clinically objective and quantitative metric for Gleason grading, allowing us to corroborate results across instruments and laboratories and feed the computer algorithms for improved accuracy.

Quantitative phase imaging for enhanced assessment of optomechanical cancer cell properties

NASA Astrophysics Data System (ADS)

Kastl, Lena; Kemper, Björn; Schnekenburger, Jürgen

2018-02-01

Optical cell stretching provides label-free investigations of cells by measuring their biomechanical properties based on deformability determination in a fiber optical two-beam trap. However, the stretching forces in this two-beam laser trap depend on the optical properties of the investigated specimen. Therefore, we characterized in parallel four cancer cell lines with varying degree of differentiation utilizing quantitative phase imaging (QPI) and optical cell stretching. The QPI data allowed enhanced assessment of the mechanical cell properties measured with the optical cell stretcher and demonstrates the high potential of cell phenotyping when both techniques are combined.

Combined multi-plane phase retrieval and super-resolution optical fluctuation imaging for 4D cell microscopy

NASA Astrophysics Data System (ADS)

Descloux, A.; Grußmayer, K. S.; Bostan, E.; Lukes, T.; Bouwens, A.; Sharipov, A.; Geissbuehler, S.; Mahul-Mellier, A.-L.; Lashuel, H. A.; Leutenegger, M.; Lasser, T.

2018-03-01

Super-resolution fluorescence microscopy provides unprecedented insight into cellular and subcellular structures. However, going `beyond the diffraction barrier' comes at a price, since most far-field super-resolution imaging techniques trade temporal for spatial super-resolution. We propose the combination of a novel label-free white light quantitative phase imaging with fluorescence to provide high-speed imaging and spatial super-resolution. The non-iterative phase retrieval relies on the acquisition of single images at each z-location and thus enables straightforward 3D phase imaging using a classical microscope. We realized multi-plane imaging using a customized prism for the simultaneous acquisition of eight planes. This allowed us to not only image live cells in 3D at up to 200 Hz, but also to integrate fluorescence super-resolution optical fluctuation imaging within the same optical instrument. The 4D microscope platform unifies the sensitivity and high temporal resolution of phase imaging with the specificity and high spatial resolution of fluorescence microscopy.

Resolving phase information of the optical local density of state with scattering near-field probes

NASA Astrophysics Data System (ADS)

Prasad, R.; Vincent, R.

2016-10-01

We theoretically discuss the link between the phase measured using a scattering optical scanning near-field microscopy (s-SNOM) and the local density of optical states (LDOS). A remarkable result is that the LDOS information is directly included in the phase of the probe. Therefore by monitoring the spatial variation of the trans-scattering phase, we locally measure the phase modulation associated with the probe and the optical paths. We demonstrate numerically that a technique involving two-phase imaging of a sample with two different sized tips should allow to obtain the image the pLDOS. For this imaging method, numerical comparison with extinction probe measurement shows crucial qualitative and quantitative improvement.

High sensitivity phase retrieval method in grating-based x-ray phase contrast imaging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wu, Zhao; Gao, Kun; Chen, Jian

2015-02-15

Purpose: Grating-based x-ray phase contrast imaging is considered as one of the most promising techniques for future medical imaging. Many different methods have been developed to retrieve phase signal, among which the phase stepping (PS) method is widely used. However, further practical implementations are hindered, due to its complex scanning mode and high radiation dose. In contrast, the reverse projection (RP) method is a novel fast and low dose extraction approach. In this contribution, the authors present a quantitative analysis of the noise properties of the refraction signals retrieved by the two methods and compare their sensitivities. Methods: Using themore » error propagation formula, the authors analyze theoretically the signal-to-noise ratios (SNRs) of the refraction images retrieved by the two methods. Then, the sensitivities of the two extraction methods are compared under an identical exposure dose. Numerical experiments are performed to validate the theoretical results and provide some quantitative insight. Results: The SNRs of the two methods are both dependent on the system parameters, but in different ways. Comparison between their sensitivities reveals that for the refraction signal, the RP method possesses a higher sensitivity, especially in the case of high visibility and/or at the edge of the object. Conclusions: Compared with the PS method, the RP method has a superior sensitivity and provides refraction images with a higher SNR. Therefore, one can obtain highly sensitive refraction images in grating-based phase contrast imaging. This is very important for future preclinical and clinical implementations.« less

Quantitative phase microscopy via optimized inversion of the phase optical transfer function.

PubMed

Jenkins, Micah H; Gaylord, Thomas K

2015-10-01

Although the field of quantitative phase imaging (QPI) has wide-ranging biomedical applicability, many QPI methods are not well-suited for such applications due to their reliance on coherent illumination and specialized hardware. By contrast, methods utilizing partially coherent illumination have the potential to promote the widespread adoption of QPI due to their compatibility with microscopy, which is ubiquitous in the biomedical community. Described herein is a new defocus-based reconstruction method that utilizes a small number of efficiently sampled micrographs to optimally invert the partially coherent phase optical transfer function under assumptions of weak absorption and slowly varying phase. Simulation results are provided that compare the performance of this method with similar algorithms and demonstrate compatibility with large phase objects. The accuracy of the method is validated experimentally using a microlens array as a test phase object. Lastly, time-lapse images of live adherent cells are obtained with an off-the-shelf microscope, thus demonstrating the new method's potential for extending QPI capability widely in the biomedical community.

Single and two-shot quantitative phase imaging using Hilbert-Huang Transform based fringe pattern analysis

NASA Astrophysics Data System (ADS)

Trusiak, Maciej; Micó, Vicente; Patorski, Krzysztof; García-Monreal, Javier; Sluzewski, Lukasz; Ferreira, Carlos

2016-08-01

In this contribution we propose two Hilbert-Huang Transform based algorithms for fast and accurate single-shot and two-shot quantitative phase imaging applicable in both on-axis and off-axis configurations. In the first scheme a single fringe pattern containing information about biological phase-sample under study is adaptively pre-filtered using empirical mode decomposition based approach. Further it is phase demodulated by the Hilbert Spiral Transform aided by the Principal Component Analysis for the local fringe orientation estimation. Orientation calculation enables closed fringes efficient analysis and can be avoided using arbitrary phase-shifted two-shot Gram-Schmidt Orthonormalization scheme aided by Hilbert-Huang Transform pre-filtering. This two-shot approach is a trade-off between single-frame and temporal phase shifting demodulation. Robustness of the proposed techniques is corroborated using experimental digital holographic microscopy studies of polystyrene micro-beads and red blood cells. Both algorithms compare favorably with the temporal phase shifting scheme which is used as a reference method.

Quantitative photoacoustic elasticity and viscosity imaging for cirrhosis detection

NASA Astrophysics Data System (ADS)

Wang, Qian; Shi, Yujiao; Yang, Fen; Yang, Sihua

2018-05-01

Elasticity and viscosity assessments are essential for understanding and characterizing the physiological and pathological states of tissue. In this work, by establishing a photoacoustic (PA) shear wave model, an approach for quantitative PA elasticity imaging based on measurement of the rise time of the thermoelastic displacement was developed. Thus, using an existing PA viscoelasticity imaging method that features a phase delay measurement, quantitative PA elasticity imaging and viscosity imaging can be obtained in a simultaneous manner. The method was tested and validated by imaging viscoelastic agar phantoms prepared at different agar concentrations, and the imaging data were in good agreement with rheometry results. Ex vivo experiments on liver pathological models demonstrated the capability for cirrhosis detection, and the results were consistent with the corresponding histological results. This method expands the scope of conventional PA imaging and has potential to become an important alternative imaging modality.

Multimodal imaging of ischemic wounds

NASA Astrophysics Data System (ADS)

Zhang, Shiwu; Gnyawali, Surya; Huang, Jiwei; Liu, Peng; Gordillo, Gayle; Sen, Chandan K.; Xu, Ronald

2012-12-01

The wound healing process involves the reparative phases of inflammation, proliferation, and remodeling. Interrupting any of these phases may result in chronically unhealed wounds, amputation, or even patient death. Quantitative assessment of wound tissue ischemia, perfusion, and inflammation provides critical information for appropriate detection, staging, and treatment of chronic wounds. However, no method is available for noninvasive, simultaneous, and quantitative imaging of these tissue parameters. We integrated hyperspectral, laser speckle, and thermographic imaging modalities into a single setup for multimodal assessment of tissue oxygenation, perfusion, and inflammation characteristics. Advanced algorithms were developed for accurate reconstruction of wound oxygenation and appropriate co-registration between different imaging modalities. The multimodal wound imaging system was validated by an ongoing clinical trials approved by OSU IRB. In the clinical trial, a wound of 3mm in diameter was introduced on a healthy subject's lower extremity and the healing process was serially monitored by the multimodal imaging setup. Our experiments demonstrated the clinical usability of multimodal wound imaging.

Non-interferometric phase retrieval using refractive index manipulation

PubMed Central

Chen, Chyong-Hua; Hsu, Hsin-Feng; Chen, Hou-Ren; Hsieh, Wen-Feng

2017-01-01

We present a novel, inexpensive and non-interferometric technique to retrieve phase images by using a liquid crystal phase shifter without including any physically moving parts. First, we derive a new equation of the intensity-phase relation with respect to the change of refractive index, which is similar to the transport of the intensity equation. The equation indicates that this technique is unneeded to consider the variation of magnifications between optical images. For proof of the concept, we use a liquid crystal mixture MLC 2144 to manufacture a phase shifter and to capture the optical images in a rapid succession by electrically tuning the applied voltage of the phase shifter. Experimental results demonstrate that this technique is capable of reconstructing high-resolution phase images and to realize the thickness profile of a microlens array quantitatively. PMID:28387382

Quantitative phase imaging using grating-based quadrature phase interferometer

NASA Astrophysics Data System (ADS)

Wu, Jigang; Yaqoob, Zahid; Heng, Xin; Cui, Xiquan; Yang, Changhuei

2007-02-01

In this paper, we report the use of holographic gratings, which act as the free-space equivalent of the 3x3 fiber-optic coupler, to perform full field phase imaging. By recording two harmonically-related gratings in the same holographic plate, we are able to obtain nontrivial phase shift between different output ports of the gratings-based Mach-Zehnder interferometer. The phase difference can be adjusted by changing the relative phase of the recording beams when recording the hologram. We have built a Mach-Zehnder interferometer using harmonically-related holographic gratings with 600 and 1200 lines/mm spacing. Two CCD cameras at the output ports of the gratings-based Mach-Zehnder interferometer are used to record the full-field quadrature interferograms, which are subsequently processed to reconstruct the phase image. The imaging system has ~12X magnification with ~420μmx315μm field-of-view. To demonstrate the capability of our system, we have successfully performed phase imaging of a pure phase object and a paramecium caudatum.

Studying the relationship between redox and cell growth using quantitative phase imaging (Conference Presentation)

NASA Astrophysics Data System (ADS)

Sridharan, Shamira; Leslie, Matthew T.; Bapst, Natalya; Smith, John; Gaskins, H. Rex; Popescu, Gabriel

2016-03-01

Quantitative phase imaging has been used in the past to study the dry mass of cells and study cell growth under various treatment conditions. However, the relationship between cellular redox and growth rates has not yet been studied in this context. This study employed the recombinant Glrx-roGFP2 redox biosensor targeted to the mitochondrial matrix or cytosolic compartments of A549 lung epithelial carcinoma cells. The Glrx-roGFP2s biosensor consists of a modified GFP protein containing internal cysteine residues sensitive to the local redox environment. The formation/dissolution of sulfide bridges contorts the internal chromophore, dictating corresponding changes in florescence emission that provide direct measures of the local redox potential. Combining 2-channel florescent imaging of the redox sensor with quantitative phase imaging allowed observation of redox homeostasis alongside measurements of cellular mass during full cycles of cellular division. The results indicate that mitochondrial redox showed a stronger inverse correlation with cell growth than cytoplasmic redox states; although redox changes are restricted to a 5% range. We are now studying the relationship between mitochondrial redox and cell growth in an isogenic series of breast cell lines built upon the MCF-10A genetic background that vary both in malignancy and metastatic potential.

A general theory of interference fringes in x-ray phase grating imaging.

PubMed

Yan, Aimin; Wu, Xizeng; Liu, Hong

2015-06-01

The authors note that the concept of the Talbot self-image distance in x-ray phase grating interferometry is indeed not well defined for polychromatic x-rays, because both the grating phase shift and the fractional Talbot distances are all x-ray wavelength-dependent. For x-ray interferometry optimization, there is a need for a quantitative theory that is able to predict if a good intensity modulation is attainable at a given grating-to-detector distance. In this work, the authors set out to meet this need. In order to apply Fourier analysis directly to the intensity fringe patterns of two-dimensional and one-dimensional phase grating interferometers, the authors start their derivation from a general phase space theory of x-ray phase-contrast imaging. Unlike previous Fourier analyses, the authors evolved the Wigner distribution to obtain closed-form expressions of the Fourier coefficients of the intensity fringes for any grating-to-detector distance, even if it is not a fractional Talbot distance. The developed theory determines the visibility of any diffraction order as a function of the grating-to-detector distance, the phase shift of the grating, and the x-ray spectrum. The authors demonstrate that the visibilities of diffraction orders can serve as the indicators of the underlying interference intensity modulation. Applying the theory to the conventional and inverse geometry configurations of single-grating interferometers, the authors demonstrated that the proposed theory provides a quantitative tool for the grating interferometer optimization with or without the Talbot-distance constraints. In this work, the authors developed a novel theory of the interference intensity fringes in phase grating x-ray interferometry. This theory provides a quantitative tool in design optimization of phase grating x-ray interferometers.

Quantitative multi-modality imaging analysis of a bioabsorbable poly-L-lactic acid stent design in the acute phase: a comparison between 2- and 3D-QCA, QCU and QMSCT-CA.

PubMed

Bruining, Nico; Tanimoto, Shuzou; Otsuka, Masato; Weustink, Annick; Ligthart, Jurgen; de Winter, Sebastiaan; van Mieghem, Carlos; Nieman, Koen; de Feyter, Pim J; van Domburg, Ron T; Serruys, Patrick W

2008-08-01

To investigate if three-dimensional (3D) based quantitative techniques are comparable to each other and to explore possible differences with respect to the reference method of 2D-QCA in the acute phase and to study whether non-invasive MSCT could potentially be applied to quantify luminal dimensions of a stented coronary segment with a novel bioabsorable drug-eluting stent made of poly-l-lactic-acid (PLLA). Quantitative imaging data derived from 16 patients enrolled at our institution in a first-in-man trial (ABSORB) receiving a biodegradable stent and who were imaged with standard coronary angiography and intravascular ultrasound were compared. Shortly, after stenting the patients also underwent a MSCT procedure. Standard 2D-QCA showed significant smaller stent lengths (p < 0.01). Although, the absolute measured stent diameters and areas by 2D-QCA tend to be smaller, the differences failed to be statistically different when compared to the 3D based quantitative modalities. Measurements made by non-invasive QMSCT-CA of implanted PLLA stents appeared to be comparable to the other 3D modalities without significant differences. Three-dimensional based quantitative analyses showed similar results quantifying luminal dimensions as compared to 2D-QCA during an evaluation of a new bioabsorbable coronary stent design in the acute phase. Furthermore, in biodegradable stents made of PLLA, non-invasive QMSCT-CA can be used to quantify luminal dimensions.

Analogous on-axis interference topographic phase microscopy (AOITPM).

PubMed

Xiu, P; Liu, Q; Zhou, X; Xu, Y; Kuang, C; Liu, X

2018-05-01

The refractive index (RI) of a sample as an endogenous contrast agent plays an important role in transparent live cell imaging. In tomographic phase microscopy (TPM), 3D quantitative RI maps can be reconstructed based on the measured projections of the RI in multiple directions. The resolution of the RI maps not only depends on the numerical aperture of the employed objective lens, but also is determined by the accuracy of the quantitative phase of the sample measured at multiple scanning illumination angles. This paper reports an analogous on-axis interference TPM, where the interference angle between the sample and reference beams is kept constant for projections in multiple directions to improve the accuracy of the phase maps and the resolution of RI tomograms. The system has been validated with both silica beads and red blood cells. Compared with conventional TPM, the proposed system acquires quantitative RI maps with higher resolution (420 nm @λ = 633 nm) and signal-to-noise ratio that can be beneficial for live cell imaging in biomedical applications. © 2018 The Authors Journal of Microscopy © 2018 Royal Microscopical Society.

Quantitative Oxygenation Venography from MRI Phase

PubMed Central

Fan, Audrey P.; Bilgic, Berkin; Gagnon, Louis; Witzel, Thomas; Bhat, Himanshu; Rosen, Bruce R.; Adalsteinsson, Elfar

2014-01-01

Purpose To demonstrate acquisition and processing methods for quantitative oxygenation venograms that map in vivo oxygen saturation (SvO2) along cerebral venous vasculature. Methods Regularized quantitative susceptibility mapping (QSM) is used to reconstruct susceptibility values and estimate SvO2 in veins. QSM with ℓ1 and ℓ2 regularization are compared in numerical simulations of vessel structures with known magnetic susceptibility. Dual-echo, flow-compensated phase images are collected in three healthy volunteers to create QSM images. Bright veins in the susceptibility maps are vectorized and used to form a three-dimensional vascular mesh, or venogram, along which to display SvO2 values from QSM. Results Quantitative oxygenation venograms that map SvO2 along brain vessels of arbitrary orientation and geometry are shown in vivo. SvO2 values in major cerebral veins lie within the normal physiological range reported by 15O positron emission tomography. SvO2 from QSM is consistent with previous MR susceptometry methods for vessel segments oriented parallel to the main magnetic field. In vessel simulations, ℓ1 regularization results in less than 10% SvO2 absolute error across all vessel tilt orientations and provides more accurate SvO2 estimation than ℓ2 regularization. Conclusion The proposed analysis of susceptibility images enables reliable mapping of quantitative SvO2 along venograms and may facilitate clinical use of venous oxygenation imaging. PMID:24006229

Holographic quantitative imaging of sample hidden by turbid medium or occluding objects

NASA Astrophysics Data System (ADS)

Bianco, V.; Miccio, L.; Merola, F.; Memmolo, P.; Gennari, O.; Paturzo, Melania; Netti, P. A.; Ferraro, P.

2015-03-01

Digital Holography (DH) numerical procedures have been developed to allow imaging through turbid media. A fluid is considered turbid when dispersed particles provoke strong light scattering, thus destroying the image formation by any standard optical system. Here we show that sharp amplitude imaging and phase-contrast mapping of object hidden behind turbid medium and/or occluding objects are possible in harsh noise conditions and with a large field-of view by Multi-Look DH microscopy. In particular, it will be shown that both amplitude imaging and phase-contrast mapping of cells hidden behind a flow of Red Blood Cells can be obtained. This allows, in a noninvasive way, the quantitative evaluation of living processes in Lab on Chip platforms where conventional microscopy techniques fail. The combination of this technique with endoscopic imaging can pave the way for the holographic blood vessel inspection, e.g. to look for settled cholesterol plaques as well as blood clots for a rapid diagnostics of blood diseases.

Integral refractive index imaging of flowing cell nuclei using quantitative phase microscopy combined with fluorescence microscopy.

PubMed

Dardikman, Gili; Nygate, Yoav N; Barnea, Itay; Turko, Nir A; Singh, Gyanendra; Javidi, Barham; Shaked, Natan T

2018-03-01

We suggest a new multimodal imaging technique for quantitatively measuring the integral (thickness-average) refractive index of the nuclei of live biological cells in suspension. For this aim, we combined quantitative phase microscopy with simultaneous 2-D fluorescence microscopy. We used 2-D fluorescence microscopy to localize the nucleus inside the quantitative phase map of the cell, as well as for measuring the nucleus radii. As verified offline by both 3-D confocal fluorescence microscopy and 2-D fluorescence microscopy while rotating the cells during flow, the nucleus of cells in suspension that are not during division can be assumed to be an ellipsoid. The entire shape of a cell in suspension can be assumed to be a sphere. Then, the cell and nucleus 3-D shapes can be evaluated based on their in-plain radii available from the 2-D phase and fluorescent measurements, respectively. Finally, the nucleus integral refractive index profile is calculated. We demonstrate the new technique on cancer cells, obtaining nucleus refractive index values that are lower than those of the cytoplasm, coinciding with recent findings. We believe that the proposed technique has the potential to be used for flow cytometry, where full 3-D refractive index tomography is too slow to be implemented during flow.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hoff, M; Rane-Levandovsky, S; Andre, J

Purpose: Traditional arterial spin labeling (ASL) acquisitions with echo planar imaging (EPI) readouts suffer from image distortion due to susceptibility effects, compromising ASL’s ability to accurately quantify cerebral blood flow (CBF) and assess disease-specific patterns associated with CBF abnormalities. Phase labeling for additional coordinate encoding (PLACE) can remove image distortion; our goal is to apply PLACE to improve the quantitative accuracy of ASL CBF in humans. Methods: Four subjects were imaged on a 3T Philips Ingenia scanner using a 16-channel receive coil with a 21/21/10cm (frequency/phase/slice direction) field-of-view. An ASL sequence with a pseudo-continuous ASL (pCASL) labeling scheme was employedmore » to acquire thirty dynamics of single-shot EPI data, with control and label datasets for all dynamics, and PLACE gradients applied on odd dynamics. Parameters included a post-labeling delay = 2s, label duration = 1.8s, flip angle = 90°, TR/TE = 5000/23.5ms, and 2.9/2.9/5.0mm (frequency/phase/slice direction) voxel size. “M0” EPI-reference images and T1-weighted spin-echo images with 0.8/1.0/3.3mm (frequency/phase/slice directions) voxel size were also acquired. Complex conjugate image products of pCASL odd and even dynamics were formed, a linear phase ramp applied, and data expanded and smoothed. Data phase was extracted to map control, label, and M0 magnitude image pixels to their undistorted locations, and images were rebinned to original size. All images were corrected for motion artifacts in FSL 5.0. pCASL images were registered to M0 images, and control and label images were subtracted to compute quantitative CBF maps. Results: pCASL image and CBF map distortions were removed by PLACE in all subjects. Corrected images conformed well to the anatomical T1-weighted reference image, and deviations in corrected CBF maps were evident. Conclusion: Eliminating pCASL distortion with PLACE can improve CBF quantification accuracy using minimal pulse sequence modifications and no additional scan time, improving ASL’s clinical applicability.« less

X-ray phase contrast tomography by tracking near field speckle

PubMed Central

Wang, Hongchang; Berujon, Sebastien; Herzen, Julia; Atwood, Robert; Laundy, David; Hipp, Alexander; Sawhney, Kawal

2015-01-01

X-ray imaging techniques that capture variations in the x-ray phase can yield higher contrast images with lower x-ray dose than is possible with conventional absorption radiography. However, the extraction of phase information is often more difficult than the extraction of absorption information and requires a more sophisticated experimental arrangement. We here report a method for three-dimensional (3D) X-ray phase contrast computed tomography (CT) which gives quantitative volumetric information on the real part of the refractive index. The method is based on the recently developed X-ray speckle tracking technique in which the displacement of near field speckle is tracked using a digital image correlation algorithm. In addition to differential phase contrast projection images, the method allows the dark-field images to be simultaneously extracted. After reconstruction, compared to conventional absorption CT images, the 3D phase CT images show greatly enhanced contrast. This new imaging method has advantages compared to other X-ray imaging methods in simplicity of experimental arrangement, speed of measurement and relative insensitivity to beam movements. These features make the technique an attractive candidate for material imaging such as in-vivo imaging of biological systems containing soft tissue. PMID:25735237

Three-dimensional coherent x-ray diffraction imaging of molten iron in mantle olivine at nanoscale resolution.

PubMed

Jiang, Huaidong; Xu, Rui; Chen, Chien-Chun; Yang, Wenge; Fan, Jiadong; Tao, Xutang; Song, Changyong; Kohmura, Yoshiki; Xiao, Tiqiao; Wang, Yong; Fei, Yingwei; Ishikawa, Tetsuya; Mao, Wendy L; Miao, Jianwei

2013-05-17

We report quantitative 3D coherent x-ray diffraction imaging of a molten Fe-rich alloy and crystalline olivine sample, synthesized at 6 GPa and 1800 °C, with nanoscale resolution. The 3D mass density map is determined and the 3D distribution of the Fe-rich and Fe-S phases in the olivine-Fe-S sample is observed. Our results indicate that the Fe-rich melt exhibits varied 3D shapes and sizes in the olivine matrix. This work has potential for not only improving our understanding of the complex interactions between Fe-rich core-forming melts and mantle silicate phases but also paves the way for quantitative 3D imaging of materials at nanoscale resolution under extreme pressures and temperatures.

Hyperspectral and differential CARS microscopy for quantitative chemical imaging in human adipocytes

PubMed Central

Di Napoli, Claudia; Pope, Iestyn; Masia, Francesco; Watson, Peter; Langbein, Wolfgang; Borri, Paola

2014-01-01

In this work, we demonstrate the applicability of coherent anti-Stokes Raman scattering (CARS) micro-spectroscopy for quantitative chemical imaging of saturated and unsaturated lipids in human stem-cell derived adipocytes. We compare dual-frequency/differential CARS (D-CARS), which enables rapid imaging and simple data analysis, with broadband hyperspectral CARS microscopy analyzed using an unsupervised phase-retrieval and factorization method recently developed by us for quantitative chemical image analysis. Measurements were taken in the vibrational fingerprint region (1200–2000/cm) and in the CH stretch region (2600–3300/cm) using a home-built CARS set-up which enables hyperspectral imaging with 10/cm resolution via spectral focussing from a single broadband 5 fs Ti:Sa laser source. Through a ratiometric analysis, both D-CARS and phase-retrieved hyperspectral CARS determine the concentration of unsaturated lipids with comparable accuracy in the fingerprint region, while in the CH stretch region D-CARS provides only a qualitative contrast owing to its non-linear behavior. When analyzing hyperspectral CARS images using the blind factorization into susceptibilities and concentrations of chemical components recently demonstrated by us, we are able to determine vol:vol concentrations of different lipid components and spatially resolve inhomogeneities in lipid composition with superior accuracy compared to state-of-the art ratiometric methods. PMID:24877002

Single-shot water-immersion microscopy platform for qualitative visualization and quantitative phase imaging of biosamples

NASA Astrophysics Data System (ADS)

Picazo-Bueno, José Ángel; Cojoc, Dan; Torre, Vincent; Micó, Vicente

2017-07-01

We present the combination of a single-shot water-immersion digital holographic microscopy with broadband illumination for simultaneous visualization of coherent and incoherent images using microbeads and different biosamples.

Evaluation of osteoarthritis progression using polarization-sensitive optical coherence tomography

NASA Astrophysics Data System (ADS)

Nassif, Nader A.; Pierce, Mark C.; Park, B. Hyle; Cense, Barry; de Boer, Johannes F.

2004-07-01

Osteoarthritis is a prevalent medical condition that presents a diagnostic and therapeutic challenge to physicians today because of the inability to assess the integrity of the articular cartilage early in the disease. Polarization sensitive optical coherence tomography (PS-OCT) is a high resolution, non-contact imaging modality that provides cross-sectional images with additional information regarding the integrity of the collagen matrix. Using PS-OCT to image provides information regarding thickness of the articular cartilage and gives an index of biochemical changes based on alterations in optical properties (i.e. birefringence) of the tissue. We demonstrate initial experiments performed on specimens collected following total knee replacement surgery. Articular cartilage was imaged using a 1310 nm PS-OCT system where both intensity and phase images were acquired. PS-OCT images were compared with histology, and the changes in tissue optical properties were characterized. Analysis of the intensity images demonstrates differences between healthy and diseased cartilage surface and thickness. Phase maps of the tissue demonstrated distinct differences between healthy and diseased tissue. PS-OCT was able to image a gradual loss of birefringence as the tissue became more diseased. In this way, determining the rate of change of the phase provides a quantitative measure of pathology. Thus, imaging and evaluation of osteoarthritis using PS-OCT can be a useful means of quantitative assessment of the disease.

TU-C-12A-02: Development of a Multiparametric Statistical Response Map for Quantitative Imaging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bosca, R; The University of Texas MD Anderson Cancer Center, Houston, TX; Mahajan, A

2014-06-15

Purpose: Quantitative imaging biomarkers (QIB) are becoming increasingly utilized in early phase clinical trials as a means of non-invasively assessing treatment response and associated response heterogeneity. The aim of this study was to develop a flexible multiparametric statistical framework to predict voxel-by-voxel response of several potential MRI QIBs. Methods: Patients with histologically proven glioblastomas (n=11) were treated with chemoradiation (with/without bevacizumab) and underwent one baseline and two mid-treatment (3–4wks) MRIs. Dynamic contrast-enhanced (3D FSPGR, 6.3sec/phase, 0.1 mmol/kg Gd-DTPA), dynamic susceptibility contrast (2D GRE-EPI, 1.5sec/phase, 0.2mmol/kg Gd-DTPA), and diffusion tensor (2D DW-EPI, b=0, 1200 s/mm{sup 2}, 27 directions) imaging acquisitions weremore » obtained during each study. Mid-treatment and pre-treatment images were rigidly aligned, and regions of partial response (PR), stable disease (SD), and progressive disease (PD) were contoured in consensus by two experienced radiation oncologists. Voxels in these categories were used to train ordinal (PR« less

Broadband quantitative phase microscopy with extended field of view using off-axis interferometric multiplexing.

PubMed

Girshovitz, Pinhas; Frenklach, Irena; Shaked, Natan T

2015-11-01

We propose a new portable imaging configuration that can double the field of view (FOV) of existing off-axis interferometric imaging setups, including broadband off-axis interferometers. This configuration is attached at the output port of the off-axis interferometer and optically creates a multiplexed interferogram on the digital camera, which is composed of two off-axis interferograms with straight fringes at orthogonal directions. Each of these interferograms contains a different FOV of the imaged sample. Due to the separation of these two FOVs in the spatial-frequency domain, they can be fully reconstructed separately, while obtaining two complex wavefronts from the sample at once. Since the optically multiplexed off-axis interferogram is recorded by the camera in a single exposure, fast dynamics can be recorded with a doubled imaging area. We used this technique for quantitative phase microscopy of biological samples with extended FOV. We demonstrate attaching the proposed module to a diffractive phase microscopy interferometer, illuminated by a broadband light source. The biological samples used for the experimental demonstrations include microscopic diatom shells, cancer cells, and flowing blood cells.

Movies of cellular and sub-cellular motion by digital holographic microscopy.

PubMed

Mann, Christopher J; Yu, Lingfeng; Kim, Myung K

2006-03-23

Many biological specimens, such as living cells and their intracellular components, often exhibit very little amplitude contrast, making it difficult for conventional bright field microscopes to distinguish them from their surroundings. To overcome this problem phase contrast techniques such as Zernike, Normarsky and dark-field microscopies have been developed to improve specimen visibility without chemically or physically altering them by the process of staining. These techniques have proven to be invaluable tools for studying living cells and furthering scientific understanding of fundamental cellular processes such as mitosis. However a drawback of these techniques is that direct quantitative phase imaging is not possible. Quantitative phase imaging is important because it enables determination of either the refractive index or optical thickness variations from the measured optical path length with sub-wavelength accuracy. Digital holography is an emergent phase contrast technique that offers an excellent approach in obtaining both qualitative and quantitative phase information from the hologram. A CCD camera is used to record a hologram onto a computer and numerical methods are subsequently applied to reconstruct the hologram to enable direct access to both phase and amplitude information. Another attractive feature of digital holography is the ability to focus on multiple focal planes from a single hologram, emulating the focusing control of a conventional microscope. A modified Mach-Zender off-axis setup in transmission is used to record and reconstruct a number of holographic amplitude and phase images of cellular and sub-cellular features. Both cellular and sub-cellular features are imaged with sub-micron, diffraction-limited resolution. Movies of holographic amplitude and phase images of living microbes and cells are created from a series of holograms and reconstructed with numerically adjustable focus, so that the moving object can be accurately tracked with a reconstruction rate of 300ms for each hologram. The holographic movies show paramecium swimming among other microbes as well as displaying some of their intracellular processes. A time lapse movie is also shown for fibroblast cells in the process of migration. Digital holography and movies of digital holography are seen to be useful new tools for visualization of dynamic processes in biological microscopy. Phase imaging digital holography is a promising technique in terms of the lack of coherent noise and the precision with which the optical thickness of a sample can be profiled, which can lead to images with an axial resolution of a few nanometres.

Critical behavior of subcellular density organization during neutrophil activation and migration.

PubMed

Baker-Groberg, Sandra M; Phillips, Kevin G; Healy, Laura D; Itakura, Asako; Porter, Juliana E; Newton, Paul K; Nan, Xiaolin; McCarty, Owen J T

2015-12-01

Physical theories of active matter continue to provide a quantitative understanding of dynamic cellular phenomena, including cell locomotion. Although various investigations of the rheology of cells have identified important viscoelastic and traction force parameters for use in these theoretical approaches, a key variable has remained elusive both in theoretical and experimental approaches: the spatiotemporal behavior of the subcellular density. The evolution of the subcellular density has been qualitatively observed for decades as it provides the source of image contrast in label-free imaging modalities (e.g., differential interference contrast, phase contrast) used to investigate cellular specimens. While these modalities directly visualize cell structure, they do not provide quantitative access to the structures being visualized. We present an established quantitative imaging approach, non-interferometric quantitative phase microscopy, to elucidate the subcellular density dynamics in neutrophils undergoing chemokinesis following uniform bacterial peptide stimulation. Through this approach, we identify a power law dependence of the neutrophil mean density on time with a critical point, suggesting a critical density is required for motility on 2D substrates. Next we elucidate a continuum law relating mean cell density, area, and total mass that is conserved during neutrophil polarization and migration. Together, our approach and quantitative findings will enable investigators to define the physics coupling cytoskeletal dynamics with subcellular density dynamics during cell migration.

Critical behavior of subcellular density organization during neutrophil activation and migration

PubMed Central

Baker-Groberg, Sandra M.; Phillips, Kevin G.; Healy, Laura D.; Itakura, Asako; Porter, Juliana E.; Newton, Paul K.; Nan, Xiaolin; McCarty, Owen J.T.

2015-01-01

Physical theories of active matter continue to provide a quantitative understanding of dynamic cellular phenomena, including cell locomotion. Although various investigations of the rheology of cells have identified important viscoelastic and traction force parameters for use in these theoretical approaches, a key variable has remained elusive both in theoretical and experimental approaches: the spatiotemporal behavior of the subcellular density. The evolution of the subcellular density has been qualitatively observed for decades as it provides the source of image contrast in label-free imaging modalities (e.g., differential interference contrast, phase contrast) used to investigate cellular specimens. While these modalities directly visualize cell structure, they do not provide quantitative access to the structures being visualized. We present an established quantitative imaging approach, non-interferometric quantitative phase microscopy, to elucidate the subcellular density dynamics in neutrophils undergoing chemokinesis following uniform bacterial peptide stimulation. Through this approach, we identify a power law dependence of the neutrophil mean density on time with a critical point, suggesting a critical density is required for motility on 2D substrates. Next we elucidate a continuum law relating mean cell density, area, and total mass that is conserved during neutrophil polarization and migration. Together, our approach and quantitative findings will enable investigators to define the physics coupling cytoskeletal dynamics with subcellular density dynamics during cell migration. PMID:26640599

Confocal reflectance quantitative phase microscope system for cellular membranes dynamics study (Conference Presentation)

NASA Astrophysics Data System (ADS)

Singh, Vijay Raj; Yaqoob, Zahid; So, Peter T. C.

2017-02-01

Quantitative phase microscopy (QPM) techniques developed so far primarily belongs to high speed transmitted light based systems that has enough sensitivity to resolve membrane fluctuations and dynamics, but has no depth resolution. Therefore, most biomechanics studies using QPM today is confined to simple cells, such as RBCs, without internal organelles. An important instrument that will greatly extend the biomedical applications of QPM is to develop next generation microscope with 3D capability and sufficient temporal resolution to study biomechanics of complex eukaryotic cells including the mechanics of their internal compartments. For eukaryotic cells, the depth sectioning capability is critical and should be sufficient to distinguish nucleic membrane fluctuations from plasma membrane fluctuations. Further, this microscope must provide high temporal resolution since typical eukaryotes membranes are substantially stiffer than RBCs. A confocal reflectance quantitative phase microscope is presented based on multi-pinhole scanning, with the capabilities of higher temporal resolution and sensitivity for nucleic and plasma membranes of eukaryotic cells. System hardware is developed based on an array of confocal pinhole generated by using the `ON' state of subset of micro-mirrors of digital micro-mirror device (DMD, from Texas Instruments) and high-speed raster scanning provides 14ms imaging speed in wide-field mode. A common path interferometer is integrated at the imaging arm for detection of specimens' quantitative phase information. Theoretical investigation of quantitative phase reconstructed from system is investigated and application of system is presented for dimensional fluctuations measurements of both cellular plasma and nucleic membranes of embryonic stem cells.

Deciphering the internal complexity of living cells with quantitative phase microscopy: a multiscale approach

NASA Astrophysics Data System (ADS)

Martinez-Torres, Cristina; Laperrousaz, Bastien; Berguiga, Lotfi; Boyer-Provera, Elise; Elezgaray, Juan; Nicolini, Franck E.; Maguer-Satta, Veronique; Arneodo, Alain; Argoul, Françoise

2015-09-01

The distribution of refractive indices (RIs) of a living cell contributes in a nonintuitive manner to its optical phase image and quite rarely can be inverted to recover its internal structure. The interpretation of the quantitative phase images of living cells remains a difficult task because (1) we still have very little knowledge on the impact of its internal macromolecular complexes on the local RI and (2) phase changes produced by light propagation through the sample are mixed with diffraction effects by the internal cell bodies. We propose to implement a two-dimensional wavelet-based contour chain detection method to distinguish internal boundaries based on their greatest optical path difference gradients. These contour chains correspond to the highest image phase contrast and follow the local RI inhomogeneities linked to the intracellular structural intricacy. Their statistics and spatial distribution are the morphological indicators suited for comparing cells of different origins and/or to follow their transformation in pathologic situations. We use this method to compare nonadherent blood cells from primary and laboratory culture origins and to assess the internal transformation of hematopoietic stem cells by the transduction of the BCR-ABL oncogene responsible for the chronic myelogenous leukemia.

Revising the lower statistical limit of x-ray grating-based phase-contrast computed tomography.

PubMed

Marschner, Mathias; Birnbacher, Lorenz; Willner, Marian; Chabior, Michael; Herzen, Julia; Noël, Peter B; Pfeiffer, Franz

2017-01-01

Phase-contrast x-ray computed tomography (PCCT) is currently investigated as an interesting extension of conventional CT, providing high soft-tissue contrast even if examining weakly absorbing specimen. Until now, the potential for dose reduction was thought to be limited compared to attenuation CT, since meaningful phase retrieval fails for scans with very low photon counts when using the conventional phase retrieval method via phase stepping. In this work, we examine the statistical behaviour of the reverse projection method, an alternative phase retrieval approach and compare the results to the conventional phase retrieval technique. We investigate the noise levels in the projections as well as the image quality and quantitative accuracy of the reconstructed tomographic volumes. The results of our study show that this method performs better in a low-dose scenario than the conventional phase retrieval approach, resulting in lower noise levels, enhanced image quality and more accurate quantitative values. Overall, we demonstrate that the lower statistical limit of the phase stepping procedure as proposed by recent literature does not apply to this alternative phase retrieval technique. However, further development is necessary to overcome experimental challenges posed by this method which would enable mainstream or even clinical application of PCCT.

Wave field restoration using three-dimensional Fourier filtering method.

PubMed

Kawasaki, T; Takai, Y; Ikuta, T; Shimizu, R

2001-11-01

A wave field restoration method in transmission electron microscopy (TEM) was mathematically derived based on a three-dimensional (3D) image formation theory. Wave field restoration using this method together with spherical aberration correction was experimentally confirmed in through-focus images of amorphous tungsten thin film, and the resolution of the reconstructed phase image was successfully improved from the Scherzer resolution limit to the information limit. In an application of this method to a crystalline sample, the surface structure of Au(110) was observed in a profile-imaging mode. The processed phase image showed quantitatively the atomic relaxation of the topmost layer.

Histopathological image analysis of chemical-induced hepatocellular hypertrophy in mice.

PubMed

Asaoka, Yoshiji; Togashi, Yuko; Mutsuga, Mayu; Imura, Naoko; Miyoshi, Tomoya; Miyamoto, Yohei

2016-04-01

Chemical-induced hepatocellular hypertrophy is frequently observed in rodents, and is mostly caused by the induction of phase I and phase II drug metabolic enzymes and peroxisomal lipid metabolic enzymes. Liver weight is a sensitive and commonly used marker for detecting hepatocellular hypertrophy, but is also increased by a number of other factors. Histopathological observations subjectively detect changes such as hepatocellular hypertrophy based on the size of a hepatocyte. Therefore, quantitative microscopic observations are required to evaluate histopathological alterations objectively. In the present study, we developed a novel quantitative method for an image analysis of hepatocellular hypertrophy using liver sections stained with hematoxylin and eosin, and demonstrated its usefulness for evaluating hepatocellular hypertrophy induced by phenobarbital (a phase I and phase II enzyme inducer) and clofibrate (a peroxisomal enzyme inducer) in mice. The algorithm of this imaging analysis was designed to recognize an individual hepatocyte through a combination of pixel-based and object-based analyses. Hepatocellular nuclei and the surrounding non-hepatocellular cells were recognized by the pixel-based analysis, while the areas of the recognized hepatocellular nuclei were then expanded until they ran against their expanding neighboring hepatocytes and surrounding non-hepatocellular cells by the object-based analysis. The expanded area of each hepatocellular nucleus was regarded as the size of an individual hepatocyte. The results of this imaging analysis showed that changes in the sizes of hepatocytes corresponded with histopathological observations in phenobarbital and clofibrate-treated mice, and revealed a correlation between hepatocyte size and liver weight. In conclusion, our novel image analysis method is very useful for quantitative evaluations of chemical-induced hepatocellular hypertrophy. Copyright © 2015 Elsevier GmbH. All rights reserved.

Quantitative shear wave optical coherence elastography (SW-OCE) with acoustic radiation force impulses (ARFI) induced by phase array transducer

NASA Astrophysics Data System (ADS)

Song, Shaozhen; Le, Nhan Minh; Wang, Ruikang K.; Huang, Zhihong

2015-03-01

Shear Wave Optical Coherence Elastography (SW-OCE) uses the speed of propagating shear waves to provide a quantitative measurement of localized shear modulus, making it a valuable technique for the elasticity characterization of tissues such as skin and ocular tissue. One of the main challenges in shear wave elastography is to induce a reliable source of shear wave; most of nowadays techniques use external vibrators which have several drawbacks such as limited wave propagation range and/or difficulties in non-invasive scans requiring precisions, accuracy. Thus, we propose linear phase array ultrasound transducer as a remote wave source, combined with the high-speed, 47,000-frame-per-second Shear-wave visualization provided by phase-sensitive OCT. In this study, we observed for the first time shear waves induced by a 128 element linear array ultrasound imaging transducer, while the ultrasound and OCT images (within the OCE detection range) were triggered simultaneously. Acoustic radiation force impulses are induced by emitting 10 MHz tone-bursts of sub-millisecond durations (between 50 μm - 100 μm). Ultrasound beam steering is achieved by programming appropriate phase delay, covering a lateral range of 10 mm and full OCT axial (depth) range in the imaging sample. Tissue-mimicking phantoms with agarose concentration of 0.5% and 1% was used in the SW-OCE measurements as the only imaging samples. The results show extensive improvements over the range of SW-OCE elasticity map; such improvements can also be seen over shear wave velocities in softer and stiffer phantoms, as well as determining the boundary of multiple inclusions with different stiffness. This approach opens up the feasibility to combine medical ultrasound imaging and SW-OCE for high-resolution localized quantitative measurement of tissue biomechanical property.

Quantitative phase retrieval with arbitrary pupil and illumination

DOE PAGES

Claus, Rene A.; Naulleau, Patrick P.; Neureuther, Andrew R.; ...

2015-10-02

We present a general algorithm for combining measurements taken under various illumination and imaging conditions to quantitatively extract the amplitude and phase of an object wave. The algorithm uses the weak object transfer function, which incorporates arbitrary pupil functions and partially coherent illumination. The approach is extended beyond the weak object regime using an iterative algorithm. Finally, we demonstrate the method on measurements of Extreme Ultraviolet Lithography (EUV) multilayer mask defects taken in an EUV zone plate microscope with both a standard zone plate lens and a zone plate implementing Zernike phase contrast.

X-ray Phase Contrast Allows Three Dimensional, Quantitative Imaging of Hydrogel Implants

DOE PAGES

Appel, Alyssa A.; Larson, Jeffrey C.; Jiang, Bin; ...

2015-10-20

Three dimensional imaging techniques are needed for the evaluation and assessment of biomaterials used for tissue engineering and drug delivery applications. Hydrogels are a particularly popular class of materials for medical applications but are difficult to image in tissue using most available imaging modalities. Imaging techniques based on X-ray Phase Contrast (XPC) have shown promise for tissue engineering applications due to their ability to provide image contrast based on multiple X-ray properties. In this manuscript we describe results using XPC to image a model hydrogel and soft tissue structure. Porous fibrin loaded poly(ethylene glycol) hydrogels were synthesized and implanted inmore » a rodent subcutaneous model. Samples were explanted and imaged with an analyzer-based XPC technique and processed and stained for histology for comparison. Both hydrogel and soft tissues structures could be identified in XPC images. Structure in skeletal muscle adjacent could be visualized and invading fibrovascular tissue could be quantified. In quantitative results, there were no differences between XPC and the gold-standard histological measurements. These results provide evidence of the significant potential of techniques based on XPC for 3D imaging of hydrogel structure and local tissue response.« less

Phase correlation imaging of unlabeled cell dynamics

NASA Astrophysics Data System (ADS)

Ma, Lihong; Rajshekhar, Gannavarpu; Wang, Ru; Bhaduri, Basanta; Sridharan, Shamira; Mir, Mustafa; Chakraborty, Arindam; Iyer, Rajashekar; Prasanth, Supriya; Millet, Larry; Gillette, Martha U.; Popescu, Gabriel

2016-09-01

We present phase correlation imaging (PCI) as a novel approach to study cell dynamics in a spatially-resolved manner. PCI relies on quantitative phase imaging time-lapse data and, as such, functions in label-free mode, without the limitations associated with exogenous markers. The correlation time map outputted in PCI informs on the dynamics of the intracellular mass transport. Specifically, we show that PCI can extract quantitatively the diffusion coefficient map associated with live cells, as well as standard Brownian particles. Due to its high sensitivity to mass transport, PCI can be applied to studying the integrity of actin polymerization dynamics. Our results indicate that the cyto-D treatment blocking the actin polymerization has a dominant effect at the large spatial scales, in the region surrounding the cell. We found that PCI can distinguish between senescent and quiescent cells, which is extremely difficult without using specific markers currently. We anticipate that PCI will be used alongside established, fluorescence-based techniques to enable valuable new studies of cell function.

Quantitative hard x-ray phase contrast imaging of micropipes in SiC

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kohn, V. G.; Argunova, T. S.; Je, J. H., E-mail: jhje@postech.ac.kr

2013-12-15

Peculiarities of quantitative hard x-ray phase contrast imaging of micropipes in SiC are discussed. The micropipe is assumed as a hollow cylinder with an elliptical cross section. The major and minor diameters can be restored using the least square fitting procedure by comparing the experimental data, i.e. the profile across the micropipe axis, with those calculated based on phase contrast theory. It is shown that one projection image gives an information which does not allow a complete determination of the elliptical cross section, if an orientation of micropipe is not known. Another problem is a weak accuracy in estimating themore » diameters, partly because of using pink synchrotron radiation, which is necessary because a monochromatic beam intensity is not sufficient to reveal the weak contrast from a very small object. The general problems of accuracy in estimating the two diameters using the least square procedure are discussed. Two experimental examples are considered to demonstrate small as well as modest accuracies in estimating the diameters.« less

Multifocus image fusion using phase congruency

NASA Astrophysics Data System (ADS)

Zhan, Kun; Li, Qiaoqiao; Teng, Jicai; Wang, Mingying; Shi, Jinhui

2015-05-01

We address the problem of fusing multifocus images based on the phase congruency (PC). PC provides a sharpness feature of a natural image. The focus measure (FM) is identified as strong PC near a distinctive image feature evaluated by the complex Gabor wavelet. The PC is more robust against noise than other FMs. The fusion image is obtained by a new fusion rule (FR), and the focused region is selected by the FR from one of the input images. Experimental results show that the proposed fusion scheme achieves the fusion performance of the state-of-the-art methods in terms of visual quality and quantitative evaluations.

Probing myocardium biomechanics using quantitative optical coherence elastography

NASA Astrophysics Data System (ADS)

Wang, Shang; Lopez, Andrew L.; Morikawa, Yuka; Tao, Ge; Li, Jiasong; Larina, Irina V.; Martin, James F.; Larin, Kirill V.

2015-03-01

We present a quantitative optical coherence elastographic method for noncontact assessment of the myocardium elasticity. The method is based on shear wave imaging optical coherence tomography (SWI-OCT), where a focused air-puff system is used to induce localized tissue deformation through a low-pressure short-duration air stream and a phase-sensitive OCT system is utilized to monitor the propagation of the induced tissue displacement with nanoscale sensitivity. The 1-D scanning of M-mode OCT imaging and the application of optical phase retrieval and mapping techniques enable the reconstruction and visualization of 2-D depth-resolved shear wave propagation in tissue with ultra-high frame rate. The feasibility of this method in quantitative elasticity measurement is demonstrated on tissue-mimicking phantoms with the estimated Young's modulus compared with uniaxial compression tests. We also performed pilot experiments on ex vivo mouse cardiac muscle tissues with normal and genetically altered cardiomyocytes. Our results indicate this noncontact quantitative optical coherence elastographic method can be a useful tool for the cardiac muscle research and studies.

[Value of quantitative iodine-based material decomposition images with gemstone spectral CT imaging in the follow-up of patients with hepatocellular carcinoma after TACE treatment].

PubMed

Xing, Gusheng; Wang, Shuang; Li, Chenrui; Zhao, Xinming; Zhou, Chunwu

2015-03-01

To investigate the value of quantitative iodine-based material decomposition images with gemstone spectral CT imaging in the follow-up of patients with hepatocellular carcinoma (HCC) after transcatheter arterial chemoebolization (TACE). Consecutive 32 HCC patients with previous TACE treatment were included in this study. For the follow-up, arterial phase (AP) and venous phase (VP) dual-phase CT scans were performed with a single-source dual-energy CT scanner (Discovery CT 750HD, GE Healthcare). Iodine concentrations were derived from iodine-based material-decomposition images in the liver parenchyma, tumors and coagulation necrosis (CN) areas. The iodine concentration difference (ICD) between the arterial-phase (AP) and venal-phase (VP) were quantitatively evaluated in different tissues.The lesion-to-normal parenchyma iodine concentration ratio (LNR) was calculated. ROC analysis was performed for the qualitative evaluation, and the area under ROC (Az) was calculated to represent the diagnostic ability of ICD and LNR. In all the 32 HCC patients, the region of interesting (ROI) for iodine concentrations included liver parenchyma (n=42), tumors (n=28) and coagulation necrosis (n=24). During the AP the iodine concentration of CNs (median value 0.088 µg/mm(3)) appeared significantly higher than that of the tumors (0.064 µg/mm(3), P=0.022) and liver parenchyma (0.048 µg/mm(3), P=0.005). But it showed no significant difference between liver parenchyma and tumors (P=0.454). During the VP the iodine concentration in hepatic parenchyma (median value 0.181 µg/mm(3)) was significantly higher than that in CNs (0.140 µg/mm(3), P=0.042). There was no significant difference between liver parenchyma and tumors, CNs and tumors (both P>0.05). The median value of ICD in CNs was 0.006 µg/mm(3), significantly lower than that of the HCC (0.201 µg/mm(3), P<0.001) and hepatic parenchyma (0.117 µg/mm(3), P<0.001). The ICDs in tumors and hepatic parenchyma showed no significant difference (P=0.829). During the AP, the LNR had no significant difference between CNs and tumors (a median value 1.805 vs. 1.310, P=0.389), and during the VP, the difference was also non-significant (the median value 0.647 vs. 0.713, P=0.660). The mean Az value of ICDs for evaluation of surviving tumor tissues was 0.804, whiles LNR measured a disappointing result in both AV images and VP images. Quantitative iodine-based material decomposition images with gemstone spectral CT imaging can improve the diagnostic efficacy of CT imaging for HCC patients after TACE treatment.

Quantitative phase imaging of biological cells using spatially low and temporally high coherent light source.

PubMed

Ahmad, Azeem; Dubey, Vishesh; Singh, Gyanendra; Singh, Veena; Mehta, Dalip Singh

2016-04-01

In this Letter, we demonstrate quantitative phase imaging of biological samples, such as human red blood cells (RBCs) and onion cells using narrow temporal frequency and wide angular frequency spectrum light source. This type of light source was synthesized by the combined effect of spatial, angular, and temporal diversity of speckle reduction technique. The importance of using low spatial and high temporal coherence light source over the broad band and narrow band light source is that it does not require any dispersion compensation mechanism for biological samples. Further, it avoids the formation of speckle or spurious fringes which arises while using narrow band light source.

Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy

NASA Astrophysics Data System (ADS)

Kemper, Björn; Bauwens, Andreas; Vollmer, Angelika; Ketelhut, Steffi; Langehanenberg, Patrik; Müthing, Johannes; Karch, Helge; von Bally, Gert

2010-05-01

Digital holographic microscopy (DHM) enables quantitative multifocus phase contrast imaging for nondestructive technical inspection and live cell analysis. Time-lapse investigations on human brain microvascular endothelial cells demonstrate the use of DHM for label-free dynamic quantitative monitoring of cell division of mother cells into daughter cells. Cytokinetic DHM analysis provides future applications in toxicology and cancer research.

DMD-based quantitative phase microscopy and optical diffraction tomography

NASA Astrophysics Data System (ADS)

Zhou, Renjie

2018-02-01

Digital micromirror devices (DMDs), which offer high speed and high degree of freedoms in steering light illuminations, have been increasingly applied to optical microscopy systems in recent years. Lately, we introduced DMDs into digital holography to enable new imaging modalities and break existing imaging limitations. In this paper, we will first present our progress in using DMDs for demonstrating laser-illumination Fourier ptychographic microscopy (FPM) with shotnoise limited detection. After that, we will present a novel common-path quantitative phase microscopy (QPM) system based on using a DMD. Building on those early developments, a DMD-based high speed optical diffraction tomography (ODT) system has been recently demonstrated, and the results will also be presented. This ODT system is able to achieve video-rate 3D refractive-index imaging, which can potentially enable observations of high-speed 3D sample structural changes.

Live cell refractometry using microfluidic devices.

PubMed

Lue, Niyom; Popescu, Gabriel; Ikeda, Takahiro; Dasari, Ramachandra R; Badizadegan, Kamran; Feld, Michael S

2006-09-15

Using Hilbert phase microscopy for extracting quantitative phase images, we measured the average refractive index associated with live cells in culture. To decouple the contributions to the phase signal from the cell refractive index and thickness, we confined the cells in microchannels. The results are confirmed by comparison with measurements of spherical cells in suspension.

Single grating x-ray imaging for dynamic biological systems

NASA Astrophysics Data System (ADS)

Morgan, Kaye S.; Paganin, David M.; Parsons, David W.; Donnelley, Martin; Yagi, Naoto; Uesugi, Kentaro; Suzuki, Yoshio; Takeuchi, Akihisa; Siu, Karen K. W.

2012-07-01

Biomedical studies are already benefiting from the excellent contrast offered by phase contrast x-ray imaging, but live imaging work presents several challenges. Living samples make it particularly difficult to achieve high resolution, sensitive phase contrast images, as exposures must be short and cannot be repeated. We therefore present a single-exposure, high-flux method of differential phase contrast imaging [1, 2, 3] in the context of imaging live airways for Cystic Fibrosis (CF) treatment assessment [4]. The CF study seeks to non-invasively observe the liquid lining the airways, which should increase in depth in response to effective treatments. Both high spatial resolution and sensitivity are required in order to track micron size changes in a liquid that is not easily differentiated from the tissue on which it lies. Our imaging method achieves these goals by using a single attenuation grating or grid as a reference pattern, and analyzing how the sample deforms the pattern to quantitatively retrieve the phase depth of the sample. The deformations are mapped at each pixel in the image using local cross-correlations comparing each 'sample and pattern' image with a reference 'pattern only' image taken before the sample is introduced. This produces a differential phase image, which may be integrated to give the sample phase depth.

Label-free tissue scanner for colorectal cancer screening

NASA Astrophysics Data System (ADS)

Kandel, Mikhail E.; Sridharan, Shamira; Liang, Jon; Luo, Zelun; Han, Kevin; Macias, Virgilia; Shah, Anish; Patel, Roshan; Tangella, Krishnarao; Kajdacsy-Balla, Andre; Guzman, Grace; Popescu, Gabriel

2017-06-01

The current practice of surgical pathology relies on external contrast agents to reveal tissue architecture, which is then qualitatively examined by a trained pathologist. The diagnosis is based on the comparison with standardized empirical, qualitative assessments of limited objectivity. We propose an approach to pathology based on interferometric imaging of "unstained" biopsies, which provides unique capabilities for quantitative diagnosis and automation. We developed a label-free tissue scanner based on "quantitative phase imaging," which maps out optical path length at each point in the field of view and, thus, yields images that are sensitive to the "nanoscale" tissue architecture. Unlike analysis of stained tissue, which is qualitative in nature and affected by color balance, staining strength and imaging conditions, optical path length measurements are intrinsically quantitative, i.e., images can be compared across different instruments and clinical sites. These critical features allow us to automate the diagnosis process. We paired our interferometric optical system with highly parallelized, dedicated software algorithms for data acquisition, allowing us to image at a throughput comparable to that of commercial tissue scanners while maintaining the nanoscale sensitivity to morphology. Based on the measured phase information, we implemented software tools for autofocusing during imaging, as well as image archiving and data access. To illustrate the potential of our technology for large volume pathology screening, we established an "intrinsic marker" for colorectal disease that detects tissue with dysplasia or colorectal cancer and flags specific areas for further examination, potentially improving the efficiency of existing pathology workflows.

Characterization of a high-energy in-line phase contrast tomosynthesis prototype.

PubMed

Wu, Di; Yan, Aimin; Li, Yuhua; Wong, Molly D; Zheng, Bin; Wu, Xizeng; Liu, Hong

2015-05-01

In this research, a high-energy in-line phase contrast tomosynthesis prototype was developed and characterized through quantitative investigations and phantom studies. The prototype system consists of an x-ray source, a motorized rotation stage, and a CMOS detector with a pixel pitch of 0.05 mm. The x-ray source was operated at 120 kVp for this study, and the objects were mounted on the rotation stage 76.2 cm (R1) from the source and 114.3 cm (R2) from the detector. The large air gap between the object and detector guarantees sufficient phase-shift effects. The quantitative evaluation of this prototype included modulation transfer function and noise power spectrum measurements conducted under both projection mode and tomosynthesis mode. Phantom studies were performed including three custom designed phantoms with complex structures: a five-layer bubble wrap phantom, a fishbone phantom, and a chicken breast phantom with embedded fibrils and mass structures extracted from an ACR phantom. In-plane images of the phantoms were acquired to investigate their image qualities through observation, intensity profile plots, edge enhancement evaluations, and/or contrast-to-noise ratio calculations. In addition, the robust phase-attenuation duality (PAD)-based phase retrieval method was applied to tomosynthesis for the first time in this research. It was utilized as a preprocessing method to fully exhibit phase contrast on the angular projection before reconstruction. The resolution and noise characteristics of this high-energy in-line phase contrast tomosynthesis prototype were successfully investigated and demonstrated. The phantom studies demonstrated that this imaging prototype can successfully remove the structure overlapping in phantom projections, obtain delineate interfaces, and achieve better contrast-to-noise ratio after applying phase retrieval to the angular projections. This research successfully demonstrated a high-energy in-line phase contrast tomosynthesis prototype. In addition, the PAD-based method of phase retrieval was combined with tomosynthesis imaging for the first time, which demonstrated its capability in significantly improving the contrast-to-noise ratios in the images.

Analysis of Vaginal Microbicide Film Hydration Kinetics by Quantitative Imaging Refractometry

PubMed Central

Rinehart, Matthew; Grab, Sheila; Rohan, Lisa; Katz, David; Wax, Adam

2014-01-01

We have developed a quantitative imaging refractometry technique, based on holographic phase microscopy, as a tool for investigating microscopic structural changes in water-soluble polymeric materials. Here we apply the approach to analyze the structural degradation of vaginal topical microbicide films due to water uptake. We implemented transmission imaging of 1-mm diameter film samples loaded into a flow chamber with a 1.5×2 mm field of view. After water was flooded into the chamber, interference images were captured and analyzed to obtain high resolution maps of the local refractive index and subsequently the volume fraction and mass density of film material at each spatial location. Here, we compare the hydration dynamics of a panel of films with varying thicknesses and polymer compositions, demonstrating that quantitative imaging refractometry can be an effective tool for evaluating and characterizing the performance of candidate microbicide film designs for anti-HIV drug delivery. PMID:24736376

Analysis of vaginal microbicide film hydration kinetics by quantitative imaging refractometry.

PubMed

Rinehart, Matthew; Grab, Sheila; Rohan, Lisa; Katz, David; Wax, Adam

2014-01-01

We have developed a quantitative imaging refractometry technique, based on holographic phase microscopy, as a tool for investigating microscopic structural changes in water-soluble polymeric materials. Here we apply the approach to analyze the structural degradation of vaginal topical microbicide films due to water uptake. We implemented transmission imaging of 1-mm diameter film samples loaded into a flow chamber with a 1.5×2 mm field of view. After water was flooded into the chamber, interference images were captured and analyzed to obtain high resolution maps of the local refractive index and subsequently the volume fraction and mass density of film material at each spatial location. Here, we compare the hydration dynamics of a panel of films with varying thicknesses and polymer compositions, demonstrating that quantitative imaging refractometry can be an effective tool for evaluating and characterizing the performance of candidate microbicide film designs for anti-HIV drug delivery.

Quantitative phase imaging using a programmable wavefront sensor

NASA Astrophysics Data System (ADS)

Soldevila, F.; Durán, V.; Clemente, P.; Lancis, J.; Tajahuerce, E.

2018-02-01

We perform phase imaging using a non-interferometric approach to measure the complex amplitude of a wavefront. We overcome the limitations in spatial resolution, optical efficiency, and dynamic range that are found in Shack-Hartmann wavefront sensing. To do so, we sample the wavefront with a high-speed spatial light modulator. A single lens forms a time-dependent light distribution on its focal plane, where a position detector is placed. Our approach is lenslet-free and does not rely on any kind of iterative or unwrap algorithm. The validity of our technique is demonstrated by performing both aberration sensing and phase imaging of transparent samples.

Regularized iterative integration combined with non-linear diffusion filtering for phase-contrast x-ray computed tomography.

PubMed

Burger, Karin; Koehler, Thomas; Chabior, Michael; Allner, Sebastian; Marschner, Mathias; Fehringer, Andreas; Willner, Marian; Pfeiffer, Franz; Noël, Peter

2014-12-29

Phase-contrast x-ray computed tomography has a high potential to become clinically implemented because of its complementarity to conventional absorption-contrast.In this study, we investigate noise-reducing but resolution-preserving analytical reconstruction methods to improve differential phase-contrast imaging. We apply the non-linear Perona-Malik filter on phase-contrast data prior or post filtered backprojected reconstruction. Secondly, the Hilbert kernel is replaced by regularized iterative integration followed by ramp filtered backprojection as used for absorption-contrast imaging. Combining the Perona-Malik filter with this integration algorithm allows to successfully reveal relevant sample features, quantitatively confirmed by significantly increased structural similarity indices and contrast-to-noise ratios. With this concept, phase-contrast imaging can be performed at considerably lower dose.

Coherent diffraction imaging of non-isolated object with apodized illumination.

PubMed

Khakurel, Krishna P; Kimura, Takashi; Joti, Yasumasa; Matsuyama, Satoshi; Yamauchi, Kazuto; Nishino, Yoshinori

2015-11-02

Coherent diffraction imaging (CDI) is an established lensless imaging method widely used at the x-ray regime applicable to the imaging of non-periodic materials. Conventional CDI can practically image isolated objects only, which hinders the broader application of the method. We present the imaging of non-isolated objects by employing recently proposed "non-scanning" apodized-illumination CDI at an optical wavelength. We realized isolated apodized illumination with a specially designed optical configuration and succeeded in imaging phase objects as well as amplitude objects. The non-scanning nature of the method is important particularly in imaging live cells and tissues, where fast imaging is required for non-isolated objects, and is an advantage over ptychography. We believe that our result of phase contrast imaging at an optical wavelength can be extended to the quantitative phase imaging of cells and tissues. The method also provides the feasibility of the lensless single-shot imaging of extended objects with x-ray free-electron lasers.

Evaluation of a New Motion-correction Algorithm Using On-rigid Registration in Respiratory-gated PET/CT Images of Liver Tumors.

PubMed

Wagatsuma, Kei; Osawa, Tatsufumi; Yokokawa, Naoki; Miwa, Kenta; Oda, Keiichi; Kudo, Yoshiro; Unno, Yasushi; Ito, Kimiteru; Ishii, Kenji

2016-01-01

The present study aimed to determine the qualitative and quantitative accuracy of the Q.Freeze algorithm in PET/CT images of liver tumors. A body phantom and hot spheres representing liver tumors contained 5.3 and 21.2 kBq/mL of a solution containing 18 F radioactivity, respectively. The phantoms were moved in the superior-inferior direction at a motion displacement of 20 mm. Conventional respiratory-gated (RG) and Q.Freeze images were sorted into 6, 10, and 13 phase-groups. The SUV ave was calculated from the background of the body phantom, and the SUV max was determined from the hot spheres of the liver tumors. Three patients with four liver tumors were also clinically assessed by whole-body and RG PET. The RG and Q.Freeze images derived from the clinical study were also sorted into 6, 10 and 13 phase-groups. Liver signal-to-noise ratio (SNR) and SUV max were determined from the RG and Q.Freeze clinical images. The SUV ave of Q.Freeze images was the same as those derived from the body phantom using RG. The liver SNR improved with Q.Freeze, and the SUVs max was not overestimated when Q.Freeze was applied in both the phantom and clinical studies. Q.Freeze did not degrade the liver SNR and SUV max even though the phase number was larger. Q.Freeze delivered qualitative and quantitative motion correction than conventional RG imaging even in 10-phase groups.

Phase-contrast Hounsfield units of fixated and non-fixated soft-tissue samples

DOE Office of Scientific and Technical Information (OSTI.GOV)

Willner, Marian; Fior, Gabriel; Marschner, Mathias

X-ray phase-contrast imaging is a novel technology that achieves high soft-tissue contrast. Although its clinical impact is still under investigation, the technique may potentially improve clinical diagnostics. In conventional attenuation-based X-ray computed tomography, radiological diagnostics are quantified by Hounsfield units. Corresponding Hounsfield units for phase-contrast imaging have been recently introduced, enabling a setup-independent comparison and standardized interpretation of imaging results. Thus far, the experimental values of few tissue types have been reported; these values have been determined from fixated tissue samples. This study presents phase-contrast Hounsfield units for various types of non-fixated human soft tissues. A large variety of tissuemore » specimens ranging from adipose, muscle and connective tissues to liver, kidney and pancreas tissues were imaged by a grating interferometer with a rotating-anode X-ray tube and a photon-counting detector. In addition, we investigated the effects of formalin fixation on the quantitative phase-contrast imaging results.« less

Phase-Contrast Hounsfield Units of Fixated and Non-Fixated Soft-Tissue Samples

PubMed Central

Willner, Marian; Fior, Gabriel; Marschner, Mathias; Birnbacher, Lorenz; Schock, Jonathan; Braun, Christian; Fingerle, Alexander A.; Noël, Peter B.; Rummeny, Ernst J.; Pfeiffer, Franz; Herzen, Julia

2015-01-01

X-ray phase-contrast imaging is a novel technology that achieves high soft-tissue contrast. Although its clinical impact is still under investigation, the technique may potentially improve clinical diagnostics. In conventional attenuation-based X-ray computed tomography, radiological diagnostics are quantified by Hounsfield units. Corresponding Hounsfield units for phase-contrast imaging have been recently introduced, enabling a setup-independent comparison and standardized interpretation of imaging results. Thus far, the experimental values of few tissue types have been reported; these values have been determined from fixated tissue samples. This study presents phase-contrast Hounsfield units for various types of non-fixated human soft tissues. A large variety of tissue specimens ranging from adipose, muscle and connective tissues to liver, kidney and pancreas tissues were imaged by a grating interferometer with a rotating-anode X-ray tube and a photon-counting detector. Furthermore, we investigated the effects of formalin fixation on the quantitative phase-contrast imaging results. PMID:26322638

Phase-contrast Hounsfield units of fixated and non-fixated soft-tissue samples

DOE PAGES

Willner, Marian; Fior, Gabriel; Marschner, Mathias; ...

2015-08-31

X-ray phase-contrast imaging is a novel technology that achieves high soft-tissue contrast. Although its clinical impact is still under investigation, the technique may potentially improve clinical diagnostics. In conventional attenuation-based X-ray computed tomography, radiological diagnostics are quantified by Hounsfield units. Corresponding Hounsfield units for phase-contrast imaging have been recently introduced, enabling a setup-independent comparison and standardized interpretation of imaging results. Thus far, the experimental values of few tissue types have been reported; these values have been determined from fixated tissue samples. This study presents phase-contrast Hounsfield units for various types of non-fixated human soft tissues. A large variety of tissuemore » specimens ranging from adipose, muscle and connective tissues to liver, kidney and pancreas tissues were imaged by a grating interferometer with a rotating-anode X-ray tube and a photon-counting detector. In addition, we investigated the effects of formalin fixation on the quantitative phase-contrast imaging results.« less

Coherent diffraction surface imaging in reflection geometry.

PubMed

Marathe, Shashidhara; Kim, S S; Kim, S N; Kim, Chan; Kang, H C; Nickles, P V; Noh, D Y

2010-03-29

We present a reflection based coherent diffraction imaging method which can be used to reconstruct a non periodic surface image from a diffraction amplitude measured in reflection geometry. Using a He-Ne laser, we demonstrated that a surface image can be reconstructed solely from the reflected intensity from a surface without relying on any prior knowledge of the sample object or the object support. The reconstructed phase image of the exit wave is particularly interesting since it can be used to obtain quantitative information of the surface depth profile or the phase change during the reflection process. We believe that this work will broaden the application areas of coherent diffraction imaging techniques using light sources with limited penetration depth.

Spectral interferometry for morphological imaging in in vitro fertilization (IVF) (Conference Presentation)

NASA Astrophysics Data System (ADS)

Zhu, Yizheng; Li, Chengshuai

2016-03-01

Morphological assessment of spermatozoa is of critical importance for in vitro fertilization (IVF), especially intracytoplasmic sperm injection (ICSI)-based IVF. In ICSI, a single sperm cell is selected and injected into an egg to achieve fertilization. The quality of the sperm cell is found to be highly correlated to IVF success. Sperm morphology, such as shape, head birefringence and motility, among others, are typically evaluated under a microscope. Current observation relies on conventional techniques such as differential interference contrast microscopy and polarized light microscopy. Their qualitative nature, however, limits the ability to provide accurate quantitative analysis. Here, we demonstrate quantitative morphological measurement of sperm cells using two types of spectral interferometric techniques, namely spectral modulation interferometry and spectral multiplexing interferometry. Both are based on spectral-domain low coherence interferometry, which is known for its exquisite phase determination ability. While spectral modulation interferometry encodes sample phase in a single spectrum, spectral multiplexing interferometry does so for sample birefringence. Therefore they are capable of highly sensitive phase and birefringence imaging. These features suit well in the imaging of live sperm cells, which are small, dynamic objects with only low to moderate levels of phase and birefringence contrast. We will introduce the operation of both techniques and demonstrate their application to measuring the phase and birefringence morphology of sperm cells.

Qualitative and Quantitative Imaging Evaluation of Renal Cell Carcinoma Subtypes with Grating-based X-ray Phase-contrast CT

NASA Astrophysics Data System (ADS)

Braunagel, Margarita; Birnbacher, Lorenz; Willner, Marian; Marschner, Mathias; De Marco, Fabio; Viermetz, Manuel; Notohamiprodjo, Susan; Hellbach, Katharina; Auweter, Sigrid; Link, Vera; Woischke, Christine; Reiser, Maximilian F.; Pfeiffer, Franz; Notohamiprodjo, Mike; Herzen, Julia

2017-03-01

Current clinical imaging methods face limitations in the detection and correct characterization of different subtypes of renal cell carcinoma (RCC), while these are important for therapy and prognosis. The present study evaluates the potential of grating-based X-ray phase-contrast computed tomography (gbPC-CT) for visualization and characterization of human RCC subtypes. The imaging results for 23 ex vivo formalin-fixed human kidney specimens obtained with phase-contrast CT were compared to the results of the absorption-based CT (gbCT), clinical CT and a 3T MRI and validated using histology. Regions of interest were placed on each specimen for quantitative evaluation. Qualitative and quantitative gbPC-CT imaging could significantly discriminate between normal kidney cortex (54 ± 4 HUp) and clear cell (42 ± 10), papillary (43 ± 6) and chromophobe RCCs (39 ± 7), p < 0.05 respectively. The sensitivity for detection of tumor areas was 100%, 50% and 40% for gbPC-CT, gbCT and clinical CT, respectively. RCC architecture like fibrous strands, pseudocapsules, necrosis or hyalinization was depicted clearly in gbPC-CT and was not equally well visualized in gbCT, clinical CT and MRI. The results show that gbPC-CT enables improved discrimination of normal kidney parenchyma and tumorous tissues as well as different soft-tissue components of RCCs without the use of contrast media.

Effect of masking phase-only holograms on the quality of reconstructed images.

PubMed

Deng, Yuanbo; Chu, Daping

2016-04-20

A phase-only hologram modulates the phase of the incident light and diffracts it efficiently with low energy loss because of the minimum absorption. Much research attention has been focused on how to generate phase-only holograms, and little work has been done to understand the effect and limitation of their partial implementation, possibly due to physical defects and constraints, in particular as in the practical situations where a phase-only hologram is confined or needs to be sliced or tiled. The present study simulates the effect of masking phase-only holograms on the quality of reconstructed images in three different scenarios with different filling factors, filling positions, and illumination intensity profiles. Quantitative analysis confirms that the width of the image point spread function becomes wider and the image quality decreases, as expected, when the filling factor decreases, and the image quality remains the same for different filling positions as well. The width of the image point spread function as derived from different filling factors shows a consistent behavior to that as measured directly from the reconstructed image, especially as the filling factor becomes small. Finally, mask profiles of different shapes and intensity distributions are shown to have more complicated effects on the image point spread function, which in turn affects the quality and textures of the reconstructed image.

Cell classification using big data analytics plus time stretch imaging (Conference Presentation)

NASA Astrophysics Data System (ADS)

Jalali, Bahram; Chen, Claire L.; Mahjoubfar, Ata

2016-09-01

We show that blood cells can be classified with high accuracy and high throughput by combining machine learning with time stretch quantitative phase imaging. Our diagnostic system captures quantitative phase images in a flow microscope at millions of frames per second and extracts multiple biophysical features from individual cells including morphological characteristics, light absorption and scattering parameters, and protein concentration. These parameters form a hyperdimensional feature space in which supervised learning and cell classification is performed. We show binary classification of T-cells against colon cancer cells, as well classification of algae cell strains with high and low lipid content. The label-free screening averts the negative impact of staining reagents on cellular viability or cell signaling. The combination of time stretch machine vision and learning offers unprecedented cell analysis capabilities for cancer diagnostics, drug development and liquid biopsy for personalized genomics.

Quantitative comparison between a multiecho sequence and a single-echo sequence for susceptibility-weighted phase imaging.

PubMed

Gilbert, Guillaume; Savard, Geneviève; Bard, Céline; Beaudoin, Gilles

2012-06-01

The aim of this study was to investigate the benefits arising from the use of a multiecho sequence for susceptibility-weighted phase imaging using a quantitative comparison with a standard single-echo acquisition. Four healthy adult volunteers were imaged on a clinical 3-T system using a protocol comprising two different three-dimensional susceptibility-weighted gradient-echo sequences: a standard single-echo sequence and a multiecho sequence. Both sequences were repeated twice in order to evaluate the local noise contribution by a subtraction of the two acquisitions. For the multiecho sequence, the phase information from each echo was independently unwrapped, and the background field contribution was removed using either homodyne filtering or the projection onto dipole fields method. The phase information from all echoes was then combined using a weighted linear regression. R2 maps were also calculated from the multiecho acquisitions. The noise standard deviation in the reconstructed phase images was evaluated for six manually segmented regions of interest (frontal white matter, posterior white matter, globus pallidus, putamen, caudate nucleus and lateral ventricle). The use of the multiecho sequence for susceptibility-weighted phase imaging led to a reduction of the noise standard deviation for all subjects and all regions of interest investigated in comparison to the reference single-echo acquisition. On average, the noise reduction ranged from 18.4% for the globus pallidus to 47.9% for the lateral ventricle. In addition, the amount of noise reduction was found to be strongly inversely correlated to the estimated R2 value (R=-0.92). In conclusion, the use of a multiecho sequence is an effective way to decrease the noise contribution in susceptibility-weighted phase images, while preserving both contrast and acquisition time. The proposed approach additionally permits the calculation of R2 maps. Copyright © 2012 Elsevier Inc. All rights reserved.

Low-dose quantitative phase contrast medical CT

NASA Astrophysics Data System (ADS)

Mittone, A.; Bravin, A.; Coan, P.

2018-02-01

X-ray computed tomography (CT) is a powerful and routinely used clinical diagnostic technique, which is well tolerated by patients, and which provides high-resolution images and volumetric information about the body. However, two important limitations still affect this examination procedure: (1) its low sensitivity with respect to soft tissues, and (2) the hazards associated with x-ray exposure. Conventional radiology is based on the detection of the different photon absorption properties that characterize biological tissues, and thus the obtainable image contrast from soft and/or similar tissues is intrinsically limited. In this scenario, x-ray phase contrast imaging (XPCI) has been extensively tested and proven to overcome some of the main issues surrounding standard x-ray imaging. In addition to the absorption signal, XPCI relies on detecting the phase shifts induced by an object. Interestingly, as the order of magnitude of the phase contrast is higher than that of absorption, XPCI can, in principle, offer higher sensitivity at lower radiation doses. However, other technical aspects may counterbalance this gain, and an optimized setup and image processing solutions need to be implemented. The work presented here describes the strategies and developments we have realized, with the aim of controlling the radiation dose for the highly sensitive and quantitative XPCI-CT. Different algorithms for the phase retrieval and CT reconstruction of the XPCI data are presented. The CT algorithms we have implemented, namely the equally sloped tomography and the dictionary learning method, allow the image quality to be preserved while reducing the number of angular projections required by a factor of five. The results applied to breast imaging report accurate reconstructions at clinically compatible doses of the 3D distribution of the refractive properties of full human organs obtained by using three different phase retrieval methods. The described methodologies and the presented results have been validated by a team of clinical radiologists and represent an important step in the exploitation of XPCI-CT for in vivo and possible clinical applications.

Analysis of objects in binary images. M.S. Thesis - Old Dominion Univ.

NASA Technical Reports Server (NTRS)

Leonard, Desiree M.

1991-01-01

Digital image processing techniques are typically used to produce improved digital images through the application of successive enhancement techniques to a given image or to generate quantitative data about the objects within that image. In support of and to assist researchers in a wide range of disciplines, e.g., interferometry, heavy rain effects on aerodynamics, and structure recognition research, it is often desirable to count objects in an image and compute their geometric properties. Therefore, an image analysis application package, focusing on a subset of image analysis techniques used for object recognition in binary images, was developed. This report describes the techniques and algorithms utilized in three main phases of the application and are categorized as: image segmentation, object recognition, and quantitative analysis. Appendices provide supplemental formulas for the algorithms employed as well as examples and results from the various image segmentation techniques and the object recognition algorithm implemented.

Quantitative characterization of edge enhancement in phase contrast x-ray imaging.

PubMed

Monnin, P; Bulling, S; Hoszowska, J; Valley, J F; Meuli, R; Verdun, F R

2004-06-01

The aim of this study was to model the edge enhancement effect in in-line holography phase contrast imaging. A simple analytical approach was used to quantify refraction and interference contrasts in terms of beam energy and imaging geometry. The model was applied to predict the peak intensity and frequency of the edge enhancement for images of cylindrical fibers. The calculations were compared with measurements, and the relationship between the spatial resolution of the detector and the amplitude of the phase contrast signal was investigated. Calculations using the analytical model were in good agreement with experimental results for nylon, aluminum and copper wires of 50 to 240 microm diameter, and with numerical simulations based on Fresnel-Kirchhoff theory. A relationship between the defocusing distance and the pixel size of the image detector was established. This analytical model is a useful tool for optimizing imaging parameters in phase contrast in-line holography, including defocusing distance, detector resolution and beam energy.

Structured illumination multimodal 3D-resolved quantitative phase and fluorescence sub-diffraction microscopy

PubMed Central

Chowdhury, Shwetadwip; Eldridge, Will J.; Wax, Adam; Izatt, Joseph A.

2017-01-01

Sub-diffraction resolution imaging has played a pivotal role in biological research by visualizing key, but previously unresolvable, sub-cellular structures. Unfortunately, applications of far-field sub-diffraction resolution are currently divided between fluorescent and coherent-diffraction regimes, and a multimodal sub-diffraction technique that bridges this gap has not yet been demonstrated. Here we report that structured illumination (SI) allows multimodal sub-diffraction imaging of both coherent quantitative-phase (QP) and fluorescence. Due to SI’s conventionally fluorescent applications, we first demonstrate the principle of SI-enabled three-dimensional (3D) QP sub-diffraction imaging with calibration microspheres. Image analysis confirmed enhanced lateral and axial resolutions over diffraction-limited QP imaging, and established striking parallels between coherent SI and conventional optical diffraction tomography. We next introduce an optical system utilizing SI to achieve 3D sub-diffraction, multimodal QP/fluorescent visualization of A549 biological cells fluorescently tagged for F-actin. Our results suggest that SI has a unique utility in studying biological phenomena with significant molecular, biophysical, and biochemical components. PMID:28663887

Linear information retrieval method in X-ray grating-based phase contrast imaging and its interchangeability with tomographic reconstruction

NASA Astrophysics Data System (ADS)

Wu, Z.; Gao, K.; Wang, Z. L.; Shao, Q. G.; Hu, R. F.; Wei, C. X.; Zan, G. B.; Wali, F.; Luo, R. H.; Zhu, P. P.; Tian, Y. C.

2017-06-01

In X-ray grating-based phase contrast imaging, information retrieval is necessary for quantitative research, especially for phase tomography. However, numerous and repetitive processes have to be performed for tomographic reconstruction. In this paper, we report a novel information retrieval method, which enables retrieving phase and absorption information by means of a linear combination of two mutually conjugate images. Thanks to the distributive law of the multiplication as well as the commutative law and associative law of the addition, the information retrieval can be performed after tomographic reconstruction, thus simplifying the information retrieval procedure dramatically. The theoretical model of this method is established in both parallel beam geometry for Talbot interferometer and fan beam geometry for Talbot-Lau interferometer. Numerical experiments are also performed to confirm the feasibility and validity of the proposed method. In addition, we discuss its possibility in cone beam geometry and its advantages compared with other methods. Moreover, this method can also be employed in other differential phase contrast imaging methods, such as diffraction enhanced imaging, non-interferometric imaging, and edge illumination.

Optical Ptychographic Microscope for Quantitative Bio-Mechanical Imaging

NASA Astrophysics Data System (ADS)

Anthony, Nicholas; Cadenazzi, Guido; Nugent, Keith; Abbey, Brian

The role that mechanical forces play in biological processes such as cell movement and death is becoming of significant interest to further develop our understanding of the inner workings of cells. The most common method used to obtain stress information is photoelasticity which maps a samples birefringence, or its direction dependent refractive indices, using polarized light. However this method only provides qualitative data and for stress information to be useful quantitative data is required. Ptychography is a method for quantitatively determining the phase of a samples complex transmission function. The technique relies upon the collection of multiple overlapping coherent diffraction patterns from laterally displaced points on the sample. The overlap of measurement points provides complementary information that significantly aids in the reconstruction of the complex wavefield exiting the sample and allows for quantitative imaging of weakly interacting specimens. Here we describe recent advances at La Trobe University Melbourne on achieving quantitative birefringence mapping using polarized light ptychography with applications in cell mechanics. Australian Synchrotron, ARC Centre of Excellence for Advanced Molecular Imaging.

Dynamic measurements of flowing cells labeled by gold nanoparticles using full-field photothermal interferometric imaging

NASA Astrophysics Data System (ADS)

Turko, Nir A.; Roitshtain, Darina; Blum, Omry; Kemper, Björn; Shaked, Natan T.

2017-06-01

We present highly dynamic photothermal interferometric phase microscopy for quantitative, selective contrast imaging of live cells during flow. Gold nanoparticles can be biofunctionalized to bind to specific cells, and stimulated for local temperature increase due to plasmon resonance, causing a rapid change of the optical phase. These phase changes can be recorded by interferometric phase microscopy and analyzed to form an image of the binding sites of the nanoparticles in the cells, gaining molecular specificity. Since the nanoparticle excitation frequency might overlap with the sample dynamics frequencies, photothermal phase imaging was performed on stationary or slowly dynamic samples. Furthermore, the computational analysis of the photothermal signals is time consuming. This makes photothermal imaging unsuitable for applications requiring dynamic imaging or real-time analysis, such as analyzing and sorting cells during fast flow. To overcome these drawbacks, we utilized an external interferometric module and developed new algorithms, based on discrete Fourier transform variants, enabling fast analysis of photothermal signals in highly dynamic live cells. Due to the self-interference module, the cells are imaged with and without excitation in video-rate, effectively increasing signal-to-noise ratio. Our approach holds potential for using photothermal cell imaging and depletion in flow cytometry.

Medicine, material science and security: the versatility of the coded-aperture approach.

PubMed

Munro, P R T; Endrizzi, M; Diemoz, P C; Hagen, C K; Szafraniec, M B; Millard, T P; Zapata, C E; Speller, R D; Olivo, A

2014-03-06

The principal limitation to the widespread deployment of X-ray phase imaging in a variety of applications is probably versatility. A versatile X-ray phase imaging system must be able to work with polychromatic and non-microfocus sources (for example, those currently used in medical and industrial applications), have physical dimensions sufficiently large to accommodate samples of interest, be insensitive to environmental disturbances (such as vibrations and temperature variations), require only simple system set-up and maintenance, and be able to perform quantitative imaging. The coded-aperture technique, based upon the edge illumination principle, satisfies each of these criteria. To date, we have applied the technique to mammography, materials science, small-animal imaging, non-destructive testing and security. In this paper, we outline the theory of coded-aperture phase imaging and show an example of how the technique may be applied to imaging samples with a practically important scale.

Flow-gated radial phase-contrast imaging in the presence of weak flow.

PubMed

Peng, Hsu-Hsia; Huang, Teng-Yi; Wang, Fu-Nien; Chung, Hsiao-Wen

2013-01-01

To implement a flow-gating method to acquire phase-contrast (PC) images of carotid arteries without use of an electrocardiography (ECG) signal to synchronize the acquisition of imaging data with pulsatile arterial flow. The flow-gating method was realized through radial scanning and sophisticated post-processing methods including downsampling, complex difference, and correlation analysis to improve the evaluation of flow-gating times in radial phase-contrast scans. Quantitatively comparable results (R = 0.92-0.96, n = 9) of flow-related parameters, including mean velocity, mean flow rate, and flow volume, with conventional ECG-gated imaging demonstrated that the proposed method is highly feasible. The radial flow-gating PC imaging method is applicable in carotid arteries. The proposed flow-gating method can potentially avoid the setting up of ECG-related equipment for brain imaging. This technique has potential use in patients with arrhythmia or weak ECG signals.

Quantitative assessment on coronary computed tomography angiography (CCTA) image quality: comparisons between genders and different tube voltage settings.

PubMed

Chian, Teo Chee; Nassir, Norziana Mat; Ibrahim, Mohd Izuan; Yusof, Ahmad Khairuddin Md; Sabarudin, Akmal

2017-02-01

This study was carried out to quantify and compare the quantitative image quality of coronary computed tomography angiography (CCTA) between genders as well as between different tube voltages scan protocols. Fifty-five cases of CCTA were collected retrospectively and all images including reformatted axial images at systolic and diastolic phases as well as images with curved multi planar reformation (cMPR) were obtained. Quantitative image quality including signal intensity, image noise, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of right coronary artery (RCA), left anterior descending artery (LAD), left circumflex artery (LCx) and left main artery (LM) were quantified using Analyze 12.0 software. Six hundred and fifty-seven coronary arteries were evaluated. There were no significant differences in any quantitative image quality parameters between genders. 100 kilovoltage peak (kVp) scanning protocol produced images with significantly higher signal intensity compared to 120 kVp scanning protocol (P<0.001) in all coronary arteries in all types of images. Higher SNR was also observed in 100 kVp scan protocol in all coronary arteries except in LCx where 120 kVp showed better SNR than 100 kVp. There were no significant differences in image quality of CCTA between genders and different tube voltages. Lower tube voltage (100 kVp) scanning protocol is recommended in clinical practice to reduce the radiation dose to patient.

Lensless transport-of-intensity phase microscopy and tomography with a color LED matrix

NASA Astrophysics Data System (ADS)

Zuo, Chao; Sun, Jiasong; Zhang, Jialin; Hu, Yan; Chen, Qian

2015-07-01

We demonstrate lens-less quantitative phase microscopy and diffraction tomography based on a compact on-chip platform, using only a CMOS image sensor and a programmable color LED array. Based on multi-wavelength transport-of- intensity phase retrieval and multi-angle illumination diffraction tomography, this platform offers high quality, depth resolved images with a lateral resolution of ˜3.7μm and an axial resolution of ˜5μm, over wide large imaging FOV of 24mm2. The resolution and FOV can be further improved by using a larger image sensors with small pixels straightforwardly. This compact, low-cost, robust, portable platform with a decent imaging performance may offer a cost-effective tool for telemedicine needs, or for reducing health care costs for point-of-care diagnostics in resource-limited environments.

The evolution of phase holographic imaging from a research idea to publicly traded company

NASA Astrophysics Data System (ADS)

Egelberg, Peter

2018-02-01

Recognizing the value and unmet need for label-free kinetic cell analysis, Phase Holograhic Imaging defines its market segment as automated, easy to use and affordable time-lapse cytometry. The process of developing new technology, meeting customer expectations, sources of corporate funding and R&D adjustments prompted by field experience will be reviewed. Additionally, it is discussed how relevant biological information can be extracted from a sequence of quantitative phase images, with negligible user assistance and parameter tweaking, to simultaneously provide cell culture characteristics such as cell growth rate, viability, division rate, mitosis duration, phagocytosis rate, migration, motility and cell-cell adherence without requiring any artificial cell manipulation.

High resolution laboratory grating-based x-ray phase-contrast CT

NASA Astrophysics Data System (ADS)

Viermetz, Manuel P.; Birnbacher, Lorenz J. B.; Fehringer, Andreas; Willner, Marian; Noel, Peter B.; Pfeiffer, Franz; Herzen, Julia

2017-03-01

Grating-based phase-contrast computed tomography (gbPC-CT) is a promising imaging method for imaging of soft tissue contrast without the need of any contrast agent. The focus of this study is the increase in spatial resolution without loss in sensitivity to allow visualization of pathologies comparable to the convincing results obtained at the synchrotron. To improve the effective pixel size a super-resolution reconstruction based on subpixel shifts involving a deconvolution of the image is applied on differential phase-contrast data. In our study we could achieve an effective pixel sizes of 28mm without any drawback in terms of sensitivity or the ability to measure quantitative data.

Three-dimensional label-free imaging and quantification of lipid droplets in live hepatocytes

NASA Astrophysics Data System (ADS)

Kim, Kyoohyun; Lee, Seoeun; Yoon, Jonghee; Heo, Jihan; Choi, Chulhee; Park, Yongkeun

2016-11-01

Lipid droplets (LDs) are subcellular organelles with important roles in lipid storage and metabolism and involved in various diseases including cancer, obesity, and diabetes. Conventional methods, however, have limited ability to provide quantitative information on individual LDs and have limited capability for three-dimensional (3-D) imaging of LDs in live cells especially for fast acquisition of 3-D dynamics. Here, we present an optical method based on 3-D quantitative phase imaging to measure the 3-D structural distribution and biochemical parameters (concentration and dry mass) of individual LDs in live cells without using exogenous labelling agents. The biochemical change of LDs under oleic acid treatment was quantitatively investigated, and 4-D tracking of the fast dynamics of LDs revealed the intracellular transport of LDs in live cells.

X-Ray Nanoscopy of a Bulk Heterojunction

NASA Astrophysics Data System (ADS)

Patil, Nilesh; Torbjørn, Eirik; Skjønsfjell, Bakken; Van den Brande, Niko; Chavez Panduro, Elvia Anabela; Claessens, Raf; Guizar-Sicairos, Manuel; Van Mele, Bruno; Breiby, Dag Werner

2016-07-01

Optimizing the morphology of bulk heterojunctions is known to significantly improve the photovoltaic performance of organic solar cells, but available quantitative imaging techniques are few and have severe limitations. We demonstrate X-ray ptychographic coherent diffractive imaging applied to all-organic blends. Specifically, the phase-separated morphology in bulk heterojunction photoactive layers for organic solar cells, prepared from a 50:50 blend of poly(3-hexylthiophene) (P3HT) and phenyl-C61-butyric acid methyl ester (PCBM) and thermally treated for different annealing times is imaged to high resolution. Moreover, using a fast-scanning calorimetry chip setup, the nano-morphological changes caused by repeated thermal annealing applied to the same sample could be monitored. X-ray ptychography resolves to better than 100 nm the phase-segregated domains of electron donor and electron acceptor materials over a large field of view within the active layers. The quantitative phase contrast images further allow us to estimate the local volume fraction of PCBM across the photovoltaically active layers. The volume fraction gradient for different regions provides insight on the PCBM diffusion across the depletion zone surrounding PCBM aggregates. Phase contrast X-ray microscopy is under rapid development, and the results presented here are promising for future studies of organic-organic blends, also under in situ conditions, e.g., for monitoring the structural stability during UV-Vis irradiation.

X-Ray Nanoscopy of a Bulk Heterojunction.

PubMed

Patil, Nilesh; Skjønsfjell, Eirik Torbjørn Bakken; Van den Brande, Niko; Chavez Panduro, Elvia Anabela; Claessens, Raf; Guizar-Sicairos, Manuel; Van Mele, Bruno; Breiby, Dag Werner

2016-01-01

Optimizing the morphology of bulk heterojunctions is known to significantly improve the photovoltaic performance of organic solar cells, but available quantitative imaging techniques are few and have severe limitations. We demonstrate X-ray ptychographic coherent diffractive imaging applied to all-organic blends. Specifically, the phase-separated morphology in bulk heterojunction photoactive layers for organic solar cells, prepared from a 50:50 blend of poly(3-hexylthiophene) (P3HT) and phenyl-C61-butyric acid methyl ester (PCBM) and thermally treated for different annealing times is imaged to high resolution. Moreover, using a fast-scanning calorimetry chip setup, the nano-morphological changes caused by repeated thermal annealing applied to the same sample could be monitored. X-ray ptychography resolves to better than 100 nm the phase-segregated domains of electron donor and electron acceptor materials over a large field of view within the active layers. The quantitative phase contrast images further allow us to estimate the local volume fraction of PCBM across the photovoltaically active layers. The volume fraction gradient for different regions provides insight on the PCBM diffusion across the depletion zone surrounding PCBM aggregates. Phase contrast X-ray microscopy is under rapid development, and the results presented here are promising for future studies of organic-organic blends, also under in situ conditions, e.g., for monitoring the structural stability during UV-Vis irradiation.

A three-image algorithm for hard x-ray grating interferometry.

PubMed

Pelliccia, Daniele; Rigon, Luigi; Arfelli, Fulvia; Menk, Ralf-Hendrik; Bukreeva, Inna; Cedola, Alessia

2013-08-12

A three-image method to extract absorption, refraction and scattering information for hard x-ray grating interferometry is presented. The method comprises a post-processing approach alternative to the conventional phase stepping procedure and is inspired by a similar three-image technique developed for analyzer-based x-ray imaging. Results obtained with this algorithm are quantitatively comparable with phase-stepping. This method can be further extended to samples with negligible scattering, where only two images are needed to separate absorption and refraction signal. Thanks to the limited number of images required, this technique is a viable route to bio-compatible imaging with x-ray grating interferometer. In addition our method elucidates and strengthens the formal and practical analogies between grating interferometry and the (non-interferometric) diffraction enhanced imaging technique.

Quantitative assessment of cancer cell morphology and motility using telecentric digital holographic microscopy and machine learning.

PubMed

Lam, Van K; Nguyen, Thanh C; Chung, Byung M; Nehmetallah, George; Raub, Christopher B

2018-03-01

The noninvasive, fast acquisition of quantitative phase maps using digital holographic microscopy (DHM) allows tracking of rapid cellular motility on transparent substrates. On two-dimensional surfaces in vitro, MDA-MB-231 cancer cells assume several morphologies related to the mode of migration and substrate stiffness, relevant to mechanisms of cancer invasiveness in vivo. The quantitative phase information from DHM may accurately classify adhesive cancer cell subpopulations with clinical relevance. To test this, cells from the invasive breast cancer MDA-MB-231 cell line were cultured on glass, tissue-culture treated polystyrene, and collagen hydrogels, and imaged with DHM followed by epifluorescence microscopy after staining F-actin and nuclei. Trends in cell phase parameters were tracked on the different substrates, during cell division, and during matrix adhesion, relating them to F-actin features. Support vector machine learning algorithms were trained and tested using parameters from holographic phase reconstructions and cell geometric features from conventional phase images, and used to distinguish between elongated and rounded cell morphologies. DHM was able to distinguish between elongated and rounded morphologies of MDA-MB-231 cells with 94% accuracy, compared to 83% accuracy using cell geometric features from conventional brightfield microscopy. This finding indicates the potential of DHM to detect and monitor cancer cell morphologies relevant to cell cycle phase status, substrate adhesion, and motility. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

High-resolution three-dimensional partially coherent diffraction imaging.

PubMed

Clark, J N; Huang, X; Harder, R; Robinson, I K

2012-01-01

The wave properties of light, particularly its coherence, are responsible for interference effects, which can be exploited in powerful imaging applications. Coherent diffractive imaging relies heavily on coherence and has recently experienced rapid growth. Coherent diffractive imaging recovers an object from its diffraction pattern by computational phasing with the potential of wavelength-limited resolution. Diminished coherence results in reconstructions that suffer from artefacts or fail completely. Here we demonstrate ab initio phasing of partially coherent diffraction patterns in three dimensions, while simultaneously determining the coherence properties of the illuminating wavefield. Both the dramatic improvements in image interpretability and the three-dimensional evaluation of the coherence will have broad implications for quantitative imaging of nanostructures and wavefield characterization with X-rays and electrons.

Modeling of optical quadrature microscopy for imaging mouse embryos

NASA Astrophysics Data System (ADS)

Warger, William C., II; DiMarzio, Charles A.

2008-02-01

Optical quadrature microscopy (OQM) has been shown to provide the optical path difference through a mouse embryo, and has led to a novel method to count the total number of cells further into development than current non-toxic imaging techniques used in the clinic. The cell counting method has the potential to provide an additional quantitative viability marker for blastocyst transfer during in vitro fertilization. OQM uses a 633 nm laser within a modified Mach-Zehnder interferometer configuration to measure the amplitude and phase of the signal beam that travels through the embryo. Four cameras preceded by multiple beamsplitters record the four interferograms that are used within a reconstruction algorithm to produce an image of the complex electric field amplitude. Here we present a model for the electric field through the primary optical components in the imaging configuration and the reconstruction algorithm to calculate the signal to noise ratio when imaging mouse embryos. The model includes magnitude and phase errors in the individual reference and sample paths, fixed pattern noise, and noise within the laser and detectors. This analysis provides the foundation for determining the imaging limitations of OQM and the basis to optimize the cell counting method in order to introduce additional quantitative viability markers.

Automated Quantitative Rare Earth Elements Mineralogy by Scanning Electron Microscopy

NASA Astrophysics Data System (ADS)

Sindern, Sven; Meyer, F. Michael

2016-09-01

Increasing industrial demand of rare earth elements (REEs) stems from the central role they play for advanced technologies and the accelerating move away from carbon-based fuels. However, REE production is often hampered by the chemical, mineralogical as well as textural complexity of the ores with a need for better understanding of their salient properties. This is not only essential for in-depth genetic interpretations but also for a robust assessment of ore quality and economic viability. The design of energy and cost-efficient processing of REE ores depends heavily on information about REE element deportment that can be made available employing automated quantitative process mineralogy. Quantitative mineralogy assigns numeric values to compositional and textural properties of mineral matter. Scanning electron microscopy (SEM) combined with a suitable software package for acquisition of backscatter electron and X-ray signals, phase assignment and image analysis is one of the most efficient tools for quantitative mineralogy. The four different SEM-based automated quantitative mineralogy systems, i.e. FEI QEMSCAN and MLA, Tescan TIMA and Zeiss Mineralogic Mining, which are commercially available, are briefly characterized. Using examples of quantitative REE mineralogy, this chapter illustrates capabilities and limitations of automated SEM-based systems. Chemical variability of REE minerals and analytical uncertainty can reduce performance of phase assignment. This is shown for the REE phases parisite and synchysite. In another example from a monazite REE deposit, the quantitative mineralogical parameters surface roughness and mineral association derived from image analysis are applied for automated discrimination of apatite formed in a breakdown reaction of monazite and apatite formed by metamorphism prior to monazite breakdown. SEM-based automated mineralogy fulfils all requirements for characterization of complex unconventional REE ores that will become increasingly important for supply of REEs in the future.

A software platform for phase contrast x-ray breast imaging research.

PubMed

Bliznakova, K; Russo, P; Mettivier, G; Requardt, H; Popov, P; Bravin, A; Buliev, I

2015-06-01

To present and validate a computer-based simulation platform dedicated for phase contrast x-ray breast imaging research. The software platform, developed at the Technical University of Varna on the basis of a previously validated x-ray imaging software simulator, comprises modules for object creation and for x-ray image formation. These modules were updated to take into account the refractive index for phase contrast imaging as well as implementation of the Fresnel-Kirchhoff diffraction theory of the propagating x-ray waves. Projection images are generated in an in-line acquisition geometry. To test and validate the platform, several phantoms differing in their complexity were constructed and imaged at 25 keV and 60 keV at the beamline ID17 of the European Synchrotron Radiation Facility. The software platform was used to design computational phantoms that mimic those used in the experimental study and to generate x-ray images in absorption and phase contrast modes. The visual and quantitative results of the validation process showed an overall good correlation between simulated and experimental images and show the potential of this platform for research in phase contrast x-ray imaging of the breast. The application of the platform is demonstrated in a feasibility study for phase contrast images of complex inhomogeneous and anthropomorphic breast phantoms, compared to x-ray images generated in absorption mode. The improved visibility of mammographic structures suggests further investigation and optimisation of phase contrast x-ray breast imaging, especially when abnormalities are present. The software platform can be exploited also for educational purposes. Copyright © 2015 Elsevier Ltd. All rights reserved.

TH-A-207B-00: Shear-Wave Imaging and a QIBA US Biomarker Update

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

Imaging of tissue elastic properties is a relatively new and powerful approach to one of the oldest and most important diagnostic tools. Imaging of shear wave speed with ultrasound is has been added to most high-end ultrasound systems. Understanding this exciting imaging mode aiding its most effective use in medicine can be a rewarding effort for medical physicists and other medical imaging and treatment professionals. Assuring consistent, quantitative measurements across the many ultrasound systems in a typical imaging department will constitute a major step toward realizing the great potential of this technique and other quantitative imaging. This session will targetmore » these two goals with two presentations. A. Basics and Current Implementations of Ultrasound Imaging of Shear Wave Speed and Elasticity - Shigao Chen, Ph.D. Learning objectives-To understand: Introduction: Importance of tissue elasticity measurement Strain vs. shear wave elastography (SWE), beneficial features of SWE The link between shear wave speed and material properties, influence of viscosity Generation of shear waves External vibration (Fibroscan) ultrasound radiation force Point push Supersonic push (Aixplorer) Comb push (GE Logiq E9) Detection of shear waves Motion detection from pulse-echo ultrasound Importance of frame rate for shear wave imaging Plane wave imaging detection How to achieve high effective frame rate using line-by-line scanners Shear wave speed calculation Time to peak Random sample consensus (RANSAC) Cross correlation Sources of bias and variation in SWE Tissue viscosity Transducer compression or internal pressure of organ Reflection of shear waves at boundaries B. Elasticity Imaging System Biomarker Qualification and User Testing of Systems – Brian Garra, M.D. Learning objectives-To understand: Goals Review the need for quantitative medical imaging Provide examples of quantitative imaging biomarkers Acquaint the participant with the purpose of the RSNA Quantitative Imaging Biomarker Alliance and the need for such an organization Review the QIBA process for creating a quantitative biomarker Summarize steps needed to verify adherence of site, operators, and imaging systems to a QIBA profile Underlying Premise and Assumptions Objective, quantifiable results are needed to enhance the value of diagnostic imaging in clinical practice Reasons for quantification Evidence based medicine requires objective, not subjective observer data Computerized decision support tools (eg CAD) generally require quantitative input. Quantitative, reproducible measures are more easily used to develop personalized molecular medical diagnostic and treatment systems What is quantitative imaging? Definition from Imaging Metrology Workshop The Quantitative Imaging Biomarker Alliance Formation 2008 Mission Structure Example Imaging Biomarkers Being Explored Biomarker Selection Groundwork Draft Protocol for imaging and data evaluation QIBA Profile Drafting Equipment and Site Validation Technical Clinical Site and Equipment QA and Compliance Checking Ultrasound Elasticity Estimation Biomarker US Elasticity Estimation Background Current Status and Problems Biomarker Selection-process and outcome US SWS for Liver Fibrosis Biomarker Work Groundwork Literature search and analysis results Phase I phantom testing-Elastic phantoms Phase II phantom testing-Viscoelastic phantoms Digital Simulated Data Protocol and Profile Drafting Protocol: based on UPICT and existing literature and standards bodies protocols Profile-Current claims, Manufacturer specific appendices What comes after the profile Profile Validation Technical validation Clinical validation QA and Compliance Possible approaches Site Operator testing Site protocol re-evaluation Imaging system Manufacturer testing and attestation User acceptance testing and periodic QA Phantom Tests Digital Phantom Based Testing Standard QA Testing Remediation Schemes Profile Evolution Towards additional applications Towards higher accuracy and precision Supported in part by NIH contract HHSN268201300071C from NIBIB. Collaboration with GE Global Research, no personal support.; S. Chen, Some technologies described in this presentation have been licensed. Mayo Clinic and Dr. Chen have financial interests these technologies.« less

TH-A-207B-01: Basics and Current Implementations of Ultrasound Imaging of Shear Wave Speed and Elasticity

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen, S.

Imaging of tissue elastic properties is a relatively new and powerful approach to one of the oldest and most important diagnostic tools. Imaging of shear wave speed with ultrasound is has been added to most high-end ultrasound systems. Understanding this exciting imaging mode aiding its most effective use in medicine can be a rewarding effort for medical physicists and other medical imaging and treatment professionals. Assuring consistent, quantitative measurements across the many ultrasound systems in a typical imaging department will constitute a major step toward realizing the great potential of this technique and other quantitative imaging. This session will targetmore » these two goals with two presentations. A. Basics and Current Implementations of Ultrasound Imaging of Shear Wave Speed and Elasticity - Shigao Chen, Ph.D. Learning objectives-To understand: Introduction: Importance of tissue elasticity measurement Strain vs. shear wave elastography (SWE), beneficial features of SWE The link between shear wave speed and material properties, influence of viscosity Generation of shear waves External vibration (Fibroscan) ultrasound radiation force Point push Supersonic push (Aixplorer) Comb push (GE Logiq E9) Detection of shear waves Motion detection from pulse-echo ultrasound Importance of frame rate for shear wave imaging Plane wave imaging detection How to achieve high effective frame rate using line-by-line scanners Shear wave speed calculation Time to peak Random sample consensus (RANSAC) Cross correlation Sources of bias and variation in SWE Tissue viscosity Transducer compression or internal pressure of organ Reflection of shear waves at boundaries B. Elasticity Imaging System Biomarker Qualification and User Testing of Systems – Brian Garra, M.D. Learning objectives-To understand: Goals Review the need for quantitative medical imaging Provide examples of quantitative imaging biomarkers Acquaint the participant with the purpose of the RSNA Quantitative Imaging Biomarker Alliance and the need for such an organization Review the QIBA process for creating a quantitative biomarker Summarize steps needed to verify adherence of site, operators, and imaging systems to a QIBA profile Underlying Premise and Assumptions Objective, quantifiable results are needed to enhance the value of diagnostic imaging in clinical practice Reasons for quantification Evidence based medicine requires objective, not subjective observer data Computerized decision support tools (eg CAD) generally require quantitative input. Quantitative, reproducible measures are more easily used to develop personalized molecular medical diagnostic and treatment systems What is quantitative imaging? Definition from Imaging Metrology Workshop The Quantitative Imaging Biomarker Alliance Formation 2008 Mission Structure Example Imaging Biomarkers Being Explored Biomarker Selection Groundwork Draft Protocol for imaging and data evaluation QIBA Profile Drafting Equipment and Site Validation Technical Clinical Site and Equipment QA and Compliance Checking Ultrasound Elasticity Estimation Biomarker US Elasticity Estimation Background Current Status and Problems Biomarker Selection-process and outcome US SWS for Liver Fibrosis Biomarker Work Groundwork Literature search and analysis results Phase I phantom testing-Elastic phantoms Phase II phantom testing-Viscoelastic phantoms Digital Simulated Data Protocol and Profile Drafting Protocol: based on UPICT and existing literature and standards bodies protocols Profile-Current claims, Manufacturer specific appendices What comes after the profile Profile Validation Technical validation Clinical validation QA and Compliance Possible approaches Site Operator testing Site protocol re-evaluation Imaging system Manufacturer testing and attestation User acceptance testing and periodic QA Phantom Tests Digital Phantom Based Testing Standard QA Testing Remediation Schemes Profile Evolution Towards additional applications Towards higher accuracy and precision Supported in part by NIH contract HHSN268201300071C from NIBIB. Collaboration with GE Global Research, no personal support.; S. Chen, Some technologies described in this presentation have been licensed. Mayo Clinic and Dr. Chen have financial interests these technologies.« less

TH-A-207B-02: QIBA Ultrasound Elasticity Imaging System Biomarker Qualification and User Testing of Systems

DOE Office of Scientific and Technical Information (OSTI.GOV)

Garra, B.

Imaging of tissue elastic properties is a relatively new and powerful approach to one of the oldest and most important diagnostic tools. Imaging of shear wave speed with ultrasound is has been added to most high-end ultrasound systems. Understanding this exciting imaging mode aiding its most effective use in medicine can be a rewarding effort for medical physicists and other medical imaging and treatment professionals. Assuring consistent, quantitative measurements across the many ultrasound systems in a typical imaging department will constitute a major step toward realizing the great potential of this technique and other quantitative imaging. This session will targetmore » these two goals with two presentations. A. Basics and Current Implementations of Ultrasound Imaging of Shear Wave Speed and Elasticity - Shigao Chen, Ph.D. Learning objectives-To understand: Introduction: Importance of tissue elasticity measurement Strain vs. shear wave elastography (SWE), beneficial features of SWE The link between shear wave speed and material properties, influence of viscosity Generation of shear waves External vibration (Fibroscan) ultrasound radiation force Point push Supersonic push (Aixplorer) Comb push (GE Logiq E9) Detection of shear waves Motion detection from pulse-echo ultrasound Importance of frame rate for shear wave imaging Plane wave imaging detection How to achieve high effective frame rate using line-by-line scanners Shear wave speed calculation Time to peak Random sample consensus (RANSAC) Cross correlation Sources of bias and variation in SWE Tissue viscosity Transducer compression or internal pressure of organ Reflection of shear waves at boundaries B. Elasticity Imaging System Biomarker Qualification and User Testing of Systems – Brian Garra, M.D. Learning objectives-To understand: Goals Review the need for quantitative medical imaging Provide examples of quantitative imaging biomarkers Acquaint the participant with the purpose of the RSNA Quantitative Imaging Biomarker Alliance and the need for such an organization Review the QIBA process for creating a quantitative biomarker Summarize steps needed to verify adherence of site, operators, and imaging systems to a QIBA profile Underlying Premise and Assumptions Objective, quantifiable results are needed to enhance the value of diagnostic imaging in clinical practice Reasons for quantification Evidence based medicine requires objective, not subjective observer data Computerized decision support tools (eg CAD) generally require quantitative input. Quantitative, reproducible measures are more easily used to develop personalized molecular medical diagnostic and treatment systems What is quantitative imaging? Definition from Imaging Metrology Workshop The Quantitative Imaging Biomarker Alliance Formation 2008 Mission Structure Example Imaging Biomarkers Being Explored Biomarker Selection Groundwork Draft Protocol for imaging and data evaluation QIBA Profile Drafting Equipment and Site Validation Technical Clinical Site and Equipment QA and Compliance Checking Ultrasound Elasticity Estimation Biomarker US Elasticity Estimation Background Current Status and Problems Biomarker Selection-process and outcome US SWS for Liver Fibrosis Biomarker Work Groundwork Literature search and analysis results Phase I phantom testing-Elastic phantoms Phase II phantom testing-Viscoelastic phantoms Digital Simulated Data Protocol and Profile Drafting Protocol: based on UPICT and existing literature and standards bodies protocols Profile-Current claims, Manufacturer specific appendices What comes after the profile Profile Validation Technical validation Clinical validation QA and Compliance Possible approaches Site Operator testing Site protocol re-evaluation Imaging system Manufacturer testing and attestation User acceptance testing and periodic QA Phantom Tests Digital Phantom Based Testing Standard QA Testing Remediation Schemes Profile Evolution Towards additional applications Towards higher accuracy and precision Supported in part by NIH contract HHSN268201300071C from NIBIB. Collaboration with GE Global Research, no personal support.; S. Chen, Some technologies described in this presentation have been licensed. Mayo Clinic and Dr. Chen have financial interests these technologies.« less

Cardiac contraction motion compensation in gated myocardial perfusion SPECT: A comparative study.

PubMed

Salehi, Narges; Rahmim, Arman; Fatemizadeh, Emad; Akbarzadeh, Afshin; Farahani, Mohammad Hossein; Farzanefar, Saeed; Ay, Mohammad Reza

2018-05-01

Cardiac contraction significantly degrades quality and quantitative accuracy of gated myocardial perfusion SPECT (MPS) images. In this study, we aimed to explore different techniques in motion-compensated temporal processing of MPS images and their impact on image quality and quantitative accuracy. 50 patients without known heart condition underwent gated MPS. 3D motion compensation methods using Motion Freezing by Cedars Sinai (MF), Log-domain Diffeomorphic Demons (LDD) and Free-Form Deformation (FFD) were applied to warp all image phases to fit the end-diastolic (ED) phase. Afterwards, myocardial wall thickness, myocardial to blood pool contrast, and image contrast-to noise ratio (CNR) were measured in summed images with no motion compensation (NoMC) and compensated images (MF, LDD and FFD). Total Perfusion Defect (TPD) was derived from Cedars-Sinai software, on the basis of sex-specific normal limits. Left ventricle (LV) lateral wall thickness was reduced after applying motion compensation (p < 0.05). Myocardial to blood pool contrast and CNR in compensated images were greater than NoMC (p < 0.05). TPD_LDD was in good agreement with the corresponding TPD_MF (p = 0.13). All methods have improved image quality and quantitative performance relative to NoMC. LDD and FFD are fully automatic and do not require any manual intervention, while MF is dependent on contour definition. In terms of diagnostic parameters LDD is in good agreement with MF which is a clinically accepted method. Further investigation along with diagnostic reference standards, in order to specify diagnostic value of each technique is recommended. Copyright © 2018 Associazione Italiana di Fisica Medica. All rights reserved.

Quantitative X-ray dark-field and phase tomography using single directional speckle scanning technique

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Hongchang, E-mail: hongchang.wang@diamond.ac.uk; Kashyap, Yogesh; Sawhney, Kawal

2016-03-21

X-ray dark-field contrast tomography can provide important supplementary information inside a sample to the conventional absorption tomography. Recently, the X-ray speckle based technique has been proposed to provide qualitative two-dimensional dark-field imaging with a simple experimental arrangement. In this letter, we deduce a relationship between the second moment of scattering angle distribution and cross-correlation degradation of speckle and establish a quantitative basis of X-ray dark-field tomography using single directional speckle scanning technique. In addition, the phase contrast images can be simultaneously retrieved permitting tomographic reconstruction, which yields enhanced contrast in weakly absorbing materials. Such complementary tomography technique can allow systematicmore » investigation of complex samples containing both soft and hard materials.« less

Phase imaging using highly coherent X-rays: radiography, tomography, diffraction topography.

PubMed

Baruchel, J; Cloetens, P; Härtwig, J; Ludwig, W; Mancini, L; Pernot, P; Schlenker, M

2000-05-01

Several hard X-rays imaging techniques greatly benefit from the coherence of the beams delivered by the modern synchrotron radiation sources. This is illustrated with examples recorded on the 'long' (145 m) ID19 'imaging' beamline of the ESRF. Phase imaging is directly related to the small angular size of the source as seen from one point of the sample ('effective divergence' approximately microradians). When using the ;propagation' technique, phase radiography and tomography are instrumentally very simple. They are often used in the 'edge detection' regime, where the jumps of density are clearly observed. The in situ damage assessment of micro-heterogeneous materials is one example of the many applications. Recently a more quantitative approach has been developed, which provides a three-dimensional density mapping of the sample ('holotomography'). The combination of diffraction topography and phase-contrast imaging constitutes a powerful tool. The observation of holes of discrete sizes in quasicrystals, and the investigation of poled ferroelectric materials, result from this combination.

Accurate modelling of single-particle cryo-EM images quantifies the benefits expected from using Zernike phase contrast

PubMed Central

Hall, R. J.; Nogales, E.; Glaeser, R. M.

2011-01-01

The use of a Zernike-type phase plate in biological cryo-electron microscopy allows the imaging, without using defocus, of what are predominantly phase objects. It is thought that such phase-plate implementations might result in higher quality images, free from the problems of CTF correction that occur when images must be recorded at extremely high values of defocus. In single-particle cryo-electron microscopy it is hoped that these improvements in image quality will facilitate work on structures that have proved difficult to study, either because of their relatively small size or because the structures are not completely homogeneous. There is still a need, however, to quantify how much improvement can be gained by using a phase plate for single-particle cryo-electron microscopy. We present a method for quantitatively modelling the images recorded with 200 keV electrons, for single particles embedded in vitreous ice. We then investigate what difference the use of a phase-plate device could have on the processing of single-particle data. We confirm that using a phase plate results in single-particle datasets in which smaller molecules can be detected, particles can be more accurately aligned and problems of heterogeneity can be more easily addressed. PMID:21463690

Portable low-coherence interferometry for quantitatively imaging fast dynamics with extended field of view

NASA Astrophysics Data System (ADS)

Shaked, Natan T.; Girshovitz, Pinhas; Frenklach, Irena

2014-06-01

We present our recent advances in the development of compact, highly portable and inexpensive wide-field interferometric modules. By a smart design of the interferometric system, including the usage of low-coherence illumination sources and common-path off-axis geometry of the interferometers, spatial and temporal noise levels of the resulting quantitative thickness profile can be sub-nanometric, while processing the phase profile in real time. In addition, due to novel experimentally-implemented multiplexing methods, we can capture low-coherence off-axis interferograms with significantly extended field of view and in faster acquisition rates. Using these techniques, we quantitatively imaged rapid dynamics of live biological cells including sperm cells and unicellular microorganisms. Then, we demonstrated dynamic profiling during lithography processes of microscopic elements, with thicknesses that may vary from several nanometers to hundreds of microns. Finally, we present new algorithms for fast reconstruction (including digital phase unwrapping) of off-axis interferograms, which allow real-time processing in more than video rate on regular single-core computers.

Applications of pathology-assisted image analysis of immunohistochemistry-based biomarkers in oncology.

PubMed

Shinde, V; Burke, K E; Chakravarty, A; Fleming, M; McDonald, A A; Berger, A; Ecsedy, J; Blakemore, S J; Tirrell, S M; Bowman, D

2014-01-01

Immunohistochemistry-based biomarkers are commonly used to understand target inhibition in key cancer pathways in preclinical models and clinical studies. Automated slide-scanning and advanced high-throughput image analysis software technologies have evolved into a routine methodology for quantitative analysis of immunohistochemistry-based biomarkers. Alongside the traditional pathology H-score based on physical slides, the pathology world is welcoming digital pathology and advanced quantitative image analysis, which have enabled tissue- and cellular-level analysis. An automated workflow was implemented that includes automated staining, slide-scanning, and image analysis methodologies to explore biomarkers involved in 2 cancer targets: Aurora A and NEDD8-activating enzyme (NAE). The 2 workflows highlight the evolution of our immunohistochemistry laboratory and the different needs and requirements of each biological assay. Skin biopsies obtained from MLN8237 (Aurora A inhibitor) phase 1 clinical trials were evaluated for mitotic and apoptotic index, while mitotic index and defects in chromosome alignment and spindles were assessed in tumor biopsies to demonstrate Aurora A inhibition. Additionally, in both preclinical xenograft models and an acute myeloid leukemia phase 1 trial of the NAE inhibitor MLN4924, development of a novel image algorithm enabled measurement of downstream pathway modulation upon NAE inhibition. In the highlighted studies, developing a biomarker strategy based on automated image analysis solutions enabled project teams to confirm target and pathway inhibition and understand downstream outcomes of target inhibition with increased throughput and quantitative accuracy. These case studies demonstrate a strategy that combines a pathologist's expertise with automated image analysis to support oncology drug discovery and development programs.

Quantitative X-ray mapping, scatter diagrams and the generation of correction maps to obtain more information about your material

NASA Astrophysics Data System (ADS)

Wuhrer, R.; Moran, K.

2014-03-01

Quantitative X-ray mapping with silicon drift detectors and multi-EDS detector systems have become an invaluable analysis technique and one of the most useful methods of X-ray microanalysis today. The time to perform an X-ray map has reduced considerably with the ability to map minor and trace elements very accurately due to the larger detector area and higher count rate detectors. Live X-ray imaging can now be performed with a significant amount of data collected in a matter of minutes. A great deal of information can be obtained from X-ray maps. This includes; elemental relationship or scatter diagram creation, elemental ratio mapping, chemical phase mapping (CPM) and quantitative X-ray maps. In obtaining quantitative x-ray maps, we are able to easily generate atomic number (Z), absorption (A), fluorescence (F), theoretical back scatter coefficient (η), and quantitative total maps from each pixel in the image. This allows us to generate an image corresponding to each factor (for each element present). These images allow the user to predict and verify where they are likely to have problems in our images, and are especially helpful to look at possible interface artefacts. The post-processing techniques to improve the quantitation of X-ray map data and the development of post processing techniques for improved characterisation are covered in this paper.

Bilateral filtering using the full noise covariance matrix applied to x-ray phase-contrast computed tomography.

PubMed

Allner, S; Koehler, T; Fehringer, A; Birnbacher, L; Willner, M; Pfeiffer, F; Noël, P B

2016-05-21

The purpose of this work is to develop an image-based de-noising algorithm that exploits complementary information and noise statistics from multi-modal images, as they emerge in x-ray tomography techniques, for instance grating-based phase-contrast CT and spectral CT. Among the noise reduction methods, image-based de-noising is one popular approach and the so-called bilateral filter is a well known algorithm for edge-preserving filtering. We developed a generalization of the bilateral filter for the case where the imaging system provides two or more perfectly aligned images. The proposed generalization is statistically motivated and takes the full second order noise statistics of these images into account. In particular, it includes a noise correlation between the images and spatial noise correlation within the same image. The novel generalized three-dimensional bilateral filter is applied to the attenuation and phase images created with filtered backprojection reconstructions from grating-based phase-contrast tomography. In comparison to established bilateral filters, we obtain improved noise reduction and at the same time a better preservation of edges in the images on the examples of a simulated soft-tissue phantom, a human cerebellum and a human artery sample. The applied full noise covariance is determined via cross-correlation of the image noise. The filter results yield an improved feature recovery based on enhanced noise suppression and edge preservation as shown here on the example of attenuation and phase images captured with grating-based phase-contrast computed tomography. This is supported by quantitative image analysis. Without being bound to phase-contrast imaging, this generalized filter is applicable to any kind of noise-afflicted image data with or without noise correlation. Therefore, it can be utilized in various imaging applications and fields.

Label-free imaging of the dynamics of cell-to-cell string-like structure bridging in the free-space by low-coherent quantitative phase microscopy

NASA Astrophysics Data System (ADS)

Yamauchi, Toyohiko; Iwai, Hidenao; Yamashita, Yutaka

2013-03-01

We succeeded in utilizing our low-coherent quantitative phase microscopy (LC-QPM) to achieve label-free and three-dimensional imaging of string-like structures bridging the free-space between live cells. In past studies, three dimensional morphology of the string-like structures between cells had been investigated by electron microscopies and fluorescence microscopies and these structures were called "membrane nanotubes" or "tunneling nanotubes." However, use of electron microscopy inevitably kills these cells and fluorescence microscopy is itself a potentially invasive method. To achieve noninvasive imaging of live cells, we applied our LC-QPM which is a reflection-type, phase resolved and full-field interference microscope employing a low-coherent light source. LC-QPM is able to visualize the three-dimensional morphology of live cells without labeling by means of low-coherence interferometry. The lateral (diffraction limit) and longitudinal (coherence-length) spatial resolution of LC-QPM were respectively 0.49 and 0.93 micrometers and the repeatability of the phase measurement was 0.02 radians (1.0 nm). We successfully obtained three-dimensional morphology of live cultured epithelial cells (cell type: HeLa, derived from cervix cancer) and were able to clearly observe the individual string-like structures interconnecting the cells. When we performed volumetric imaging, a 80 micrometer by 60 micrometer by 6.5 micrometer volume was scanned every 5.67 seconds and 70 frames of a three-dimensional movie were recorded for a duration of 397 seconds. Moreover, the optical phase images gave us detailed information about the three-dimensional morphology of the string-like structure at sub-wavelength resolution. We believe that our LC-QPM will be a useful tool for the study of three-dimensional morphology of live cells.

Characterization of a high-energy in-line phase contrast tomosynthesis prototype

PubMed Central

Wu, Di; Yan, Aimin; Li, Yuhua; Wong, Molly D.; Zheng, Bin; Wu, Xizeng; Liu, Hong

2015-01-01

Purpose: In this research, a high-energy in-line phase contrast tomosynthesis prototype was developed and characterized through quantitative investigations and phantom studies. Methods: The prototype system consists of an x-ray source, a motorized rotation stage, and a CMOS detector with a pixel pitch of 0.05 mm. The x-ray source was operated at 120 kVp for this study, and the objects were mounted on the rotation stage 76.2 cm (R1) from the source and 114.3 cm (R2) from the detector. The large air gap between the object and detector guarantees sufficient phase-shift effects. The quantitative evaluation of this prototype included modulation transfer function and noise power spectrum measurements conducted under both projection mode and tomosynthesis mode. Phantom studies were performed including three custom designed phantoms with complex structures: a five-layer bubble wrap phantom, a fishbone phantom, and a chicken breast phantom with embedded fibrils and mass structures extracted from an ACR phantom. In-plane images of the phantoms were acquired to investigate their image qualities through observation, intensity profile plots, edge enhancement evaluations, and/or contrast-to-noise ratio calculations. In addition, the robust phase-attenuation duality (PAD)-based phase retrieval method was applied to tomosynthesis for the first time in this research. It was utilized as a preprocessing method to fully exhibit phase contrast on the angular projection before reconstruction. Results: The resolution and noise characteristics of this high-energy in-line phase contrast tomosynthesis prototype were successfully investigated and demonstrated. The phantom studies demonstrated that this imaging prototype can successfully remove the structure overlapping in phantom projections, obtain delineate interfaces, and achieve better contrast-to-noise ratio after applying phase retrieval to the angular projections. Conclusions: This research successfully demonstrated a high-energy in-line phase contrast tomosynthesis prototype. In addition, the PAD-based method of phase retrieval was combined with tomosynthesis imaging for the first time, which demonstrated its capability in significantly improving the contrast-to-noise ratios in the images. PMID:25979035

Preliminary evaluation of cryogenic two-phase flow imaging using electrical capacitance tomography

NASA Astrophysics Data System (ADS)

Xie, Huangjun; Yu, Liu; Zhou, Rui; Qiu, Limin; Zhang, Xiaobin

2017-09-01

The potential application of the 2-D eight-electrode electrical capacitance tomography (ECT) to the inversion imaging of the liquid nitrogen-vaporous nitrogen (LN2-VN2) flow in the tube is theoretically evaluated. The phase distribution of the computational domain is obtained using the simultaneous iterative reconstruction technique with variable iterative step size. The detailed mathematical derivations for the calculations are presented. The calculated phase distribution for the two detached LN2 column case shows the comparable results with the water-air case, regardless of the much reduced dielectric permittivity of LN2 compared with water. The inversion images of total eight different LN2-VN2 flow patterns are presented and quantitatively evaluated by calculating the relative void fraction error and the correlation coefficient. The results demonstrate that the developed reconstruction technique for ECT has the capacity to reconstruct the phase distribution of the complex LN2-VN2 flow, while the accuracy of the inversion images is significantly influenced by the size of the discrete phase. The influence of the measurement noise on the image quality is also considered in the calculations.

Image-guided plasma therapy of cutaneous wound

NASA Astrophysics Data System (ADS)

Zhang, Zhiwu; Ren, Wenqi; Yu, Zelin; Zhang, Shiwu; Yue, Ting; Xu, Ronald

2014-02-01

The wound healing process involves the reparative phases of inflammation, proliferation, and remodeling. Interrupting any of these phases may result in chronically unhealed wounds, amputation, or even patient death. Despite the clinical significance in chronic wound management, no effective methods have been developed for quantitative image-guided treatment. We integrated a multimodal imaging system with a cold atmospheric plasma probe for image-guided treatment of chronic wound. Multimodal imaging system offers a non-invasive, painless, simultaneous and quantitative assessment of cutaneous wound healing. Cold atmospheric plasma accelerates the wound healing process through many mechanisms including decontamination, coagulation and stimulation of the wound healing. The therapeutic effect of cold atmospheric plasma is studied in vivo under the guidance of a multimodal imaging system. Cutaneous wounds are created on the dorsal skin of the nude mice. During the healing process, the sample wound is treated by cold atmospheric plasma at different controlled dosage, while the control wound is healed naturally. The multimodal imaging system integrating a multispectral imaging module and a laser speckle imaging module is used to collect the information of cutaneous tissue oxygenation (i.e. oxygen saturation, StO2) and blood perfusion simultaneously to assess and guide the plasma therapy. Our preliminary tests show that cold atmospheric plasma in combination with multimodal imaging guidance has the potential to facilitate the healing of chronic wounds.

Segmentation and classification of cell cycle phases in fluorescence imaging.

PubMed

Ersoy, Ilker; Bunyak, Filiz; Chagin, Vadim; Cardoso, M Christina; Palaniappan, Kannappan

2009-01-01

Current chemical biology methods for studying spatiotemporal correlation between biochemical networks and cell cycle phase progression in live-cells typically use fluorescence-based imaging of fusion proteins. Stable cell lines expressing fluorescently tagged protein GFP-PCNA produce rich, dynamically varying sub-cellular foci patterns characterizing the cell cycle phases, including the progress during the S-phase. Variable fluorescence patterns, drastic changes in SNR, shape and position changes and abundance of touching cells require sophisticated algorithms for reliable automatic segmentation and cell cycle classification. We extend the recently proposed graph partitioning active contours (GPAC) for fluorescence-based nucleus segmentation using regional density functions and dramatically improve its efficiency, making it scalable for high content microscopy imaging. We utilize surface shape properties of GFP-PCNA intensity field to obtain descriptors of foci patterns and perform automated cell cycle phase classification, and give quantitative performance by comparing our results to manually labeled data.

Halo-free Phase Contrast Microscopy

NASA Astrophysics Data System (ADS)

Nguyen, Tan H.; Kandel, Mikhail; Shakir, Haadi M.; Best-Popescu, Catherine; Arikkath, Jyothi; Do, Minh N.; Popescu, Gabriel

2017-03-01

We present a new approach for retrieving halo-free phase contrast microscopy (hfPC) images by upgrading the conventional PC microscope with an external interferometric module, which generates sufficient data for reversing the halo artifact. Acquiring four independent intensity images, our approach first measures haloed phase maps of the sample. We solve for the halo-free sample transmission function by using a physical model of the image formation under partial spatial coherence. Using this halo-free sample transmission, we can numerically generate artifact-free PC images. Furthermore, this transmission can be further used to obtain quantitative information about the sample, e.g., the thickness with known refractive indices, dry mass of live cells during their cycles. We tested our hfPC method on various control samples, e.g., beads, pillars and validated its potential for biological investigation by imaging live HeLa cells, red blood cells, and neurons.

Tomographic phase microscopy and its biological applications

NASA Astrophysics Data System (ADS)

Choi, Wonshik

2012-12-01

Conventional interferometric microscopy techniques such as digital holographic microscopy and quantitative phase microscopy are often classified as 3D imaging techniques because a recorded complex field image can be numerically propagated to a different depth. In a strict sense, however, a single complex field image contains only 2D information on a specimen. The measured 2D image is only a subset of the 3D structure. For the 3D mapping of an object, multiple independent 2D images are to be taken, for example at multiple incident angles or wavelengths, and then combined by the so-called optical diffraction tomography (ODT). In this Letter, tomographic phase microscopy (TPM) is reviewed that experimentally realizes the concept of the ODT for the 3D mapping of biological cells in their native state, and some of its interesting biological and biomedical applications are introduced. [Figure not available: see fulltext.

Quantitative shear-wave optical coherence elastography with a programmable phased array ultrasound as the wave source.

PubMed

Song, Shaozhen; Le, Nhan Minh; Huang, Zhihong; Shen, Tueng; Wang, Ruikang K

2015-11-01

The purpose of this study is to implement a beam-steering ultrasound as the wave source for shear-wave optical coherence elastography (SW-OCE) to achieve an extended range of elastic imaging of the tissue sample. We introduce a linear phased array ultrasound transducer (LPAUT) as the remote and programmable wave source and a phase-sensitive optical coherence tomography (OCT) as the sensitive shear-wave detector. The LPAUT is programmed to launch acoustic radiation force impulses (ARFI) focused at desired locations within the range of OCT imaging, upon which the elasticity map of the entire OCT B-scan cross section is recovered by spatial compounding of the elastic maps derived from each launch of AFRIs. We also propose a directional filter to separate the shear-wave propagation at different directions in order to reduce the effect of tissue heterogeneity on the shear-wave propagation within tissue. The feasibility of this proposed approach is then demonstrated by determining the stiffness of tissue-mimicking phantoms with agarose concentrations of 0.5% and 1% and also by imaging the Young's modulus of retinal and choroidal tissues within a porcine eye ball ex vivo. The approach opens up opportunities to combine medical ultrasound imaging and SW-OCE for high-resolution localized quantitative assessment of tissue biomechanical property.

Actinic imaging and evaluation of phase structures on EUV lithography masks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mochi, Iacopo; Goldberg, Kenneth; Huh, Sungmin

2010-09-28

The authors describe the implementation of a phase-retrieval algorithm to reconstruct phase and complex amplitude of structures on EUV lithography masks. Many native defects commonly found on EUV reticles are difficult to detect and review accurately because they have a strong phase component. Understanding the complex amplitude of mask features is essential for predictive modeling of defect printability and defect repair. Besides printing in a stepper, the most accurate way to characterize such defects is with actinic inspection, performed at the design, EUV wavelength. Phase defect and phase structures show a distinct through-focus behavior that enables qualitative evaluation of themore » object phase from two or more high-resolution intensity measurements. For the first time, phase of structures and defects on EUV masks were quantitatively reconstructed based on aerial image measurements, using a modified version of a phase-retrieval algorithm developed to test optical phase shifting reticles.« less

Optimal visualization of focal nodular hyperplasia: quantitative and qualitative evaluation of single and multiphasic arterial phase acquisition at 1.5 T MR imaging.

PubMed

Rousseau, Caroline; Ronot, Maxime; Vilgrain, Valérie; Zins, Marc

2016-05-01

To evaluate the qualitative and quantitative benefit of multiple arterial phase acquisitions for the depiction of hypervascularity in FNH explored MR imaging using an extracellular contrast agent. Between 2007 and 2014, all patients who underwent MR imaging for the exploration of FNH were included. The protocol included a single or a triple arterial phase ("single" and "triple" group, respectively). Arterial phases were visually divided into four types: (1) angiographic, (2) early, (3) late, and (4) portal. Signal intensity on arterial phase images was visually recorded as intense, moderate, or low for each lesion. Lesion-to-liver contrast (LLC) and relative lesion enhancement (RE) were calculated and compared between the two groups using the Mann-Whitney test. Thirty-five women were included (mean 45-year old, range 20-66), with 50 FNH (mean size 30 mm). Single and triple groups included 20 patients (30 FNH) and 15 patients (20 FNH), respectively. Signal intensity was intense in all lesions in the triple group and in 22/30 (73%) in the single group (p = 0.041). Intense signals were more frequently found in the early arterial phase (p < 0.001). RE was not significantly different (1.78 ± 0.84 vs. 1.98 ± 1.81 p = 0.430, in the single and triple groups, respectively) but LLC was significantly higher in the triple group (0.32 ± 0.10 vs. 0.22 ± 0.10, p = 0.005). LLC was significantly higher in the first two arterial phases in the triple group (p < 0.001). Acquisition of three arterial phases improves the visualization of hypervascularity of FNH, as lesions show high visual signal intensity and contrast. Optimal visualization is obtained in the early arterial phase.

Quantitative assessment on coronary computed tomography angiography (CCTA) image quality: comparisons between genders and different tube voltage settings

PubMed Central

Chian, Teo Chee; Nassir, Norziana Mat; Ibrahim, Mohd Izuan; Yusof, Ahmad Khairuddin Md

2017-01-01

Background This study was carried out to quantify and compare the quantitative image quality of coronary computed tomography angiography (CCTA) between genders as well as between different tube voltages scan protocols. Methods Fifty-five cases of CCTA were collected retrospectively and all images including reformatted axial images at systolic and diastolic phases as well as images with curved multi planar reformation (cMPR) were obtained. Quantitative image quality including signal intensity, image noise, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of right coronary artery (RCA), left anterior descending artery (LAD), left circumflex artery (LCx) and left main artery (LM) were quantified using Analyze 12.0 software. Results Six hundred and fifty-seven coronary arteries were evaluated. There were no significant differences in any quantitative image quality parameters between genders. 100 kilovoltage peak (kVp) scanning protocol produced images with significantly higher signal intensity compared to 120 kVp scanning protocol (P<0.001) in all coronary arteries in all types of images. Higher SNR was also observed in 100 kVp scan protocol in all coronary arteries except in LCx where 120 kVp showed better SNR than 100 kVp. Conclusions There were no significant differences in image quality of CCTA between genders and different tube voltages. Lower tube voltage (100 kVp) scanning protocol is recommended in clinical practice to reduce the radiation dose to patient. PMID:28275559

Atomic force microscope image contrast mechanisms on supported lipid bilayers.

PubMed

Schneider, J; Dufrêne, Y F; Barger, W R; Lee, G U

2000-08-01

This work presents a methodology to measure and quantitatively interpret force curves on supported lipid bilayers in water. We then use this method to correlate topographic imaging contrast in atomic force microscopy (AFM) images of phase-separated Langmuir-Blodgett bilayers with imaging load. Force curves collected on pure monolayers of both distearoylphosphatidylethanolamine (DSPE) and monogalactosylethanolamine (MGDG) and dioleoylethanolamine (DOPE) deposited at similar surface pressures onto a monolayer of DSPE show an abrupt breakthrough event at a repeatable, material-dependent force. The breakthrough force for DSPE and MGDG is sizable, whereas the breakthrough force for DOPE is too small to measure accurately. Contact-mode AFM images on 1:1 mixed monolayers of DSPE/DOPE and MGDG/DOPE have a high topographic contrast at loads between the breakthrough force of each phase, and a low topographic contrast at loads above the breakthrough force of both phases. Frictional contrast is inverted and magnified at loads above the breakthrough force of both phases. These results emphasize the important role that surface forces and mechanics can play in imaging multicomponent biomembranes with AFM.

High-resolution dynamic imaging and quantitative analysis of lung cancer xenografts in nude mice using clinical PET/CT

PubMed Central

Wang, Ying Yi; Wang, Kai; Xu, Zuo Yu; Song, Yan; Wang, Chu Nan; Zhang, Chong Qing; Sun, Xi Lin; Shen, Bao Zhong

2017-01-01

Considering the general application of dedicated small-animal positron emission tomography/computed tomography is limited, an acceptable alternative in many situations might be clinical PET/CT. To estimate the feasibility of using clinical PET/CT with [F-18]-fluoro-2-deoxy-D-glucose for high-resolution dynamic imaging and quantitative analysis of cancer xenografts in nude mice. Dynamic clinical PET/CT scans were performed on xenografts for 60 min after injection with [F-18]-fluoro-2-deoxy-D-glucose. Scans were reconstructed with or without SharpIR method in two phases. And mice were sacrificed to extracting major organs and tumors, using ex vivo γ-counting as a reference. Strikingly, we observed that the image quality and the correlation between the all quantitive data from clinical PET/CT and the ex vivo counting was better with the SharpIR reconstructions than without. Our data demonstrate that clinical PET/CT scanner with SharpIR reconstruction is a valuable tool for imaging small animals in preclinical cancer research, offering dynamic imaging parameters, good image quality and accurate data quatification. PMID:28881772

High-resolution dynamic imaging and quantitative analysis of lung cancer xenografts in nude mice using clinical PET/CT.

PubMed

Wang, Ying Yi; Wang, Kai; Xu, Zuo Yu; Song, Yan; Wang, Chu Nan; Zhang, Chong Qing; Sun, Xi Lin; Shen, Bao Zhong

2017-08-08

Considering the general application of dedicated small-animal positron emission tomography/computed tomography is limited, an acceptable alternative in many situations might be clinical PET/CT. To estimate the feasibility of using clinical PET/CT with [F-18]-fluoro-2-deoxy-D-glucose for high-resolution dynamic imaging and quantitative analysis of cancer xenografts in nude mice. Dynamic clinical PET/CT scans were performed on xenografts for 60 min after injection with [F-18]-fluoro-2-deoxy-D-glucose. Scans were reconstructed with or without SharpIR method in two phases. And mice were sacrificed to extracting major organs and tumors, using ex vivo γ-counting as a reference. Strikingly, we observed that the image quality and the correlation between the all quantitive data from clinical PET/CT and the ex vivo counting was better with the SharpIR reconstructions than without. Our data demonstrate that clinical PET/CT scanner with SharpIR reconstruction is a valuable tool for imaging small animals in preclinical cancer research, offering dynamic imaging parameters, good image quality and accurate data quatification.

How a phase image of a cell with nucleus refractive index smaller than that of the cytoplasm should look like?: A Comment on two papers by Steelman et al. and Schürmann et al. Read the Responses to this Comment: e201800091 and e201800095.

PubMed

Yurkin, Maxim A

2018-05-02

In recent papers Steelman et al. ("Is the nuclear refractive index lower than cytoplasm? Validation of phase measurements and implications for light scattering technologies") and Schürmann et al. ("Cell nuclei have lower refractive index and mass density than cytoplasm") obtained quantitative phase images of whole cells of various types and corresponding isolated nuclei and concluded that the refractive index (RI) of the nucleus is significantly smaller than that of the cytoplasm. The comment shows that this conclusion and assumptions used in retrieving the RI necessarily imply a characteristic dip in the center of the whole-cell phase images. This dip is not present in any of the phase images in the discussed papers, which is a strong argument against the conclusion of smaller nucleus RI. It is also discussed whether a different processing of the phase images can help to clarify this issue. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Feasibility of quantitative regional ventilation and perfusion mapping with phase-resolved functional lung (PREFUL) MRI in healthy volunteers and COPD, CTEPH, and CF patients.

PubMed

Voskrebenzev, Andreas; Gutberlet, Marcel; Klimeš, Filip; Kaireit, Till F; Schönfeld, Christian; Rotärmel, Alexander; Wacker, Frank; Vogel-Claussen, Jens

2018-04-01

In this feasibility study, a phase-resolved functional lung imaging postprocessing method for extraction of dynamic perfusion (Q) and ventilation (V) parameters using a conventional 1H lung MRI Fourier decomposition acquisition is introduced. Time series of coronal gradient-echo MR images with a temporal resolution of 288 to 324 ms of two healthy volunteers, one patient with chronic thromboembolic hypertension, one patient with cystic fibrosis, and one patient with chronic obstructive pulmonary disease were acquired at 1.5 T. Using a sine model to estimate cardiac and respiratory phases of each image, all images were sorted to reconstruct full cardiac and respiratory cycles. Time to peak (TTP), V/Q maps, and fractional ventilation flow-volume loops were calculated. For the volunteers, homogenous ventilation and perfusion TTP maps (V-TTP, Q-TTP) were obtained. The chronic thromboembolic hypertension patient showed increased perfusion TTP in hypoperfused regions in visual agreement with dynamic contrast-enhanced MRI, which improved postpulmonary endaterectomy surgery. Cystic fibrosis and chronic obstructive pulmonary disease patients showed a pattern of increased V-TTP and Q-TTP in regions of hypoventilation and decreased perfusion. Fractional ventilation flow-volume loops of the chronic obstructive pulmonary disease patient were smaller in comparison with the healthy volunteer, and showed regional differences in visual agreement with functional small airways disease and emphysema on CT. This study shows the feasibility of phase-resolved functional lung imaging to gain quantitative information regarding regional lung perfusion and ventilation without the need for ultrafast imaging, which will be advantageous for future clinical translation. Magn Reson Med 79:2306-2314, 2018. © 2017 International Society for Magnetic Resonance in Medicine. © 2017 International Society for Magnetic Resonance in Medicine.

Low cost label-free live cell imaging for biological samples

NASA Astrophysics Data System (ADS)

Seniya, C.; Towers, C. E.; Towers, D. P.

2017-02-01

This paper reports the progress to develop a practical phase measuring microscope offering new capabilities in terms of phase measurement accuracy and quantification of cell:cell interactions over the longer term. A novel, low cost phase interference microscope for imaging live cells (label-free) is described. The method combines the Zernike phase contrast approach with a dual mirror design to enable phase modulation between the scattered and un-scattered optical fields. Two designs are proposed and demonstrated, one of which retains the common path nature of Zernike's original microscopy concept. In both setups the phase shift is simple to control via a piezoelectric driven mirror in the back focal plane of the imaging system. The approach is significantly cheaper to implement than those based on spatial light modulators (SLM) at approximately 20% of the cost. A quantitative assessment of the performance of a set of phase shifting algorithms is also presented, specifically with regard to broad bandwidth illumination in phase contrast microscopy. The simulation results show that the phase measurement accuracy is strongly dependent on the algorithm selected and the optical path difference in the sample.

Optimization of propagation-based x-ray phase-contrast tomography for breast cancer imaging

NASA Astrophysics Data System (ADS)

Baran, P.; Pacile, S.; Nesterets, Y. I.; Mayo, S. C.; Dullin, C.; Dreossi, D.; Arfelli, F.; Thompson, D.; Lockie, D.; McCormack, M.; Taba, S. T.; Brun, F.; Pinamonti, M.; Nickson, C.; Hall, C.; Dimmock, M.; Zanconati, F.; Cholewa, M.; Quiney, H.; Brennan, P. C.; Tromba, G.; Gureyev, T. E.

2017-03-01

The aim of this study was to optimise the experimental protocol and data analysis for in-vivo breast cancer x-ray imaging. Results are presented of the experiment at the SYRMEP beamline of Elettra Synchrotron using the propagation-based phase-contrast mammographic tomography method, which incorporates not only absorption, but also x-ray phase information. In this study the images of breast tissue samples, of a size corresponding to a full human breast, with radiologically acceptable x-ray doses were obtained, and the degree of improvement of the image quality (from the diagnostic point of view) achievable using propagation-based phase-contrast image acquisition protocols with proper incorporation of x-ray phase retrieval into the reconstruction pipeline was investigated. Parameters such as the x-ray energy, sample-to-detector distance and data processing methods were tested, evaluated and optimized with respect to the estimated diagnostic value using a mastectomy sample with a malignant lesion. The results of quantitative evaluation of images were obtained by means of radiological assessment carried out by 13 experienced specialists. A comparative analysis was performed between the x-ray and the histological images of the specimen. The results of the analysis indicate that, within the investigated range of parameters, both the objective image quality characteristics and the subjective radiological scores of propagation-based phase-contrast images of breast tissues monotonically increase with the strength of phase contrast which in turn is directly proportional to the product of the radiation wavelength and the sample-to-detector distance. The outcomes of this study serve to define the practical imaging conditions and the CT reconstruction procedures appropriate for low-dose phase-contrast mammographic imaging of live patients at specially designed synchrotron beamlines.

Living cell dry mass measurement using quantitative phase imaging with quadriwave lateral shearing interferometry: an accuracy and sensitivity discussion.

PubMed

Aknoun, Sherazade; Savatier, Julien; Bon, Pierre; Galland, Frédéric; Abdeladim, Lamiae; Wattellier, Benoit; Monneret, Serge

2015-01-01

Single-cell dry mass measurement is used in biology to follow cell cycle, to address effects of drugs, or to investigate cell metabolism. Quantitative phase imaging technique with quadriwave lateral shearing interferometry (QWLSI) allows measuring cell dry mass. The technique is very simple to set up, as it is integrated in a camera-like instrument. It simply plugs onto a standard microscope and uses a white light illumination source. Its working principle is first explained, from image acquisition to automated segmentation algorithm and dry mass quantification. Metrology of the whole process, including its sensitivity, repeatability, reliability, sources of error, over different kinds of samples and under different experimental conditions, is developed. We show that there is no influence of magnification or spatial light coherence on dry mass measurement; effect of defocus is more critical but can be calibrated. As a consequence, QWLSI is a well-suited technique for fast, simple, and reliable cell dry mass study, especially for live cells.

Off-axis low coherence digital holographic interferometry for quantitative phase imaging with an LED

NASA Astrophysics Data System (ADS)

Guo, Rongli; Wang, Fan; Hu, Xiaoying; Yang, Wenqian

2017-11-01

Off-axis digital holographic interferometry with the light source of a light emitting diode (LED) is presented and its application for quantitative phase imaging in a large range with low noise is demonstrated. The scheme is implemented in a grating based Mach-Zehnder interferometer. To achieve off-axis interferometry, firstly, the collimated beam emitted from an LED is diffracted into multiple orders by a grating and they are split into two copies by a beam splitter; secondly, in the object arm the zero order of one copy is filtered in the Fourier plane and is reshaped to illuminate the sample, while in the reference arm one of its first order of another copy is selected to serve as the reference beam, and then an off-axis hologram can be obtained at the image plane. The main advantage stemming from an LED illumination is its high spatial phase resolution, due to the subdued speckle effect. The off-axis geometry enables one-shot recording of the hologram in the millisecond scale. The utility of the proposed setup is illustrated with measurements of a resolution target and part of a wing of green-lacewing, and dynamic evaporation process of an ethanol film.

Electron holography—basics and applications

NASA Astrophysics Data System (ADS)

Lichte, Hannes; Lehmann, Michael

2008-01-01

Despite the huge progress achieved recently by means of the corrector for aberrations, allowing now a true atomic resolution of 0.1 nm, hence making it an unrivalled tool for nanoscience, transmission electron microscopy (TEM) suffers from a severe drawback: in a conventional electron micrograph only a poor phase contrast can be achieved, i.e. phase structures are virtually invisible. Therefore, conventional TEM is nearly blind for electric and magnetic fields, which are pure phase objects. Since such fields provoked by the atomic structure, e.g. of semiconductors and ferroelectrics, largely determine the solid state properties, hence the importance for high technology applications, substantial object information is missing. Electron holography in TEM offers the solution: by superposition with a coherent reference wave, a hologram is recorded, from which the image wave can be completely reconstructed by amplitude and phase. Now the object is displayed quantitatively in two separate images: one representing the amplitude, the other the phase. From the phase image, electric and magnetic fields can be determined quantitatively in the range from micrometre down to atomic dimensions by all wave optical methods that one can think of, both in real space and in Fourier space. Electron holography is pure wave optics. Therefore, we discuss the basics of coherence and interference, the implementation into a TEM, the path of rays for recording holograms as well as the limits in lateral and signal resolution. We outline the methods of reconstructing the wave by numerical image processing and procedures for extracting the object properties of interest. Furthermore, we present a broad spectrum of applications both at mesoscopic and atomic dimensions. This paper gives an overview of the state of the art pointing at the needs for further development. It is also meant as encouragement for those who refrain from holography, thinking that it can only be performed by specialists in highly specialized laboratories. In fact, a modern TEM built for atomic resolution and equipped with a field emitter or a Schottky emitter, well aligned by a skilled operator, can deliver good holograms. Running commercially available image processing software and mathematics programs on a laptop-computer is sufficient for reconstruction of the amplitude and phase images and extracting desirable object information.

Quantitative phase imaging characterization of tumor-associated blood vessel formation on a chip

NASA Astrophysics Data System (ADS)

Guo, Peng; Huang, Jing; Moses, Marsha A.

2018-02-01

Angiogenesis, the formation of new blood vessels from existing ones, is a biological process that has an essential role in solid tumor growth, development, and progression. Recent advances in Lab-on-a-Chip technology has created an opportunity for scientists to observe endothelial cell (EC) behaviors during the dynamic process of angiogenesis using a simple and economical in vitro platform that recapitulates in vivo blood vessel formation. Here, we use quantitative phase imaging (QPI) microscopy to continuously and non-invasively characterize the dynamic process of tumor cell-induced angiogenic sprout formation on a microfluidic chip. The live tumor cell-induced angiogenic sprouts are generated by multicellular endothelial sprouting into 3 dimensional (3D) Matrigel using human umbilical vein endothelial cells (HUVECs). By using QPI, we quantitatively measure a panel of cellular morphological and behavioral parameters of each individual EC participating in this sprouting. In this proof-of-principle study, we demonstrate that QPI is a powerful tool that can provide real-time quantitative analysis of biological processes in in vitro 3D biomimetic devices, which, in turn, can improve our understanding of the biology underlying functional tissue engineering.

Improved cancer risk stratification and diagnosis via quantitative phase microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Liu, Yang; Uttam, Shikhar; Pham, Hoa V.; Hartman, Douglas J.

2017-02-01

Pathology remains the gold standard for cancer diagnosis and in some cases prognosis, in which trained pathologists examine abnormality in tissue architecture and cell morphology characteristic of cancer cells with a bright-field microscope. The limited resolution of conventional microscope can result in intra-observer variation, missed early-stage cancers, and indeterminate cases that often result in unnecessary invasive procedures in the absence of cancer. Assessment of nanoscale structural characteristics via quantitative phase represents a promising strategy for identifying pre-cancerous or cancerous cells, due to its nanoscale sensitivity to optical path length, simple sample preparation (i.e., label-free) and low cost. I will present the development of quantitative phase microscopy system in transmission and reflection configuration to detect the structural changes in nuclear architecture, not be easily identifiable by conventional pathology. Specifically, we will present the use of transmission-mode quantitative phase imaging to improve diagnostic accuracy of urine cytology and the nuclear dry mass is progressively correlate with negative, atypical, suspicious and positive cytological diagnosis. In a second application, we will present the use of reflection-mode quantitative phase microscopy for depth-resolved nanoscale nuclear architecture mapping (nanoNAM) of clinically prepared formalin-fixed, paraffin-embedded tissue sections. We demonstrated that the quantitative phase microscopy system detects a gradual increase in the density alteration of nuclear architecture during malignant transformation in animal models of colon carcinogenesis and in human patients with ulcerative colitis, even in tissue that appears histologically normal according to pathologists. We evaluated the ability of nanoNAM to predict "future" cancer progression in patients with ulcerative colitis.

Lensless digital holography with diffuse illumination through a pseudo-random phase mask.

PubMed

Bernet, Stefan; Harm, Walter; Jesacher, Alexander; Ritsch-Marte, Monika

2011-12-05

Microscopic imaging with a setup consisting of a pseudo-random phase mask, and an open CMOS camera, without an imaging objective, is demonstrated. The pseudo random phase mask acts as a diffuser for an incoming laser beam, scattering a speckle pattern to a CMOS chip, which is recorded once as a reference. A sample which is afterwards inserted somewhere in the optical beam path changes the speckle pattern. A single (non-iterative) image processing step, comparing the modified speckle pattern with the previously recorded one, generates a sharp image of the sample. After a first calibration the method works in real-time and allows quantitative imaging of complex (amplitude and phase) samples in an extended three-dimensional volume. Since no lenses are used, the method is free from lens abberations. Compared to standard inline holography the diffuse sample illumination improves the axial sectioning capability by increasing the effective numerical aperture in the illumination path, and it suppresses the undesired so-called twin images. For demonstration, a high resolution spatial light modulator (SLM) is programmed to act as the pseudo-random phase mask. We show experimental results, imaging microscopic biological samples, e.g. insects, within an extended volume at a distance of 15 cm with a transverse and longitudinal resolution of about 60 μm and 400 μm, respectively.

Coherent imaging with incoherent light in digital holographic microscopy

NASA Astrophysics Data System (ADS)

Chmelik, Radim

2012-01-01

Digital holographic microscope (DHM) allows for imaging with a quantitative phase contrast. In this way it becomes an important instrument, a completely non-invasive tool for a contrast intravital observation of living cells and a cell drymass density distribution measurement. A serious drawback of current DHMs is highly coherent illumination which makes the lateral resolution worse and impairs the image quality by a coherence noise and a parasitic interference. An uncompromising solution to this problem can be found in the Leith concept of incoherent holography. An off-axis hologram can be formed with arbitrary degree of light coherence in systems equipped with an achromatic interferometer and thus the resolution and the image quality typical for an incoherent-light wide-field microscopy can be achieved. In addition, advanced imaging modes based on limited coherence can be utilized. The typical example is a coherence-gating effect which provides a finite axial resolution and makes DHM image similar to that of a confocal microscope. These possibilities were described theoretically using the formalism of three-dimensional coherent transfer functions and proved experimentally by the coherence-controlled holographic microscope which is DHM based on the Leith achromatic interferometer. Quantitative-phase-contrast imaging is demonstrated with incoherent light by the living cancer cells observation and their motility evaluation. The coherence-gating effect was proved by imaging of model samples through a scattering layer and living cells inside an opalescent medium.

Quantitative orientation-independent differential interference contrast (DIC) microscopy

NASA Astrophysics Data System (ADS)

Shribak, Michael; LaFountain, James; Biggs, David; Inoué, Shinya

2007-02-01

We describe a new DIC technique, which records phase gradients within microscopic specimens independently of their orientation. The proposed system allows the generation of images representing the distribution of dry mass (optical path difference) in the specimen. Unlike in other forms of interference microscopes, this approach does not require a narrow illuminating cone. The orientation-independent differential interference contrast (OI-DIC) system can also be combined with orientation-independent polarization (OI-Pol) measurements to yield two complementary images: one showing dry mass distribution (which is proportional to refractive index) and the other showing distribution of birefringence (due to structural or internal anisotropy). With a model specimen used for this work -- living spermatocytes from the crane fly, Nephrotoma suturalis --- the OI-DIC image clearly reveals the detailed shape of the chromosomes while the polarization image quantitatively depicts the distribution of the birefringent microtubules in the spindle, both without any need for staining or other modifications of the cell. We present examples of a pseudo-color combined image incorporating both orientation-independent DIC and polarization images of a spermatocyte at diakinesis and metaphase of meiosis I. Those images provide clear evidence that the proposed technique can reveal fine architecture and molecular organization in live cells without perturbation associated with staining or fluorescent labeling. The phase image was obtained using optics having a numerical aperture 1.4, thus achieving a level of resolution never before achieved with any interference microscope.

Quantitative electron density characterization of soft tissue substitute plastic materials using grating-based x-ray phase-contrast imaging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sarapata, A.; Chabior, M.; Zanette, I.

2014-10-15

Many scientific research areas rely on accurate electron density characterization of various materials. For instance in X-ray optics and radiation therapy, there is a need for a fast and reliable technique to quantitatively characterize samples for electron density. We present how a precise measurement of electron density can be performed using an X-ray phase-contrast grating interferometer in a radiographic mode of a homogenous sample in a controlled geometry. A batch of various plastic materials was characterized quantitatively and compared with calculated results. We found that the measured electron densities closely match theoretical values. The technique yields comparable results between amore » monochromatic and a polychromatic X-ray source. Measured electron densities can be further used to design dedicated X-ray phase contrast phantoms and the additional information on small angle scattering should be taken into account in order to exclude unsuitable materials.« less

Texture analysis of ultrahigh field T2*-weighted MR images of the brain: application to Huntington's disease.

PubMed

Doan, Nhat Trung; van den Bogaard, Simon J A; Dumas, Eve M; Webb, Andrew G; van Buchem, Mark A; Roos, Raymund A C; van der Grond, Jeroen; Reiber, Johan H C; Milles, Julien

2014-03-01

To develop a framework for quantitative detection of between-group textural differences in ultrahigh field T2*-weighted MR images of the brain. MR images were acquired using a three-dimensional (3D) T2*-weighted gradient echo sequence on a 7 Tesla MRI system. The phase images were high-pass filtered to remove phase wraps. Thirteen textural features were computed for both the magnitude and phase images of a region of interest based on 3D Gray-Level Co-occurrence Matrix, and subsequently evaluated to detect between-group differences using a Mann-Whitney U-test. We applied the framework to study textural differences in subcortical structures between premanifest Huntington's disease (HD), manifest HD patients, and controls. In premanifest HD, four phase-based features showed a difference in the caudate nucleus. In manifest HD, 7 magnitude-based features showed a difference in the pallidum, 6 phase-based features in the caudate nucleus, and 10 phase-based features in the putamen. After multiple comparison correction, significant differences were shown in the putamen in manifest HD by two phase-based features (both adjusted P values=0.04). This study provides the first evidence of textural heterogeneity of subcortical structures in HD. Texture analysis of ultrahigh field T2*-weighted MR images can be useful for noninvasive monitoring of neurodegenerative diseases. Copyright © 2013 Wiley Periodicals, Inc.

Development of Simultaneous Beta-and-Coincidence-Gamma Imager for Plant Imaging Research

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tai, Yuan-Chuan

2016-09-30

The goal of this project is to develop a novel imaging system that can simultaneously acquire beta and coincidence gamma images of positron sources in thin objects such as leaves of plants. This hybrid imager can be used to measure carbon assimilation in plants quantitatively and in real-time after C-11 labeled carbon-dioxide is administered. A better understanding of carbon assimilation, particularly under the increasingly elevated atmospheric CO 2 level, is extremely critical for plant scientists who study food crop and biofuel production. Phase 1 of this project is focused on the technology development with 3 specific aims: (1) develop amore » hybrid detector that can detect beta and gamma rays simultaneously; (2) develop an imaging system that can differentiate these two types of radiation and acquire beta and coincidence gamma images in real-time; (3) develop techniques to quantify radiotracer distribution using beta and gamma images. Phase 2 of this project is to apply technologies developed in phase 1 to study plants using positron-emitting radionuclide such as 11C to study carbon assimilation in biofuel plants.« less

Registration of 2D to 3D joint images using phase-based mutual information

NASA Astrophysics Data System (ADS)

Dalvi, Rupin; Abugharbieh, Rafeef; Pickering, Mark; Scarvell, Jennie; Smith, Paul

2007-03-01

Registration of two dimensional to three dimensional orthopaedic medical image data has important applications particularly in the area of image guided surgery and sports medicine. Fluoroscopy to computer tomography (CT) registration is an important case, wherein digitally reconstructed radiographs derived from the CT data are registered to the fluoroscopy data. Traditional registration metrics such as intensity-based mutual information (MI) typically work well but often suffer from gross misregistration errors when the image to be registered contains a partial view of the anatomy visible in the target image. Phase-based MI provides a robust alternative similarity measure which, in addition to possessing the general robustness and noise immunity that MI provides, also employs local phase information in the registration process which makes it less susceptible to the aforementioned errors. In this paper, we propose using the complex wavelet transform for computing image phase information and incorporating that into a phase-based MI measure for image registration. Tests on a CT volume and 6 fluoroscopy images of the knee are presented. The femur and the tibia in the CT volume were individually registered to the fluoroscopy images using intensity-based MI, gradient-based MI and phase-based MI. Errors in the coordinates of fiducials present in the bone structures were used to assess the accuracy of the different registration schemes. Quantitative results demonstrate that the performance of intensity-based MI was the worst. Gradient-based MI performed slightly better, while phase-based MI results were the best consistently producing the lowest errors.

WE-H-207A-02: Attenuation Correction in 4D-PET Using a Single-Phase Attenuation Map

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kalantari, F; Wang, J

2016-06-15

Purpose: 4D-PET imaging has been proposed as a potential solution to the respiratory motion effect in thoracic region. CT-based attenuation correction (AC) is an essential step toward quantitative imaging for PET. However, due to the temporal difference of 4D-PET and a single breath-hold CT, motion artifacts are observed in the attenuation-corrected PET images that can lead to error in tumor shape and uptake. We introduce a practical method for aligning single-phase CT to all other 4D-PET phases using a penalized non-rigid demons registration. Methods: Individual 4D-PET frames were reconstructed without AC. Non-rigid Demons registration was used to derive deformation vectormore » fields (DVFs) between the PET matched with CT phase and other 4D-PET images. While attenuated PET images provide enough useful data for organ borders such as lung and liver, tumors are not distinguishable from background due to loss of contrast. To preserve tumor shape in different phases, from CT image an ROI covering tumor was excluded from non-rigid transformation. Mean DVF of the central region of the tumor was assigned to all voxels in the ROI. This process mimics a rigid transformation of tumor along with a non-rigid transformation of other organs. 4D XCAT phantom with spherical tumors in lung with diameters ranging from 10 to 40 mm was used to evaluate the algorithm. Results: Motion related induced artifacts in attenuation-corrected 4D-PET images were significantly reduced. For tumors smaller than 20 mm, non-rigid transformation was capable to provide quantitative results. However, for larger tumors, where tumor self-attenuation is considerable, our combined method yields superior results. Conclusion: We introduced a practical method for deforming a single CT to match all 4D-PET images for accurate AC. Although 4D-PET data include insignificant anatomical information, we showed that they are still useful to estimate DVFs for aligning attenuation map and accurate AC.« less

From synchrotron radiation to lab source: advanced speckle-based X-ray imaging using abrasive paper

NASA Astrophysics Data System (ADS)

Wang, Hongchang; Kashyap, Yogesh; Sawhney, Kawal

2016-02-01

X-ray phase and dark-field imaging techniques provide complementary and inaccessible information compared to conventional X-ray absorption or visible light imaging. However, such methods typically require sophisticated experimental apparatus or X-ray beams with specific properties. Recently, an X-ray speckle-based technique has shown great potential for X-ray phase and dark-field imaging using a simple experimental arrangement. However, it still suffers from either poor resolution or the time consuming process of collecting a large number of images. To overcome these limitations, in this report we demonstrate that absorption, dark-field, phase contrast, and two orthogonal differential phase contrast images can simultaneously be generated by scanning a piece of abrasive paper in only one direction. We propose a novel theoretical approach to quantitatively extract the above five images by utilising the remarkable properties of speckles. Importantly, the technique has been extended from a synchrotron light source to utilise a lab-based microfocus X-ray source and flat panel detector. Removing the need to raster the optics in two directions significantly reduces the acquisition time and absorbed dose, which can be of vital importance for many biological samples. This new imaging method could potentially provide a breakthrough for numerous practical imaging applications in biomedical research and materials science.

Advanced ballistic range technology

NASA Technical Reports Server (NTRS)

Yates, Leslie A.

1993-01-01

Experimental interferograms, schlieren, and shadowgraphs are used for quantitative and qualitative flow-field studies. These images are created by passing light through a flow field, and the recorded intensity patterns are functions of the phase shift and angular deflection of the light. As part of the grant NCC2-583, techniques and software have been developed for obtaining phase shifts from finite-fringe interferograms and for constructing optical images from Computational Fluid Dynamics (CFD) solutions. During the period from 1 Nov. 1992 - 30 Jun. 1993, research efforts have been concentrated in improving these techniques.

Asymmetric masks for laboratory-based X-ray phase-contrast imaging with edge illumination.

PubMed

Endrizzi, Marco; Astolfo, Alberto; Vittoria, Fabio A; Millard, Thomas P; Olivo, Alessandro

2016-05-05

We report on an asymmetric mask concept that enables X-ray phase-contrast imaging without requiring any movement in the system during data acquisition. The method is compatible with laboratory equipment, namely a commercial detector and a rotating anode tube. The only motion required is that of the object under investigation which is scanned through the imaging system. Two proof-of-principle optical elements were designed, fabricated and experimentally tested. Quantitative measurements on samples of known shape and composition were compared to theory with good agreement. The method is capable of measuring the attenuation, refraction and (ultra-small-angle) X-ray scattering, does not have coherence requirements and naturally adapts to all those situations in which the X-ray image is obtained by scanning a sample through the imaging system.

AI-augmented time stretch microscopy

NASA Astrophysics Data System (ADS)

Mahjoubfar, Ata; Chen, Claire L.; Lin, Jiahao; Jalali, Bahram

2017-02-01

Cell reagents used in biomedical analysis often change behavior of the cells that they are attached to, inhibiting their native signaling. On the other hand, label-free cell analysis techniques have long been viewed as challenging either due to insufficient accuracy by limited features, or because of low throughput as a sacrifice of improved precision. We present a recently developed artificial-intelligence augmented microscope, which builds upon high-throughput time stretch quantitative phase imaging (TS-QPI) and deep learning to perform label-free cell classification with record high-accuracy. Our system captures quantitative optical phase and intensity images simultaneously by frequency multiplexing, extracts multiple biophysical features of the individual cells from these images fused, and feeds these features into a supervised machine learning model for classification. The enhanced performance of our system compared to other label-free assays is demonstrated by classification of white blood T-cells versus colon cancer cells and lipid accumulating algal strains for biofuel production, which is as much as five-fold reduction in inaccuracy. This system obtains the accuracy required in practical applications such as personalized drug development, while the cells remain intact and the throughput is not sacrificed. Here, we introduce a data acquisition scheme based on quadrature phase demodulation that enables interruptionless storage of TS-QPI cell images. Our proof of principle demonstration is capable of saving 40 TB of cell images in about four hours, i.e. pictures of every single cell in 10 mL of a sample.

Simultaneous fast scanning XRF, dark field, phase-, and absorption contrast tomography

NASA Astrophysics Data System (ADS)

Medjoubi, Kadda; Bonissent, Alain; Leclercq, Nicolas; Langlois, Florent; Mercère, Pascal; Somogyi, Andrea

2013-09-01

Scanning hard X-ray nanoprobe imaging provides a unique tool for probing specimens with high sensitivity and large penetration depth. Moreover, the combination of complementary techniques such as X-ray fluorescence, absorption, phase contrast and dark field imaging gives complete quantitative information on the sample structure, composition and chemistry. The multi-technique "FLYSCAN" data acquisition scheme developed at Synchrotron SOLEIL permits to perform fast continuous scanning imaging and as such makes scanning tomography techniques feasible in a time-frame well-adapted to typical user experiments. Here we present the recent results of simultaneous fast scanning multi-technique tomography performed at Soleil. This fast scanning scheme will be implemented at the Nanoscopium beamline for large field of view 2D and 3D multimodal imaging.

Photothermal nanoparticles as molecular specificity agents in interferometric phase microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Shaked, Natan T.

2017-02-01

I review our latest advances in wide-field interferometric imaging of biological cells with molecular specificity, obtained by time-modulated photothermal excitation of gold nanoparticles. Heat emitted from the nanoparticles affects the measured phase signal via both the nanoparticle surrounding refractive-index and thickness changes. These nanoparticles can be bio-functionalized to bind certain biological cell components; thus, they can be used for biomedical imaging with molecular specificity, as new nanoscopy labels, and for photothermal therapy. Predicting the ideal nanoparticle parameters requires a model that computes the thermal and phase distributions around the particle, enabling more efficient phase imaging of plasmonic nanoparticles, and sparing trial and error experiments of using unsuitable nanoparticles. We thus developed a new model for predicting phase signatures from photothermal nanoparticles with arbitrary parameters. We also present a dual-modality technique based on wide-field photothermal interferometric phase imaging and simultaneous ablation to selectively deplete specific cell populations labelled by plasmonic nanoparticles. We experimentally demonstrated our ability to detect and specifically ablate in vitro cancer cells over-expressing epidermal growth factor receptors (EGFRs), labelled with plasmonic nanoparticles, in the presence of either EGFR under-expressing cancer cells or white blood cells. This demonstration established an initial model for depletion of circulating tumour cells in blood. The proposed system is able to image in wide field the label-free quantitative phase profile together with the photothermal phase profile of the sample, and provides the ability of both detection and ablation of chosen cells after their selective imaging.

Probing the limits of the rigid-intensity-shift model in differential-phase-contrast scanning transmission electron microscopy

NASA Astrophysics Data System (ADS)

Clark, L.; Brown, H. G.; Paganin, D. M.; Morgan, M. J.; Matsumoto, T.; Shibata, N.; Petersen, T. C.; Findlay, S. D.

2018-04-01

The rigid-intensity-shift model of differential-phase-contrast imaging assumes that the phase gradient imposed on the transmitted probe by the sample causes the diffraction pattern intensity to shift rigidly by an amount proportional to that phase gradient. This behavior is seldom realized exactly in practice. Through a combination of experimental results, analytical modeling and numerical calculations, using as case studies electron microscope imaging of the built-in electric field in a p-n junction and nanoscale domains in a magnetic alloy, we explore the breakdown of rigid-intensity-shift behavior and how this depends on the magnitude of the phase gradient and the relative scale of features in the phase profile and the probe size. We present guidelines as to when the rigid-intensity-shift model can be applied for quantitative phase reconstruction using segmented detectors, and propose probe-shaping strategies to further improve the accuracy.

Assessment of left ventricular mechanical dyssynchrony by phase analysis of gated-SPECT myocardial perfusion imaging and tissue Doppler imaging: comparison between QGS and ECTb software packages.

PubMed

Rastgou, Fereydoon; Shojaeifard, Maryam; Amin, Ahmad; Ghaedian, Tahereh; Firoozabadi, Hasan; Malek, Hadi; Yaghoobi, Nahid; Bitarafan-Rajabi, Ahmad; Haghjoo, Majid; Amouzadeh, Hedieh; Barati, Hossein

2014-12-01

Recently, the phase analysis of gated single-photon emission computed tomography (SPECT) myocardial perfusion imaging (MPI) has become feasible via several software packages for the evaluation of left ventricular mechanical dyssynchrony. We compared two quantitative software packages, quantitative gated SPECT (QGS) and Emory cardiac toolbox (ECTb), with tissue Doppler imaging (TDI) as the conventional method for the evaluation of left ventricular mechanical dyssynchrony. Thirty-one patients with severe heart failure (ejection fraction ≤35%) and regular heart rhythm, who referred for gated-SPECT MPI, were enrolled. TDI was performed within 3 days after MPI. Dyssynchrony parameters derived from gated-SPECT MPI were analyzed by QGS and ECTb and were compared with the Yu index and septal-lateral wall delay measured by TDI. QGS and ECTb showed a good correlation for assessment of phase histogram bandwidth (PHB) and phase standard deviation (PSD) (r = 0.664 and r = 0.731, P < .001, respectively). However, the mean value of PHB and PSD by ECTb was significantly higher than that of QGS. No significant correlation was found between ECTb and QGS and the Yu index. Nevertheless, PHB, PSD, and entropy derived from QGS revealed a significant (r = 0.424, r = 0.478, r = 0.543, respectively; P < .02) correlation with septal-lateral wall delay. Despite a good correlation between QGS and ECTb software packages, different normal cut-off values of PSD and PHB should be defined for each software package. There was only a modest correlation between phase analysis of gated-SPECT MPI and TDI data, especially in the population of heart failure patients with both narrow and wide QRS complex.

Noncontact quantitative biomechanical characterization of cardiac muscle using shear wave imaging optical coherence tomography

PubMed Central

Wang, Shang; Lopez, Andrew L.; Morikawa, Yuka; Tao, Ge; Li, Jiasong; Larina, Irina V.; Martin, James F.; Larin, Kirill V.

2014-01-01

We report on a quantitative optical elastographic method based on shear wave imaging optical coherence tomography (SWI-OCT) for biomechanical characterization of cardiac muscle through noncontact elasticity measurement. The SWI-OCT system employs a focused air-puff device for localized loading of the cardiac muscle and utilizes phase-sensitive OCT to monitor the induced tissue deformation. Phase information from the optical interferometry is used to reconstruct 2-D depth-resolved shear wave propagation inside the muscle tissue. Cross-correlation of the displacement profiles at various spatial locations in the propagation direction is applied to measure the group velocity of the shear waves, based on which the Young’s modulus of tissue is quantified. The quantitative feature and measurement accuracy of this method is demonstrated from the experiments on tissue-mimicking phantoms with the verification using uniaxial compression test. The experiments are performed on ex vivo cardiac muscle tissue from mice with normal and genetically altered myocardium. Our results indicate this optical elastographic technique is useful as a noncontact tool to assist the cardiac muscle studies. PMID:25071943

"One-Stop Shop": Free-Breathing Dynamic Contrast-Enhanced Magnetic Resonance Imaging of the Kidney Using Iterative Reconstruction and Continuous Golden-Angle Radial Sampling.

PubMed

Riffel, Philipp; Zoellner, Frank G; Budjan, Johannes; Grimm, Robert; Block, Tobias K; Schoenberg, Stefan O; Hausmann, Daniel

2016-11-01

The purpose of the present study was to evaluate a recently introduced technique for free-breathing dynamic contrast-enhanced renal magnetic resonance imaging (MRI) applying a combination of radial k-space sampling, parallel imaging, and compressed sensing. The technique allows retrospective reconstruction of 2 motion-suppressed sets of images from the same acquisition: one with lower temporal resolution but improved image quality for subjective image analysis, and one with high temporal resolution for quantitative perfusion analysis. In this study, 25 patients underwent a kidney examination, including a prototypical fat-suppressed, golden-angle radial stack-of-stars T1-weighted 3-dimensional spoiled gradient-echo examination (GRASP) performed after contrast agent administration during free breathing. Images were reconstructed at temporal resolutions of 55 spokes per frame (6.2 seconds) and 13 spokes per frame (1.5 seconds). The GRASP images were evaluated by 2 blinded radiologists. First, the reconstructions with low temporal resolution underwent subjective image analysis: the radiologists assessed the best arterial phase and the best renal phase and rated image quality score for each patient on a 5-point Likert-type scale.In addition, the diagnostic confidence was rated according to a 3-point Likert-type scale. Similarly, respiratory motion artifacts and streak artifacts were rated according to a 3-point Likert-type scale.Then, the reconstructions with high temporal resolution were analyzed with a voxel-by-voxel deconvolution approach to determine the renal plasma flow, and the results were compared with values reported in previous literature. Reader 1 and reader 2 rated the overall image quality score for the best arterial phase and the best renal phase with a median image quality score of 4 (good image quality) for both phases, respectively. A high diagnostic confidence (median score of 3) was observed. There were no respiratory motion artifacts in any of the patients. Streak artifacts were present in all of the patients, but did not compromise diagnostic image quality.The estimated renal plasma flow was slightly higher (295 ± 78 mL/100 mL per minute) than reported in previous MRI-based studies, but also closer to the physiologically expected value. Dynamic, motion-suppressed contrast-enhanced renal MRI can be performed in high diagnostic quality during free breathing using a combination of golden-angle radial sampling, parallel imaging, and compressed sensing. Both morphologic and quantitative functional information can be acquired within a single acquisition.

Diffusion-weighted MR imaging findings of kidneys in patients with early phase of obstruction.

PubMed

Bozgeyik, Zulkif; Kocakoc, Ercan; Sonmezgoz, Fitnet

2009-04-01

Diffusion-weighted (DW) magnetic resonance (MR) imaging is an MR technique used to show molecular diffusion. The apparent diffusion coefficient (ADC), as a quantitative parameter calculated from the DW MR images. The purpose of this study is to evaluate the ability of DW MR imaging in early phase of obstruction due to urolithiasis. Twenty-six patients with acute dilatation of the pelvicalyceal system detected by intravenous urography were included in this study. MR imaging was performed using a 1.5 T whole-body superconducting MR scanner. DW imaging can be performed using single-shot spin-echo, echo-planar imaging (EPI) sequences with the following diffusion gradient b values: 100, 600, 1000 s/mm(2). Circular region of interest (ROI) was placed in the renal parenchyma for the measurement of ADC values in the normal and obstructed kidney. For statistical analyses, Paired t test were used. In spite of obstructed kidneys had the lower ADC values compared to normal kidneys, these alterations were statistically insignificant. We did not observe significantly different ADC values of early phase of obstructed kidneys compared to normal kidneys.

3D quantitative phase imaging of neural networks using WDT

NASA Astrophysics Data System (ADS)

Kim, Taewoo; Liu, S. C.; Iyer, Raj; Gillette, Martha U.; Popescu, Gabriel

2015-03-01

White-light diffraction tomography (WDT) is a recently developed 3D imaging technique based on a quantitative phase imaging system called spatial light interference microscopy (SLIM). The technique has achieved a sub-micron resolution in all three directions with high sensitivity granted by the low-coherence of a white-light source. Demonstrations of the technique on single cell imaging have been presented previously; however, imaging on any larger sample, including a cluster of cells, has not been demonstrated using the technique. Neurons in an animal body form a highly complex and spatially organized 3D structure, which can be characterized by neuronal networks or circuits. Currently, the most common method of studying the 3D structure of neuron networks is by using a confocal fluorescence microscope, which requires fluorescence tagging with either transient membrane dyes or after fixation of the cells. Therefore, studies on neurons are often limited to samples that are chemically treated and/or dead. WDT presents a solution for imaging live neuron networks with a high spatial and temporal resolution, because it is a 3D imaging method that is label-free and non-invasive. Using this method, a mouse or rat hippocampal neuron culture and a mouse dorsal root ganglion (DRG) neuron culture have been imaged in order to see the extension of processes between the cells in 3D. Furthermore, the tomogram is compared with a confocal fluorescence image in order to investigate the 3D structure at synapses.

Shape based segmentation of MRIs of the bones in the knee using phase and intensity information

NASA Astrophysics Data System (ADS)

Fripp, Jurgen; Bourgeat, Pierrick; Crozier, Stuart; Ourselin, Sébastien

2007-03-01

The segmentation of the bones from MR images is useful for performing subsequent segmentation and quantitative measurements of cartilage tissue. In this paper, we present a shape based segmentation scheme for the bones that uses texture features derived from the phase and intensity information in the complex MR image. The phase can provide additional information about the tissue interfaces, but due to the phase unwrapping problem, this information is usually discarded. By using a Gabor filter bank on the complex MR image, texture features (including phase) can be extracted without requiring phase unwrapping. These texture features are then analyzed using a support vector machine classifier to obtain probability tissue matches. The segmentation of the bone is fully automatic and performed using a 3D active shape model based approach driven using gradient and texture information. The 3D active shape model is automatically initialized using a robust affine registration. The approach is validated using a database of 18 FLASH MR images that are manually segmented, with an average segmentation overlap (Dice similarity coefficient) of 0.92 compared to 0.9 obtained using the classifier only.

Dual-wavelength common-path digital holographic microscopy for quantitative phase imaging of biological cells

NASA Astrophysics Data System (ADS)

Di, Jianglei; Song, Yu; Xi, Teli; Zhang, Jiwei; Li, Ying; Ma, Chaojie; Wang, Kaiqiang; Zhao, Jianlin

2017-11-01

Biological cells are usually transparent with a small refractive index gradient. Digital holographic interferometry can be used in the measurement of biological cells. We propose a dual-wavelength common-path digital holographic microscopy for the quantitative phase imaging of biological cells. In the proposed configuration, a parallel glass plate is inserted in the light path to create the lateral shearing, and two lasers with different wavelengths are used as the light source to form the dual-wavelength composite digital hologram. The information of biological cells for different wavelengths is separated and extracted in the Fourier domain of the hologram, and then combined to a shorter wavelength in the measurement process. This method could improve the system's temporal stability and reduce speckle noises simultaneously. Mouse osteoblastic cells and peony pollens are measured to show the feasibility of this method.

Motion-gated acquisition for in vivo optical imaging

PubMed Central

Gioux, Sylvain; Ashitate, Yoshitomo; Hutteman, Merlijn; Frangioni, John V.

2009-01-01

Wide-field continuous wave fluorescence imaging, fluorescence lifetime imaging, frequency domain photon migration, and spatially modulated imaging have the potential to provide quantitative measurements in vivo. However, most of these techniques have not yet been successfully translated to the clinic due to challenging environmental constraints. In many circumstances, cardiac and respiratory motion greatly impair image quality and∕or quantitative processing. To address this fundamental problem, we have developed a low-cost, field-programmable gate array–based, hardware-only gating device that delivers a phase-locked acquisition window of arbitrary delay and width that is derived from an unlimited number of pseudo-periodic and nonperiodic input signals. All device features can be controlled manually or via USB serial commands. The working range of the device spans the extremes of mouse electrocardiogram (1000 beats per minute) to human respiration (4 breaths per minute), with timing resolution ⩽0.06%, and jitter ⩽0.008%, of the input signal period. We demonstrate the performance of the gating device, including dramatic improvements in quantitative measurements, in vitro using a motion simulator and in vivo using near-infrared fluorescence angiography of beating pig heart. This gating device should help to enable the clinical translation of promising new optical imaging technologies. PMID:20059276

Automated thermal mapping techniques using chromatic image analysis

NASA Technical Reports Server (NTRS)

Buck, Gregory M.

1989-01-01

Thermal imaging techniques are introduced using a chromatic image analysis system and temperature sensitive coatings. These techniques are used for thermal mapping and surface heat transfer measurements on aerothermodynamic test models in hypersonic wind tunnels. Measurements are made on complex vehicle configurations in a timely manner and at minimal expense. The image analysis system uses separate wavelength filtered images to analyze surface spectral intensity data. The system was initially developed for quantitative surface temperature mapping using two-color thermographic phosphors but was found useful in interpreting phase change paint and liquid crystal data as well.

Grand Tour outer planet missions definition phase. Part 1: Quantitative imaging of the outer planets and their satellites

NASA Technical Reports Server (NTRS)

Belton, M. J. S.; Aksnes, K.; Davies, M. E.; Hartmann, W. K.; Millis, R. L.; Owen, T. C.; Reilly, T. H.; Sagan, C.; Suomi, V. E.; Collins, S. A., Jr.

1972-01-01

A recommended imaging system is outlined for use aboard the Outer Planet Grand Tour Explorer. The system features the high angular resolution capacity necessary to accommodate large encounter distances, and to satisfy the demand for a reasonable amount of time coverage. Specifications for all components within the system are provided in detail.

Wideband digital frequency detector with subtraction-based phase comparator for frequency modulation atomic force microscopy.

PubMed

Mitani, Yuji; Kubo, Mamoru; Muramoto, Ken-ichiro; Fukuma, Takeshi

2009-08-01

We have developed a wideband digital frequency detector for high-speed frequency modulation atomic force microscopy (FM-AFM). We used a subtraction-based phase comparator (PC) in a phase-locked loop circuit instead of a commonly used multiplication-based PC, which has enhanced the detection bandwidth to 100 kHz. The quantitative analysis of the noise performance revealed that the internal noise from the developed detector is small enough to provide the theoretically limited noise performance in FM-AFM experiments in liquid. FM-AFM imaging of mica in liquid was performed with the developed detector, showing its stability and applicability to true atomic-resolution imaging in liquid.

Quantifying lung morphology with respiratory-gated micro-CT in a murine model of emphysema

NASA Astrophysics Data System (ADS)

Ford, N. L.; Martin, E. L.; Lewis, J. F.; Veldhuizen, R. A. W.; Holdsworth, D. W.; Drangova, M.

2009-04-01

Non-invasive micro-CT imaging techniques have been developed to investigate lung structure in free-breathing rodents. In this study, we investigate the utility of retrospectively respiratory-gated micro-CT imaging in an emphysema model to determine if anatomical changes could be observed in the image-derived quantitative analysis at two respiratory phases. The emphysema model chosen was a well-characterized, genetically altered model (TIMP-3 knockout mice) that exhibits a homogeneous phenotype. Micro-CT scans of the free-breathing, anaesthetized mice were obtained in 50 s and retrospectively respiratory sorted and reconstructed, providing 3D images representing peak inspiration and end expiration with 0.15 mm isotropic voxel spacing. Anatomical measurements included the volume and CT density of the lungs and the volume of the major airways, along with the diameters of the trachea, left bronchus and right bronchus. From these measurements, functional parameters such as functional residual capacity and tidal volume were calculated. Significant differences between the wild-type and TIMP-3 knockout groups were observed for measurements of CT density over the entire lung, indicating increased air content in the lungs of TIMP-3 knockout mice. These results demonstrate retrospective respiratory-gated micro-CT, providing images at multiple respiratory phases that can be analyzed quantitatively to investigate anatomical changes in murine models of emphysema.

Phase measurements of EUV mask defects

DOE PAGES

Claus, Rene A.; Wang, Yow-Gwo; Wojdyla, Antoine; ...

2015-02-22

Extreme Ultraviolet (EUV) Lithography mask defects were examined on the actinic mask imaging system, SHARP, at Lawrence Berkeley National Laboratory. Also, a quantitative phase retrieval algorithm based on the Weak Object Transfer Function was applied to the measured through-focus aerial images to examine the amplitude and phase of the defects. The accuracy of the algorithm was demonstrated by comparing the results of measurements using a phase contrast zone plate and a standard zone plate. Using partially coherent illumination to measure frequencies that would otherwise fall outside the numerical aperture (NA), it was shown that some defects are smaller than themore » conventional resolution of the microscope. We found that the programmed defects of various sizes were measured and shown to have both an amplitude and a phase component that the algorithm is able to recover.« less

Spectrally resolved laser interference microscopy

NASA Astrophysics Data System (ADS)

Butola, Ankit; Ahmad, Azeem; Dubey, Vishesh; Senthilkumaran, P.; Singh Mehta, Dalip

2018-07-01

We developed a new quantitative phase microscopy technique, namely, spectrally resolved laser interference microscopy (SR-LIM), with which it is possible to quantify multi-spectral phase information related to biological specimens without color crosstalk using a color CCD camera. It is a single shot technique where sequential switched on/off of red, green, and blue (RGB) wavelength light sources are not required. The method is implemented using a three-wavelength interference microscope and a customized compact grating based imaging spectrometer fitted at the output port. The results of the USAF resolution chart while employing three different light sources, namely, a halogen lamp, light emitting diodes, and lasers, are discussed and compared. The broadband light sources like the halogen lamp and light emitting diodes lead to stretching in the spectrally decomposed images, whereas it is not observed in the case of narrow-band light sources, i.e. lasers. The proposed technique is further successfully employed for single-shot quantitative phase imaging of human red blood cells at three wavelengths simultaneously without color crosstalk. Using the present technique, one can also use a monochrome camera, even though the experiments are performed using multi-color light sources. Finally, SR-LIM is not only limited to RGB wavelengths, it can be further extended to red, near infra-red, and infra-red wavelengths, which are suitable for various biological applications.

Digital micromirror device-based common-path quantitative phase imaging.

PubMed

Zheng, Cheng; Zhou, Renjie; Kuang, Cuifang; Zhao, Guangyuan; Yaqoob, Zahid; So, Peter T C

2017-04-01

We propose a novel common-path quantitative phase imaging (QPI) method based on a digital micromirror device (DMD). The DMD is placed in a plane conjugate to the objective back-aperture plane for the purpose of generating two plane waves that illuminate the sample. A pinhole is used in the detection arm to filter one of the beams after sample to create a reference beam. Additionally, a transmission-type liquid crystal device, placed at the objective back-aperture plane, eliminates the specular reflection noise arising from all the "off" state DMD micromirrors, which is common in all DMD-based illuminations. We have demonstrated high sensitivity QPI, which has a measured spatial and temporal noise of 4.92 nm and 2.16 nm, respectively. Experiments with calibrated polystyrene beads illustrate the desired phase measurement accuracy. In addition, we have measured the dynamic height maps of red blood cell membrane fluctuations, showing the efficacy of the proposed system for live cell imaging. Most importantly, the DMD grants the system convenience in varying the interference fringe period on the camera to easily satisfy the pixel sampling conditions. This feature also alleviates the pinhole alignment complexity. We envision that the proposed DMD-based common-path QPI system will allow for system miniaturization and automation for a broader adaption.

Digital micromirror device-based common-path quantitative phase imaging

PubMed Central

Zheng, Cheng; Zhou, Renjie; Kuang, Cuifang; Zhao, Guangyuan; Yaqoob, Zahid; So, Peter T. C.

2017-01-01

We propose a novel common-path quantitative phase imaging (QPI) method based on a digital micromirror device (DMD). The DMD is placed in a plane conjugate to the objective back-aperture plane for the purpose of generating two plane waves that illuminate the sample. A pinhole is used in the detection arm to filter one of the beams after sample to create a reference beam. Additionally, a transmission-type liquid crystal device, placed at the objective back-aperture plane, eliminates the specular reflection noise arising from all the “off” state DMD micromirrors, which is common in all DMD-based illuminations. We have demonstrated high sensitivity QPI, which has a measured spatial and temporal noise of 4.92 nm and 2.16 nm, respectively. Experiments with calibrated polystyrene beads illustrate the desired phase measurement accuracy. In addition, we have measured the dynamic height maps of red blood cell membrane fluctuations, showing the efficacy of the proposed system for live cell imaging. Most importantly, the DMD grants the system convenience in varying the interference fringe period on the camera to easily satisfy the pixel sampling conditions. This feature also alleviates the pinhole alignment complexity. We envision that the proposed DMD-based common-path QPI system will allow for system miniaturization and automation for a broader adaption. PMID:28362789

ESR imaging investigations of two-phase systems.

PubMed

Herrmann, Werner; Stösser, Reinhard; Borchert, Hans-Hubert

2007-06-01

The possibilities of electron spin resonance (ESR) and electron spin resonance imaging (ESRI) for investigating the properties of the spin probes TEMPO and TEMPOL in two-phase systems have been examined in the systems water/n-octanol, Miglyol/Miglyol, and Precirol/Miglyol. Phases and regions of the phase boundary could be mapped successfully by means of the isotropic hyperfine coupling constants, and, moreover, the quantification of rotational and lateral diffusion of the spin probes was possible. For the quantitative treatment of the micropolarity, a simplified empirical model was established on the basis of the Nernst distribution and the experimentally determined isotropic hyperfine coupling constants. The model does not only describe the summarized micropolarities of coexisting phases, but also the region of the phase boundary, where solvent molecules of different polarities and tendencies to form hydrogen bonds compete to interact with the NO group of the spin probe. Copyright 2007 John Wiley & Sons, Ltd.

Topography and refractometry of sperm cells using spatial light interference microscopy

NASA Astrophysics Data System (ADS)

Liu, Lina; Kandel, Mikhail E.; Rubessa, Marcello; Schreiber, Sierra; Wheeler, Mathew B.; Popescu, Gabriel

2018-02-01

Characterization of spermatozoon viability is a common test in treating infertility. Recently, it has been shown that label-free, phase-sensitive imaging can provide a valuable alternative for this type of assay. We employ spatial light interference microscopy (SLIM) to perform high-accuracy single-cell phase imaging and decouple the average thickness and refractive index information for the population. This procedure was enabled by quantitative-phase imaging cells on media of two different refractive indices and using a numerical tool to remove the curvature from the cell tails. This way, we achieved ensemble averaging of topography and refractometry of 100 cells in each of the two groups. The results show that the thickness profile of the cell tail goes down to 150 nm and the refractive index can reach values of 1.6 close to the head.

Combined use of X-ray fluorescence microscopy, phase contrast imaging for high resolution quantitative iron mapping in inflamed cells

NASA Astrophysics Data System (ADS)

Gramaccioni, C.; Procopio, A.; Farruggia, G.; Malucelli, E.; Iotti, S.; Notargiacomo, A.; Fratini, M.; Yang, Y.; Pacureanu, A.; Cloetens, P.; Bohic, S.; Massimi, L.; Cutone, A.; Valenti, P.; Rosa, L.; Berlutti, F.; Lagomarsino, S.

2017-06-01

X-ray fluorescence microscopy (XRFM) is a powerful technique to detect and localize elements in cells. To derive information useful for biology and medicine, it is essential not only to localize, but also to map quantitatively the element concentration. Here we applied quantitative XRFM to iron in phagocytic cells. Iron, a primary component of living cells, can become toxic when present in excess. In human fluids, free iron is maintained at 10-18 M concentration thanks to iron binding proteins as lactoferrin (Lf). The iron homeostasis, involving the physiological ratio of iron between tissues/secretions and blood, is strictly regulated by ferroportin, the sole protein able to export iron from cells to blood. Inflammatory processes induced by lipopolysaccharide (LPS) or bacterial pathoge inhibit ferroportin synthesis in epithelial and phagocytic cells thus hindering iron export, increasing intracellular iron and bacterial multiplication. In this respect, Lf is emerging as an important regulator of both iron and inflammatory homeostasis. Here we studied phagocytic cells inflamed by bacterial LPS and untreated or treated with milk derived bovine Lf. Quantitative mapping of iron concentration and mass fraction at high spatial resolution is obtained combining X-ray fluorescence microscopy, atomic force microscopy and synchrotron phase contrast imaging.

Three-dimensional morphological imaging of human induced pluripotent stem cells by using low-coherence quantitative phase microscopy

NASA Astrophysics Data System (ADS)

Yamauchi, Toyohiko; Kakuno, Yumi; Goto, Kentaro; Fukami, Tadashi; Sugiyama, Norikazu; Iwai, Hidenao; Mizuguchi, Yoshinori; Yamashita, Yutaka

2014-03-01

There is an increasing need for non-invasive imaging techniques in the field of stem cell research. Label-free techniques are the best choice for assessment of stem cells because the cells remain intact after imaging and can be used for further studies such as differentiation induction. To develop a high-resolution label-free imaging system, we have been working on a low-coherence quantitative phase microscope (LC-QPM). LC-QPM is a Linnik-type interference microscope equipped with nanometer-resolution optical-path-length control and capable of obtaining three-dimensional volumetric images. The lateral and vertical resolutions of our system are respectively 0.5 and 0.93 μm and this performance allows capturing sub-cellular morphological features of live cells without labeling. Utilizing LC-QPM, we reported on three-dimensional imaging of membrane fluctuations, dynamics of filopodia, and motions of intracellular organelles. In this presentation, we report three-dimensional morphological imaging of human induced pluripotent stem cells (hiPS cells). Two groups of monolayer hiPS cell cultures were prepared so that one group was cultured in a suitable culture medium that kept the cells undifferentiated, and the other group was cultured in a medium supplemented with retinoic acid, which forces the stem cells to differentiate. The volumetric images of the 2 groups show distinctive differences, especially in surface roughness. We believe that our LC-QPM system will prove useful in assessing many other stem cell conditions.

SU-G-JeP3-14: Positioning and Dosimetric Uncertainties in Image-Guided Radiation Therapy with Respiratory Gating

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ali, I; Hossain, S; Algan, O

Purpose: To investigate quantitatively positioning and dosimetric uncertainties due to 4D-CT intra-phase motion in the internal-target-volume (ITV) associated with radiation therapy using respiratory-gating for patients setup with image-guidance-radiation-therapy (IGRT) using free-breathing or average-phase CT-images. Methods: A lung phantom with an embedded tissue-equivalent target is imaged with CT while it is stationary and moving. Four-sets of structures are outlined: (a) the actual target on CT-images of the stationary-target, (b) ITV on CT-images for the free-moving phantom, (c) ITV’s from the ten different phases (10–100%) and (d) ITV on the CT-images generated from combining 3 phases: 40%–50%–60%. The variations in volume, lengthmore » and center-position of the ITV’s and their effects on dosimetry during dose delivery for patients setup with image-guidance are investigated. Results: Intra-phase motion due to breathing affects the volume, center position and length of the ITVs from different respiratory-phases. The ITV’s vary by about 10% from one phase to another. The largest ITV is measured on the free breathing CT images and the smallest is on the stationary CT-images. The ITV lengths vary by about 4mm where it may shrink or elongated depending on the motion-phase. The center position of the ITV varies between the different motion-phases which shifts upto 10mm from the stationary-position which is nearly equal to motion-amplitude. This causes systematic shifts during dose delivery with beam gating using certain phases (40%–50%–60%) for patients setup with IGRT using free-breathing or average-phase CT-images. The dose coverage of the ITV depends on the margins used for treatment-planning-volume where margins larger than the motion-amplitudes are needed to ensure dose coverage of the ITV. Conclusion: Volume, length, and center position of the ITV’s change between the different motion phases. Large systematic shifts are induced by respiratory-gating with ITVs on certain phases when patients are setup with IGRT using free-breathing or average-phase CT-images.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Claus, Rene A.; Wang, Yow-Gwo; Wojdyla, Antoine

Extreme Ultraviolet (EUV) Lithography mask defects were examined on the actinic mask imaging system, SHARP, at Lawrence Berkeley National Laboratory. Also, a quantitative phase retrieval algorithm based on the Weak Object Transfer Function was applied to the measured through-focus aerial images to examine the amplitude and phase of the defects. The accuracy of the algorithm was demonstrated by comparing the results of measurements using a phase contrast zone plate and a standard zone plate. Using partially coherent illumination to measure frequencies that would otherwise fall outside the numerical aperture (NA), it was shown that some defects are smaller than themore » conventional resolution of the microscope. We found that the programmed defects of various sizes were measured and shown to have both an amplitude and a phase component that the algorithm is able to recover.« less

Non-destructive phase contrast hard x-ray imaging to reveal the three-dimensional microstructure of soft and hard tissues

NASA Astrophysics Data System (ADS)

Khimchenko, Anna; Schulz, Georg; Deyhle, Hans; Hieber, Simone E.; Hasan, Samiul; Bikis, Christos; Schulz, Joachim; Costeur, Loïc.; Müller, Bert

2016-04-01

X-ray imaging in the absorption contrast mode is an established method of visualising calcified tissues such as bone and teeth. Physically soft tissues such as brain or muscle are often imaged using magnetic resonance imaging (MRI). However, the spatial resolution of MRI is insufficient for identifying individual biological cells within three-dimensional tissue. X-ray grating interferometry (XGI) has advantages for the investigation of soft tissues or the simultaneous three-dimensional visualisation of soft and hard tissues. Since laboratory microtomography (μCT) systems have better accessibility than tomography set-ups at synchrotron radiation facilities, a great deal of effort has been invested in optimising XGI set-ups for conventional μCT systems. In this conference proceeding, we present how a two-grating interferometer is incorporated into a commercially available nanotom m (GE Sensing and Inspection Technologies GmbH) μCT system to extend its capabilities toward phase contrast. We intend to demonstrate superior contrast in spiders (Hogna radiata (Fam. Lycosidae) and Xysticus erraticus (Fam. Thomisidae)), as well as the simultaneous visualisation of hard and soft tissues. XGI is an imaging modality that provides quantitative data, and visualisation is an important part of biomimetics; consequently, hard X-ray imaging provides a sound basis for bioinspiration, bioreplication and biomimetics and allows for the quantitative comparison of biofabricated products with their natural counterparts.

Chemotaxis of cancer cells in three-dimensional environment monitored label-free by quantitative phase digital holographic microscopy

NASA Astrophysics Data System (ADS)

Kemper, Björn; Schnekenburger, Jürgen; Ketelhut, Steffi

2017-02-01

We investigated the capabilities of digital holographic microscopy (DHM) for label-free quantification of the response of living single cells to chemical stimuli in 3D assays. Fibro sarcoma cells were observed in a collagen matrix inside 3D chemotaxis chambers with a Mach-Zehnder interferometer-based DHM setup. From the obtained series of quantitative phase images, the migration trajectories of single cells were retrieved by automated cell tracking and subsequently analyzed for maximum migration distance and motility. Our results demonstrate DHM as a highly reliable and efficient tool for label-free quantification of chemotaxis in 2D and 3D environments.

Single shot white light interference microscopy with colour fringe analysis for quantitative phase imaging of biological cells

NASA Astrophysics Data System (ADS)

Srivastava, Vishal; Mehta, D. S.

2013-02-01

To quantitatively obtain the phase map of Onion and human red blood cell (RBC) from white light interferogram we used Hilbert transform color fringe analysis technique. The three Red, Blue and Green color components are decomposed from single white light interferogram and Refractive index profile for Red, Blue and Green colour were computed in a completely non-invasive manner for Onion and human RBC. The present technique might be useful for non-invasive determination of the refractive index variation within cells and tissues and morphological features of sample with ease of operation and low cost.

Synthetic aperture tomographic phase microscopy for 3D imaging of live cells in translational motion

PubMed Central

Lue, Niyom; Choi, Wonshik; Popescu, Gabriel; Badizadegan, Kamran; Dasari, Ramachandra R.; Feld, Michael S.

2009-01-01

We present a technique for 3D imaging of live cells in translational motion without need of axial scanning of objective lens. A set of transmitted electric field images of cells at successive points of transverse translation is taken with a focused beam illumination. Based on Hyugens’ principle, angular plane waves are synthesized from E-field images of a focused beam. For a set of synthesized angular plane waves, we apply a filtered back-projection algorithm and obtain 3D maps of refractive index of live cells. This technique, which we refer to as synthetic aperture tomographic phase microscopy, can potentially be combined with flow cytometry or microfluidic devices, and will enable high throughput acquisition of quantitative refractive index data from large numbers of cells. PMID:18825263

Quantitative Assessment of Regional Wall Motion Abnormalities Using Dual-Energy Digital Subtraction Intravenous Ventriculography

NASA Astrophysics Data System (ADS)

McCollough, Cynthia H.

Healthy portions of the left ventricle (LV) can often compensate for regional dysfunction, thereby masking regional disease when global indices of LV function are employed. Thus, quantitation of regional function provides a more useful method of assessing LV function, especially in diseases that have regional effects such as coronary artery disease. This dissertation studied the ability of a phase -matched dual-energy digital subtraction angiography (DE -DSA) technique to quantitate changes in regional LV systolic volume. The potential benefits and a theoretical description of the DE imaging technique are detailed. A correlated noise reduction algorithm is also presented which raises the signal-to-noise ratio of DE images by a factor of 2 -4. Ten open-chest dogs were instrumented with transmural ultrasonic crystals to assess regional LV function in terms of systolic normalized-wall-thickening rate (NWTR) and percent-systolic-thickening (PST). A pneumatic occluder was placed on the left-anterior-descending (LAD) coronary artery to temporarily reduce myocardial blood flow, thereby changing regional LV function in the LAD bed. DE-DSA intravenous left ventriculograms were obtained at control and four levels of graded myocardial ischemia, as determined by reductions in PST. Phase-matched images displaying changes in systolic contractile function were created by subtracting an end-systolic (ES) control image from ES images acquired at each level of myocardial ischemia. The resulting wall-motion difference signal (WMD), which represents a change in regional systolic volume between the control and ischemic states, was quantitated by videodensitometry and compared with changes in NWTR and PST. Regression analysis of 56 data points from 10 animals shows a linear relationship between WMD and both NWTR and PST: WMD = -2.46 NWTR + 13.9, r = 0.64, p < 0.001; WMD = -2.11 PST + 18.4, r = 0.54, p < 0.001. Thus, changes in regional ES LV volume between rest and ischemic states, as measured using the described imaging technique, appear linearly related to changes in wall-thickening, as measured using transmural ultrasonic crystals. This type of image analysis may prove useful in a variety of clinical and research applications and further investigation is proposed.

Investigation of fluorocarbon blowing agents in insulating polymer foams by 19F NMR imaging.

PubMed

Fyfe, C A; Mei, Z; Grondey, H

1996-01-01

Currently, there is no reliable and readily accessible technique with which the distribution and diffusion of blowing agents in rigid insulating foams can be detected and monitored. In this paper, we demonstrate that 19F NMR microscopic imaging together with 19F solid-state MAS NMR spectroscopy is ideally suited for such measurements and yield quantitatively reliable information that will be critical to the development and fabrication of optimized insulating materials with alternative blowing agents. Polystyrene (PS) and polyurethane (PU) foam samples were investigated with the objective of determining quantitatively the amount of blowing agents in the gaseous phase and dissolved in the polymer phase, and to determine and monitor the distribution of the blowing agents in aged foams as a function of time and temperature. The concentrations of the gaseous blowing agents in the cells and dissolved in the solid were simultaneously and quantitatively measured by 19F MAS NMR spectroscopy. An unfaced 1-yr-old PS foam filled with CH3CF2Cl has about 13% of total HCFCs dissolved in the solid; while there is about 24% of HCFCs in the solid of a faced 3-mos-old PU foam filled with CH3CCl2F. The data from 19F NMR imaging demonstrate that the distributions of the blowing agents in an aged foam are quite uniform around the center part (2 cm away from any edge) of a foam board; however, a gradient in blowing agent concentration was found as a function of distance from the initial factory cut edge. The effective diffusion coefficients of the blowing agents can be directly calculated from the imaging data. Quantitative diffusion constants and activation barriers were determined. Additionally, a foam treated with a second blowing agent was monitored with chemical shift selective imaging and the diffusion of the second gas into the foam and the out-diffusion of the original gas were determined.

Evaluation of chemotherapy response in ovarian cancer treatment using quantitative CT image biomarkers: a preliminary study

NASA Astrophysics Data System (ADS)

Qiu, Yuchen; Tan, Maxine; McMeekin, Scott; Thai, Theresa; Moore, Kathleen; Ding, Kai; Liu, Hong; Zheng, Bin

2015-03-01

The purpose of this study is to identify and apply quantitative image biomarkers for early prediction of the tumor response to the chemotherapy among the ovarian cancer patients participated in the clinical trials of testing new drugs. In the experiment, we retrospectively selected 30 cases from the patients who participated in Phase I clinical trials of new drug or drug agents for ovarian cancer treatment. Each case is composed of two sets of CT images acquired pre- and post-treatment (4-6 weeks after starting treatment). A computer-aided detection (CAD) scheme was developed to extract and analyze the quantitative image features of the metastatic tumors previously tracked by the radiologists using the standard Response Evaluation Criteria in Solid Tumors (RECIST) guideline. The CAD scheme first segmented 3-D tumor volumes from the background using a hybrid tumor segmentation scheme. Then, for each segmented tumor, CAD computed three quantitative image features including the change of tumor volume, tumor CT number (density) and density variance. The feature changes were calculated between the matched tumors tracked on the CT images acquired pre- and post-treatments. Finally, CAD predicted patient's 6-month progression-free survival (PFS) using a decision-tree based classifier. The performance of the CAD scheme was compared with the RECIST category. The result shows that the CAD scheme achieved a prediction accuracy of 76.7% (23/30 cases) with a Kappa coefficient of 0.493, which is significantly higher than the performance of RECIST prediction with a prediction accuracy and Kappa coefficient of 60% (17/30) and 0.062, respectively. This study demonstrated the feasibility of analyzing quantitative image features to improve the early predicting accuracy of the tumor response to the new testing drugs or therapeutic methods for the ovarian cancer patients.

Phase imaging microscopy for the diagnostics of plasma-cell interaction

NASA Astrophysics Data System (ADS)

Ohene, Yolanda; Marinov, Ilya; de Laulanié, Lucie; Dupuy, Corinne; Wattelier, Benoit; Starikovskaia, Svetlana

2015-06-01

Phase images of biological specimens were obtained by the method of Quadriwave Lateral Shearing Interferometry (QWLSI). The QWLSI technique produces, at high resolution, phase images of the cells having been exposed to a plasma treatment and enables the quantitative analysis of the changes in the surface area of the cells over time. Morphological changes in the HTori normal thyroid cells were demonstrated using this method. There was a comparison of the cell behaviour between control cells, cells treated by plasma of a nanosecond dielectric barrier discharge, including cells pre-treated by catalase, and cells treated with an equivalent amount of H2O2. The major changes in the cell membrane morphology were observed at only 5 min after the plasma treatment. The primary role of reactive oxygen species (ROS) in this degradation is suggested. Deformation and condensation of the cell nucleus were observed 2-3 h after the treatment and are supposedly related to apoptosis induction. The coupling of the phase QWLSI with immunofluorescence imaging would give a deeper insight into the mechanisms of plasma induced cell death.

Facing the phase problem in Coherent Diffractive Imaging via Memetic Algorithms.

PubMed

Colombo, Alessandro; Galli, Davide Emilio; De Caro, Liberato; Scattarella, Francesco; Carlino, Elvio

2017-02-09

Coherent Diffractive Imaging is a lensless technique that allows imaging of matter at a spatial resolution not limited by lens aberrations. This technique exploits the measured diffraction pattern of a coherent beam scattered by periodic and non-periodic objects to retrieve spatial information. The diffracted intensity, for weak-scattering objects, is proportional to the modulus of the Fourier Transform of the object scattering function. Any phase information, needed to retrieve its scattering function, has to be retrieved by means of suitable algorithms. Here we present a new approach, based on a memetic algorithm, i.e. a hybrid genetic algorithm, to face the phase problem, which exploits the synergy of deterministic and stochastic optimization methods. The new approach has been tested on simulated data and applied to the phasing of transmission electron microscopy coherent electron diffraction data of a SrTiO 3 sample. We have been able to quantitatively retrieve the projected atomic potential, and also image the oxygen columns, which are not directly visible in the relevant high-resolution transmission electron microscopy images. Our approach proves to be a new powerful tool for the study of matter at atomic resolution and opens new perspectives in those applications in which effective phase retrieval is necessary.

Correction of phase errors in quantitative water-fat imaging using a monopolar time-interleaved multi-echo gradient echo sequence.

PubMed

Ruschke, Stefan; Eggers, Holger; Kooijman, Hendrik; Diefenbach, Maximilian N; Baum, Thomas; Haase, Axel; Rummeny, Ernst J; Hu, Houchun H; Karampinos, Dimitrios C

2017-09-01

To propose a phase error correction scheme for monopolar time-interleaved multi-echo gradient echo water-fat imaging that allows accurate and robust complex-based quantification of the proton density fat fraction (PDFF). A three-step phase correction scheme is proposed to address a) a phase term induced by echo misalignments that can be measured with a reference scan using reversed readout polarity, b) a phase term induced by the concomitant gradient field that can be predicted from the gradient waveforms, and c) a phase offset between time-interleaved echo trains. Simulations were carried out to characterize the concomitant gradient field-induced PDFF bias and the performance estimating the phase offset between time-interleaved echo trains. Phantom experiments and in vivo liver and thigh imaging were performed to study the relevance of each of the three phase correction steps on PDFF accuracy and robustness. The simulation, phantom, and in vivo results showed in agreement with the theory an echo time-dependent PDFF bias introduced by the three phase error sources. The proposed phase correction scheme was found to provide accurate PDFF estimation independent of the employed echo time combination. Complex-based time-interleaved water-fat imaging was found to give accurate and robust PDFF measurements after applying the proposed phase error correction scheme. Magn Reson Med 78:984-996, 2017. © 2016 International Society for Magnetic Resonance in Medicine. © 2016 International Society for Magnetic Resonance in Medicine.

Halo-free phase contrast microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Nguyen, Tan H.; Kandel, Mikhail E.; Shakir, Haadi M.; Best, Catherine; Do, Minh N.; Popescu, Gabriel

2017-02-01

The phase contrast (PC) method is one of the most impactful developments in the four-century long history of microscopy. It allows for intrinsic, nondestructive contrast of transparent specimens, such as live cells. However, PC is plagued by the halo artifact, a result of insufficient spatial coherence in the illumination field, which limits its applicability. We present a new approach for retrieving halo-free phase contrast microscopy (hfPC) images by upgrading the conventional PC microscope with an external interferometric module, which generates sufficient data for reversing the halo artifact. Measuring four independent intensity images, our approach first measures haloed phase maps of the sample. We solve for the halo-free sample transmission function by using a physical model of the image formation under partial spatial coherence. Using this halo-free sample transmission, we can numerically generate artifact-free PC images. Furthermore, this transmission can be further used to obtain quantitative information about the sample, e.g., the thickness with known refractive indices, dry mass of live cells during their cycles. We tested our hfPC method on various control samples, e.g., beads, pillars and validated its potential for biological investigation by imaging live HeLa cells, red blood cells, and neurons.

Attenuation correction in 4D-PET using a single-phase attenuation map and rigidity-adaptive deformable registration

PubMed Central

Kalantari, Faraz; Wang, Jing

2017-01-01

Purpose Four-dimensional positron emission tomography (4D-PET) imaging is a potential solution to the respiratory motion effect in the thoracic region. Computed tomography (CT)-based attenuation correction (AC) is an essential step toward quantitative imaging for PET. However, due to the temporal difference between 4D-PET and a single attenuation map from CT, typically available in routine clinical scanning, motion artifacts are observed in the attenuation-corrected PET images, leading to errors in tumor shape and uptake. We introduced a practical method to align single-phase CT with all other 4D-PET phases for AC. Methods A penalized non-rigid Demons registration between individual 4D-PET frames without AC provides the motion vectors to be used for warping single-phase attenuation map. The non-rigid Demons registration was used to derive deformation vector fields (DVFs) between PET matched with the CT phase and other 4D-PET images. While attenuated PET images provide useful data for organ borders such as those of the lung and the liver, tumors cannot be distinguished from the background due to loss of contrast. To preserve the tumor shape in different phases, an ROI-covering tumor was excluded from non-rigid transformation. Instead the mean DVF of the central region of the tumor was assigned to all voxels in the ROI. This process mimics a rigid transformation of the tumor along with a non-rigid transformation of other organs. A 4D-XCAT phantom with spherical lung tumors, with diameters ranging from 10 to 40 mm, was used to evaluate the algorithm. The performance of the proposed hybrid method for attenuation map estimation was compared to 1) the Demons non-rigid registration only and 2) a single attenuation map based on quantitative parameters in individual PET frames. Results Motion-related artifacts were significantly reduced in the attenuation-corrected 4D-PET images. When a single attenuation map was used for all individual PET frames, the normalized root mean square error (NRMSE) values in tumor region were 49.3% (STD: 8.3%), 50.5% (STD: 9.3%), 51.8% (STD: 10.8%) and 51.5% (STD: 12.1%) for 10-mm, 20-mm, 30-mm and 40-mm tumors respectively. These errors were reduced to 11.9% (STD: 2.9%), 13.6% (STD: 3.9%), 13.8% (STD: 4.8%), and 16.7% (STD: 9.3%) by our proposed method for deforming the attenuation map. The relative errors in total lesion glycolysis (TLG) values were −0.25% (STD: 2.87%) and 3.19% (STD: 2.35%) for 30-mm and 40-mm tumors respectively in proposed method. The corresponding values for Demons method were 25.22% (STD: 14.79%) and 18.42% (STD: 7.06%). Our proposed hybrid method outperforms the Demons method especially for larger tumors. For tumors smaller than 20 mm, non-rigid transformation could also provide quantitative results. Conclusion Although non-AC 4D-PET frames include insignificant anatomical information, they are still useful to estimate the DVFs to align the attenuation map for accurate AC. The proposed hybrid method can recover the AC-related artifacts and provide quantitative AC-PET images. PMID:27987223

Two-dimensional phase unwrapping using robust derivative estimation and adaptive integration.

PubMed

Strand, Jarle; Taxt, Torfinn

2002-01-01

The adaptive integration (ADI) method for two-dimensional (2-D) phase unwrapping is presented. The method uses an algorithm for noise robust estimation of partial derivatives, followed by a noise robust adaptive integration process. The ADI method can easily unwrap phase images with moderate noise levels, and the resulting images are congruent modulo 2pi with the observed, wrapped, input images. In a quantitative evaluation, both the ADI and the BLS methods (Strand et al.) were better than the least-squares methods of Ghiglia and Romero (GR), and of Marroquin and Rivera (MRM). In a qualitative evaluation, the ADI, the BLS, and a conjugate gradient version of the MRM method (MRMCG), were all compared using a synthetic image with shear, using 115 magnetic resonance images, and using 22 fiber-optic interferometry images. For the synthetic image and the interferometry images, the ADI method gave consistently visually better results than the other methods. For the MR images, the MRMCG method was best, and the ADI method second best. The ADI method was less sensitive to the mask definition and the block size than the BLS method, and successfully unwrapped images with shears that were not marked in the masks. The computational requirements of the ADI method for images of nonrectangular objects were comparable to only two iterations of many least-squares-based methods (e.g., GR). We believe the ADI method provides a powerful addition to the ensemble of tools available for 2-D phase unwrapping.

Quantitative light-induced fluorescence technology for quantitative evaluation of tooth wear

NASA Astrophysics Data System (ADS)

Kim, Sang-Kyeom; Lee, Hyung-Suk; Park, Seok-Woo; Lee, Eun-Song; de Josselin de Jong, Elbert; Jung, Hoi-In; Kim, Baek-Il

2017-12-01

Various technologies used to objectively determine enamel thickness or dentin exposure have been suggested. However, most methods have clinical limitations. This study was conducted to confirm the potential of quantitative light-induced fluorescence (QLF) using autofluorescence intensity of occlusal surfaces of worn teeth according to enamel grinding depth in vitro. Sixteen permanent premolars were used. Each tooth was gradationally ground down at the occlusal surface in the apical direction. QLF-digital and swept-source optical coherence tomography images were acquired at each grinding depth (in steps of 100 μm). All QLF images were converted to 8-bit grayscale images to calculate the fluorescence intensity. The maximum brightness (MB) values of the same sound regions in grayscale images before (MB) and phased values after (MB) the grinding process were calculated. Finally, 13 samples were evaluated. MB increased over the grinding depth range with a strong correlation (r=0.994, P<0.001). In conclusion, the fluorescence intensity of the teeth and grinding depth was strongly correlated in the QLF images. Therefore, QLF technology may be a useful noninvasive tool used to monitor the progression of tooth wear and to conveniently estimate enamel thickness.

Lossless and lossy compression of quantitative phase images of red blood cells obtained by digital holographic imaging.

PubMed

Jaferzadeh, Keyvan; Gholami, Samaneh; Moon, Inkyu

2016-12-20

In this paper, we evaluate lossless and lossy compression techniques to compress quantitative phase images of red blood cells (RBCs) obtained by an off-axis digital holographic microscopy (DHM). The RBC phase images are numerically reconstructed from their digital holograms and are stored in 16-bit unsigned integer format. In the case of lossless compression, predictive coding of JPEG lossless (JPEG-LS), JPEG2000, and JP3D are evaluated, and compression ratio (CR) and complexity (compression time) are compared against each other. It turns out that JP2k can outperform other methods by having the best CR. In the lossy case, JP2k and JP3D with different CRs are examined. Because some data is lost in a lossy way, the degradation level is measured by comparing different morphological and biochemical parameters of RBC before and after compression. Morphological parameters are volume, surface area, RBC diameter, sphericity index, and the biochemical cell parameter is mean corpuscular hemoglobin (MCH). Experimental results show that JP2k outperforms JP3D not only in terms of mean square error (MSE) when CR increases, but also in compression time in the lossy compression way. In addition, our compression results with both algorithms demonstrate that with high CR values the three-dimensional profile of RBC can be preserved and morphological and biochemical parameters can still be within the range of reported values.

Nuclear medicine and quantitative imaging research (quantitative studies in radiopharmaceutical science): Comprehensive progress report, April 1, 1986-December 31, 1988

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cooper, M.D.; Beck, R.N.

1988-06-01

This document describes several years research to improve PET imaging and diagnostic techniques in man. This program addresses the problems involving the basic science and technology underlying the physical and conceptual tools of radioactive tracer methodology as they relate to the measurement of structural and functional parameters of physiologic importance in health and disease. The principal tool is quantitative radionuclide imaging. The overall objective of this program is to further the development and transfer of radiotracer methodology from basic theory to routine clinical practice in order that individual patients and society as a whole will receive the maximum net benefitmore » from the new knowledge gained. The focus of the research is on the development of new instruments and radiopharmaceuticals, and the evaluation of these through the phase of clinical feasibility. The reports in the study were processed separately for the data bases. (TEM)« less

Cortical phase changes in Alzheimer's disease at 7T MRI: a novel imaging marker.

PubMed

van Rooden, Sanneke; Versluis, Maarten J; Liem, Michael K; Milles, Julien; Maier, Andrea B; Oleksik, Ania M; Webb, Andrew G; van Buchem, Mark A; van der Grond, Jeroen

2014-01-01

Postmortem studies have indicated the potential of high-field magnetic resonance imaging (MRI) to visualize amyloid depositions in the cerebral cortex. The aim of this study is to test this hypothesis in patients with Alzheimer's disease (AD). T2*-weighted MRI was performed in 16 AD patients and 15 control subjects. All magnetic resonance images were scored qualitatively by visual assessment, and quantitatively by measuring phase shifts in the cortical gray matter and hippocampus. Statistical analysis was performed to assess differences between groups. Patients with AD demonstrated an increased phase shift in the cortex in the temporoparietal, frontal, and parietal regions (P < .005), and this was associated with individual Mini-Mental State Examination scores (r = -0.54, P < .05). Increased cortical phase shift in AD patients demonstrated on 7-tesla T2*-weighted MRI is a potential new biomarker for AD, which may reflect amyloid pathology in the early stages. Copyright © 2014 The Alzheimer's Association. Published by Elsevier Inc. All rights reserved.

Partial scan artifact reduction (PSAR) for the assessment of cardiac perfusion in dynamic phase-correlated CT.

PubMed

Stenner, Philip; Schmidt, Bernhard; Bruder, Herbert; Allmendinger, Thomas; Haberland, Ulrike; Flohr, Thomas; Kachelriess, Marc

2009-12-01

Cardiac CT achieves its high temporal resolution by lowering the scan range from 2pi to pi plus fan angle (partial scan). This, however, introduces CT-value variations, depending on the angular position of the pi range. These partial scan artifacts are of the order of a few HU and prevent the quantitative evaluation of perfusion measurements. The authors present the new algorithm partial scan artifact reduction (PSAR) that corrects a dynamic phase-correlated scan without a priori information. In general, a full scan does not suffer from partial scan artifacts since all projections in [0, 2pi] contribute to the data. To maintain the optimum temporal resolution and the phase correlation, PSAR creates an artificial full scan pn(AF) by projectionwise averaging a set of neighboring partial scans pn(P) from the same perfusion examination (typically N approximately 30 phase-correlated partial scans distributed over 20 s and n = 1, ..., N). Corresponding to the angular range of each partial scan, the authors extract virtual partial scans pn(V) from the artificial full scan pn(AF). A standard reconstruction yields the corresponding images fn(P), fn(AF), and fn(V). Subtracting the virtual partial scan image fn(V) from the artificial full scan image fn(AF) yields an artifact image that can be used to correct the original partial scan image: fn(C) = fn(P) - fn(V) + fn(AF), where fn(C) is the corrected image. The authors evaluated the effects of scattered radiation on the partial scan artifacts using simulated and measured water phantoms and found a strong correlation. The PSAR algorithm has been validated with a simulated semianthropomorphic heart phantom and with measurements of a dynamic biological perfusion phantom. For the stationary phantoms, real full scans have been performed to provide theoretical reference values. The improvement in the root mean square errors between the full and the partial scans with respect to the errors between the full and the corrected scans is up to 54% for the simulations and 90% for the measurements. The phase-correlated data now appear accurate enough for a quantitative analysis of cardiac perfusion.

Direct observation of dopant distribution in GaAs compound semiconductors using phase-shifting electron holography and Lorentz microscopy.

PubMed

Sasaki, Hirokazu; Otomo, Shinya; Minato, Ryuichiro; Yamamoto, Kazuo; Hirayama, Tsukasa

2014-06-01

Phase-shifting electron holography and Lorentz microscopy were used to map dopant distributions in GaAs compound semiconductors with step-like dopant concentration. Transmission electron microscope specimens were prepared using a triple beam focused ion beam (FIB) system, which combines a Ga ion beam, a scanning electron microscope, and an Ar ion beam to remove the FIB damaged layers. The p-n junctions were clearly observed in both under-focused and over-focused Lorentz microscopy images. A phase image was obtained by using a phase-shifting reconstruction method to simultaneously achieve high sensitivity and high spatial resolution. Differences in dopant concentrations between 1 × 10(19) cm(-3) and 1 × 10(18) cm(-3) regions were clearly observed by using phase-shifting electron holography. We also interpreted phase profiles quantitatively by considering inactive layers induced by ion implantation during the FIB process. The thickness of an inactive layer at different dopant concentration area can be measured from the phase image. © The Author 2014. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Optofluidic bioimaging platform for quantitative phase imaging of lab on a chip devices using digital holographic microscopy.

PubMed

Pandiyan, Vimal Prabhu; John, Renu

2016-01-20

We propose a versatile 3D phase-imaging microscope platform for real-time imaging of optomicrofluidic devices based on the principle of digital holographic microscopy (DHM). Lab-on-chip microfluidic devices fabricated on transparent polydimethylsiloxane (PDMS) and glass substrates have attained wide popularity in biological sensing applications. However, monitoring, visualization, and characterization of microfluidic devices, microfluidic flows, and the biochemical kinetics happening in these devices is difficult due to the lack of proper techniques for real-time imaging and analysis. The traditional bright-field microscopic techniques fail in imaging applications, as the microfluidic channels and the fluids carrying biological samples are transparent and not visible in bright light. Phase-based microscopy techniques that can image the phase of the microfluidic channel and changes in refractive indices due to the fluids and biological samples present in the channel are ideal for imaging the fluid flow dynamics in a microfluidic channel at high resolutions. This paper demonstrates three-dimensional imaging of a microfluidic device with nanometric depth precisions and high SNR. We demonstrate imaging of microelectrodes of nanometric thickness patterned on glass substrate and the microfluidic channel. Three-dimensional imaging of a transparent PDMS optomicrofluidic channel, fluid flow, and live yeast cell flow in this channel has been demonstrated using DHM. We also quantify the average velocity of fluid flow through the channel. In comparison to any conventional bright-field microscope, the 3D depth information in the images illustrated in this work carry much information about the biological system under observation. The results demonstrated in this paper prove the high potential of DHM in imaging optofluidic devices; detection of pathogens, cells, and bioanalytes on lab-on-chip devices; and in studying microfluidic dynamics in real time based on phase changes.

Precisely detecting atomic position of atomic intensity images.

PubMed

Wang, Zhijun; Guo, Yaolin; Tang, Sai; Li, Junjie; Wang, Jincheng; Zhou, Yaohe

2015-03-01

We proposed a quantitative method to detect atomic position in atomic intensity images from experiments such as high-resolution transmission electron microscopy, atomic force microscopy, and simulation such as phase field crystal modeling. The evaluation of detection accuracy proves the excellent performance of the method. This method provides a chance to precisely determine atomic interactions based on the detected atomic positions from the atomic intensity image, and hence to investigate the related physical, chemical and electrical properties. Copyright © 2014 Elsevier B.V. All rights reserved.

Comparison of three‐dimensional analysis and stereological techniques for quantifying lithium‐ion battery electrode microstructures

PubMed Central

TAIWO, OLUWADAMILOLA O.; FINEGAN, DONAL P.; EASTWOOD, DAVID S.; FIFE, JULIE L.; BROWN, LEON D.; DARR, JAWWAD A.; LEE, PETER D.; BRETT, DANIEL J.L.

2016-01-01

Summary Lithium‐ion battery performance is intrinsically linked to electrode microstructure. Quantitative measurement of key structural parameters of lithium‐ion battery electrode microstructures will enable optimization as well as motivate systematic numerical studies for the improvement of battery performance. With the rapid development of 3‐D imaging techniques, quantitative assessment of 3‐D microstructures from 2‐D image sections by stereological methods appears outmoded; however, in spite of the proliferation of tomographic imaging techniques, it remains significantly easier to obtain two‐dimensional (2‐D) data sets. In this study, stereological prediction and three‐dimensional (3‐D) analysis techniques for quantitative assessment of key geometric parameters for characterizing battery electrode microstructures are examined and compared. Lithium‐ion battery electrodes were imaged using synchrotron‐based X‐ray tomographic microscopy. For each electrode sample investigated, stereological analysis was performed on reconstructed 2‐D image sections generated from tomographic imaging, whereas direct 3‐D analysis was performed on reconstructed image volumes. The analysis showed that geometric parameter estimation using 2‐D image sections is bound to be associated with ambiguity and that volume‐based 3‐D characterization of nonconvex, irregular and interconnected particles can be used to more accurately quantify spatially‐dependent parameters, such as tortuosity and pore‐phase connectivity. PMID:26999804

Comparison of three-dimensional analysis and stereological techniques for quantifying lithium-ion battery electrode microstructures.

PubMed

Taiwo, Oluwadamilola O; Finegan, Donal P; Eastwood, David S; Fife, Julie L; Brown, Leon D; Darr, Jawwad A; Lee, Peter D; Brett, Daniel J L; Shearing, Paul R

2016-09-01

Lithium-ion battery performance is intrinsically linked to electrode microstructure. Quantitative measurement of key structural parameters of lithium-ion battery electrode microstructures will enable optimization as well as motivate systematic numerical studies for the improvement of battery performance. With the rapid development of 3-D imaging techniques, quantitative assessment of 3-D microstructures from 2-D image sections by stereological methods appears outmoded; however, in spite of the proliferation of tomographic imaging techniques, it remains significantly easier to obtain two-dimensional (2-D) data sets. In this study, stereological prediction and three-dimensional (3-D) analysis techniques for quantitative assessment of key geometric parameters for characterizing battery electrode microstructures are examined and compared. Lithium-ion battery electrodes were imaged using synchrotron-based X-ray tomographic microscopy. For each electrode sample investigated, stereological analysis was performed on reconstructed 2-D image sections generated from tomographic imaging, whereas direct 3-D analysis was performed on reconstructed image volumes. The analysis showed that geometric parameter estimation using 2-D image sections is bound to be associated with ambiguity and that volume-based 3-D characterization of nonconvex, irregular and interconnected particles can be used to more accurately quantify spatially-dependent parameters, such as tortuosity and pore-phase connectivity. © 2016 The Authors. Journal of Microscopy published by John Wiley & Sons Ltd on behalf of Royal Microscopical Society.

Why phase errors affect the electron function more than amplitude errors.

PubMed

Lattman, Eaton; DeRosier, David

2008-03-01

If Fexp(ialpha) are the set of structure factors for a structure f, the amplitudes can be converted to those of an uncorrelated structure g (amplitude swapping) by multiplying each F by the positive number G/F. Correspondingly, the image f is convoluted with k, the Fourier transform of G/F; k has a large peak at the origin, so that f * k approximately f. For swapped phases, the image f is convoluted with l, the Fourier transform of exp(iDeltaalpha), where Deltaalpha, the phase difference between F and G, is a random variable; l does not have a large peak at the origin, so that f * l does not resemble f. The paper provides quantitative descriptions of these arguments.

Topography and refractometry of sperm cells using spatial light interference microscopy.

PubMed

Liu, Lina; Kandel, Mikhail E; Rubessa, Marcello; Schreiber, Sierra; Wheeler, Mathew B; Popescu, Gabriel

2018-02-01

Characterization of spermatozoon viability is a common test in treating infertility. Recently, it has been shown that label-free, phase-sensitive imaging can provide a valuable alternative for this type of assay. We employ spatial light interference microscopy (SLIM) to perform high-accuracy single-cell phase imaging and decouple the average thickness and refractive index information for the population. This procedure was enabled by quantitative-phase imaging cells on media of two different refractive indices and using a numerical tool to remove the curvature from the cell tails. This way, we achieved ensemble averaging of topography and refractometry of 100 cells in each of the two groups. The results show that the thickness profile of the cell tail goes down to 150 nm and the refractive index can reach values of 1.6 close to the head. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Segmentation-free image processing and analysis of precipitate shapes in 2D and 3D

NASA Astrophysics Data System (ADS)

Bales, Ben; Pollock, Tresa; Petzold, Linda

2017-06-01

Segmentation based image analysis techniques are routinely employed for quantitative analysis of complex microstructures containing two or more phases. The primary advantage of these approaches is that spatial information on the distribution of phases is retained, enabling subjective judgements of the quality of the segmentation and subsequent analysis process. The downside is that computing micrograph segmentations with data from morphologically complex microstructures gathered with error-prone detectors is challenging and, if no special care is taken, the artifacts of the segmentation will make any subsequent analysis and conclusions uncertain. In this paper we demonstrate, using a two phase nickel-base superalloy microstructure as a model system, a new methodology for analysis of precipitate shapes using a segmentation-free approach based on the histogram of oriented gradients feature descriptor, a classic tool in image analysis. The benefits of this methodology for analysis of microstructure in two and three-dimensions are demonstrated.

Intensity non-uniformity correction using N3 on 3-T scanners with multichannel phased array coils

PubMed Central

Boyes, Richard G.; Gunter, Jeff L.; Frost, Chris; Janke, Andrew L.; Yeatman, Thomas; Hill, Derek L.G.; Bernstein, Matt A.; Thompson, Paul M.; Weiner, Michael W.; Schuff, Norbert; Alexander, Gene E.; Killiany, Ronald J.; DeCarli, Charles; Jack, Clifford R.; Fox, Nick C.

2008-01-01

Measures of structural brain change based on longitudinal MR imaging are increasingly important but can be degraded by intensity non-uniformity. This non-uniformity can be more pronounced at higher field strengths, or when using multichannel receiver coils. We assessed the ability of the non-parametric non-uniform intensity normalization (N3) technique to correct non-uniformity in 72 volumetric brain MR scans from the preparatory phase of the Alzheimer’s Disease Neuroimaging Initiative (ADNI). Normal elderly subjects (n = 18) were scanned on different 3-T scanners with a multichannel phased array receiver coil at baseline, using magnetization prepared rapid gradient echo (MP-RAGE) and spoiled gradient echo (SPGR) pulse sequences, and again 2 weeks later. When applying N3, we used five brain masks of varying accuracy and four spline smoothing distances (d = 50, 100, 150 and 200 mm) to ascertain which combination of parameters optimally reduces the non-uniformity. We used the normalized white matter intensity variance (standard deviation/mean) to ascertain quantitatively the correction for a single scan; we used the variance of the normalized difference image to assess quantitatively the consistency of the correction over time from registered scan pairs. Our results showed statistically significant (p < 0.01) improvement in uniformity for individual scans and reduction in the normalized difference image variance when using masks that identified distinct brain tissue classes, and when using smaller spline smoothing distances (e.g., 50-100 mm) for both MP-RAGE and SPGR pulse sequences. These optimized settings may assist future large-scale studies where 3-T scanners and phased array receiver coils are used, such as ADNI, so that intensity non-uniformity does not influence the power of MR imaging to detect disease progression and the factors that influence it. PMID:18063391

Intraindividual Crossover Comparison of Gadoxetic Acid Dose for Liver MRI in Normal Volunteers.

PubMed

Motosugi, Utaroh; Bannas, Peter; Hernando, Diego; Salmani Rahimi, Mahdi; Holmes, James H; Reeder, Scott B

2016-01-01

We performed a quantitative intraindividual comparison of the performance of 0.025- and 0.05-mmol/kg doses for gadoxetic acid-enhanced liver magnetic resonance (MR) imaging. Eleven healthy volunteers underwent liver MR imaging twice, once with a 0.025- and once with a 0.05-mmol/kg dose of gadoxetic acid. MR spectroscopy and 3-dimensional gradient-echo T1-weighted images (3D-GRE) were obtained before and 3, 10, and 20 min after injection of the contrast medium to measure T1 and T2 values and signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) performance. During the dynamic phase, highly time-resolved 3D-GRE was used to estimate the relative CNR (CNRrel) of the hepatic artery and portal vein (PV) to the liver. We used paired t-tests to compare the results of different doses. During the hepatobiliary phase, we observed shorter T1 values and higher SNRs of the liver (P < 0.001) and higher liver-to-PV and liver-to-muscle CNRs (P < 0.002) using 0.05 mmol/kg compared to 0.025 mmol/kg. Increasing the dose to 0.05 mmol/kg yielded a greater T1-shortening effect at 10 min delay even compared with 0.025 mmol/kg at 20 min (P < 0.001). During the dynamic phase, the peak CNRrel for the hepatic artery and portal vein were higher using 0.05 mmol/kg (P = 0.007 to 0.035). Use of gadoxetic acid at a dose of 0.05 mmol/kg leads to significantly higher SNR and CNR performance than with 0.025 mmol/kg. Quantitatively, a 10-min delay may be feasible for hepatobiliary-phase imaging when using 0.05 mmol/kg of gadoxetic acid.

Image-derived and arterial blood sampled input functions for quantitative PET imaging of the angiotensin II subtype 1 receptor in the kidney

DOE Office of Scientific and Technical Information (OSTI.GOV)

Feng, Tao; Tsui, Benjamin M. W.; Li, Xin

Purpose: The radioligand {sup 11}C-KR31173 has been introduced for positron emission tomography (PET) imaging of the angiotensin II subtype 1 receptor in the kidney in vivo. To study the biokinetics of {sup 11}C-KR31173 with a compartmental model, the input function is needed. Collection and analysis of arterial blood samples are the established approach to obtain the input function but they are not feasible in patients with renal diseases. The goal of this study was to develop a quantitative technique that can provide an accurate image-derived input function (ID-IF) to replace the conventional invasive arterial sampling and test the method inmore » pigs with the goal of translation into human studies. Methods: The experimental animals were injected with [{sup 11}C]KR31173 and scanned up to 90 min with dynamic PET. Arterial blood samples were collected for the artery derived input function (AD-IF) and used as a gold standard for ID-IF. Before PET, magnetic resonance angiography of the kidneys was obtained to provide the anatomical information required for derivation of the recovery coefficients in the abdominal aorta, a requirement for partial volume correction of the ID-IF. Different image reconstruction methods, filtered back projection (FBP) and ordered subset expectation maximization (OS-EM), were investigated for the best trade-off between bias and variance of the ID-IF. The effects of kidney uptakes on the quantitative accuracy of ID-IF were also studied. Biological variables such as red blood cell binding and radioligand metabolism were also taken into consideration. A single blood sample was used for calibration in the later phase of the input function. Results: In the first 2 min after injection, the OS-EM based ID-IF was found to be biased, and the bias was found to be induced by the kidney uptake. No such bias was found with the FBP based image reconstruction method. However, the OS-EM based image reconstruction was found to reduce variance in the subsequent phase of the ID-IF. The combined use of FBP and OS-EM resulted in reduced bias and noise. After performing all the necessary corrections, the areas under the curves (AUCs) of the AD-IF were close to that of the AD-IF (average AUC ratio =1 ± 0.08) during the early phase. When applied in a two-tissue-compartmental kinetic model, the average difference between the estimated model parameters from ID-IF and AD-IF was 10% which was within the error of the estimation method. Conclusions: The bias of radioligand concentration in the aorta from the OS-EM image reconstruction is significantly affected by radioligand uptake in the adjacent kidney and cannot be neglected for quantitative evaluation. With careful calibrations and corrections, the ID-IF derived from quantitative dynamic PET images can be used as the input function of the compartmental model to quantify the renal kinetics of {sup 11}C-KR31173 in experimental animals and the authors intend to evaluate this method in future human studies.« less

Quantitative Susceptibility Mapping of Human Brain Reflects Spatial Variation in Tissue Composition

PubMed Central

Li, Wei; Wu, Bing; Liu, Chunlei

2011-01-01

Image phase from gradient echo MRI provides a unique contrast that reflects brain tissue composition variations, such as iron and myelin distribution. Phase imaging is emerging as a powerful tool for the investigation of functional brain anatomy and disease diagnosis. However, the quantitative value of phase is compromised by its nonlocal and orientation dependent properties. There is an increasing need for reliable quantification of magnetic susceptibility, the intrinsic property of tissue. In this study, we developed a novel and accurate susceptibility mapping method that is also phase-wrap insensitive. The proposed susceptibility mapping method utilized two complementary equations: (1) the Fourier relationship of phase and magnetic susceptibility; and (2) the first-order partial derivative of the first equation in the spatial frequency domain. In numerical simulation, this method reconstructed the susceptibility map almost free of streaking artifact. Further, the iterative implementation of this method allowed for high quality reconstruction of susceptibility maps of human brain in vivo. The reconstructed susceptibility map provided excellent contrast of iron-rich deep nuclei and white matter bundles from surrounding tissues. Further, it also revealed anisotropic magnetic susceptibility in brain white matter. Hence, the proposed susceptibility mapping method may provide a powerful tool for the study of brain physiology and pathophysiology. Further elucidation of anisotropic magnetic susceptibility in vivo may allow us to gain more insight into the white matter microarchitectures. PMID:21224002

X-ray computed tomography of wood-adhesive bondlines: Attenuation and phase-contrast effects

DOE PAGES

Paris, Jesse L.; Kamke, Frederick A.; Xiao, Xianghui

2015-07-29

Microscale X-ray computed tomography (XCT) is discussed as a technique for identifying 3D adhesive distribution in wood-adhesive bondlines. Visualization and material segmentation of the adhesives from the surrounding cellular structures require sufficient gray-scale contrast in the reconstructed XCT data. Commercial wood-adhesive polymers have similar chemical characteristics and density to wood cell wall polymers and therefore do not provide good XCT attenuation contrast in their native form. Here, three different adhesive types, namely phenol formaldehyde, polymeric diphenylmethane diisocyanate, and a hybrid polyvinyl acetate, are tagged with iodine such that they yield sufficient X-ray attenuation contrast. However, phase-contrast effects at material edgesmore » complicate image quality and segmentation in XCT data reconstructed with conventional filtered backprojection absorption contrast algorithms. A quantitative phase retrieval algorithm, which isolates and removes the phase-contrast effect, was demonstrated. The paper discusses and illustrates the balance between material X-ray attenuation and phase-contrast effects in all quantitative XCT analyses of wood-adhesive bondlines.« less

X-ray computed tomography of wood-adhesive bondlines: Attenuation and phase-contrast effects

DOE Office of Scientific and Technical Information (OSTI.GOV)

Paris, Jesse L.; Kamke, Frederick A.; Xiao, Xianghui

Microscale X-ray computed tomography (XCT) is discussed as a technique for identifying 3D adhesive distribution in wood-adhesive bondlines. Visualization and material segmentation of the adhesives from the surrounding cellular structures require sufficient gray-scale contrast in the reconstructed XCT data. Commercial wood-adhesive polymers have similar chemical characteristics and density to wood cell wall polymers and therefore do not provide good XCT attenuation contrast in their native form. Here, three different adhesive types, namely phenol formaldehyde, polymeric diphenylmethane diisocyanate, and a hybrid polyvinyl acetate, are tagged with iodine such that they yield sufficient X-ray attenuation contrast. However, phase-contrast effects at material edgesmore » complicate image quality and segmentation in XCT data reconstructed with conventional filtered backprojection absorption contrast algorithms. A quantitative phase retrieval algorithm, which isolates and removes the phase-contrast effect, was demonstrated. The paper discusses and illustrates the balance between material X-ray attenuation and phase-contrast effects in all quantitative XCT analyses of wood-adhesive bondlines.« less

Echo-Planar Imaging-Based, J-Resolved Spectroscopic Imaging for Improved Metabolite Detection in Prostate Cancer

DTIC Science & Technology

2016-12-01

tiple dimensions (20). Hu et al. employed pseudo-random phase-encoding blips during the EPSI readout to create nonuniform sampling along the spatial...resolved MRSI with Nonuniform Undersampling and Compressed Sensing 514 30.5 Prior-knowledge Fitting for Metabolite Quantitation 515 30.6 Future Directions... NONUNIFORM UNDERSAMPLING AND COMPRESSED SENSING Nonuniform undersampling (NUS) of k-space and subsequent reconstruction using compressed sensing (CS

Quantitative 3D investigation of Neuronal network in mouse spinal cord model

NASA Astrophysics Data System (ADS)

Bukreeva, I.; Campi, G.; Fratini, M.; Spanò, R.; Bucci, D.; Battaglia, G.; Giove, F.; Bravin, A.; Uccelli, A.; Venturi, C.; Mastrogiacomo, M.; Cedola, A.

2017-01-01

The investigation of the neuronal network in mouse spinal cord models represents the basis for the research on neurodegenerative diseases. In this framework, the quantitative analysis of the single elements in different districts is a crucial task. However, conventional 3D imaging techniques do not have enough spatial resolution and contrast to allow for a quantitative investigation of the neuronal network. Exploiting the high coherence and the high flux of synchrotron sources, X-ray Phase-Contrast multiscale-Tomography allows for the 3D investigation of the neuronal microanatomy without any aggressive sample preparation or sectioning. We investigated healthy-mouse neuronal architecture by imaging the 3D distribution of the neuronal-network with a spatial resolution of 640 nm. The high quality of the obtained images enables a quantitative study of the neuronal structure on a subject-by-subject basis. We developed and applied a spatial statistical analysis on the motor neurons to obtain quantitative information on their 3D arrangement in the healthy-mice spinal cord. Then, we compared the obtained results with a mouse model of multiple sclerosis. Our approach paves the way to the creation of a “database” for the characterization of the neuronal network main features for a comparative investigation of neurodegenerative diseases and therapies.

Deep Learning Automates the Quantitative Analysis of Individual Cells in Live-Cell Imaging Experiments.

PubMed

Van Valen, David A; Kudo, Takamasa; Lane, Keara M; Macklin, Derek N; Quach, Nicolas T; DeFelice, Mialy M; Maayan, Inbal; Tanouchi, Yu; Ashley, Euan A; Covert, Markus W

2016-11-01

Live-cell imaging has opened an exciting window into the role cellular heterogeneity plays in dynamic, living systems. A major critical challenge for this class of experiments is the problem of image segmentation, or determining which parts of a microscope image correspond to which individual cells. Current approaches require many hours of manual curation and depend on approaches that are difficult to share between labs. They are also unable to robustly segment the cytoplasms of mammalian cells. Here, we show that deep convolutional neural networks, a supervised machine learning method, can solve this challenge for multiple cell types across the domains of life. We demonstrate that this approach can robustly segment fluorescent images of cell nuclei as well as phase images of the cytoplasms of individual bacterial and mammalian cells from phase contrast images without the need for a fluorescent cytoplasmic marker. These networks also enable the simultaneous segmentation and identification of different mammalian cell types grown in co-culture. A quantitative comparison with prior methods demonstrates that convolutional neural networks have improved accuracy and lead to a significant reduction in curation time. We relay our experience in designing and optimizing deep convolutional neural networks for this task and outline several design rules that we found led to robust performance. We conclude that deep convolutional neural networks are an accurate method that require less curation time, are generalizable to a multiplicity of cell types, from bacteria to mammalian cells, and expand live-cell imaging capabilities to include multi-cell type systems.

Deep Learning Automates the Quantitative Analysis of Individual Cells in Live-Cell Imaging Experiments

DOE PAGES

Van Valen, David A.; Kudo, Takamasa; Lane, Keara M.; ...

2016-11-04

Live-cell imaging has opened an exciting window into the role cellular heterogeneity plays in dynamic, living systems. A major critical challenge for this class of experiments is the problem of image segmentation, or determining which parts of a microscope image correspond to which individual cells. Current approaches require many hours of manual curation and depend on approaches that are difficult to share between labs. They are also unable to robustly segment the cytoplasms of mammalian cells. Here, we show that deep convolutional neural networks, a supervised machine learning method, can solve this challenge for multiple cell types across the domainsmore » of life. We demonstrate that this approach can robustly segment fluorescent images of cell nuclei as well as phase images of the cytoplasms of individual bacterial and mammalian cells from phase contrast images without the need for a fluorescent cytoplasmic marker. These networks also enable the simultaneous segmentation and identification of different mammalian cell types grown in co-culture. A quantitative comparison with prior methods demonstrates that convolutional neural networks have improved accuracy and lead to a significant reduction in curation time. We relay our experience in designing and optimizing deep convolutional neural networks for this task and outline several design rules that we found led to robust performance. We conclude that deep convolutional neural networks are an accurate method that require less curation time, are generalizable to a multiplicity of cell types, from bacteria to mammalian cells, and expand live-cell imaging capabilities to include multi-cell type systems.« less

Deep Learning Automates the Quantitative Analysis of Individual Cells in Live-Cell Imaging Experiments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Van Valen, David A.; Kudo, Takamasa; Lane, Keara M.

Live-cell imaging has opened an exciting window into the role cellular heterogeneity plays in dynamic, living systems. A major critical challenge for this class of experiments is the problem of image segmentation, or determining which parts of a microscope image correspond to which individual cells. Current approaches require many hours of manual curation and depend on approaches that are difficult to share between labs. They are also unable to robustly segment the cytoplasms of mammalian cells. Here, we show that deep convolutional neural networks, a supervised machine learning method, can solve this challenge for multiple cell types across the domainsmore » of life. We demonstrate that this approach can robustly segment fluorescent images of cell nuclei as well as phase images of the cytoplasms of individual bacterial and mammalian cells from phase contrast images without the need for a fluorescent cytoplasmic marker. These networks also enable the simultaneous segmentation and identification of different mammalian cell types grown in co-culture. A quantitative comparison with prior methods demonstrates that convolutional neural networks have improved accuracy and lead to a significant reduction in curation time. We relay our experience in designing and optimizing deep convolutional neural networks for this task and outline several design rules that we found led to robust performance. We conclude that deep convolutional neural networks are an accurate method that require less curation time, are generalizable to a multiplicity of cell types, from bacteria to mammalian cells, and expand live-cell imaging capabilities to include multi-cell type systems.« less

Deep Learning Automates the Quantitative Analysis of Individual Cells in Live-Cell Imaging Experiments

PubMed Central

Van Valen, David A.; Lane, Keara M.; Quach, Nicolas T.; Maayan, Inbal

2016-01-01

Live-cell imaging has opened an exciting window into the role cellular heterogeneity plays in dynamic, living systems. A major critical challenge for this class of experiments is the problem of image segmentation, or determining which parts of a microscope image correspond to which individual cells. Current approaches require many hours of manual curation and depend on approaches that are difficult to share between labs. They are also unable to robustly segment the cytoplasms of mammalian cells. Here, we show that deep convolutional neural networks, a supervised machine learning method, can solve this challenge for multiple cell types across the domains of life. We demonstrate that this approach can robustly segment fluorescent images of cell nuclei as well as phase images of the cytoplasms of individual bacterial and mammalian cells from phase contrast images without the need for a fluorescent cytoplasmic marker. These networks also enable the simultaneous segmentation and identification of different mammalian cell types grown in co-culture. A quantitative comparison with prior methods demonstrates that convolutional neural networks have improved accuracy and lead to a significant reduction in curation time. We relay our experience in designing and optimizing deep convolutional neural networks for this task and outline several design rules that we found led to robust performance. We conclude that deep convolutional neural networks are an accurate method that require less curation time, are generalizable to a multiplicity of cell types, from bacteria to mammalian cells, and expand live-cell imaging capabilities to include multi-cell type systems. PMID:27814364

Velocity Measurement in Carotid Artery: Quantitative Comparison of Time-Resolved 3D Phase-Contrast MRI and Image-based Computational Fluid Dynamics

PubMed Central

Sarrami-Foroushani, Ali; Nasr Esfahany, Mohsen; Nasiraei Moghaddam, Abbas; Saligheh Rad, Hamidreza; Firouznia, Kavous; Shakiba, Madjid; Ghanaati, Hossein; Wilkinson, Iain David; Frangi, Alejandro Federico

2015-01-01

Background: Understanding hemodynamic environment in vessels is important for realizing the mechanisms leading to vascular pathologies. Objectives: Three-dimensional velocity vector field in carotid bifurcation is visualized using TR 3D phase-contrast magnetic resonance imaging (TR 3D PC MRI) and computational fluid dynamics (CFD). This study aimed to present a qualitative and quantitative comparison of the velocity vector field obtained by each technique. Subjects and Methods: MR imaging was performed on a 30-year old male normal subject. TR 3D PC MRI was performed on a 3 T scanner to measure velocity in carotid bifurcation. 3D anatomical model for CFD was created using images obtained from time-of-flight MR angiography. Velocity vector field in carotid bifurcation was predicted using CFD and PC MRI techniques. A statistical analysis was performed to assess the agreement between the two methods. Results: Although the main flow patterns were the same for the both techniques, CFD showed a greater resolution in mapping the secondary and circulating flows. Overall root mean square (RMS) errors for all the corresponding data points in PC MRI and CFD were 14.27% in peak systole and 12.91% in end diastole relative to maximum velocity measured at each cardiac phase. Bland-Altman plots showed a very good agreement between the two techniques. However, this study was not aimed to validate any of methods, instead, the consistency was assessed to accentuate the similarities and differences between Time-resolved PC MRI and CFD. Conclusion: Both techniques provided quantitatively consistent results of in vivo velocity vector fields in right internal carotid artery (RCA). PC MRI represented a good estimation of main flow patterns inside the vasculature, which seems to be acceptable for clinical use. However, limitations of each technique should be considered while interpreting results. PMID:26793288

Technical Note: Synchrotron-based high-energy x-ray phase sensitive microtomography for biomedical research

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Huiqiang; Wu, Xizeng, E-mail: xwu@uabmc.edu, E-mail: tqxiao@sinap.ac.cn; Xiao, Tiqiao, E-mail: xwu@uabmc.edu, E-mail: tqxiao@sinap.ac.cn

Purpose: Propagation-based phase-contrast CT (PPCT) utilizes highly sensitive phase-contrast technology applied to x-ray microtomography. Performing phase retrieval on the acquired angular projections can enhance image contrast and enable quantitative imaging. In this work, the authors demonstrate the validity and advantages of a novel technique for high-resolution PPCT by using the generalized phase-attenuation duality (PAD) method of phase retrieval. Methods: A high-resolution angular projection data set of a fish head specimen was acquired with a monochromatic 60-keV x-ray beam. In one approach, the projection data were directly used for tomographic reconstruction. In two other approaches, the projection data were preprocessed bymore » phase retrieval based on either the linearized PAD method or the generalized PAD method. The reconstructed images from all three approaches were then compared in terms of tissue contrast-to-noise ratio and spatial resolution. Results: The authors’ experimental results demonstrated the validity of the PPCT technique based on the generalized PAD-based method. In addition, the results show that the authors’ technique is superior to the direct PPCT technique as well as the linearized PAD-based PPCT technique in terms of their relative capabilities for tissue discrimination and characterization. Conclusions: This novel PPCT technique demonstrates great potential for biomedical imaging, especially for applications that require high spatial resolution and limited radiation exposure.« less

Photothermal technique in cell microscopy studies

NASA Astrophysics Data System (ADS)

Lapotko, Dmitry; Chebot'ko, Igor; Kutchinsky, Georgy; Cherenkevitch, Sergey

1995-01-01

Photothermal (PT) method is applied for a cell imaging and quantitative studies. The techniques for cell monitoring, imaging and cell viability test are developed. The method and experimental set up for optical and PT-image acquisition and analysis is described. Dual- pulsed laser set up combined with phase contrast illumination of a sample provides visualization of temperature field or absorption structure of a sample with spatial resolution 0.5 micrometers . The experimental optics, hardware and software are designed using the modular principle, so the whole set up can be adjusted for various experiments: PT-response monitoring or photothermal spectroscopy studies. Sensitivity of PT-method provides the imaging of the structural elements of live (non-stained) white blood cells. The results of experiments with normal and subnormal blood cells (red blood cells, lymphocytes, neutrophyles and lymphoblasts) are reported. Obtained PT-images are different from optical analogs and deliver additional information about cell structure. The quantitative analysis of images was used for cell population comparative diagnostic. The viability test for red blood cell differentiation is described. During the study of neutrophyles in norma and sarcoidosis disease the differences in PT-images of cells were found.

SU-E-I-91: Quantitative Assessment of Early Hepatocellular Carcinoma and Cavernous Hemangioma of Live Using In-Line Phase-Contrast X-Ray Imaging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Duan, J

Purpose: To investigate the potential utility of in-line phase-contrast imaging (ILPCI) technique with synchrotron radiation in detecting early hepatocellular carcinoma and cavernous hemangioma of live using in vitro model system. Methods: Without contrast agents, three typical early hepatocellular carcinoma specimens and three typical cavernous hemangioma of live specimens were imaged using ILPCI. To quantitatively discriminate early hepatocellular carcinoma tissues and cavernous hemangioma tissues, the projection images texture feature based on gray level co-occurrence matrix (GLCM) were extracted. The texture parameters of energy, inertia, entropy, correlation, sum average, sum entropy, difference average, difference entropy and inverse difference moment, were obtained respectively.more » Results: In the ILPCI planar images of early hepatocellular carcinoma specimens, vessel trees were clearly visualized on the micrometer scale. Obvious distortion deformation was presented, and the vessel mostly appeared as a ‘dry stick’. Liver textures appeared not regularly. In the ILPCI planar images of cavernous hemangioma of live specimens, typical vessels had not been found compared with the early hepatocellular carcinoma planar images. The planar images of cavernous hemangioma of live specimens clearly displayed the dilated hepatic sinusoids with the diameter of less than 100 microns, but all of them were overlapped with each other. The texture parameters of energy, inertia, entropy, correlation, sum average, sum entropy, and difference average, showed a statistically significant between the two types specimens image (P<0.01), except the texture parameters of difference entropy and inverse difference moment(P>0.01). Conclusion: The results indicate that there are obvious changes in morphological levels including vessel structures and liver textures. The study proves that this imaging technique has a potential value in evaluating early hepatocellular carcinoma and cavernous hemangioma of live.« less

Quantitative oxygen extraction fraction from 7-Tesla MRI phase: reproducibility and application in multiple sclerosis.

PubMed

Fan, Audrey P; Govindarajan, Sindhuja T; Kinkel, R Philip; Madigan, Nancy K; Nielsen, A Scott; Benner, Thomas; Tinelli, Emanuele; Rosen, Bruce R; Adalsteinsson, Elfar; Mainero, Caterina

2015-01-01

Quantitative oxygen extraction fraction (OEF) in cortical veins was studied in patients with multiple sclerosis (MS) and healthy subjects via magnetic resonance imaging (MRI) phase images at 7 Tesla (7 T). Flow-compensated, three-dimensional gradient-echo scans were acquired for absolute OEF quantification in 23 patients with MS and 14 age-matched controls. In patients, we collected T2*-weighted images for characterization of white matter, deep gray matter, and cortical lesions, and also assessed cognitive function. Variability of OEF across readers and scan sessions was evaluated in a subset of volunteers. OEF was averaged from 2 to 3 pial veins in the sensorimotor, parietal, and prefrontal cortical regions for each subject (total of ~10 vessels). We observed good reproducibility of mean OEF, with intraobserver coefficient of variation (COV)=2.1%, interobserver COV=5.2%, and scan-rescan COV=5.9%. Patients exhibited a 3.4% reduction in cortical OEF relative to controls (P=0.0025), which was not different across brain regions. Although oxygenation did not relate with measures of structural tissue damage, mean OEF correlated with a global measure of information processing speed. These findings suggest that cortical OEF from 7-T MRI phase is a reproducible metabolic biomarker that may be sensitive to different pathologic processes than structural MRI in patients with MS.

X-ray Phase Contrast Imaging of Calcified Tissue and Biomaterial Structure in Bioreactor Engineered Tissues

DOE Office of Scientific and Technical Information (OSTI.GOV)

Appel, Alyssa A.; Larson, Jeffery C.; Garson, III, Alfred B.

2014-11-04

Tissues engineered in bioreactor systems have been used clinically to replace damaged tissues and organs. In addition, these systems are under continued development for many tissue engineering applications. The ability to quantitatively assess material structure and tissue formation is critical for evaluating bioreactor efficacy and for preimplantation assessment of tissue quality. These techniques allow for the nondestructive and longitudinal monitoring of large engineered tissues within the bioreactor systems and will be essential for the translation of these strategies to viable clinical therapies. X-ray Phase Contrast (XPC) imaging techniques have shown tremendous promise for a number of biomedical applications owing tomore » their ability to provide image contrast based on multiple X-ray properties, including absorption, refraction, and scatter. In this research, mesenchymal stem cell-seeded alginate hydrogels were prepared and cultured under osteogenic conditions in a perfusion bioreactor. The constructs were imaged at various time points using XPC microcomputed tomography (µCT). Imaging was performed with systems using both synchrotron- and tube-based X-ray sources. XPC µCT allowed for simultaneous three-dimensional (3D) quantification of hydrogel size and mineralization, as well as spatial information on hydrogel structure and mineralization. Samples were processed for histological evaluation and XPC showed similar features to histology and quantitative analysis consistent with the histomorphometry. Furthermore, these results provide evidence of the significant potential of techniques based on XPC for noninvasive 3D imaging engineered tissues grown in bioreactors.« less

Nuclear medicine and quantitative imaging research (instrumentation and quantitative methods of evaluation)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Beck, R.N.; Cooper, M.D.

1990-09-01

This report summarizes goals and accomplishments of the research program supported under DOE Grant No. FG02-86ER60418 entitled Instrumentation and Quantitative Methods of Evaluation, with R. Beck, P. I. and M. Cooper, Co-P.I. during the period January 15, 1990 through September 1, 1990. This program addresses the problems involving the basic science and technology underlying the physical and conceptual tools of radioactive tracer methodology as they relate to the measurement of structural and functional parameters of physiologic importance in health and disease. The principal tool is quantitative radionuclide imaging. The overall objective of this program is to further the development andmore » transfer of radiotracer methodology from basic theory to routine clinical practice in order that individual patients and society as a whole will receive the maximum net benefit from the new knowledge gained. The focus of the research is on the development of new instruments and radiopharmaceuticals, and the evaluation of these through the phase of clinical feasibility. 7 figs.« less

Comparison of different numerical treatments for x-ray phase tomography of soft tissue from differential phase projections

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pelliccia, Daniele; Vaz, Raquel; Svalbe, Imants

X-ray imaging of soft tissue is made difficult by their low absorbance. The use of x-ray phase imaging and tomography can significantly enhance the detection of these tissues and several approaches have been proposed to this end. Methods such as analyzer-based imaging or grating interferometry produce differential phase projections that can be used to reconstruct the 3D distribution of the sample refractive index. We report on the quantitative comparison of three different methods to obtain x-ray phase tomography with filtered back-projection from differential phase projections in the presence of noise. The three procedures represent different numerical approaches to solve themore » same mathematical problem, namely phase retrieval and filtered back-projection. It is found that obtaining individual phase projections and subsequently applying a conventional filtered back-projection algorithm produces the best results for noisy experimental data, when compared with other procedures based on the Hilbert transform. The algorithms are tested on simulated phantom data with added noise and the predictions are confirmed by experimental data acquired using a grating interferometer. The experiment is performed on unstained adult zebrafish, an important model organism for biomedical studies. The method optimization described here allows resolution of weak soft tissue features, such as muscle fibers.« less

Heat release and flame structure measurements of self-excited acoustically-driven premixed methane flames

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kopp-Vaughan, Kristin M.; Tuttle, Steven G.; Renfro, Michael W.

An open-open organ pipe burner (Rijke tube) with a bluff-body ring was used to create a self-excited, acoustically-driven, premixed methane-air conical flame, with equivalence ratios ranging from 0.85 to 1.05. The feed tube velocities corresponded to Re = 1780-4450. Coupled oscillations in pressure, velocity, and heat release from the flame are naturally encouraged at resonant frequencies in the Rijke tube combustor. This coupling creates sustainable self-excited oscillations in flame front area and shape. The period of the oscillations occur at the resonant frequency of the combustion chamber when the flame is placed {proportional_to}1/4 of the distance from the bottom ofmore » the tube. In this investigation, the shape of these acoustically-driven flames is measured by employing both OH planar laser-induced fluorescence (PLIF) and chemiluminescence imaging and the images are correlated to simultaneously measured pressure in the combustor. Past research on acoustically perturbed flames has focused on qualitative flame area and heat release relationships under imposed velocity perturbations at imposed frequencies. This study reports quantitative empirical fits with respect to pressure or phase angle in a self-generated pressure oscillation. The OH-PLIF images were single temporal shots and the chemiluminescence images were phase averaged on chip, such that 15 exposures were used to create one image. Thus, both measurements were time resolved during the flame oscillation. Phase-resolved area and heat release variations throughout the pressure oscillation were computed. A relation between flame area and the phase angle before the pressure maximum was derived for all flames in order to quantitatively show that the Rayleigh criterion was satisfied in the combustor. Qualitative trends in oscillating flame area were found with respect to feed tube flow rates. A logarithmic relation was found between the RMS pressure and both the normalized average area and heat release rate for all flames. (author)« less

Phase-sensitive dual-inversion recovery for accelerated carotid vessel wall imaging.

PubMed

Bonanno, Gabriele; Brotman, David; Stuber, Matthias

2015-03-01

Dual-inversion recovery (DIR) is widely used for magnetic resonance vessel wall imaging. However, optimal contrast may be difficult to obtain and is subject to RR variability. Furthermore, DIR imaging is time-inefficient and multislice acquisitions may lead to prolonged scanning times. Therefore, an extension of phase-sensitive (PS) DIR is proposed for carotid vessel wall imaging. The statistical distribution of the phase signal after DIR is probed to segment carotid lumens and suppress their residual blood signal. The proposed PS-DIR technique was characterized over a broad range of inversion times. Multislice imaging was then implemented by interleaving the acquisition of 3 slices after DIR. Quantitative evaluation was then performed in healthy adult subjects and compared with conventional DIR imaging. Single-slice PS-DIR provided effective blood-signal suppression over a wide range of inversion times, enhancing wall-lumen contrast and vessel wall conspicuity for carotid arteries. Multislice PS-DIR imaging with effective blood-signal suppression is enabled. A variant of the PS-DIR method has successfully been implemented and tested for carotid vessel wall imaging. This technique removes timing constraints related to inversion recovery, enhances wall-lumen contrast, and enables a 3-fold increase in volumetric coverage at no extra cost in scanning time.

2D and 3D X-ray phase retrieval of multi-material objects using a single defocus distance.

PubMed

Beltran, M A; Paganin, D M; Uesugi, K; Kitchen, M J

2010-03-29

A method of tomographic phase retrieval is developed for multi-material objects whose components each has a distinct complex refractive index. The phase-retrieval algorithm, based on the Transport-of-Intensity equation, utilizes propagation-based X-ray phase contrast images acquired at a single defocus distance for each tomographic projection. The method requires a priori knowledge of the complex refractive index for each material present in the sample, together with the total projected thickness of the object at each orientation. The requirement of only a single defocus distance per projection simplifies the experimental setup and imposes no additional dose compared to conventional tomography. The algorithm was implemented using phase contrast data acquired at the SPring-8 Synchrotron facility in Japan. The three-dimensional (3D) complex refractive index distribution of a multi-material test object was quantitatively reconstructed using a single X-ray phase-contrast image per projection. The technique is robust in the presence of noise, compared to conventional absorption based tomography.

Visualisation and quantitative analysis of the rodent malaria liver stage by real time imaging.

PubMed

Ploemen, Ivo H J; Prudêncio, Miguel; Douradinha, Bruno G; Ramesar, Jai; Fonager, Jannik; van Gemert, Geert-Jan; Luty, Adrian J F; Hermsen, Cornelus C; Sauerwein, Robert W; Baptista, Fernanda G; Mota, Maria M; Waters, Andrew P; Que, Ivo; Lowik, Clemens W G M; Khan, Shahid M; Janse, Chris J; Franke-Fayard, Blandine M D

2009-11-18

The quantitative analysis of Plasmodium development in the liver in laboratory animals in cultured cells is hampered by low parasite infection rates and the complicated methods required to monitor intracellular development. As a consequence, this important phase of the parasite's life cycle has been poorly studied compared to blood stages, for example in screening anti-malarial drugs. Here we report the use of a transgenic P. berghei parasite, PbGFP-Luc(con), expressing the bioluminescent reporter protein luciferase to visualize and quantify parasite development in liver cells both in culture and in live mice using real-time luminescence imaging. The reporter-parasite based quantification in cultured hepatocytes by real-time imaging or using a microplate reader correlates very well with established quantitative RT-PCR methods. For the first time the liver stage of Plasmodium is visualized in whole bodies of live mice and we were able to discriminate as few as 1-5 infected hepatocytes per liver in mice using 2D-imaging and to identify individual infected hepatocytes by 3D-imaging. The analysis of liver infections by whole body imaging shows a good correlation with quantitative RT-PCR analysis of extracted livers. The luminescence-based analysis of the effects of various drugs on in vitro hepatocyte infection shows that this method can effectively be used for in vitro screening of compounds targeting Plasmodium liver stages. Furthermore, by analysing the effect of primaquine and tafenoquine in vivo we demonstrate the applicability of real time imaging to assess parasite drug sensitivity in the liver. The simplicity and speed of quantitative analysis of liver-stage development by real-time imaging compared to the PCR methodologies, as well as the possibility to analyse liver development in live mice without surgery, opens up new possibilities for research on Plasmodium liver infections and for validating the effect of drugs and vaccines on the liver stage of Plasmodium.

Visualisation and Quantitative Analysis of the Rodent Malaria Liver Stage by Real Time Imaging

PubMed Central

Douradinha, Bruno G.; Ramesar, Jai; Fonager, Jannik; van Gemert, Geert-Jan; Luty, Adrian J. F.; Hermsen, Cornelus C.; Sauerwein, Robert W.; Baptista, Fernanda G.; Mota, Maria M.; Waters, Andrew P.; Que, Ivo; Lowik, Clemens W. G. M.; Khan, Shahid M.; Janse, Chris J.; Franke-Fayard, Blandine M. D.

2009-01-01

The quantitative analysis of Plasmodium development in the liver in laboratory animals in cultured cells is hampered by low parasite infection rates and the complicated methods required to monitor intracellular development. As a consequence, this important phase of the parasite's life cycle has been poorly studied compared to blood stages, for example in screening anti-malarial drugs. Here we report the use of a transgenic P. berghei parasite, PbGFP-Luccon, expressing the bioluminescent reporter protein luciferase to visualize and quantify parasite development in liver cells both in culture and in live mice using real-time luminescence imaging. The reporter-parasite based quantification in cultured hepatocytes by real-time imaging or using a microplate reader correlates very well with established quantitative RT-PCR methods. For the first time the liver stage of Plasmodium is visualized in whole bodies of live mice and we were able to discriminate as few as 1–5 infected hepatocytes per liver in mice using 2D-imaging and to identify individual infected hepatocytes by 3D-imaging. The analysis of liver infections by whole body imaging shows a good correlation with quantitative RT-PCR analysis of extracted livers. The luminescence-based analysis of the effects of various drugs on in vitro hepatocyte infection shows that this method can effectively be used for in vitro screening of compounds targeting Plasmodium liver stages. Furthermore, by analysing the effect of primaquine and tafenoquine in vivo we demonstrate the applicability of real time imaging to assess parasite drug sensitivity in the liver. The simplicity and speed of quantitative analysis of liver-stage development by real-time imaging compared to the PCR methodologies, as well as the possibility to analyse liver development in live mice without surgery, opens up new possibilities for research on Plasmodium liver infections and for validating the effect of drugs and vaccines on the liver stage of Plasmodium. PMID:19924309

The study on the parallel processing based time series correlation analysis of RBC membrane flickering in quantitative phase imaging

NASA Astrophysics Data System (ADS)

Lee, Minsuk; Won, Youngjae; Park, Byungjun; Lee, Seungrag

2017-02-01

Not only static characteristics but also dynamic characteristics of the red blood cell (RBC) contains useful information for the blood diagnosis. Quantitative phase imaging (QPI) can capture sample images with subnanometer scale depth resolution and millisecond scale temporal resolution. Various researches have been used QPI for the RBC diagnosis, and recently many researches has been developed to decrease the process time of RBC information extraction using QPI by the parallel computing algorithm, however previous studies are interested in the static parameters such as morphology of the cells or simple dynamic parameters such as root mean square (RMS) of the membrane fluctuations. Previously, we presented a practical blood test method using the time series correlation analysis of RBC membrane flickering with QPI. However, this method has shown that there is a limit to the clinical application because of the long computation time. In this study, we present an accelerated time series correlation analysis of RBC membrane flickering using the parallel computing algorithm. This method showed consistent fractal scaling exponent results of the surrounding medium and the normal RBC with our previous research.

SU-E-J-215: Towards MR-Only Image Guided Identification of Calcifications and Brachytherapy Seeds: Application to Prostate and Breast LDR Implant Dosimetry

DOE Office of Scientific and Technical Information (OSTI.GOV)

Elzibak, A; Fatemi-Ardekani, A; Soliman, A

Purpose: To identify and analyze the appearance of calcifications and brachytherapy seeds on magnitude and phase MRI images and to investigate whether they can be distinguished from each other on corrected phase images for application to prostate and breast low dose rate (LDR) implant dosimetry. Methods: An agar-based gel phantom containing two LDR brachytherapy seeds (Advantage Pd-103, IsoAid, 0.8mm diameter, 4.5mm length) and two spherical calcifications (large: 7mm diameter and small: 4mm diameter) was constructed and imaged on a 3T Philips MR scanner using a 16-channel head coil and a susceptibility weighted imaging (SWI) sequence (2mm slices, 320mm FOV, TR/more » TE= 26.5/5.3ms, 15 degree flip angle). The phase images were unwrapped and corrected using a 32×32, 2D Hanning high pass filter to remove background phase noise. Appearance of the seeds and calcifications was assessed visually and quantitatively using Osirix (http://www.osirix-viewer.com/). Results: As expected, calcifications and brachytherapy seeds appeared dark (hypointense) relative to the surrounding gel on the magnitude MRI images. The diameter of each seed without the surrounding artifact was measured to be 0.1 cm on the magnitude image, while diameters of 0.79 and 0.37 cm were measured for the larger and smaller calcifications, respectively. On the corrected phase images, the brachytherapy seeds and the calcifications appeared bright (hyperintense). The diameter of the seeds was larger on the phase images (0.17 cm) likely due to the dipole effect. Conclusion: MRI has the best soft tissue contrast for accurate organ delineation leading to most accurate implant dosimetry. This work demonstrated that phase images can potentially be useful in identifying brachytherapy seeds and calcifications in the prostate and breast due to their bright appearance, which helps in their visualization and quantification for accurate dosimetry using MR-only. Future work includes optimizing phase filters to best identify and delineate seeds and calcifications.« less

High-brightness laser imaging with tunable speckle reduction enabled by electroactive micro-optic diffusers.

PubMed

Farrokhi, Hamid; Rohith, Thazhe Madam; Boonruangkan, Jeeranan; Han, Seunghwoi; Kim, Hyunwoong; Kim, Seung-Woo; Kim, Young-Jin

2017-11-10

High coherence of lasers is desirable in high-speed, high-resolution, and wide-field imaging. However, it also causes unavoidable background speckle noise thus degrades the image quality in traditional microscopy and more significantly in interferometric quantitative phase imaging (QPI). QPI utilizes optical interference for high-precision measurement of the optical properties where the speckle can severely distort the information. To overcome this, we demonstrated a light source system having a wide tunability in the spatial coherence over 43% by controlling the illumination angle, scatterer's size, and the rotational speed of an electroactive-polymer rotational micro-optic diffuser. Spatially random phase modulation was implemented for the lower speckle imaging with over a 50% speckle reduction without a significant degradation in the temporal coherence. Our coherence control technique will provide a unique solution for a low-speckle, full-field, and coherent imaging in optically scattering media in the fields of healthcare sciences, material sciences and high-precision engineering.

Visible/Infrared Optical Depths of Cirrus as Seen by Satellite and Scanning Lidar

NASA Technical Reports Server (NTRS)

Wylie, Donald; Wolf, Walt; Piironen, Paivi; Eloranta, Edwin

1996-01-01

The High Spectral Resolution Lidar (HSRL) and the Volume Imaging Lidar (VIL) were combined to produce a quantitative image of the visible optical depth of cirrus clouds. The HSRL was used to calibrate the VIL signal into backscatter cross sections of particulates. The backscatter cross sections were related to extinction by a constant backscatter phase function determined from the HSRL data. This produced a three dimensional image of visual extinction in the cirrus clouds over a one hour period. Two lidar images were constructed from one hour VIL cross section records.

[Clinical application of high-pitch excretory phase images during dual-source CT urography with stellar photon detector].

PubMed

Sun, Hao; Xue, Hua-dan; Jin, Zheng-yu; Wang, Xuan; Chen, Yu; He, Yong-lan; Zhang, Da-ming; Zhu, Liang; Wang, Yun; Qi, Bing; Xu, Kai; Wang, Ming

2014-10-01

To retrospectively evaluate the clinical feasibility of high-pitch excretory phase images during dual-source CT urography with Stellar photon detector. Totally 100 patients received dual-source CT high-pitch urinary excretory phase scanning with Stellar photon detector [80 kV, ref.92 mAs, CARE Dose 4D and CARE kV, pitch of 3.0, filter back projection reconstruction algorithm (FBP)] (group A). Another 100 patients received dual-source CT high-pitch urinary excretory phase scanning with common detector(100 kV, ref.140 mAs, CARE Dose 4D, pitch of 3.0, FBP) (group B). Quantitative measurement of CT value of urinary segments (Hounsfield units), image noise (Hounsfield units), and effective radiation dose (millisievert) were compared using independent-samples t test between two groups. Urinary system subjective opacification scores were compared using Mann-Whitney U test between two groups. There was no significant difference in subjective opacification score of intrarenal collecting system and ureters between two groups (all P>0.05). The group A images yielded significantly higher CT values of all urinary segments (all P<0.01). There was no significant difference in image noise (P>0.05). The effective radiation dose of group A (1.1 mSv) was significantly lower than that of group B (3.79 mSv) (P<0.01). High-pitch low-tube-voltage during excretory phase dual-source CT urography with Stellar photon detector is feasible, with acceptable image noise and lower radiation dose.

Dynamic speckle illumination wide-field reflection phase microscopy

PubMed Central

Choi, Youngwoon; Hosseini, Poorya; Choi, Wonshik; Dasari, Ramachandra R.; So, Peter T. C.; Yaqoob, Zahid

2014-01-01

We demonstrate a quantitative reflection-phase microscope based on time-varying speckle-field illumination. Due to the short spatial coherence length of the speckle field, the proposed imaging system features superior lateral resolution, 520 nm, as well as high-depth selectivity, 1.03 µm. Off-axis interferometric detection enables wide-field and single-shot imaging appropriate for high-speed measurements. In addition, the measured phase sensitivity of this method, which is the smallest measurable axial motion, is more than 40 times higher than that available using a transmission system. We demonstrate the utility of our method by successfully distinguishing the motion of the top surface from that of the bottom in red blood cells. The proposed method will be useful for studying membrane dynamics in complex eukaryotic cells. PMID:25361156

Application values of 99mTc-methoxyisobutylisonitrile imaging for differentiating benign and malignant thymic masses.

PubMed

Lu, Chenghui; Wang, Xufu; Liu, Bin; Liu, Xinfeng; Wang, Guoming; Zhang, Qin

2017-08-01

The aim of the present study was to investigate the application value of 99m Tc-methoxyisobutylisonitrile (MIBI) imaging to differentiate between benign and malignant thymic masses. A total of 32 patients with space-occupying mediastinal masses were enrolled and early and delayed-phase images were collected following injection with the imaging agent. The tumor to background ratio (T/N) values at the different phases were also recorded. The sensitivity of the qualitative analysis to distinguish between benign and malignant thymic masses was 95.24%, with specificity as 90.91%. The T/N values in the early and delayed phases were not significantly different in the group with benign thymic masses, but demonstrated statistical significant differences in the groups with low- and intermediate-grade malignant thymic masses. The T/N values at the above early and delayed phase were significantly different between the benign and low-grade malignancy groups, as well as between low- and moderate-grade malignancy groups. Those between the benign and moderate-grade malignancy groups demonstrated no significant difference. 99m Tc-MIBI imaging was able to differentiate between benign and malignant thymic masses, and the simultaneous semi-quantitative analysis of the T/N values of the tumors may be able to initially determine the degree of malignancy of thymoma.

Patient-specific coronary blood supply territories for quantitative perfusion analysis

PubMed Central

Zakkaroff, Constantine; Biglands, John D.; Greenwood, John P.; Plein, Sven; Boyle, Roger D.; Radjenovic, Aleksandra; Magee, Derek R.

2018-01-01

Abstract Myocardial perfusion imaging, coupled with quantitative perfusion analysis, provides an important diagnostic tool for the identification of ischaemic heart disease caused by coronary stenoses. The accurate mapping between coronary anatomy and under-perfused areas of the myocardium is important for diagnosis and treatment. However, in the absence of the actual coronary anatomy during the reporting of perfusion images, areas of ischaemia are allocated to a coronary territory based on a population-derived 17-segment (American Heart Association) AHA model of coronary blood supply. This work presents a solution for the fusion of 2D Magnetic Resonance (MR) myocardial perfusion images and 3D MR angiography data with the aim to improve the detection of ischaemic heart disease. The key contribution of this work is a novel method for the mediated spatiotemporal registration of perfusion and angiography data and a novel method for the calculation of patient-specific coronary supply territories. The registration method uses 4D cardiac MR cine series spanning the complete cardiac cycle in order to overcome the under-constrained nature of non-rigid slice-to-volume perfusion-to-angiography registration. This is achieved by separating out the deformable registration problem and solving it through phase-to-phase registration of the cine series. The use of patient-specific blood supply territories in quantitative perfusion analysis (instead of the population-based model of coronary blood supply) has the potential of increasing the accuracy of perfusion analysis. Quantitative perfusion analysis diagnostic accuracy evaluation with patient-specific territories against the AHA model demonstrates the value of the mediated spatiotemporal registration in the context of ischaemic heart disease diagnosis. PMID:29392098

Differential phase optical coherence probe for depth-resolved detection of photothermal response in tissue.

PubMed

Telenkov, Sergey A; Dave, Digant P; Sethuraman, Shriram; Akkin, Taner; Milner, Thomas E

2004-01-07

We describe a differential phase low-coherence interferometric probe for non-invasive, quantitative imaging of photothermal phenomena in biological materials. Our detection method utilizes principles of optical coherence tomography with differential phase measurement of interference fringe signals. A dual-channel optical low-coherence probe is used to analyse laser-induced thermoelastic and thermorefractive effects in tissue with micrometre axial resolution and nanometre sensitivity. We demonstrate an application of the technique using tissue phantoms and ex-vivo tissue specimens of rodent dorsal skin.

Blue intensity matters for cell cycle profiling in fluorescence DAPI-stained images.

PubMed

Ferro, Anabela; Mestre, Tânia; Carneiro, Patrícia; Sahumbaiev, Ivan; Seruca, Raquel; Sanches, João M

2017-05-01

In the past decades, there has been an amazing progress in the understanding of the molecular mechanisms of the cell cycle. This has been possible largely due to a better conceptualization of the cycle itself, but also as a consequence of technological advances. Herein, we propose a new fluorescence image-based framework targeted at the identification and segmentation of stained nuclei with the purpose to determine DNA content in distinct cell cycle stages. The method is based on discriminative features, such as total intensity and area, retrieved from in situ stained nuclei by fluorescence microscopy, allowing the determination of the cell cycle phase of both single and sub-population of cells. The analysis framework was built on a modified k-means clustering strategy and refined with a Gaussian mixture model classifier, which enabled the definition of highly accurate classification clusters corresponding to G1, S and G2 phases. Using the information retrieved from area and fluorescence total intensity, the modified k-means (k=3) cluster imaging framework classified 64.7% of the imaged nuclei, as being at G1 phase, 12.0% at G2 phase and 23.2% at S phase. Performance of the imaging framework was ascertained with normal murine mammary gland cells constitutively expressing the Fucci2 technology, exhibiting an overall sensitivity of 94.0%. Further, the results indicate that the imaging framework has a robust capacity to both identify a given DAPI-stained nucleus to its correct cell cycle phase, as well as to determine, with very high probability, true negatives. Importantly, this novel imaging approach is a non-disruptive method that allows an integrative and simultaneous quantitative analysis of molecular and morphological parameters, thus awarding the possibility of cell cycle profiling in cytological and histological samples.

MRI technique for the snapshot imaging of quantitative velocity maps using RARE

NASA Astrophysics Data System (ADS)

Shiko, G.; Sederman, A. J.; Gladden, L. F.

2012-03-01

A quantitative PGSE-RARE pulse sequence was developed and successfully applied to the in situ dissolution of two pharmaceutical formulations dissolving over a range of timescales. The new technique was chosen over other existing fast velocity imaging techniques because it is T2 weighted, not T2∗ weighted, and is, therefore, robust for imaging time-varying interfaces and flow in magnetically heterogeneous systems. The complex signal was preserved intact by separating odd and even echoes to obtain two phase maps which are then averaged in post-processing. Initially, the validity of the technique was shown when imaging laminar flow in a pipe. Subsequently, the dissolution of two drugs was followed in situ, where the technique enables the imaging and quantification of changes in the form of the tablet and the flow field surrounding it at high spatial and temporal resolution. First, the complete 3D velocity field around an eroding salicylic acid tablet was acquired at a resolution of 98 × 49 μm2, within 20 min, and monitored over ˜13 h. The tablet was observed to experience a heterogeneous flow field and, hence a heterogeneous shear field, which resulted in the non-symmetric erosion of the tablet. Second, the dissolution of a fast dissolving immediate release tablet was followed using one-shot 2D velocity images acquired every 5.2 s at a resolution of 390 × 390 μm2. The quantitative nature of the technique and fast acquisition times provided invaluable information on the dissolution behaviour of this tablet, which had not been attainable previously with conventional quantitative MRI techniques.

X-ray propagation microscopy of biological cells using waveguides as a quasipoint source

DOE Office of Scientific and Technical Information (OSTI.GOV)

Giewekemeyer, K.; Krueger, S. P.; Kalbfleisch, S.

2011-02-15

We have used x-ray waveguides as highly confining optical elements for nanoscale imaging of unstained biological cells using the simple geometry of in-line holography. The well-known twin-image problem is effectively circumvented by a simple and fast iterative reconstruction. The algorithm which combines elements of the classical Gerchberg-Saxton scheme and the hybrid-input-output algorithm is optimized for phase-contrast samples, well-justified for imaging of cells at multi-keV photon energies. The experimental scheme allows for a quantitative phase reconstruction from a single holographic image without detailed knowledge of the complex illumination function incident on the sample, as demonstrated for freeze-dried cells of the eukaryoticmore » amoeba Dictyostelium discoideum. The accessible resolution range is explored by simulations, indicating that resolutions on the order of 20 nm are within reach applying illumination times on the order of minutes at present synchrotron sources.« less

Multicomponent chemical imaging of pharmaceutical solid dosage forms with broadband CARS microscopy.

PubMed

Hartshorn, Christopher M; Lee, Young Jong; Camp, Charles H; Liu, Zhen; Heddleston, John; Canfield, Nicole; Rhodes, Timothy A; Hight Walker, Angela R; Marsac, Patrick J; Cicerone, Marcus T

2013-09-03

We compare a coherent Raman imaging modality, broadband coherent anti-Stokes Raman scattering (BCARS) microscopy, with spontaneous Raman microscopy for quantitative and qualitative assessment of multicomponent pharmaceuticals. Indomethacin was used as a model active pharmaceutical ingredient (API) and was analyzed in a tabulated solid dosage form, embedded within commonly used excipients. In comparison with wide-field spontaneous Raman chemical imaging, BCARS acquired images 10× faster, at higher spatiochemical resolution and with spectra of much higher SNR, eliminating the need for multivariate methods to identify chemical components. The significant increase in spatiochemical resolution allowed identification of an unanticipated API phase that was missed by the spontaneous wide-field method and bulk Raman spectroscopy. We confirmed the presence of the unanticipated API phase using confocal spontaneous Raman, which provided spatiochemical resolution similar to BCARS but at 100× slower acquisition times.

SU-E-J-260: Quantitative Image Feature Analysis of Multiphase Liver CT for Hepatocellular Carcinoma (HCC) in Radiation Therapy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Choi, W; Wang, J; Lu, W

Purpose: To identify the effective quantitative image features (radiomics features) for prediction of response, survival, recurrence and metastasis of hepatocellular carcinoma (HCC) in radiotherapy. Methods: Multiphase contrast enhanced liver CT images were acquired in 16 patients with HCC on pre and post radiation therapy (RT). In this study, arterial phase CT images were selected to analyze the effectiveness of image features for the prediction of treatment outcome of HCC to RT. Response evaluated by RECIST criteria, survival, local recurrence (LR), distant metastasis (DM) and liver metastasis (LM) were examined. A radiation oncologist manually delineated the tumor and normal liver onmore » pre and post CT scans, respectively. Quantitative image features were extracted to characterize the intensity distribution (n=8), spatial patterns (texture, n=36), and shape (n=16) of the tumor and liver, respectively. Moreover, differences between pre and post image features were calculated (n=120). A total of 360 features were extracted and then analyzed by unpaired student’s t-test to rank the effectiveness of features for the prediction of response. Results: The five most effective features were selected for prediction of each outcome. Significant predictors for tumor response and survival are changes in tumor shape (Second Major Axes Length, p= 0.002; Eccentricity, p=0.0002), for LR, liver texture (Standard Deviation (SD) of High Grey Level Run Emphasis and SD of Entropy, both p=0.005) on pre and post CT images, for DM, tumor texture (SD of Entropy, p=0.01) on pre CT image and for LM, liver (Mean of Cluster Shade, p=0.004) and tumor texture (SD of Entropy, p=0.006) on pre CT image. Intensity distribution features were not significant (p>0.09). Conclusion: Quantitative CT image features were found to be potential predictors of the five endpoints of HCC in RT. This work was supported in part by the National Cancer Institute Grant R01CA172638.« less

Quantitative MRI of kidneys in renal disease.

PubMed

Kline, Timothy L; Edwards, Marie E; Garg, Ishan; Irazabal, Maria V; Korfiatis, Panagiotis; Harris, Peter C; King, Bernard F; Torres, Vicente E; Venkatesh, Sudhakar K; Erickson, Bradley J

2018-03-01

To evaluate the reproducibility and utility of quantitative magnetic resonance imaging (MRI) sequences for the assessment of kidneys in young adults with normal renal function (eGFR ranged from 90 to 130 mL/min/1.73 m 2 ) and patients with early renal disease (autosomal dominant polycystic kidney disease). This prospective case-control study was performed on ten normal young adults (18-30 years old) and ten age- and sex-matched patients with early renal parenchymal disease (autosomal dominant polycystic kidney disease). All subjects underwent a comprehensive kidney MRI protocol, including qualitative imaging: T1w, T2w, FIESTA, and quantitative imaging: 2D cine phase contrast of the renal arteries, and parenchymal diffusion weighted imaging (DWI), magnetization transfer imaging (MTI), blood oxygen level dependent (BOLD) imaging, and magnetic resonance elastography (MRE). The normal controls were imaged on two separate occasions ≥24 h apart (range 24-210 h) to assess reproducibility of the measurements. Quantitative MR imaging sequences were found to be reproducible. The mean ± SD absolute percent difference between quantitative parameters measured ≥24 h apart were: MTI-derived ratio = 4.5 ± 3.6%, DWI-derived apparent diffusion coefficient (ADC) = 6.5 ± 3.4%, BOLD-derived R2* = 7.4 ± 5.9%, and MRE-derived tissue stiffness = 7.6 ± 3.3%. Compared with controls, the ADPKD patient's non-cystic renal parenchyma (NCRP) had statistically significant differences with regard to quantitative parenchymal measures: lower MTI percent ratios (16.3 ± 4.4 vs. 23.8 ± 1.2, p < 0.05), higher ADCs (2.46 ± 0.20 vs. 2.18 ± 0.10 × 10 -3  mm 2 /s, p < 0.05), lower R2*s (14.9 ± 1.7 vs. 18.1 ± 1.6 s -1 , p < 0.05), and lower tissue stiffness (3.2 ± 0.3 vs. 3.8 ± 0.5 kPa, p < 0.05). Excellent reproducibility of the quantitative measurements was obtained in all cases. Significantly different quantitative MR parenchymal measurement parameters between ADPKD patients and normal controls were obtained by MT, DWI, BOLD, and MRE indicating the potential for detecting and following renal disease at an earlier stage than the conventional qualitative imaging techniques.

A new fringeline-tracking approach for color Doppler ultrasound imaging phase unwrapping

NASA Astrophysics Data System (ADS)

Saad, Ashraf A.; Shapiro, Linda G.

2008-03-01

Color Doppler ultrasound imaging is a powerful non-invasive diagnostic tool for many clinical applications that involve examining the anatomy and hemodynamics of human blood vessels. These clinical applications include cardio-vascular diseases, obstetrics, and abdominal diseases. Since its commercial introduction in the early eighties, color Doppler ultrasound imaging has been used mainly as a qualitative tool with very little attempts to quantify its images. Many imaging artifacts hinder the quantification of the color Doppler images, the most important of which is the aliasing artifact that distorts the blood flow velocities measured by the color Doppler technique. In this work we will address the color Doppler aliasing problem and present a recovery methodology for the true flow velocities from the aliased ones. The problem is formulated as a 2D phase-unwrapping problem, which is a well-defined problem with solid theoretical foundations for other imaging domains, including synthetic aperture radar and magnetic resonance imaging. This paper documents the need for a phase unwrapping algorithm for use in color Doppler ultrasound image analysis. It describes a new phase-unwrapping algorithm that relies on the recently developed cutline detection approaches. The algorithm is novel in its use of heuristic information provided by the ultrasound imaging modality to guide the phase unwrapping process. Experiments have been performed on both in-vitro flow-phantom data and in-vivo human blood flow data. Both data types were acquired under a controlled acquisition protocol developed to minimize the distortion of the color Doppler data and hence to simplify the phase-unwrapping task. In addition to the qualitative assessment of the results, a quantitative assessment approach was developed to measure the success of the results. The results of our new algorithm have been compared on ultrasound data to those from other well-known algorithms, and it outperforms all of them.

Resolving High Amplitude Surface Motion with Diffusing Light

NASA Technical Reports Server (NTRS)

Wright, W.; Budakian, R.; Putterman, Seth J.

1996-01-01

A new technique has been developed for the purpose of imaging high amplitude surface motion. With this method one can quantitatively measure the transition to ripple wave turbulence. In addition, one can measure the phase of the turbulent state. These experiments reveal strong coherent structures in turbulent range of motion.

Image registration and analysis for quantitative myocardial perfusion: application to dynamic circular cardiac CT.

PubMed

Isola, A A; Schmitt, H; van Stevendaal, U; Begemann, P G; Coulon, P; Boussel, L; Grass, M

2011-09-21

Large area detector computed tomography systems with fast rotating gantries enable volumetric dynamic cardiac perfusion studies. Prospectively, ECG-triggered acquisitions limit the data acquisition to a predefined cardiac phase and thereby reduce x-ray dose and limit motion artefacts. Even in the case of highly accurate prospective triggering and stable heart rate, spatial misalignment of the cardiac volumes acquired and reconstructed per cardiac cycle may occur due to small motion pattern variations from cycle to cycle. These misalignments reduce the accuracy of the quantitative analysis of myocardial perfusion parameters on a per voxel basis. An image-based solution to this problem is elastic 3D image registration of dynamic volume sequences with variable contrast, as it is introduced in this contribution. After circular cone-beam CT reconstruction of cardiac volumes covering large areas of the myocardial tissue, the complete series is aligned with respect to a chosen reference volume. The results of the registration process and the perfusion analysis with and without registration are evaluated quantitatively in this paper. The spatial alignment leads to improved quantification of myocardial perfusion for three different pig data sets.

Single image super-resolution based on compressive sensing and improved TV minimization sparse recovery

NASA Astrophysics Data System (ADS)

Vishnukumar, S.; Wilscy, M.

2017-12-01

In this paper, we propose a single image Super-Resolution (SR) method based on Compressive Sensing (CS) and Improved Total Variation (TV) Minimization Sparse Recovery. In the CS framework, low-resolution (LR) image is treated as the compressed version of high-resolution (HR) image. Dictionary Training and Sparse Recovery are the two phases of the method. K-Singular Value Decomposition (K-SVD) method is used for dictionary training and the dictionary represents HR image patches in a sparse manner. Here, only the interpolated version of the LR image is used for training purpose and thereby the structural self similarity inherent in the LR image is exploited. In the sparse recovery phase the sparse representation coefficients with respect to the trained dictionary for LR image patches are derived using Improved TV Minimization method. HR image can be reconstructed by the linear combination of the dictionary and the sparse coefficients. The experimental results show that the proposed method gives better results quantitatively as well as qualitatively on both natural and remote sensing images. The reconstructed images have better visual quality since edges and other sharp details are preserved.

SU-C-9A-06: The Impact of CT Image Used for Attenuation Correction in 4D-PET

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cui, Y; Bowsher, J; Yan, S

2014-06-01

Purpose: To evaluate the appropriateness of using 3D non-gated CT image for attenuation correction (AC) in a 4D-PET (gated PET) imaging protocol used in radiotherapy treatment planning simulation. Methods: The 4D-PET imaging protocol in a Siemens PET/CT simulator (Biograph mCT, Siemens Medical Solutions, Hoffman Estates, IL) was evaluated. CIRS Dynamic Thorax Phantom (CIRS Inc., Norfolk, VA) with a moving glass sphere (8 mL) in the middle of its thorax portion was used in the experiments. The glass was filled with {sup 18}F-FDG and was in a longitudinal motion derived from a real patient breathing pattern. Varian RPM system (Varian Medicalmore » Systems, Palo Alto, CA) was used for respiratory gating. Both phase-gating and amplitude-gating methods were tested. The clinical imaging protocol was modified to use three different CT images for AC in 4D-PET reconstruction: first is to use a single-phase CT image to mimic actual clinical protocol (single-CT-PET); second is to use the average intensity projection CT (AveIP-CT) derived from 4D-CT scanning (AveIP-CT-PET); third is to use 4D-CT image to do the phase-matched AC (phase-matching- PET). Maximum SUV (SUVmax) and volume of the moving target (glass sphere) with threshold of 40% SUVmax were calculated for comparison between 4D-PET images derived with different AC methods. Results: The SUVmax varied 7.3%±6.9% over the breathing cycle in single-CT-PET, compared to 2.5%±2.8% in AveIP-CT-PET and 1.3%±1.2% in phasematching PET. The SUVmax in single-CT-PET differed by up to 15% from those in phase-matching-PET. The target volumes measured from single- CT-PET images also presented variations up to 10% among different phases of 4D PET in both phase-gating and amplitude-gating experiments. Conclusion: Attenuation correction using non-gated CT in 4D-PET imaging is not optimal process for quantitative analysis. Clinical 4D-PET imaging protocols should consider phase-matched 4D-CT image if available to achieve better accuracy.« less

3D motion picture of transparent gas flow by parallel phase-shifting digital holography

NASA Astrophysics Data System (ADS)

Awatsuji, Yasuhiro; Fukuda, Takahito; Wang, Yexin; Xia, Peng; Kakue, Takashi; Nishio, Kenzo; Matoba, Osamu

2018-03-01

Parallel phase-shifting digital holography is a technique capable of recording three-dimensional (3D) motion picture of dynamic object, quantitatively. This technique can record single hologram of an object with an image sensor having a phase-shift array device and reconstructs the instantaneous 3D image of the object with a computer. In this technique, a single hologram in which the multiple holograms required for phase-shifting digital holography are multiplexed by using space-division multiplexing technique pixel by pixel. We demonstrate 3D motion picture of dynamic and transparent gas flow recorded and reconstructed by the technique. A compressed air duster was used to generate the gas flow. A motion picture of the hologram of the gas flow was recorded at 180,000 frames/s by parallel phase-shifting digital holography. The phase motion picture of the gas flow was reconstructed from the motion picture of the hologram. The Abel inversion was applied to the phase motion picture and then the 3D motion picture of the gas flow was obtained.

Nuclear medicine and imaging research (Instrumentation and quantitative methods of evaluation)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Beck, R.N.; Cooper, M.D.

1989-09-01

This program addresses the problems involving the basic science and technology underlying the physical and conceptual tools of radioactive tracer methodology as they relate to the measurement of structural and functional parameters of physiologic importance in health and disease. The principal tool is quantitative radionuclide imaging. The overall objective of this program is to further the development and transfer of radiotracer methodology from basic theory to routine clinical practice in order that individual patients and society as a whole will receive the maximum net benefit from the new knowledge gained. The focus of the research is on the development ofmore » new instruments and radiopharmaceuticals, and the evaluation of these through the phase of clinical feasibility.« less

Computerized Liquid Crystal Phase Identification by Neural Networks Analysis of Polarizing Microscopy Textures

NASA Astrophysics Data System (ADS)

Karaszi, Zoltan; Konya, Andrew; Dragan, Feodor; Jakli, Antal; CPIP/LCI; CS Dept. of Kent State University Collaboration

Polarizing optical microscopy (POM) is traditionally the best-established method of studying liquid crystals, and using POM started already with Otto Lehman in 1890. An expert, who is familiar with the science of optics of anisotropic materials and typical textures of liquid crystals, can identify phases with relatively large confidence. However, for unambiguous identification usually other expensive and time-consuming experiments are needed. Replacement of the subjective and qualitative human eye-based liquid crystal texture analysis with quantitative computerized image analysis technique started only recently and were used to enhance the detection of smooth phase transitions, determine order parameter and birefringence of specific liquid crystal phases. We investigate if the computer can recognize and name the phase where the texture was taken. To judge the potential of reliable image recognition based on this procedure, we used 871 images of liquid crystal textures belonging to five main categories: Nematic, Smectic A, Smectic C, Cholesteric and Crystal, and used a Neural Network Clustering Technique included in the data mining software package in Java ``WEKA''. A neural network trained on a set of 827 LC textures classified the remaining 44 textures with 80% accuracy.

X-ray phase contrast tomography from whole organ down to single cells

NASA Astrophysics Data System (ADS)

Krenkel, Martin; Töpperwien, Mareike; Bartels, Matthias; Lingor, Paul; Schild, Detlev; Salditt, Tim

2014-09-01

We use propagation based hard x-ray phase contrast tomography to explore the three dimensional structure of neuronal tissues from the organ down to sub-cellular level, based on combinations of synchrotron radiation and laboratory sources. To this end a laboratory based microfocus tomography setup has been built in which the geometry was optimized for phase contrast imaging and tomography. By utilizing phase retrieval algorithms, quantitative reconstructions can be obtained that enable automatic renderings without edge artifacts. A high brightness liquid metal microfocus x-ray source in combination with a high resolution detector yielding a resolution down to 1.5 μm. To extend the method to nanoscale resolution we use a divergent x-ray waveguide beam geometry at the synchrotron. Thus, the magnification can be easily tuned by placing the sample at different defocus distances. Due to the small Fresnel numbers in this geometry the measured images are of holographic nature which poses a challenge in phase retrieval.

Use of three-dimensional time-resolved phase-contrast magnetic resonance imaging with vastly undersampled isotropic projection reconstruction to assess renal blood flow in a renal cell carcinoma patient treated with sunitinib: a case report.

PubMed

Takayama, Tatsuya; Takehara, Yasuo; Sugiyama, Masataka; Sugiyama, Takayuki; Ishii, Yasuo; Johnson, Kevin E; Wieben, Oliver; Wakayama, Tetsuya; Sakahara, Harumi; Ozono, Seiichiro

2014-08-14

New imaging modalities to assess the efficacy of drugs that have molecular targets remain under development. Here, we describe for the first time the use of time-resolved three-dimensional phase-contrast magnetic resonance imaging to monitor changes in blood supply to a tumor during sunitinib treatment in a patient with localized renal cell carcinoma. A 43-year-old Japanese woman with a tumor-bearing but functional single kidney presented at our hospital in July 2012. Computed tomography and magnetic resonance imaging revealed a cT1aN0M0 renal cell carcinoma embedded in the upper central region of the left kidney. She was prescribed sunitinib as neoadjuvant therapy for 8 months, and then underwent partial nephrectomy. Tumor monitoring during this time was done using time-resolved three-dimensional phase-contrast magnetic resonance imaging, a recent technique which specifically measures blood flow in the various vessels of the kidney. This imaging allowed visualization of the redistribution of renal blood flow during treatment, and showed that flow to the tumor was decreased and flows to other areas increased. Of note, this change occurred in the absence of any change in tumor size. The ability of time-resolved three-dimensional phase-contrast magnetic resonance imaging to provide quantitative information on blood supply to tumors may be useful in monitoring the efficacy of sunitinib treatment.

High-speed, two-dimensional synchrotron white-beam x-ray radiography of spray breakup and atomization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Halls, Benjamin R.; Radke, Christopher D.; Reuter, Benjamin J.

High-speed, two-dimensional synchrotron x-ray radiography and phase-contrast imaging are demonstrated in propulsion sprays. Measurements are performed at the 7-BM beamline at the Advanced Photon Source user facility at Argonne National Laboratory using a recently developed broadband x-ray white beam. This novel enhancement allows for high speed, high fidelity x-ray imaging for the community at large. Quantitative path-integrated liquid distributions and spatio-temporal dynamics of the sprays were imaged with a LuAG:Ce scintillator optically coupled to a high-speed CMOS camera. Images are collected with a microscope objective at frame rates of 20 kHz and with a macro lens at 120 kHz, achievingmore » spatial resolutions of 12 μm and 65 μm, respectively. Imaging with and without potassium iodide (KI) as a contrast-enhancing agent is compared, and the effects of broadband attenuation and spatial beam characteristics are determined through modeling and experimental calibration. In addition, phase contrast is used to differentiate liquid streams with varying concentrations of KI. The experimental approach is applied to different spray conditions, including quantitative measurements of mass distribution during primary atomization and qualitative visualization of turbulent binary fluid mixing. High-speed, two-dimensional synchrotron white-beam x-ray radiography of spray breakup and atomization. Available from: https://www.researchgate.net/publication/312567827_High-speed_two-dimensional_synchrotron_white-beam_x-ray_radiography_of_spray_breakup_and_atomization [accessed Aug 31, 2017].« less

Phase contrast imaging X-ray computed tomography: quantitative characterization of human patellar cartilage matrix with topological and geometrical features

NASA Astrophysics Data System (ADS)

Nagarajan, Mahesh B.; Coan, Paola; Huber, Markus B.; Diemoz, Paul C.; Wismüller, Axel

2014-03-01

Current assessment of cartilage is primarily based on identification of indirect markers such as joint space narrowing and increased subchondral bone density on x-ray images. In this context, phase contrast CT imaging (PCI-CT) has recently emerged as a novel imaging technique that allows a direct examination of chondrocyte patterns and their correlation to osteoarthritis through visualization of cartilage soft tissue. This study investigates the use of topological and geometrical approaches for characterizing chondrocyte patterns in the radial zone of the knee cartilage matrix in the presence and absence of osteoarthritic damage. For this purpose, topological features derived from Minkowski Functionals and geometric features derived from the Scaling Index Method (SIM) were extracted from 842 regions of interest (ROI) annotated on PCI-CT images of healthy and osteoarthritic specimens of human patellar cartilage. The extracted features were then used in a machine learning task involving support vector regression to classify ROIs as healthy or osteoarthritic. Classification performance was evaluated using the area under the receiver operating characteristic (ROC) curve (AUC). The best classification performance was observed with high-dimensional geometrical feature vectors derived from SIM (0.95 ± 0.06) which outperformed all Minkowski Functionals (p < 0.001). These results suggest that such quantitative analysis of chondrocyte patterns in human patellar cartilage matrix involving SIM-derived geometrical features can distinguish between healthy and osteoarthritic tissue with high accuracy.

Quantitative phase measurement for wafer-level optics

NASA Astrophysics Data System (ADS)

Qu, Weijuan; Wen, Yongfu; Wang, Zhaomin; Yang, Fang; Huang, Lei; Zuo, Chao

2015-07-01

Wafer-level-optics now is widely used in smart phone camera, mobile video conferencing or in medical equipment that require tiny cameras. Extracting quantitative phase information has received increased interest in order to quantify the quality of manufactured wafer-level-optics, detect defective devices before packaging, and provide feedback for manufacturing process control, all at the wafer-level for high-throughput microfabrication. We demonstrate two phase imaging methods, digital holographic microscopy (DHM) and Transport-of-Intensity Equation (TIE) to measure the phase of the wafer-level lenses. DHM is a laser-based interferometric method based on interference of two wavefronts. It can perform a phase measurement in a single shot. While a minimum of two measurements of the spatial intensity of the optical wave in closely spaced planes perpendicular to the direction of propagation are needed to do the direct phase retrieval by solving a second-order differential equation, i.e., with a non-iterative deterministic algorithm from intensity measurements using the Transport-of-Intensity Equation (TIE). But TIE is a non-interferometric method, thus can be applied to partial-coherence light. We demonstrated the capability and disability for the two phase measurement methods for wafer-level optics inspection.

Imaging shock waves in diamond with both high temporal and spatial resolution at an XFEL

DOE PAGES

Schropp, Andreas; Hoppe, Robert; Meier, Vivienne; ...

2015-06-18

The advent of hard x-ray free-electron lasers (XFELs) has opened up a variety of scientific opportunities in areas as diverse as atomic physics, plasma physics, nonlinear optics in the x-ray range, and protein crystallography. In this article, we access a new field of science by measuring quantitatively the local bulk properties and dynamics of matter under extreme conditions, in this case by using the short XFEL pulse to image an elastic compression wave in diamond. The elastic wave was initiated by an intense optical laser pulse and was imaged at different delay times after the optical pump pulse using magnifiedmore » x-ray phase-contrast imaging. The temporal evolution of the shock wave can be monitored, yielding detailed information on shock dynamics, such as the shock velocity, the shock front width, and the local compression of the material. The method provides a quantitative perspective on the state of matter in extreme conditions.« less

Imaging Shock Waves in Diamond with Both High Temporal and Spatial Resolution at an XFEL.

PubMed

Schropp, Andreas; Hoppe, Robert; Meier, Vivienne; Patommel, Jens; Seiboth, Frank; Ping, Yuan; Hicks, Damien G; Beckwith, Martha A; Collins, Gilbert W; Higginbotham, Andrew; Wark, Justin S; Lee, Hae Ja; Nagler, Bob; Galtier, Eric C; Arnold, Brice; Zastrau, Ulf; Hastings, Jerome B; Schroer, Christian G

2015-06-18

The advent of hard x-ray free-electron lasers (XFELs) has opened up a variety of scientific opportunities in areas as diverse as atomic physics, plasma physics, nonlinear optics in the x-ray range, and protein crystallography. In this article, we access a new field of science by measuring quantitatively the local bulk properties and dynamics of matter under extreme conditions, in this case by using the short XFEL pulse to image an elastic compression wave in diamond. The elastic wave was initiated by an intense optical laser pulse and was imaged at different delay times after the optical pump pulse using magnified x-ray phase-contrast imaging. The temporal evolution of the shock wave can be monitored, yielding detailed information on shock dynamics, such as the shock velocity, the shock front width, and the local compression of the material. The method provides a quantitative perspective on the state of matter in extreme conditions.

Resolution enhancement by extrapolation of coherent diffraction images: a quantitative study on the limits and a numerical study of nonbinary and phase objects.

PubMed

Latychevskaia, T; Chushkin, Y; Fink, H-W

2016-10-01

In coherent diffractive imaging, the resolution of the reconstructed object is limited by the numerical aperture of the experimental setup. We present here a theoretical and numerical study for achieving super-resolution by postextrapolation of coherent diffraction images, such as diffraction patterns or holograms. We demonstrate that a diffraction pattern can unambiguously be extrapolated from only a fraction of the entire pattern and that the ratio of the extrapolated signal to the originally available signal is linearly proportional to the oversampling ratio. Although there could be in principle other methods to achieve extrapolation, we devote our discussion to employing iterative phase retrieval methods and demonstrate their limits. We present two numerical studies; namely, the extrapolation of diffraction patterns of nonbinary and that of phase objects together with a discussion of the optimal extrapolation procedure. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

Quantitative non-invasive intracellular imaging of Plasmodium falciparum infected human erythrocytes

NASA Astrophysics Data System (ADS)

Edward, Kert; Farahi, Faramarz

2014-05-01

Malaria is a virulent pathological condition which results in over a million annual deaths. The parasitic agent Plasmodium falciparum has been extensively studied in connection with this epidemic but much remains unknown about its development inside the red blood cell host. Optical and fluorescence imaging are among the two most common procedures for investigating infected erythrocytes but both require the introduction of exogenous contrast agents. In this letter, we present a procedure for the non-invasive in situ imaging of malaria infected red blood cells. The procedure is based on the utilization of simultaneously acquired quantitative phase and independent topography data to extract intracellular information. Our method allows for the identification of the developmental stages of the parasite and facilitates in situ analysis of the morphological changes associated with the progression of this disease. This information may assist in the development of efficacious treatment therapies for this condition.

From university research to commercial product (Conference Presentation)

NASA Astrophysics Data System (ADS)

Mathuis, Philip

2016-03-01

Ovizio Imaging Systems, a quantitative microscopic imaging spin-off of the Université Libre de Bruxelles, Belgium, was founded in the beginning of 2010 by Philip Mathuis, Serge Jooris, Prof. Frank Dubois and Dr. Catherine Yourassowky. The company has launched a range of specialized microscopy instruments for quantitative imaging mainly focused on the bioprocessing and diagnostics fields within the life sciences market. During my talk I will present the story of how an idea, emerged from the research labs of the University made it to a manufactured and sold product. The talk will look at many aspects of entrepreneurship and setting up a company, finding the funding for the project, attracting people, industrialization and product design and commercialization. It will also be focused on choices one has to make during the start-up phase and methodologies that can be applied in many different settings.

Quantitative nanoscopy: Tackling sampling limitations in (S)TEM imaging of polymers and composites.

PubMed

Gnanasekaran, Karthikeyan; Snel, Roderick; de With, Gijsbertus; Friedrich, Heiner

2016-01-01

Sampling limitations in electron microscopy questions whether the analysis of a bulk material is representative, especially while analyzing hierarchical morphologies that extend over multiple length scales. We tackled this problem by automatically acquiring a large series of partially overlapping (S)TEM images with sufficient resolution, subsequently stitched together to generate a large-area map using an in-house developed acquisition toolbox (TU/e Acquisition ToolBox) and stitching module (TU/e Stitcher). In addition, we show that quantitative image analysis of the large scale maps provides representative information that can be related to the synthesis and process conditions of hierarchical materials, which moves electron microscopy analysis towards becoming a bulk characterization tool. We demonstrate the power of such an analysis by examining two different multi-phase materials that are structured over multiple length scales. Copyright © 2015 Elsevier B.V. All rights reserved.

Lunar mineral feedstocks from rocks and soils: X-ray digital imaging in resource evaluation

NASA Technical Reports Server (NTRS)

Chambers, John G.; Patchen, Allan; Taylor, Lawrence A.; Higgins, Stefan J.; Mckay, David S.

1994-01-01

The rocks and soils of the Moon provide raw materials essential to the successful establishment of a lunar base. Efficient exploitation of these resources requires accurate characterization of mineral abundances, sizes/shapes, and association of 'ore' and 'gangue' phases, as well as the technology to generate high-yield/high-grade feedstocks. Only recently have x-ray mapping and digital imaging techniques been applied to lunar resource evaluation. The topics covered include inherent differences between lunar basalts and soils and quantitative comparison of rock-derived and soil-derived ilmenite concentrates. It is concluded that x-ray digital-imaging characterization of lunar raw materials provides a quantitative comparison that is unattainable by traditional petrographic techniques. These data are necessary for accurately determining mineral distributions of soil and crushed rock material. Application of these techniques will provide an important link to choosing the best raw material for mineral beneficiation.

Nonlocal maximum likelihood estimation method for denoising multiple-coil magnetic resonance images.

PubMed

Rajan, Jeny; Veraart, Jelle; Van Audekerke, Johan; Verhoye, Marleen; Sijbers, Jan

2012-12-01

Effective denoising is vital for proper analysis and accurate quantitative measurements from magnetic resonance (MR) images. Even though many methods were proposed to denoise MR images, only few deal with the estimation of true signal from MR images acquired with phased-array coils. If the magnitude data from phased array coils are reconstructed as the root sum of squares, in the absence of noise correlations and subsampling, the data is assumed to follow a non central-χ distribution. However, when the k-space is subsampled to increase the acquisition speed (as in GRAPPA like methods), noise becomes spatially varying. In this note, we propose a method to denoise multiple-coil acquired MR images. Both the non central-χ distribution and the spatially varying nature of the noise is taken into account in the proposed method. Experiments were conducted on both simulated and real data sets to validate and to demonstrate the effectiveness of the proposed method. Copyright © 2012 Elsevier Inc. All rights reserved.

Wavelength-multiplexing surface plasmon holographic microscopy.

PubMed

Zhang, Jiwei; Dai, Siqing; Zhong, Jinzhan; Xi, Teli; Ma, Chaojie; Li, Ying; Di, Jianglei; Zhao, Jianlin

2018-05-14

Surface plasmon holographic microscopy (SPHM), which combines surface plasmon microscopy with digital holographic microscopy, can be applied for amplitude- and phase-contrast surface plasmon resonance (SPR) imaging. In this paper, we propose an improved SPHM with the wavelength multiplexing technique based on two laser sources and a common-path hologram recording configuration. Through recording and reconstructing the SPR images at two wavelengths simultaneously employing the improved SPHM, tiny variation of dielectric refractive index in near field is quantitatively monitored with an extended measurement range while maintaining the high sensitivity. Moreover, imaging onion tissues is performed to demonstrate that the detection sensitivities of two wavelengths can compensate for each other in SPR imaging. The proposed wavelength-multiplexing SPHM presents simple structure, high temporal stability and inherent capability of phase curvature compensation, as well as shows great potentials for further applications in monitoring diverse dynamic processes related with refractive index variations and imaging biological tissues with low-contrast refractive index distributions in the near field.

Real time blood testing using quantitative phase imaging.

PubMed

Pham, Hoa V; Bhaduri, Basanta; Tangella, Krishnarao; Best-Popescu, Catherine; Popescu, Gabriel

2013-01-01

We demonstrate a real-time blood testing system that can provide remote diagnosis with minimal human intervention in economically challenged areas. Our instrument combines novel advances in label-free optical imaging with parallel computing. Specifically, we use quantitative phase imaging for extracting red blood cell morphology with nanoscale sensitivity and NVIDIA's CUDA programming language to perform real time cellular-level analysis. While the blood smear is translated through focus, our system is able to segment and analyze all the cells in the one megapixel field of view, at a rate of 40 frames/s. The variety of diagnostic parameters measured from each cell (e.g., surface area, sphericity, and minimum cylindrical diameter) are currently not available with current state of the art clinical instruments. In addition, we show that our instrument correctly recovers the red blood cell volume distribution, as evidenced by the excellent agreement with the cell counter results obtained on normal patients and those with microcytic and macrocytic anemia. The final data outputted by our instrument represent arrays of numbers associated with these morphological parameters and not images. Thus, the memory necessary to store these data is of the order of kilobytes, which allows for their remote transmission via, for example, the cellular network. We envision that such a system will dramatically increase access for blood testing and furthermore, may pave the way to digital hematology.

X-ray Phase Contrast Allows Three Dimensional, Quantitative Imaging of Hydrogel Implants

PubMed Central

Appel, Alyssa A.; Larson, Jeffery C.; Jiang, Bin; Zhong, Zhong; Anastasio, Mark A.; Brey, Eric M.

2015-01-01

Three dimensional imaging techniques are needed for the evaluation and assessment of biomaterials used for tissue engineering and drug delivery applications. Hydrogels are a particularly popular class of materials for medical applications but are difficult to image in tissue using most available imaging modalities. Imaging techniques based on X-ray Phase Contrast (XPC) have shown promise for tissue engineering applications due to their ability to provide image contrast based on multiple X-ray properties. In this manuscript, we investigate the use of XPC for imaging a model hydrogel and soft tissue structure. Porous fibrin loaded poly(ethylene glycol) hydrogels were synthesized and implanted in a rodent subcutaneous model. Samples were explanted and imaged with an analyzer-based XPC technique and processed and stained for histology for comparison. Both hydrogel and soft tissues structures could be identified in XPC images. Structure in skeletal muscle adjacent could be visualized and invading fibrovascular tissue could be quantified. There were no differences between invading tissue measurements from XPC and the gold-standard histology. These results provide evidence of the significant potential of techniques based on XPC for 3D imaging of hydrogel structure and local tissue response. PMID:26487123

Neurient: An Algorithm for Automatic Tracing of Confluent Neuronal Images to Determine Alignment

PubMed Central

Mitchel, J.A.; Martin, I.S.

2013-01-01

A goal of neural tissue engineering is the development and evaluation of materials that guide neuronal growth and alignment. However, the methods available to quantitatively evaluate the response of neurons to guidance materials are limited and/or expensive, and may require manual tracing to be performed by the researcher. We have developed an open source, automated Matlab-based algorithm, building on previously published methods, to trace and quantify alignment of fluorescent images of neurons in culture. The algorithm is divided into three phases, including computation of a lookup table which contains directional information for each image, location of a set of seed points which may lie along neurite centerlines, and tracing neurites starting with each seed point and indexing into the lookup table. This method was used to obtain quantitative alignment data for complex images of densely cultured neurons. Complete automation of tracing allows for unsupervised processing of large numbers of images. Following image processing with our algorithm, available metrics to quantify neurite alignment include angular histograms, percent of neurite segments in a given direction, and mean neurite angle. The alignment information obtained from traced images can be used to compare the response of neurons to a range of conditions. This tracing algorithm is freely available to the scientific community under the name Neurient, and its implementation in Matlab allows a wide range of researchers to use a standardized, open source method to quantitatively evaluate the alignment of dense neuronal cultures. PMID:23384629

Measuring dynamic membrane fluctuations in cell membrane using quantitative phase imaging (Conference Presentation)

NASA Astrophysics Data System (ADS)

Lee, SangYun; Kim, Kyoohyun; Park, YongKeun

2017-02-01

There is a strong correlation between the dynamic membrane fluctuations and the biomechanical properties of living cells. The dynamic membrane fluctuation consists of submicron displacements, and can be altered by changing the cells' pathophysiological conditions. These results have significant relevance to the understanding of RBC biophysics and pathology, as follows. RBCs must withstand large mechanical deformations during repeated passages through the microvasculature and the fenestrated walls of the splenic sinusoids. This essential ability is diminished with senescence, resulting in physiological destruction of the aging RBCs. Pathological destruction of the red cells, however, occurs in cells affected by a host of diseases such as spherocytosis, malaria, and Sickle cell disease, as RBCs depart from their normal discoid shape and lose their deformability. Therefore, quantifying the RBC deformability insight into a variety of problems regarding the interplay of cell structure, dynamics, and function. Furthermore, the ability to monitor mechanical properties of RBCs is of vital interest in monitoring disease progression or response to treatment as molecular and pharmaceutical approaches for treatment of chronic diseases. Here, we present the measurements of dynamic membrane fluctuations in live cells using quantitative phase imaging techniques. Measuring both the 3-D refractive index maps and the dynamic phase images of live cells are simultaneously measured, from which dynamic membrane fluctuation and deformability of cells are precisely calculated. We also present its applications to various diseases ranging from sickle cell diseases, babesiosis, and to diabetes.

Quantitative imaging with Fucci and mathematics to uncover temporal dynamics of cell cycle progression.

PubMed

Saitou, Takashi; Imamura, Takeshi

2016-01-01

Cell cycle progression is strictly coordinated to ensure proper tissue growth, development, and regeneration of multicellular organisms. Spatiotemporal visualization of cell cycle phases directly helps us to obtain a deeper understanding of controlled, multicellular, cell cycle progression. The fluorescent ubiquitination-based cell cycle indicator (Fucci) system allows us to monitor, in living cells, the G1 and the S/G2/M phases of the cell cycle in red and green fluorescent colors, respectively. Since the discovery of Fucci technology, it has found numerous applications in the characterization of the timing of cell cycle phase transitions under diverse conditions and various biological processes. However, due to the complexity of cell cycle dynamics, understanding of specific patterns of cell cycle progression is still far from complete. In order to tackle this issue, quantitative approaches combined with mathematical modeling seem to be essential. Here, we review several studies that attempted to integrate Fucci technology and mathematical models to obtain quantitative information regarding cell cycle regulatory patterns. Focusing on the technological development of utilizing mathematics to retrieve meaningful information from the Fucci producing data, we discuss how the combined methods advance a quantitative understanding of cell cycle regulation. © 2015 Japanese Society of Developmental Biologists.

Radiologic-Pathologic Analysis of Contrast-enhanced and Diffusion-weighted MR Imaging in Patients with HCC after TACE: Diagnostic Accuracy of 3D Quantitative Image Analysis

PubMed Central

Chapiro, Julius; Wood, Laura D.; Lin, MingDe; Duran, Rafael; Cornish, Toby; Lesage, David; Charu, Vivek; Schernthaner, Rüdiger; Wang, Zhijun; Tacher, Vania; Savic, Lynn Jeanette; Kamel, Ihab R.

2014-01-01

Purpose To evaluate the diagnostic performance of three-dimensional (3Dthree-dimensional) quantitative enhancement-based and diffusion-weighted volumetric magnetic resonance (MR) imaging assessment of hepatocellular carcinoma (HCChepatocellular carcinoma) lesions in determining the extent of pathologic tumor necrosis after transarterial chemoembolization (TACEtransarterial chemoembolization). Materials and Methods This institutional review board–approved retrospective study included 17 patients with HCChepatocellular carcinoma who underwent TACEtransarterial chemoembolization before surgery. Semiautomatic 3Dthree-dimensional volumetric segmentation of target lesions was performed at the last MR examination before orthotopic liver transplantation or surgical resection. The amount of necrotic tumor tissue on contrast material–enhanced arterial phase MR images and the amount of diffusion-restricted tumor tissue on apparent diffusion coefficient (ADCapparent diffusion coefficient) maps were expressed as a percentage of the total tumor volume. Visual assessment of the extent of tumor necrosis and tumor response according to European Association for the Study of the Liver (EASLEuropean Association for the Study of the Liver) criteria was performed. Pathologic tumor necrosis was quantified by using slide-by-slide segmentation. Correlation analysis was performed to evaluate the predictive values of the radiologic techniques. Results At histopathologic examination, the mean percentage of tumor necrosis was 70% (range, 10%–100%). Both 3Dthree-dimensional quantitative techniques demonstrated a strong correlation with tumor necrosis at pathologic examination (R2 = 0.9657 and R2 = 0.9662 for quantitative EASLEuropean Association for the Study of the Liver and quantitative ADCapparent diffusion coefficient, respectively) and a strong intermethod agreement (R2 = 0.9585). Both methods showed a significantly lower discrepancy with pathologically measured necrosis (residual standard error [RSEresidual standard error] = 6.38 and 6.33 for quantitative EASLEuropean Association for the Study of the Liver and quantitative ADCapparent diffusion coefficient, respectively), when compared with non-3Dthree-dimensional techniques (RSEresidual standard error = 12.18 for visual assessment). Conclusion This radiologic-pathologic correlation study demonstrates the diagnostic accuracy of 3Dthree-dimensional quantitative MR imaging techniques in identifying pathologically measured tumor necrosis in HCChepatocellular carcinoma lesions treated with TACEtransarterial chemoembolization. © RSNA, 2014 Online supplemental material is available for this article. PMID:25028783

Pilot study of quantitative analysis of background enhancement on breast MR images: association with menstrual cycle and mammographic breast density.

PubMed

Scaranelo, Anabel M; Carrillo, Maria Claudia; Fleming, Rachel; Jacks, Lindsay M; Kulkarni, Supriya R; Crystal, Pavel

2013-06-01

To perform semiautomated quantitative analysis of the background enhancement (BE) in a cohort of patients with newly diagnosed breast cancer and to correlate it with mammographic breast density and menstrual cycle. Informed consent was waived after the research ethics board approved this study. Results of 177 consecutive preoperative breast magnetic resonance (MR) examinations performed from February to December 2009 were reviewed; 147 female patients (median age, 48 years; range, 26-86 years) were included. Ordinal values of BE and breast density were described by two independent readers by using the Breast Imaging Reporting and Data System lexicon. The BE coefficient (BEC) was calculated thus: (SI2 · 100/SI1) - 100, where SI is signal intensity, SI2 is the SI enhancement measured in the largest anteroposterior dimension in the axial plane 1 minute after the contrast agent injection, and SI1is the SI before contrast agent injection. BEC was used for the quantitative analysis of BE. Menstrual cycle status was based on the last menstrual period. The Wilcoxon rank-sum or Kruskal-Wallis test was used to compare quantitative assessment groups. Cohen weighted κ was used to evaluate agreement. Of 147 patients, 68 (46%) were premenopausal and 79 (54%) were postmenopausal. The quantitative BEC was associated with the menstrual status (BEC in premenopausal women, 31.48 ± 20.68 [standard deviation]; BEC in postmenopausal women, 25.65 ± 16.74; P = .02). The percentage of overall BE was higher when the MR imaging was performed in women in the inadequate phase of the cycle (<35 days, not 7-14 days; mean BEC, 35.7) compared with women in the postmenopausal group (P = .001). Premenopausal women had significantly higher BEC when compared with postmenopausal women (P = .03). There was no significant difference in the percentage of BE between breast density groups. Premenopausal women with breast cancer, and specifically women in the inadequate phase of the cycle, presented with higher quantitative BE than postmenopausal women. No association was found between BE and breast density.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Meisner, Ludmila, E-mail: llm@ispms.tsc.ru; Meisner, Stanislav, E-mail: msn@ispms.tsc.ru; Mironov, Yurii, E-mail: myp@ispms.tsc.ru

The paper considers the effects arising on X-ray diffraction patterns taken in different diffraction geometries and how these effects can be interpreted to judge structural states in NiTi near-surface regions after electron and ion beam treatment. It is shown that qualitative and quantitative analysis of phase composition, lattice parameters of main phases, elastic stress states, and their in-depth variation requires X-ray diffraction patterns in both symmetric Bragg–Brentano and asymmetric Lambot–Vassamilleta geometries with variation in X-ray wavelengths and imaging conditions (with and with no β-filter). These techniques of structural phase analysis are more efficient when the thickness of modified NiTi surfacemore » layers is 1–10 μm (after electron beam treatment) and requires special imaging conditions when the thickness of modified NiTi surface layers is no greater than 1 μm (after ion beam treatment)« less

Tissue refractometry using Hilbert phase microscopy.

PubMed

Lue, Niyom; Bewersdorf, Joerg; Lessard, Mark D; Badizadegan, Kamran; Dasari, Ramachandra R; Feld, Michael S; Popescu, Gabriel

2007-12-15

We present, for the first time to our knowledge, quantitative phase images associated with unstained 5 mum thick tissue slices of mouse brain, spleen, and liver. The refractive properties of the tissue are retrieved in terms of the average refractive index and its spatial variation. We find that the average refractive index varies significantly with tissue type, such that the brain is characterized by the lowest value and the liver by the highest. The spatial power spectra of the phase images reveal power law behavior with different exponents for each tissue type. This approach opens a new possibility for stain-free characterization of tissues, where the diagnostic power is provided by the intrinsic refractive properties of the biological structure. We present results obtained for liver tissue affected by a lysosomal storage disease and show that our technique can quantify structural changes during this disease development.

Super-resolution differential interference contrast microscopy by structured illumination.

PubMed

Chen, Jianling; Xu, Yan; Lv, Xiaohua; Lai, Xiaomin; Zeng, Shaoqun

2013-01-14

We propose a structured illumination differential interference contrast (SI-DIC) microscopy, breaking the diffraction resolution limit of differential interference contrast (DIC) microscopy. SI-DIC extends the bandwidth of coherent transfer function of the DIC imaging system, thus the resolution is improved. With 0.8 numerical aperture condenser and objective, the reconstructed SI-DIC image of 53 nm polystyrene beads reveals lateral resolution of approximately 190 nm, doubling that of the conventional DIC image. We also demonstrate biological observations of label-free cells with improved spatial resolution. The SI-DIC microscopy can provide sub-diffraction resolution and high contrast images with marker-free specimens, and has the potential for achieving sub-diffraction resolution quantitative phase imaging.

Quantitative Magnetic Resonance Thermometry and Its Use with MR-Guided Focused Ultrasound

NASA Astrophysics Data System (ADS)

Pauly, Kim

2014-03-01

Focused ultrasound (FUS) uses a large area array, typically outside the body, that is geometrically or electronically focused to a point deep in the body. Such focusing provides amplification of the ultrasound intensity, thereby allowing heating of tissue to the point of coagulation at the focus, without damage to the intervening tissue. Guidance of FUS treatments deep in the body can be done quantitatively with magnetic resonance (MR) thermometry, termed MRgFUS. The physics behind MR thermometry lie in the changes in hydrogen bonding with temperature. As tissue temperature rises, hydrogen bonds break, allowing the return of the electron cloud to shield water protons, reducing the magnetic field seen by the protons, and the resonant frequency. The change in resonant frequency is -0.01 ppm per degree C and is the same for all aqueous tissues. The result of the shift in proton resonant frequency is seen in the phase of gradient echo images. Subtraction of the phase of images acquired before and during heating allows the removal of background phase from other sources, yielding quantitative temperature maps. Temperature standard deviations less than 1 degree C are readily achievable and thermal dose maps are easily calculated. Thermal dose is found from a conversion of the whole temperature-time curve to an equivalent number of minutes at 43 degrees C. A thermal dose of 240 minutes is often taken as the threshold for tissue damage. MR thermometry is complicated by the motion of the target tissue and/or motion of other organs such as occurs during respiration. More sophisticated algorithms than the simple baseline subtraction take advantage of the facts that motion can be repetitive (in the case of respiratory motion) and/or the fact that the focal region in MRgFUS is small, allowing for extraction of the heat from the phase profile without subtraction of a background phase.

High-throughput label-free screening of euglena gracilis with optofluidic time-stretch quantitative phase microscopy

NASA Astrophysics Data System (ADS)

Guo, Baoshan; Lei, Cheng; Ito, Takuro; Yaxiaer, Yalikun; Kobayashi, Hirofumi; Jiang, Yiyue; Tanaka, Yo; Ozeki, Yasuyuki; Goda, Keisuke

2017-02-01

The development of reliable, sustainable, and economical sources of alternative fuels is an important, but challenging goal for the world. As an alternative to liquid fossil fuels, microalgal biofuel is expected to play a key role in reducing the detrimental effects of global warming since microalgae absorb atmospheric CO2 via photosynthesis. Unfortunately, conventional analytical methods only provide population-averaged lipid contents and fail to characterize a diverse population of microalgal cells with single-cell resolution in a noninvasive and interference-free manner. Here we demonstrate high-throughput label-free single-cell screening of lipid-producing microalgal cells with optofluidic time-stretch quantitative phase microscopy. In particular, we use Euglena gracilis - an attractive microalgal species that produces wax esters (suitable for biodiesel and aviation fuel after refinement) within lipid droplets. Our optofluidic time-stretch quantitative phase microscope is based on an integration of a hydrodynamic-focusing microfluidic chip, an optical time-stretch phase-contrast microscope, and a digital image processor equipped with machine learning. As a result, it provides both the opacity and phase contents of every single cell at a high throughput of 10,000 cells/s. We characterize heterogeneous populations of E. gracilis cells under two different culture conditions to evaluate their lipid production efficiency. Our method holds promise as an effective analytical tool for microalgaebased biofuel production.

MRI technique for the snapshot imaging of quantitative velocity maps using RARE.

PubMed

Shiko, G; Sederman, A J; Gladden, L F

2012-03-01

A quantitative PGSE-RARE pulse sequence was developed and successfully applied to the in situ dissolution of two pharmaceutical formulations dissolving over a range of timescales. The new technique was chosen over other existing fast velocity imaging techniques because it is T(2) weighted, not T(2)(∗) weighted, and is, therefore, robust for imaging time-varying interfaces and flow in magnetically heterogeneous systems. The complex signal was preserved intact by separating odd and even echoes to obtain two phase maps which are then averaged in post-processing. Initially, the validity of the technique was shown when imaging laminar flow in a pipe. Subsequently, the dissolution of two drugs was followed in situ, where the technique enables the imaging and quantification of changes in the form of the tablet and the flow field surrounding it at high spatial and temporal resolution. First, the complete 3D velocity field around an eroding salicylic acid tablet was acquired at a resolution of 98×49 μm(2), within 20 min, and monitored over ∼13 h. The tablet was observed to experience a heterogeneous flow field and, hence a heterogeneous shear field, which resulted in the non-symmetric erosion of the tablet. Second, the dissolution of a fast dissolving immediate release tablet was followed using one-shot 2D velocity images acquired every 5.2 s at a resolution of 390×390 μm(2). The quantitative nature of the technique and fast acquisition times provided invaluable information on the dissolution behaviour of this tablet, which had not been attainable previously with conventional quantitative MRI techniques. Copyright © 2012 Elsevier Inc. All rights reserved.

Dynamic quantitative phase images of pond life, insect wings, and in vitro cell cultures

NASA Astrophysics Data System (ADS)

Creath, Katherine

2010-08-01

This paper presents images and data of live biological samples taken with a novel Linnik interference microscope. The specially designed optical system enables instantaneous and 3D video measurements of dynamic motions within and among live cells without the need for contrast agents. This "label-free", vibration insensitive imaging system enables measurement of biological objects in reflection using harmless light levels with current magnifications of 10X (NA 0.3) and 20X (NA 0.5) and wavelengths of 660 nm and 785 nm over fields of view from several hundred microns up to a millimeter. At the core of the instrument is a phasemeasurement camera (PMC) enabling simultaneous measurement of multiple interference patterns utilizing a pixelated phase mask taking advantage of the polarization properties of light. Utilizing this technology enables the creation of phase image movies in real time at video rates so that dynamic motions and volumetric changes can be tracked. Objects are placed on a reflective surface in liquid under a coverslip. Phase values are converted to optical thickness data enabling volumetric, motion and morphological studies. Data from a number of different mud puddle organisms such as paramecium, flagellates and rotifers will be presented, as will measurements of flying ant wings and cultures of human breast cancer cells. These data highlight examples of monitoring different biological processes and motions. The live presentation features 4D phase movies of these examples.

Automated analysis of individual sperm cells using stain-free interferometric phase microscopy and machine learning.

PubMed

Mirsky, Simcha K; Barnea, Itay; Levi, Mattan; Greenspan, Hayit; Shaked, Natan T

2017-09-01

Currently, the delicate process of selecting sperm cells to be used for in vitro fertilization (IVF) is still based on the subjective, qualitative analysis of experienced clinicians using non-quantitative optical microscopy techniques. In this work, a method was developed for the automated analysis of sperm cells based on the quantitative phase maps acquired through use of interferometric phase microscopy (IPM). Over 1,400 human sperm cells from 8 donors were imaged using IPM, and an algorithm was designed to digitally isolate sperm cell heads from the quantitative phase maps while taking into consideration both the cell 3D morphology and contents, as well as acquire features describing sperm head morphology. A subset of these features was used to train a support vector machine (SVM) classifier to automatically classify sperm of good and bad morphology. The SVM achieves an area under the receiver operating characteristic curve of 88.59% and an area under the precision-recall curve of 88.67%, as well as precisions of 90% or higher. We believe that our automatic analysis can become the basis for objective and automatic sperm cell selection in IVF. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

Quantitative holographic interferometry applied to combustion and compressible flow research

NASA Astrophysics Data System (ADS)

Bryanston-Cross, Peter J.; Towers, D. P.

1993-03-01

The application of holographic interferometry to phase object analysis is described. Emphasis has been given to a method of extracting quantitative information automatically from the interferometric fringe data. To achieve this a carrier frequency has been added to the holographic data. This has made it possible, firstly to form a phase map using a fast Fourier transform (FFT) algorithm. Then to `solve,' or unwrap, this image to give a contiguous density map using a minimum weight spanning tree (MST) noise immune algorithm, known as fringe analysis (FRAN). Applications of this work to a burner flame and a compressible flow are presented. In both cases the spatial frequency of the fringes exceed the resolvable limit of conventional digital framestores. Therefore, a flatbed scanner with a resolution of 3200 X 2400 pixels has been used to produce very high resolution digital images from photographs. This approach has allowed the processing of data despite the presence of caustics, generated by strong thermal gradients at the edge of the combustion field. A similar example is presented from the analysis of a compressible transonic flow in the shock wave and trailing edge regions.

Quantitative Cell Cycle Analysis Based on an Endogenous All-in-One Reporter for Cell Tracking and Classification.

PubMed

Zerjatke, Thomas; Gak, Igor A; Kirova, Dilyana; Fuhrmann, Markus; Daniel, Katrin; Gonciarz, Magdalena; Müller, Doris; Glauche, Ingmar; Mansfeld, Jörg

2017-05-30

Cell cycle kinetics are crucial to cell fate decisions. Although live imaging has provided extensive insights into this relationship at the single-cell level, the limited number of fluorescent markers that can be used in a single experiment has hindered efforts to link the dynamics of individual proteins responsible for decision making directly to cell cycle progression. Here, we present fluorescently tagged endogenous proliferating cell nuclear antigen (PCNA) as an all-in-one cell cycle reporter that allows simultaneous analysis of cell cycle progression, including the transition into quiescence, and the dynamics of individual fate determinants. We also provide an image analysis pipeline for automated segmentation, tracking, and classification of all cell cycle phases. Combining the all-in-one reporter with labeled endogenous cyclin D1 and p21 as prime examples of cell-cycle-regulated fate determinants, we show how cell cycle and quantitative protein dynamics can be simultaneously extracted to gain insights into G1 phase regulation and responses to perturbations. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

Refractive index tomograms and dynamic membrane fluctuations of red blood cells from patients with diabetes mellitus.

PubMed

Lee, SangYun; Park, HyunJoo; Kim, Kyoohyun; Sohn, YongHak; Jang, Seongsoo; Park, YongKeun

2017-04-21

In this paper, we present the optical characterisations of diabetic red blood cells (RBCs) in a non-invasive manner employing three-dimensional (3-D) quantitative phase imaging. By measuring 3-D refractive index tomograms and 2-D time-series phase images, the morphological (volume, surface area and sphericity), biochemical (haemoglobin concentration and content) and mechanical (membrane fluctuation) parameters were quantitatively retrieved at the individual cell level. With simultaneous measurements of individual cell properties, systematic correlative analyses on retrieved RBC parameters were also performed. Our measurements show there exist no statistically significant alterations in morphological and biochemical parameters of diabetic RBCs, compared to those of healthy (non-diabetic) RBCs. In contrast, membrane deformability of diabetic RBCs is significantly lower than that of healthy, non-diabetic RBCs. Interestingly, non-diabetic RBCs exhibit strong correlations between the elevated glycated haemoglobin in RBC cytoplasm and decreased cell deformability, whereas diabetic RBCs do not show correlations. Our observations strongly support the idea that slow and irreversible glycation of haemoglobin and membrane proteins of RBCs by hyperglycaemia significantly compromises RBC deformability in diabetic patients.

Scintigraphic evaluation of digital circulation during the developmental and acute phases of equine laminitis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Trout, D.R.

1987-01-01

Using nuclear isotopic imaging, digital circulation was sequentially evaluated at 24-hour intervals in 11 control horses and in 9 horses affected with acute laminitis, created by administration of a high-starch ration. Following intra-arterial injection of /sup 99m/Tc macroaggregated albumin into the brachiocephalic trunk, a gamma camera and dedicated nuclear medicine computer were used to acquire static images of the right front foot. Dynamic vascular-phase and static interstitial-phase images were also obtained after jugular vein injection of /sup 99m/Tc diethylenetriamine pentaacetic acid. These procedures were performed on standing horses, using either minimal or no tranquilization. The images were quantitatively analyzed formore » parameters indicative of circulation to the foot as a whole and to specific regions of interest within the foot. There was no evidence of reduced total blood flow to the lamellae during either the developmental or acute phases of laminitis. Although total flow tended to increase throughout the peripheral/external regions of the foot, statistically significant elevations were consistently present only within the lamellae. Changes indicative of decreased total blood flow were noted in the central/internal regions of the foot. These alterations usually occurred coincident with or after the onset of clinical lameness.« less

Model-based quantification of image quality

NASA Technical Reports Server (NTRS)

Hazra, Rajeeb; Miller, Keith W.; Park, Stephen K.

1989-01-01

In 1982, Park and Schowengerdt published an end-to-end analysis of a digital imaging system quantifying three principal degradation components: (1) image blur - blurring caused by the acquisition system, (2) aliasing - caused by insufficient sampling, and (3) reconstruction blur - blurring caused by the imperfect interpolative reconstruction. This analysis, which measures degradation as the square of the radiometric error, includes the sample-scene phase as an explicit random parameter and characterizes the image degradation caused by imperfect acquisition and reconstruction together with the effects of undersampling and random sample-scene phases. In a recent paper Mitchell and Netravelli displayed the visual effects of the above mentioned degradations and presented subjective analysis about their relative importance in determining image quality. The primary aim of the research is to use the analysis of Park and Schowengerdt to correlate their mathematical criteria for measuring image degradations with subjective visual criteria. Insight gained from this research can be exploited in the end-to-end design of optical systems, so that system parameters (transfer functions of the acquisition and display systems) can be designed relative to each other, to obtain the best possible results using quantitative measurements.

Imaging Local Polarization in Ferroelectric Thin Films by Coherent X-Ray Bragg Projection Ptychography

NASA Astrophysics Data System (ADS)

Hruszkewycz, S. O.; Highland, M. J.; Holt, M. V.; Kim, Dongjin; Folkman, C. M.; Thompson, Carol; Tripathi, A.; Stephenson, G. B.; Hong, Seungbum; Fuoss, P. H.

2013-04-01

We used x-ray Bragg projection ptychography (BPP) to map spatial variations of ferroelectric polarization in thin film PbTiO3, which exhibited a striped nanoscale domain pattern on a high-miscut (001) SrTiO3 substrate. By converting the reconstructed BPP phase image to picometer-scale ionic displacements in the polar unit cell, a quantitative polarization map was made that was consistent with other characterization. The spatial resolution of 5.7 nm demonstrated here establishes BPP as an important tool for nanoscale ferroelectric domain imaging, especially in complex environments accessible with hard x rays.

Dynamic three-dimensional phase-contrast technique in MRI: application to complex flow analysis around the artificial heart valve

NASA Astrophysics Data System (ADS)

Kim, Soo Jeong; Lee, Dong Hyuk; Song, Inchang; Kim, Nam Gook; Park, Jae-Hyeung; Kim, JongHyo; Han, Man Chung; Min, Byong Goo

1998-07-01

Phase-contrast (PC) method of magnetic resonance imaging (MRI) has bee used for quantitative measurements of flow velocity and volume flow rate. It is a noninvasive technique which provides an accurate two-dimensional velocity image. Moreover, Phase Contrast Cine magnetic resonance imaging combines the flow dependent contrast of PC-MRI with the ability of cardiac cine imaging to produce images throughout the cardiac cycle. However, the accuracy of the data acquired from the single through-plane velocity encoding can be reduced by the effect of flow direction, because in many practical cases flow directions are not uniform throughout the whole region of interest. In this study, we present dynamic three-dimensional velocity vector mapping method using PC-MRI which can visualize the complex flow pattern through 3D volume rendered images displayed dynamically. The direction of velocity mapping can be selected along any three orthogonal axes. By vector summation, the three maps can be combined to form a velocity vector map that determines the velocity regardless of the flow direction. At the same time, Cine method is used to observe the dynamic change of flow. We performed a phantom study to evaluate the accuracy of the suggested PC-MRI in continuous and pulsatile flow measurement. Pulsatile flow wave form is generated by the ventricular assistant device (VAD), HEMO-PULSA (Biomedlab, Seoul, Korea). We varied flow velocity, pulsatile flow wave form, and pulsing rate. The PC-MRI-derived velocities were compared with Doppler-derived results. The velocities of the two measurements showed a significant linear correlation. Dynamic three-dimensional velocity vector mapping was carried out for two cases. First, we applied to the flow analysis around the artificial heart valve in a flat phantom. We could observe the flow pattern around the valve through the 3-dimensional cine image. Next, it is applied to the complex flow inside the polymer sac that is used as ventricle in totally implantable artificial heart (TAH). As a result we could observe the flow pattern around the valves of the sac, though complex flow can not be detected correctly in the conventional phase contrast method. In addition, we could calculate the cardiac output from TAH sac by quantitative measurement of the volume of flow across the outlet valve.

3-D Ultrafast Doppler Imaging Applied to the Noninvasive and Quantitative Imaging of Blood Vessels in Vivo

PubMed Central

Provost, J.; Papadacci, C.; Demene, C.; Gennisson, J-L.; Tanter, M.; Pernot, M.

2016-01-01

Ultrafast Doppler Imaging was introduced as a technique to quantify blood flow in an entire 2-D field of view, expanding the field of application of ultrasound imaging to the highly sensitive anatomical and functional mapping of blood vessels. We have recently developed 3-D Ultrafast Ultrasound Imaging, a technique that can produce thousands of ultrasound volumes per second, based on three-dimensional plane and diverging wave emissions, and demonstrated its clinical feasibility in human subjects in vivo. In this study, we show that non-invasive 3-D Ultrafast Power Doppler, Pulsed Doppler, and Color Doppler Imaging can be used to perform quantitative imaging of blood vessels in humans when using coherent compounding of three-dimensional tilted plane waves. A customized, programmable, 1024-channel ultrasound system was designed to perform 3-D Ultrafast Imaging. Using a 32X32, 3-MHz matrix phased array (Vermon, France), volumes were beamformed by coherently compounding successive tilted plane wave emissions. Doppler processing was then applied in a voxel-wise fashion. 3-D Ultrafast Power Doppler Imaging was first validated by imaging Tygon tubes of varying diameter and its in vivo feasibility was demonstrated by imaging small vessels in the human thyroid. Simultaneous 3-D Color and Pulsed Doppler Imaging using compounded emissions were also applied in the carotid artery and the jugular vein in one healthy volunteer. PMID:26276956

Updates in MRI characterization of the thymus in myasthenic patients.

PubMed

Popa, G A; Preda, E M; Scheau, C; Vilciu, C; Lupescu, I G

2012-06-12

To evaluate the imaging appearance of the thymus in the myasthenic patients by using chemical shift magnetic resonance imaging, and, to correlate the chemical shift ratio (CSR) with pathologic findings after surgical excision. In the past year, a total of 11 myasthenic patients (4 males, 7 females; age range of 26-65 years), have been investigated by MRI centered at the thymic lodge. Our protocol included a Dual-Echo technique, T1-weighted In-phase/Opposed-phase MR images in all patients. A chemical shift ratio (CSR) was calculated by comparing the signal intensity of the thymus gland with that of the chest wall muscle for quantitative analysis. For this purpose, we have used standard region-of-interest electronic cursors at a slice level of the maximum axial surface of the thymus. We have identified two patients groups: a thymic hyperplasia group and a thymic tumoral group. With the decrease in the signal intensity of the thymus gland at chemical shift, the MR imaging was evident only in the hyperplasia group. The mean CSR in the hyperplasia group was considerably lower than that in the tumor group, 0,4964 ± 0,1841, compared with 1,0398 ± 0,0244. The difference in CSR between the hyperplasia and tumor groups was statistically significant (P=0,0028). MR imaging using T1-weighted In-phase/Opposed-phase images could be a useful diagnostic tool in the preoperative assessment of the thymic lodge and may help differentiate thymic hyperplasia from tumors of the thymus gland.

Use of MRI in Differentiation of Papillary Renal Cell Carcinoma Subtypes: Qualitative and Quantitative Analysis.

PubMed

Doshi, Ankur M; Ream, Justin M; Kierans, Andrea S; Bilbily, Matthew; Rusinek, Henry; Huang, William C; Chandarana, Hersh

2016-03-01

The purpose of this study was to determine whether qualitative and quantitative MRI feature analysis is useful for differentiating type 1 from type 2 papillary renal cell carcinoma (PRCC). This retrospective study included 21 type 1 and 17 type 2 PRCCs evaluated with preoperative MRI. Two radiologists independently evaluated various qualitative features, including signal intensity, heterogeneity, and margin. For the quantitative analysis, a radiology fellow and a medical student independently drew 3D volumes of interest over the entire tumor on T2-weighted HASTE images, apparent diffusion coefficient parametric maps, and nephrographic phase contrast-enhanced MR images to derive first-order texture metrics. Qualitative and quantitative features were compared between the groups. For both readers, qualitative features with greater frequency in type 2 PRCC included heterogeneous enhancement, indistinct margin, and T2 heterogeneity (all, p < 0.035). Indistinct margins and heterogeneous enhancement were independent predictors (AUC, 0.822). Quantitative analysis revealed that apparent diffusion coefficient, HASTE, and contrast-enhanced entropy were greater in type 2 PRCC (p < 0.05; AUC, 0.682-0.716). A combined quantitative and qualitative model had an AUC of 0.859. Qualitative features within the model had interreader concordance of 84-95%, and the quantitative data had intraclass coefficients of 0.873-0.961. Qualitative and quantitative features can help discriminate between type 1 and type 2 PRCC. Quantitative analysis may capture useful information that complements the qualitative appearance while benefiting from high interobserver agreement.

Nanoscale imaging of fundamental li battery chemistry: solid-electrolyte interphase formation and preferential growth of lithium metal nanoclusters.

PubMed

Sacci, Robert L; Black, Jennifer M; Balke, Nina; Dudney, Nancy J; More, Karren L; Unocic, Raymond R

2015-03-11

The performance characteristics of Li-ion batteries are intrinsically linked to evolving nanoscale interfacial electrochemical reactions. To probe the mechanisms of solid electrolyte interphase (SEI) formation and to track Li nucleation and growth mechanisms from a standard organic battery electrolyte (LiPF6 in EC:DMC), we used in situ electrochemical scanning transmission electron microscopy (ec-S/TEM) to perform controlled electrochemical potential sweep measurements while simultaneously imaging site-specific structures resulting from electrochemical reactions. A combined quantitative electrochemical measurement and STEM imaging approach is used to demonstrate that chemically sensitive annular dark field STEM imaging can be used to estimate the density of the evolving SEI and to identify Li-containing phases formed in the liquid cell. We report that the SEI is approximately twice as dense as the electrolyte as determined from imaging and electron scattering theory. We also observe site-specific locations where Li nucleates and grows on the surface and edge of the glassy carbon electrode. Lastly, this report demonstrates the investigative power of quantitative nanoscale imaging combined with electrochemical measurements for studying fluid-solid interfaces and their evolving chemistries.

Quantitative evaluation of hidden defects in cast iron components using ultrasound activated lock-in vibrothermography.

PubMed

Montanini, R; Freni, F; Rossi, G L

2012-09-01

This paper reports one of the first experimental results on the application of ultrasound activated lock-in vibrothermography for quantitative assessment of buried flaws in complex cast parts. The use of amplitude modulated ultrasonic heat generation allowed selective response of defective areas within the part, as the defect itself is turned into a local thermal wave emitter. Quantitative evaluation of hidden damages was accomplished by estimating independently both the area and the depth extension of the buried flaws, while x-ray 3D computed tomography was used as reference for sizing accuracy assessment. To retrieve flaw's area, a simple yet effective histogram-based phase image segmentation algorithm with automatic pixels classification has been developed. A clear correlation was found between the thermal (phase) signature measured by the infrared camera on the target surface and the actual mean cross-section area of the flaw. Due to the very fast cycle time (<30 s/part), the method could potentially be applied for 100% quality control of casting components.

Quantitative, Comparable Coherent Anti-Stokes Raman Scattering (CARS) Spectroscopy: Correcting Errors in Phase Retrieval

PubMed Central

Camp, Charles H.; Lee, Young Jong; Cicerone, Marcus T.

2017-01-01

Coherent anti-Stokes Raman scattering (CARS) microspectroscopy has demonstrated significant potential for biological and materials imaging. To date, however, the primary mechanism of disseminating CARS spectroscopic information is through pseudocolor imagery, which explicitly neglects a vast majority of the hyperspectral data. Furthermore, current paradigms in CARS spectral processing do not lend themselves to quantitative sample-to-sample comparability. The primary limitation stems from the need to accurately measure the so-called nonresonant background (NRB) that is used to extract the chemically-sensitive Raman information from the raw spectra. Measurement of the NRB on a pixel-by-pixel basis is a nontrivial task; thus, reference NRB from glass or water are typically utilized, resulting in error between the actual and estimated amplitude and phase. In this manuscript, we present a new methodology for extracting the Raman spectral features that significantly suppresses these errors through phase detrending and scaling. Classic methods of error-correction, such as baseline detrending, are demonstrated to be inaccurate and to simply mask the underlying errors. The theoretical justification is presented by re-developing the theory of phase retrieval via the Kramers-Kronig relation, and we demonstrate that these results are also applicable to maximum entropy method-based phase retrieval. This new error-correction approach is experimentally applied to glycerol spectra and tissue images, demonstrating marked consistency between spectra obtained using different NRB estimates, and between spectra obtained on different instruments. Additionally, in order to facilitate implementation of these approaches, we have made many of the tools described herein available free for download. PMID:28819335

Atomic scale imaging of competing polar states in a Ruddlesden-Popper layered oxide.

PubMed

Stone, Greg; Ophus, Colin; Birol, Turan; Ciston, Jim; Lee, Che-Hui; Wang, Ke; Fennie, Craig J; Schlom, Darrell G; Alem, Nasim; Gopalan, Venkatraman

2016-08-31

Layered complex oxides offer an unusually rich materials platform for emergent phenomena through many built-in design knobs such as varied topologies, chemical ordering schemes and geometric tuning of the structure. A multitude of polar phases are predicted to compete in Ruddlesden-Popper (RP), An+1BnO3n+1, thin films by tuning layer dimension (n) and strain; however, direct atomic-scale evidence for such competing states is currently absent. Using aberration-corrected scanning transmission electron microscopy with sub-Ångstrom resolution in Srn+1TinO3n+1 thin films, we demonstrate the coexistence of antiferroelectric, ferroelectric and new ordered and low-symmetry phases. We also directly image the atomic rumpling of the rock salt layer, a critical feature in RP structures that is responsible for the competing phases; exceptional quantitative agreement between electron microscopy and density functional theory is demonstrated. The study shows that layered topologies can enable multifunctionality through highly competitive phases exhibiting diverse phenomena in a single structure.

Atomic scale imaging of competing polar states in a Ruddlesden–Popper layered oxide

PubMed Central

Stone, Greg; Ophus, Colin; Birol, Turan; Ciston, Jim; Lee, Che-Hui; Wang, Ke; Fennie, Craig J.; Schlom, Darrell G.; Alem, Nasim; Gopalan, Venkatraman

2016-01-01

Layered complex oxides offer an unusually rich materials platform for emergent phenomena through many built-in design knobs such as varied topologies, chemical ordering schemes and geometric tuning of the structure. A multitude of polar phases are predicted to compete in Ruddlesden–Popper (RP), An+1BnO3n+1, thin films by tuning layer dimension (n) and strain; however, direct atomic-scale evidence for such competing states is currently absent. Using aberration-corrected scanning transmission electron microscopy with sub-Ångstrom resolution in Srn+1TinO3n+1 thin films, we demonstrate the coexistence of antiferroelectric, ferroelectric and new ordered and low-symmetry phases. We also directly image the atomic rumpling of the rock salt layer, a critical feature in RP structures that is responsible for the competing phases; exceptional quantitative agreement between electron microscopy and density functional theory is demonstrated. The study shows that layered topologies can enable multifunctionality through highly competitive phases exhibiting diverse phenomena in a single structure. PMID:27578622

Off-axis digital holographic camera for quantitative phase microscopy.

PubMed

Monemhaghdoust, Zahra; Montfort, Frédéric; Emery, Yves; Depeursinge, Christian; Moser, Christophe

2014-06-01

We propose and experimentally demonstrate a digital holographic camera which can be attached to the camera port of a conventional microscope for obtaining digital holograms in a self-reference configuration, under short coherence illumination and in a single shot. A thick holographic grating filters the beam containing the sample information in two dimensions through diffraction. The filtered beam creates the reference arm of the interferometer. The spatial filtering method, based on the high angular selectivity of the thick grating, reduces the alignment sensitivity to angular displacements compared with pinhole based Fourier filtering. The addition of a thin holographic grating alters the coherence plane tilt introduced by the thick grating so as to create high-visibility interference over the entire field of view. The acquired full-field off-axis holograms are processed to retrieve the amplitude and phase information of the sample. The system produces phase images of cheek cells qualitatively similar to phase images extracted with a standard commercial DHM.

Atomic scale imaging of competing polar states in a Ruddlesden-Popper layered oxide

NASA Astrophysics Data System (ADS)

Stone, Greg; Ophus, Colin; Birol, Turan; Ciston, Jim; Lee, Che-Hui; Wang, Ke; Fennie, Craig J.; Schlom, Darrell G.; Alem, Nasim; Gopalan, Venkatraman

2016-08-01

Layered complex oxides offer an unusually rich materials platform for emergent phenomena through many built-in design knobs such as varied topologies, chemical ordering schemes and geometric tuning of the structure. A multitude of polar phases are predicted to compete in Ruddlesden-Popper (RP), An+1BnO3n+1, thin films by tuning layer dimension (n) and strain; however, direct atomic-scale evidence for such competing states is currently absent. Using aberration-corrected scanning transmission electron microscopy with sub-Ångstrom resolution in Srn+1TinO3n+1 thin films, we demonstrate the coexistence of antiferroelectric, ferroelectric and new ordered and low-symmetry phases. We also directly image the atomic rumpling of the rock salt layer, a critical feature in RP structures that is responsible for the competing phases; exceptional quantitative agreement between electron microscopy and density functional theory is demonstrated. The study shows that layered topologies can enable multifunctionality through highly competitive phases exhibiting diverse phenomena in a single structure.

Tissue refractometry using Hilbert phase microscopy

PubMed Central

Lue, Niyom; Bewersdorf, Joerg; Lessard, Mark D.; Badizadegan, Kamran; Dasari, Ramachandra R.; Feld, Michael S.; Popescu, Gabriel

2009-01-01

We present, for the first time to our knowledge, quantitative phase images associated with unstained 5 μm thick tissue slices of mouse brain, spleen, and liver. The refractive properties of the tissue are retrieved in terms of the average refractive index and its spatial variation. We find that the average refractive index varies significantly with tissue type, such that the brain is characterized by the lowest value and the liver by the highest. The spatial power spectra of the phase images reveal power law behavior with different exponents for each tissue type. This approach opens a new possibility for stain-free characterization of tissues, where the diagnostic power is provided by the intrinsic refractive properties of the biological structure. We present results obtained for liver tissue affected by a lysosomal storage disease and show that our technique can quantify structural changes during this disease development. PMID:18087529

Quantitative description of the T1 morphology and strengthening mechanisms in an age-hardenable Al-Li-Cu alloy

NASA Astrophysics Data System (ADS)

Dorin, Thomas; Deschamps, Alexis; De Geuser, Frédéric; Weyland, Matthew

In the Al-Cu-Li system, the main strengthening precipitate is the T1 phase (Al2CuLi). In order to understand the strengthening related to the formation of this phase, we first present an investigation of the morphology of the T1 phase in an AA2198 alloy using Transmission Electron Microscopy (TEM) and Differential Scanning Calorimetry (DSC) in relation with the evolution of micro-hardness. In parallel, we present an investigation of the interaction between T1 precipitates and dislocations using High Angle Annular Dark Field (HAADF) imaging in an atomic resolution Scanning Transmission Electron Microscope (STEM). The atomic scale imaging of precipitates makes it possible to quantify the density of shearing events, which turns out to be insufficient to account for the imposed plastic strain. We discuss the implications of this result in terms of precipitate-dislocation interactions.

Qualitative and Quantitative Gadoxetic Acid-enhanced MR Imaging Helps Subtype Hepatocellular Adenomas.

PubMed

Tse, Justin R; Naini, Bita V; Lu, David S K; Raman, Steven S

2016-04-01

To determine which clinical variables and gadoxetic acid disodium (Gd-EOB-DTPA)-enhanced magnetic resonance (MR) imaging features are associated with histologically proved hepatocellular adenoma (HCA) genotypic subtypes. In this institutional review board-approved and Health Insurance Portability and Accountability Act-compliant study, clinical information and MR images of 49 histologically proved HCAs from January 2002 to December 2013 (21 patients; mean age, 39 years; age range, 15-59 years) were retrospectively reviewed by two radiologists. Qualitative and quantitative imaging features, including the signal intensity ratio relative to liver in each phase, were studied. HCA tissues were stained with subtype-specific markers and subclassified by a pathologist. Clinical and imaging data were correlated with pathologic findings and compared by using Fisher exact or t test, with a Bonferroni correction for multiple comparisons. Forty-nine HCAs were subclassified into 14 inflammatory, 20 hepatocyte nuclear factor (HNF)-1α-mutated, one β-catenin-activated, and 14 unclassified lesions. Intralesional steatosis was exclusively seen in HNF-1α-mutated lesions. Marked hyperintensity on T2-weighted images was seen in 12 of 14 (86%) inflammatory lesions compared with four of 21 (19%) HNF-1α-mutated, seven of 14 (50%) unclassified, and zero of one (0%) β-catenin-activated lesion. Two large lesions (one β-catenin-activated and one unclassified) transformed into hepatocellular carcinomas and were the only lesions to enhance with marked heterogeneity. In the hepatobiliary phase, all HCA subtypes were hypoenhancing compared with surrounding liver parenchyma, and they reached their nadir signal intensity by 10 minutes after the administration of contrast material before plateauing. HNF-1α-mutated lesions had the lowest lesion signal intensity ratio of 0.47 ± 0.09, compared with 0.73 ± 0.18 for inflammatory lesions (P = .0004), 0.82 for the β-catenin-activated lesion, and 0.73 ± 0.06 for the unclassified lesion (P = .00002). In this study, all HCA subtypes were hypoenhancing at Gd-EOB-DTPA-enhanced MR imaging in the hepatobiliary phase and reached their nadir signal intensity at 10 minutes. HNF-1α-mutated lesions could be distinguished from other subtypes by having the lowest lesion signal intensity ratio.

32-channel 3 Tesla receive-only phased-array head coil with soccer-ball element geometry.

PubMed

Wiggins, G C; Triantafyllou, C; Potthast, A; Reykowski, A; Nittka, M; Wald, L L

2006-07-01

A 32-channel 3T receive-only phased-array head coil was developed for human brain imaging. The helmet-shaped array was designed to closely fit the head with individual overlapping circular elements arranged in patterns of hexagonal and pentagonal symmetry similar to that of a soccer ball. The signal-to-noise ratio (SNR) and noise amplification (g-factor) in accelerated imaging applications were quantitatively evaluated in phantom and human images and compared with commercially available head coils. The 32-channel coil showed SNR gains of up to 3.5-fold in the cortex and 1.4-fold in the corpus callosum compared to a (larger) commercial eight-channel head coil. The experimentally measured g-factor performance of the helmet array showed significant improvement compared to the eight-channel array (peak g-factor 59% and 26% of the eight-channel values for four- and fivefold acceleration). The performance of the arrays is demonstrated in high-resolution and highly accelerated brain images. Copyright (c) 2006 Wiley-Liss, Inc.

Three-phase general border detection method for dermoscopy images using non-uniform illumination correction.

PubMed

Norton, Kerri-Ann; Iyatomi, Hitoshi; Celebi, M Emre; Ishizaki, Sumiko; Sawada, Mizuki; Suzaki, Reiko; Kobayashi, Ken; Tanaka, Masaru; Ogawa, Koichi

2012-08-01

Computer-aided diagnosis of dermoscopy images has shown great promise in developing a quantitative, objective way of classifying skin lesions. An important step in the classification process is lesion segmentation. Many studies have been successful in segmenting melanocytic skin lesions (MSLs), but few have focused on non-melanocytic skin lesions (NoMSLs), as the wide variety of lesions makes accurate segmentation difficult. We developed an automatic segmentation program for detecting borders of skin lesions in dermoscopy images. The method consists of a pre-processing phase, general lesion segmentation phase, including illumination correction, and bright region segmentation phase. We tested our method on a set of 107 NoMSLs and a set of 319 MSLs. Our method achieved precision/recall scores of 84.5% and 88.5% for NoMSLs, and 93.9% and 93.8% for MSLs, in comparison with manual extractions from four or five dermatologists. The accuracy of our method was competitive or better than five recently published methods. Our new method is the first method for detecting borders of both non-melanocytic and melanocytic skin lesions. © 2011 John Wiley & Sons A/S.

Optical Phase Measurements of Disorder Strength Link Microstructure to Cell Stiffness.

PubMed

Eldridge, Will J; Steelman, Zachary A; Loomis, Brianna; Wax, Adam

2017-02-28

There have been sustained efforts on the part of cell biologists to understand the mechanisms by which cells respond to mechanical stimuli. To this end, many rheological tools have been developed to characterize cellular stiffness. However, measurement of cellular viscoelastic properties has been limited in scope by the nature of most microrheological methods, which require direct mechanical contact, applied at the single-cell level. In this article, we describe, to our knowledge, a new analysis approach for quantitative phase imaging that relates refractive index variance to disorder strength, a parameter that is linked to cell stiffness. Significantly, both disorder strength and cell stiffness are measured with the same phase imaging system, presenting a unique alternative for label-free, noncontact, single-shot imaging of cellular rheologic properties. To demonstrate the potential applicability of the technique, we measure phase disorder strength and shear stiffness across five cellular populations with varying mechanical properties and demonstrate an inverse relationship between these two parameters. The existence of this relationship suggests that predictions of cell mechanical properties can be obtained from examining the disorder strength of cell structure using this, to our knowledge, novel, noncontact technique. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Abdominal 4D flow MR imaging in a breath hold: combination of spiral sampling and dynamic compressed sensing for highly accelerated acquisition.

PubMed

Dyvorne, Hadrien; Knight-Greenfield, Ashley; Jajamovich, Guido; Besa, Cecilia; Cui, Yong; Stalder, Aurélien; Markl, Michael; Taouli, Bachir

2015-04-01

To develop a highly accelerated phase-contrast cardiac-gated volume flow measurement (four-dimensional [4D] flow) magnetic resonance (MR) imaging technique based on spiral sampling and dynamic compressed sensing and to compare this technique with established phase-contrast imaging techniques for the quantification of blood flow in abdominal vessels. This single-center prospective study was compliant with HIPAA and approved by the institutional review board. Ten subjects (nine men, one woman; mean age, 51 years; age range, 30-70 years) were enrolled. Seven patients had liver disease. Written informed consent was obtained from all participants. Two 4D flow acquisitions were performed in each subject, one with use of Cartesian sampling with respiratory tracking and the other with use of spiral sampling and a breath hold. Cartesian two-dimensional (2D) cine phase-contrast images were also acquired in the portal vein. Two observers independently assessed vessel conspicuity on phase-contrast three-dimensional angiograms. Quantitative flow parameters were measured by two independent observers in major abdominal vessels. Intertechnique concordance was quantified by using Bland-Altman and logistic regression analyses. There was moderate to substantial agreement in vessel conspicuity between 4D flow acquisitions in arteries and veins (κ = 0.71 and 0.61, respectively, for observer 1; κ = 0.71 and 0.44 for observer 2), whereas more artifacts were observed with spiral 4D flow (κ = 0.30 and 0.20). Quantitative measurements in abdominal vessels showed good equivalence between spiral and Cartesian 4D flow techniques (lower bound of the 95% confidence interval: 63%, 77%, 60%, and 64% for flow, area, average velocity, and peak velocity, respectively). For portal venous flow, spiral 4D flow was in better agreement with 2D cine phase-contrast flow (95% limits of agreement: -8.8 and 9.3 mL/sec, respectively) than was Cartesian 4D flow (95% limits of agreement: -10.6 and 14.6 mL/sec). The combination of highly efficient spiral sampling with dynamic compressed sensing results in major acceleration for 4D flow MR imaging, which allows comprehensive assessment of abdominal vessel hemodynamics in a single breath hold.

Quantitative tracking of tumor cells in phase-contrast microscopy exploiting halo artifact pattern

NASA Astrophysics Data System (ADS)

Kang, Mi-Sun; Song, Soo-Min; Lee, Hana; Kim, Myoung-Hee

2012-03-01

Tumor cell morphology is closely related to its invasiveness characteristics and migratory behaviors. An invasive tumor cell has a highly irregular shape, whereas a spherical cell is non-metastatic. Thus, quantitative analysis of cell features is crucial to determine tumor malignancy or to test the efficacy of anticancer treatment. We use phase-contrast microscopy to analyze single cell morphology and to monitor its change because it enables observation of long-term activity of living cells without photobleaching and phototoxicity, which is common in other fluorescence-labeled microscopy. Despite this advantage, there are image-level drawbacks to phase-contrast microscopy, such as local light effect and contrast interference ring, among others. Thus, we first applied a local filter to compensate for non-uniform illumination. Then, we used intensity distribution information to detect the cell boundary. In phase-contrast microscopy images, the cell normally appears as a dark region surrounded by a bright halo. As the halo artifact around the cell body is minimal and has an asymmetric diffusion pattern, we calculated the cross-sectional plane that intersected the center of each cell and was orthogonal to the first principal axis. Then, we extracted the dark cell region by level set. However, a dense population of cultured cells still rendered single-cell analysis difficult. Finally, we measured roundness and size to classify tumor cells into malignant and benign groups. We validated segmentation accuracy by comparing our findings with manually obtained results.

Quantitative T1 and T2* carotid atherosclerotic plaque imaging using a three-dimensional multi-echo phase-sensitive inversion recovery sequence: a feasibility study.

PubMed

Fujiwara, Yasuhiro; Maruyama, Hirotoshi; Toyomaru, Kanako; Nishizaka, Yuri; Fukamatsu, Masahiro

2018-06-01

Magnetic resonance imaging (MRI) is widely used to detect carotid atherosclerotic plaques. Although it is important to evaluate vulnerable carotid plaques containing lipids and intra-plaque hemorrhages (IPHs) using T 1 -weighted images, the image contrast changes depending on the imaging settings. Moreover, to distinguish between a thrombus and a hemorrhage, it is useful to evaluate the iron content of the plaque using both T 1 -weighted and T 2 *-weighted images. Therefore, a quantitative evaluation of carotid atherosclerotic plaques using T 1 and T 2 * values may be necessary for the accurate evaluation of plaque components. The purpose of this study was to determine whether the multi-echo phase-sensitive inversion recovery (mPSIR) sequence can improve T 1 contrast while simultaneously providing accurate T 1 and T 2 * values of an IPH. T 1 and T 2 * values measured using mPSIR were compared to values from conventional methods in phantom and in vivo studies. In the phantom study, the T 1 and T 2 * values estimated using mPSIR were linearly correlated with those of conventional methods. In the in vivo study, mPSIR demonstrated higher T 1 contrast between the IPH phantom and sternocleidomastoid muscle than the conventional method. Moreover, the T 1 and T 2 * values of the blood vessel wall and sternocleidomastoid muscle estimated using mPSIR were correlated with values measured by conventional methods and with values reported previously. The mPSIR sequence improved T 1 contrast while simultaneously providing accurate T 1 and T 2 * values of the neck region. Although further study is required to evaluate the clinical utility, mPSIR may improve carotid atherosclerotic plaque detection and provide detailed information about plaque components.

Utility of Intermediate-Delay Washout CT Images for Differentiation of Malignant and Benign Adrenal Lesions: A Multivariate Analysis.

PubMed

Ng, Chaan S; Altinmakas, Emre; Wei, Wei; Ghosh, Payel; Li, Xiao; Grubbs, Elizabeth G; Perrier, Nancy D; Lee, Jeffrey E; Prieto, Victor G; Hobbs, Brian P

2018-06-27

The objective of this study was to identify features that impact the diagnostic performance of intermediate-delay washout CT for distinguishing malignant from benign adrenal lesions. This retrospective study evaluated 127 pathologically proven adrenal lesions (82 malignant, 45 benign) in 126 patients who had undergone portal venous phase and intermediate-delay washout CT (1-3 minutes after portal venous phase) with or without unenhanced images. Unenhanced images were available for 103 lesions. Quantitatively, lesion CT attenuation on unenhanced (UA) and delayed (DL) images, absolute and relative percentage of enhancement washout (APEW and RPEW, respectively), descriptive CT features (lesion size, margin characteristics, heterogeneity or homogeneity, fat, calcification), patient demographics, and medical history were evaluated for association with lesion status using multiple logistic regression with stepwise model selection. Area under the ROC curve (A z ) was calculated from both univariate and multivariate analyses. The predictive diagnostic performance of multivariate evaluations was ascertained through cross-validation. A z for DL, APEW, RPEW, and UA was 0.751, 0.795, 0.829, and 0.839, respectively. Multivariate analyses yielded the following significant CT quantitative features and associated A z when combined: RPEW and DL (A z = 0.861) when unenhanced images were not available and APEW and UA (A z = 0.889) when unenhanced images were available. Patient demographics and presence of a prior malignancy were additional significant factors, increasing A z to 0.903 and 0.927, respectively. The combined predictive classifier, without and with UA available, yielded 85.7% and 87.3% accuracies with cross-validation, respectively. When appropriately combined with other CT features, washout derived from intermediate-delay CT with or without additional clinical data has potential utility in differentiating malignant from benign adrenal lesions.

Poster - Thur Eve - 16: Four-dimensional x-ray computed tomography and hyperpolarized 3 He magnetic resonance imaging of gas distribution in lung cancer.

PubMed

Mathew, L; Castillo, R; Castillo, E; Yaremko, B; Rodrigues, G; Etemad-Rezai, R; Guerrero, T; Parraga, G

2012-07-01

Dynamic imaging methods such as four-dimensional computed tomography (4DCT) and static imaging methods such as noble gas magnetic resonance imaging (MRI) deliver direct and regional measurements of lung function even in lung cancer patients in whom global lung function measurements are dominated by tumour burden. The purpose of this study was to directly compare quantitative measurements of gas distribution from static hyperpolarized 3 He MRI and dynamic 4DCT in a small group of lung cancer patients. MRI and 4DCT were performed in 11 subjects prior to radiation therapy. MRI was performed at 3.0T in breath-hold after inhalation 1L of hyperpolarized 3 He gas. Gas distribution in 3 He MRI was quantified using a semi-automated segmentation algorithm to generate percent-ventilated volume (PVV), reflecting the volume of gas in the lung normalized to the thoracic cavity volume. 4DCT pulmonary function maps were generated using deformable image registration of six expiratory phase images. The correspondence between identical tissue elements at inspiratory and expiratory phases was used to estimate regional gas distribution and PVV was quantified from these images. After accounting for differences in lung volumes between 3 He MRI (1.9±0.5L ipsilateral, 2.3±0.7 contralateral) and 4DCT (1.2±0.3L ipsilateral, 1.3±0.4L contralateral) during image acquisition, there was no statistically significant difference in PVV between 3 He MRI (72±11% ipsilateral, 79±12% contralateral) and 4DCT (74±3% ipsilateral, 75±4% contralateral). Our results indicate quantitative agreement in the regional distribution of inhaled gas in both static and dynamic imaging methods. PVV may be considered as a regional surrogate measurement of lung function or ventilation. © 2012 American Association of Physicists in Medicine.

SU-E-J-219: Quantitative Evaluation of Motion Effects On Accuracy of Image-Guided Radiotherapy with Fiducial Markers Using CT Imaging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ali, I; Oyewale, S; Ahmad, S

2014-06-01

Purpose: To investigate quantitatively patient motion effects on the localization accuracy of image-guided radiation with fiducial markers using axial CT (ACT), helical CT (HCT) and cone-beam CT (CBCT) using modeling and experimental phantom studies. Methods: Markers with different lengths (2.5 mm, 5 mm, 10 mm, and 20 mm) were inserted in a mobile thorax phantom which was imaged using ACT, HCT and CBCT. The phantom moved with sinusoidal motion with amplitudes ranging 0–20 mm and a frequency of 15 cycles-per-minute. Three parameters that include: apparent marker lengths, center position and distance between the centers of the markers were measured inmore » the different CT images of the mobile phantom. A motion mathematical model was derived to predict the variations in the previous three parameters and their dependence on the motion in the different imaging modalities. Results: In CBCT, the measured marker lengths increased linearly with increase in motion amplitude. For example, the apparent length of the 10 mm marker was about 20 mm when phantom moved with amplitude of 5 mm. Although the markers have elongated, the center position and the distance between markers remained at the same position for different motion amplitudes in CBCT. These parameters were not affected by motion frequency and phase in CBCT. In HCT and ACT, the measured marker length, center and distance between markers varied irregularly with motion parameters. The apparent lengths of the markers varied with inverse of the phantom velocity which depends on motion frequency and phase. Similarly the center position and distance between markers varied inversely with phantom speed. Conclusion: Motion may lead to variations in maker length, center position and distance between markers using CT imaging. These effects should be considered in patient setup using image-guided radiation therapy based on fiducial markers matching using 2D-radiographs or volumetric CT imaging.« less

Imaging properties and its improvements of scanning/imaging x-ray microscope

DOE Office of Scientific and Technical Information (OSTI.GOV)

Takeuchi, Akihisa, E-mail: take@spring8.or.jp; Uesugi, Kentaro; Suzuki, Yoshio

A scanning / imaging X-ray microscope (SIXM) system has been developed at SPring-8. The SIXM consists of a scanning X-ray microscope with a one-dimensional (1D) X-ray focusing device and an imaging (full-field) X-ray microscope with a 1D X-ray objective. The motivation of the SIXM system is to realize a quantitative and highly-sensitive multimodal 3D X-ray tomography by taking advantages of both the scanning X-ray microscope using multi-pixel detector and the imaging X-ray microscope. Data acquisition process of a 2D image is completely different between in the horizontal direction and in the vertical direction; a 1D signal is obtained with themore » linear-scanning while the other dimensional signal is obtained with the imaging optics. Such condition have caused a serious problem on the imaging properties that the imaging quality in the vertical direction has been much worse than that in the horizontal direction. In this paper, two approaches to solve this problem will be presented. One is introducing a Fourier transform method for phase retrieval from one phase derivative image, and the other to develop and employ a 1D diffuser to produce an asymmetrical coherent illumination.« less

Dynamic quantitative analysis of adherent cell cultures by means of lens-free video microscopy

NASA Astrophysics Data System (ADS)

Allier, C.; Vincent, R.; Navarro, F.; Menneteau, M.; Ghenim, L.; Gidrol, X.; Bordy, T.; Hervé, L.; Cioni, O.; Bardin, S.; Bornens, M.; Usson, Y.; Morales, S.

2018-02-01

We present our implementation of lens-free video microscopy setup for the monitoring of adherent cell cultures. We use a multi-wavelength LED illumination together with a dedicated holographic reconstruction algorithm that allows for an efficient removal of twin images from the reconstructed phase image for densities up to those of confluent cell cultures (>500 cells/mm2). We thereby demonstrate that lens-free video microscopy, with a large field of view ( 30 mm2) can enable us to capture the images of thousands of cells simultaneously and directly inside the incubator. It is then possible to trace and quantify single cells along several cell cycles. We thus prove that lens-free microscopy is a quantitative phase imaging technique enabling estimation of several metrics at the single cell level as a function of time, for example the area, dry mass, maximum thickness, major axis length and aspect ratio of each cell. Combined with cell tracking, it is then possible to extract important parameters such as the initial cell dry mass (just after cell division), the final cell dry mass (just before cell division), the average cell growth rate, and the cell cycle duration. As an example, we discuss the monitoring of a HeLa cell cultures which provided us with a data-set featuring more than 10 000 cell cycle tracks and more than 2x106 cell morphological measurements in a single time-lapse.

A biomechanical modeling-guided simultaneous motion estimation and image reconstruction technique (SMEIR-Bio) for 4D-CBCT reconstruction

NASA Astrophysics Data System (ADS)

Huang, Xiaokun; Zhang, You; Wang, Jing

2018-02-01

Reconstructing four-dimensional cone-beam computed tomography (4D-CBCT) images directly from respiratory phase-sorted traditional 3D-CBCT projections can capture target motion trajectory, reduce motion artifacts, and reduce imaging dose and time. However, the limited numbers of projections in each phase after phase-sorting decreases CBCT image quality under traditional reconstruction techniques. To address this problem, we developed a simultaneous motion estimation and image reconstruction (SMEIR) algorithm, an iterative method that can reconstruct higher quality 4D-CBCT images from limited projections using an inter-phase intensity-driven motion model. However, the accuracy of the intensity-driven motion model is limited in regions with fine details whose quality is degraded due to insufficient projection number, which consequently degrades the reconstructed image quality in corresponding regions. In this study, we developed a new 4D-CBCT reconstruction algorithm by introducing biomechanical modeling into SMEIR (SMEIR-Bio) to boost the accuracy of the motion model in regions with small fine structures. The biomechanical modeling uses tetrahedral meshes to model organs of interest and solves internal organ motion using tissue elasticity parameters and mesh boundary conditions. This physics-driven approach enhances the accuracy of solved motion in the organ’s fine structures regions. This study used 11 lung patient cases to evaluate the performance of SMEIR-Bio, making both qualitative and quantitative comparisons between SMEIR-Bio, SMEIR, and the algebraic reconstruction technique with total variation regularization (ART-TV). The reconstruction results suggest that SMEIR-Bio improves the motion model’s accuracy in regions containing small fine details, which consequently enhances the accuracy and quality of the reconstructed 4D-CBCT images.

Novel dental dynamic depth profilometric imaging using simultaneous frequency-domain infrared photothermal radiometry and laser luminescence

NASA Astrophysics Data System (ADS)

Nicolaides, Lena; Mandelis, Andreas

2000-01-01

A high-spatial-resolution dynamic experimental imaging setup, which can provide simultaneous measurements of laser- induced frequency-domain infrared photothermal radiometric and luminescence signals from defects in teeth, has been developed for the first time. The major findings of this work are: (1) radiometric images are complementary to (anticorrelated with) luminescence images, as a result of the nature of the two physical signal generation processes; (2) the radiometric amplitude exhibits much superior dynamic (signal resolution) range to luminescence in distinguishing between intact and cracked sub-surface structures in the enamel; (3) the radiometric signal (amplitude and phase) produces dental images with much better defect localization, delineation, and resolution; (4) radiometric images (amplitude and phase) at a fixed modulation frequency are depth profilometric, whereas luminescence images are not; and (5) luminescence frequency responses from enamel and hydroxyapatite exhibit two relaxation lifetimes, the longer of which (approximately ms) is common to all and is not sensitive to the defect state and overall quality of the enamel. Simultaneous radiometric and luminescence frequency scans for the purpose of depth profiling were performed and a quantitative theoretical two-lifetime rate model of dental luminescence was advanced.

Non-destructive evaluation of teeth restored with different composite resins using synchrotron based micro-imaging.

PubMed

Fatima, A; Kulkarni, V K; Banda, N R; Agrawal, A K; Singh, B; Sarkar, P S; Tripathi, S; Shripathi, T; Kashyap, Y; Sinha, A

2016-01-01

Application of high resolution synchrotron micro-imaging in microdefects studies of restored dental samples. The purpose of this study was to identify and compare the defects in restorations done by two different resin systems on teeth samples using synchrotron based micro-imaging techniques namely Phase Contrast Imaging (PCI) and micro-computed tomography (MCT). With this aim acquired image quality was also compared with routinely used RVG (Radiovisiograph). Crowns of human teeth samples were fractured mechanically involving only enamel and dentin, without exposure of pulp chamber and were divided into two groups depending on the restorative composite materials used. Group A samples were restored using a submicron Hybrid composite material and Group B samples were restored using a Nano-Hybrid restorative composite material. Synchrotron based PCI and MCT was performed with the aim of visualization of tooth structure, composite resin and their interface. The quantitative and qualitative comparison of phase contrast and absorption contrast images along with MCT on the restored teeth samples shows comparatively large number of voids in Group A samples. Quality assessment of dental restorations using synchrotron based micro-imaging suggests Nano-Hybrid resin restorations (Group B) are better than Group A.

High-throughput, label-free, single-cell, microalgal lipid screening by machine-learning-equipped optofluidic time-stretch quantitative phase microscopy.

PubMed

Guo, Baoshan; Lei, Cheng; Kobayashi, Hirofumi; Ito, Takuro; Yalikun, Yaxiaer; Jiang, Yiyue; Tanaka, Yo; Ozeki, Yasuyuki; Goda, Keisuke

2017-05-01

The development of reliable, sustainable, and economical sources of alternative fuels to petroleum is required to tackle the global energy crisis. One such alternative is microalgal biofuel, which is expected to play a key role in reducing the detrimental effects of global warming as microalgae absorb atmospheric CO 2 via photosynthesis. Unfortunately, conventional analytical methods only provide population-averaged lipid amounts and fail to characterize a diverse population of microalgal cells with single-cell resolution in a non-invasive and interference-free manner. Here high-throughput label-free single-cell screening of lipid-producing microalgal cells with optofluidic time-stretch quantitative phase microscopy was demonstrated. In particular, Euglena gracilis, an attractive microalgal species that produces wax esters (suitable for biodiesel and aviation fuel after refinement), within lipid droplets was investigated. The optofluidic time-stretch quantitative phase microscope is based on an integration of a hydrodynamic-focusing microfluidic chip, an optical time-stretch quantitative phase microscope, and a digital image processor equipped with machine learning. As a result, it provides both the opacity and phase maps of every single cell at a high throughput of 10,000 cells/s, enabling accurate cell classification without the need for fluorescent staining. Specifically, the dataset was used to characterize heterogeneous populations of E. gracilis cells under two different culture conditions (nitrogen-sufficient and nitrogen-deficient) and achieve the cell classification with an error rate of only 2.15%. The method holds promise as an effective analytical tool for microalgae-based biofuel production. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

SU-F-R-21: The Stability of Radiomics Features On 4D FDG-PET/CT Images

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ma, C

2016-06-15

Purpose: The aim of our study was to perform a stability analysis of 4D PET-derived features in non-small cell lung carcinoma (NSCLC) based on six different respiratory phases. Methods: The 4D FDG-PET/CT respiratory phases were labeled as T0%, T17%, T33%,T50%, T67%, T83% phases, with the T0% phase approximately corresponding to the normal end-inspiration. Lesions were manually delineated based on fused PET-CT, using a standardized clinical delineation protocol. Six texture parameters were analyzed. Results: Results showed that the majority of assessed features had a low stability such as Homogeneity (0.385–0.416), Dissimilarity (3.707–3.861), Angular two moments (0.013–0.019), Contrast (39.782–49.562), Entropy(4.683–5.002) and Inversemore » differential moment (0.317–0.362) on different respiratory phases. Conclusion: This study suggest that further research of quantitative PET imaging features is warranted with respect to respiratory motion.« less

An Assessment of Imaging Informatics for Precision Medicine in Cancer.

PubMed

Chennubhotla, C; Clarke, L P; Fedorov, A; Foran, D; Harris, G; Helton, E; Nordstrom, R; Prior, F; Rubin, D; Saltz, J H; Shalley, E; Sharma, A

2017-08-01

Objectives: Precision medicine requires the measurement, quantification, and cataloging of medical characteristics to identify the most effective medical intervention. However, the amount of available data exceeds our current capacity to extract meaningful information. We examine the informatics needs to achieve precision medicine from the perspective of quantitative imaging and oncology. Methods: The National Cancer Institute (NCI) organized several workshops on the topic of medical imaging and precision medicine. The observations and recommendations are summarized herein. Results: Recommendations include: use of standards in data collection and clinical correlates to promote interoperability; data sharing and validation of imaging tools; clinician's feedback in all phases of research and development; use of open-source architecture to encourage reproducibility and reusability; use of challenges which simulate real-world situations to incentivize innovation; partnership with industry to facilitate commercialization; and education in academic communities regarding the challenges involved with translation of technology from the research domain to clinical utility and the benefits of doing so. Conclusions: This article provides a survey of the role and priorities for imaging informatics to help advance quantitative imaging in the era of precision medicine. While these recommendations were drawn from oncology, they are relevant and applicable to other clinical domains where imaging aids precision medicine. Georg Thieme Verlag KG Stuttgart.

Chemoenzymatic method for glycomics: isolation, identification, and quantitation

PubMed Central

Yang, Shuang; Rubin, Abigail; Eshghi, Shadi Toghi; Zhang, Hui

2015-01-01

Over the past decade, considerable progress has been made with respect to the analytical methods for analysis of glycans from biological sources. Regardless of the specific methods that are used, glycan analysis includes isolation, identification, and quantitation. Derivatization is indispensable to increase their identification. Derivatization of glycans can be performed by permethylation or carbodiimide coupling / esterification. By introducing a fluorophore or chromophore at their reducing end, glycans can be separated by electrophoresis or chromatography. The fluorogenically labeled glycans can be quantitated using fluorescent detection. The recently developed approaches using solid-phase such as glycoprotein immobilization for glycan extraction and on-tissue glycan mass spectrometry imaging demonstrate advantages over methods performed in solution. Derivatization of sialic acids is favorably implemented on the solid support using carbodiimide coupling, and the released glycans can be further modified at the reducing end or permethylated for quantitative analysis. In this review, methods for glycan isolation, identification, and quantitation are discussed. PMID:26390280

Impact of CT attenuation correction method on quantitative respiratory-correlated (4D) PET/CT imaging

PubMed Central

Lee, Tzu-Cheng; Alessio, Adam M.; Wollenweber, Scott D.; Stearns, Charles W.; Bowen, Stephen R.; Kinahan, Paul E.

2015-01-01

Purpose: Respiratory-correlated positron emission tomography (PET/CT) 4D PET/CT is used to mitigate errors from respiratory motion; however, the optimal CT attenuation correction (CTAC) method for 4D PET/CT is unknown. The authors performed a phantom study to evaluate the quantitative performance of CTAC methods for 4D PET/CT in the ground truth setting. Methods: A programmable respiratory motion phantom with a custom movable insert designed to emulate a lung lesion and lung tissue was used for this study. The insert was driven by one of five waveforms: two sinusoidal waveforms or three patient-specific respiratory waveforms. 3DPET and 4DPET images of the phantom under motion were acquired and reconstructed with six CTAC methods: helical breath-hold (3DHEL), helical free-breathing (3DMOT), 4D phase-averaged (4DAVG), 4D maximum intensity projection (4DMIP), 4D phase-matched (4DMATCH), and 4D end-exhale (4DEXH) CTAC. Recovery of SUVmax, SUVmean, SUVpeak, and segmented tumor volume was evaluated as RCmax, RCmean, RCpeak, and RCvol, representing percent difference relative to the static ground truth case. Paired Wilcoxon tests and Kruskal–Wallis ANOVA were used to test for significant differences. Results: For 4DPET imaging, the maximum intensity projection CTAC produced significantly more accurate recovery coefficients than all other CTAC methods (p < 0.0001 over all metrics). Over all motion waveforms, ratios of 4DMIP CTAC recovery were 0.2 ± 5.4, −1.8 ± 6.5, −3.2 ± 5.0, and 3.0 ± 5.9 for RCmax, RCpeak, RCmean, and RCvol. In comparison, recovery coefficients for phase-matched CTAC were −8.4 ± 5.3, −10.5 ± 6.2, −7.6 ± 5.0, and −13.0 ± 7.7 for RCmax, RCpeak, RCmean, and RCvol. When testing differences between phases over all CTAC methods and waveforms, end-exhale phases were significantly more accurate (p = 0.005). However, these differences were driven by the patient-specific respiratory waveforms; when testing patient and sinusoidal waveforms separately, patient waveforms were significantly different between phases (p < 0.0001) while the sinusoidal waveforms were not significantly different (p = 0.98). When considering only the subset of 4DMATCH images that corresponded to the end-exhale image phase, 4DEXH, mean and interquartile range were similar to 4DMATCH but variability was considerably reduced. Conclusions: Comparative advantages in accuracy and precision of SUV metrics and segmented volumes were demonstrated with the use of the maximum intensity projection and end-exhale CT attenuation correction. While respiratory phase-matched CTAC should in theory provide optimal corrections, image artifacts and differences in implementation of 4DCT and 4DPET sorting can degrade the benefit of this approach. These results may be useful to guide the implementation, analysis, and development of respiratory-correlated thoracic PET/CT in the radiation oncology and diagnostic settings. PMID:25563252

Ultrasound capsule endoscopy: sounding out the future

PubMed Central

Stewart, Fraser; Lay, Holly; Cummins, Gerard; Newton, Ian P.; Desmulliez, Marc P. Y.; Steele, Robert J. C.; Näthke, Inke; Cochran, Sandy

2017-01-01

Video capsule endoscopy (VCE) has been of immense benefit in the diagnosis and management of gastrointestinal (GI) disorders since its introduction in 2001. However, it suffers from a number of well recognized deficiencies. Amongst these is the limited capability of white light imaging, which is restricted to analysis of the mucosal surface. Current capsule endoscopes are dependent on visual manifestation of disease and limited in regards to transmural imaging and detection of deeper pathology. Ultrasound capsule endoscopy (USCE) has the potential to overcome surface only imaging and provide transmural scans of the GI tract. The integration of high frequency microultrasound (µUS) into capsule endoscopy would allow high resolution transmural images and provide a means of both qualitative and quantitative assessment of the bowel wall. Quantitative ultrasound (QUS) can provide data in an objective and measurable manner, potentially reducing lengthy interpretation times by incorporation into an automated diagnostic process. The research described here is focused on the development of USCE and other complementary diagnostic and therapeutic modalities. Presently investigations have entered a preclinical phase with laboratory investigations running concurrently. PMID:28567381

Surface plasmon resonance microscopy: achieving a quantitative optical response

PubMed Central

Peterson, Alexander W.; Halter, Michael; Plant, Anne L.; Elliott, John T.

2016-01-01

Surface plasmon resonance (SPR) imaging allows real-time label-free imaging based on index of refraction, and changes in index of refraction at an interface. Optical parameter analysis is achieved by application of the Fresnel model to SPR data typically taken by an instrument in a prism based configuration. We carry out SPR imaging on a microscope by launching light into a sample, and collecting reflected light through a high numerical aperture microscope objective. The SPR microscope enables spatial resolution that approaches the diffraction limit, and has a dynamic range that allows detection of subnanometer to submicrometer changes in thickness of biological material at a surface. However, unambiguous quantitative interpretation of SPR changes using the microscope system could not be achieved using the Fresnel model because of polarization dependent attenuation and optical aberration that occurs in the high numerical aperture objective. To overcome this problem, we demonstrate a model to correct for polarization diattenuation and optical aberrations in the SPR data, and develop a procedure to calibrate reflectivity to index of refraction values. The calibration and correction strategy for quantitative analysis was validated by comparing the known indices of refraction of bulk materials with corrected SPR data interpreted with the Fresnel model. Subsequently, we applied our SPR microscopy method to evaluate the index of refraction for a series of polymer microspheres in aqueous media and validated the quality of the measurement with quantitative phase microscopy. PMID:27782542

Capturing the crystalline phase of two-dimensional nanocrystal superlattices in action.

PubMed

Jiang, Zhang; Lin, Xiao-Min; Sprung, Michael; Narayanan, Suresh; Wang, Jin

2010-03-10

Critical photonic, electronic, and magnetic applications of two-dimensional nanocrystal superlattices often require nanostructures in perfect single-crystal phases with long-range order and limited defects. Here we discovered a crystalline phase with quasi-long-range positional order for two-dimensional nanocrystal superlattice domains self-assembled at the liquid-air interface during droplet evaporation, using in situ time-resolved X-ray scattering along with rigorous theories on two dimensional crystal structures. Surprisingly, it was observed that drying these superlattice domains preserved only an orientational order but not a long-range positional order, also supported by quantitative analysis of transmission electron microscopy images.

Quantitative evaluation of 3D images produced from computer-generated holograms

NASA Astrophysics Data System (ADS)

Sheerin, David T.; Mason, Ian R.; Cameron, Colin D.; Payne, Douglas A.; Slinger, Christopher W.

1999-08-01

Advances in computing and optical modulation techniques now make it possible to anticipate the generation of near real- time, reconfigurable, high quality, three-dimensional images using holographic methods. Computer generated holography (CGH) is the only technique which holds promise of producing synthetic images having the full range of visual depth cues. These realistic images will be viewable by several users simultaneously, without the need for headtracking or special glasses. Such a data visualization tool will be key to speeding up the manufacture of new commercial and military equipment by negating the need for the production of physical 3D models in the design phase. DERA Malvern has been involved in designing and testing fixed CGH in order to understand the connection between the complexity of the CGH, the algorithms used to design them, the processes employed in their implementation and the quality of the images produced. This poster describes results from CGH containing up to 108 pixels. The methods used to evaluate the reconstructed images are discussed and quantitative measures of image fidelity made. An understanding of the effect of the various system parameters upon final image quality enables a study of the possible system trade-offs to be carried out. Such an understanding of CGH production and resulting image quality is key to effective implementation of a reconfigurable CGH system currently under development at DERA.

SU-E-J-226: Propagation of Pancreas Target Contours On Respiratory Correlated CT Images Using Deformable Image Registration

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, F; Yorke, E; Mageras, G

2014-06-01

Purpose: Respiratory Correlated CT (RCCT) scans to assess intra-fraction motion among pancreatic cancer patients undergoing radiotherapy allow for dose sparing of normal tissues, in particular for the duodenum. Contour propagation of the gross tumor volume (GTV) from one reference respiratory phase to 9 other phases is time consuming. Deformable image registration (DIR) has been successfully used for high contrast disease sites but lower contrast for pancreatic tumors may compromise accuracy. This study evaluates the accuracy of Fast Free Form (FFF) registration-based contour propagation of the GTV on RCCT scans of pancreas cancer patients. Methods: Twenty-four pancreatic cancer patients were retrospectivelymore » studied; 20 had tumors in the pancreatic head/neck, 4 in the body/tail. Patients were simulated with RCCT and images were sorted into 10 respiratory phases. A radiation oncologist manually delineated the GTV for 5 phases (0%, 30%, 50%, 70% and 90%). The FFF algorithm was used to map deformations between the EE (50%) phase and each of the other 4 phases. The resultant deformation fields served to propagate GTV contours from EE to the other phases. The Dice Similarity Coefficient (DSC), which measures agreement between the DIR-propagated and manually-delineated GTVs, was used to quantitatively examine DIR accuracy. Results: Average DSC over all scans and patients is 0.82 and standard deviation is 0.09 (DSC range 0.97–0.57). For GTV volumes above and below the median volume of 20.2 cc, a Wilcoxon rank-sum test shows significantly different DSC (p=0.0000002). For the GTVs above the median volume, average +/− SD is 0.85 +/− 0.07; and for the GTVs below, the average +/− SD is 0.75 +/−0.08. Conclusion: For pancreatic tumors, the FFF DIR algorithm accurately propagated the GTV between the images in different phases of RCCT, with improved performance for larger tumors.« less

3D visualization of two-phase flow in the micro-tube by a simple but effective method

NASA Astrophysics Data System (ADS)

Fu, X.; Zhang, P.; Hu, H.; Huang, C. J.; Huang, Y.; Wang, R. Z.

2009-08-01

The present study provides a simple but effective method for 3D visualization of the two-phase flow in the micro-tube. An isosceles right-angle prism combined with a mirror located 45° bevel to the prism is employed to synchronously obtain the front and side views of the flow patterns with a single camera, where the locations of the prism and the micro-tube for clear imaging should satisfy a fixed relationship which is specified in the present study. The optical design is proven successfully by the tough visualization work at the cryogenic temperature range. The image deformation due to the refraction and geometrical configuration of the test section is quantitatively investigated. It is calculated that the image is enlarged by about 20% in inner diameter compared to the real object, which is validated by the experimental results. Meanwhile, the image deformation by adding a rectangular optical correction box outside the circular tube is comparatively investigated. It is calculated that the image is reduced by about 20% in inner diameter with a rectangular optical correction box compared to the real object. The 3D re-construction process based on the two views is conducted through three steps, which shows that the 3D visualization method can easily be applied for two-phase flow research in micro-scale channels and improves the measurement accuracy of some important parameters of the two-phase flow such as void fraction, spatial distribution of bubbles, etc.

Phase measurement error in summation of electron holography series.

PubMed

McLeod, Robert A; Bergen, Michael; Malac, Marek

2014-06-01

Off-axis electron holography is a method for the transmission electron microscope (TEM) that measures the electric and magnetic properties of a specimen. The electrostatic and magnetic potentials modulate the electron wavefront phase. The error in measurement of the phase therefore determines the smallest observable changes in electric and magnetic properties. Here we explore the summation of a hologram series to reduce the phase error and thereby improve the sensitivity of electron holography. Summation of hologram series requires independent registration and correction of image drift and phase wavefront drift, the consequences of which are discussed. Optimization of the electro-optical configuration of the TEM for the double biprism configuration is examined. An analytical model of image and phase drift, composed of a combination of linear drift and Brownian random-walk, is derived and experimentally verified. The accuracy of image registration via cross-correlation and phase registration is characterized by simulated hologram series. The model of series summation errors allows the optimization of phase error as a function of exposure time and fringe carrier frequency for a target spatial resolution. An experimental example of hologram series summation is provided on WS2 fullerenes. A metric is provided to measure the object phase error from experimental results and compared to analytical predictions. The ultimate experimental object root-mean-square phase error is 0.006 rad (2π/1050) at a spatial resolution less than 0.615 nm and a total exposure time of 900 s. The ultimate phase error in vacuum adjacent to the specimen is 0.0037 rad (2π/1700). The analytical prediction of phase error differs with the experimental metrics by +7% inside the object and -5% in the vacuum, indicating that the model can provide reliable quantitative predictions. Crown Copyright © 2014. Published by Elsevier B.V. All rights reserved.

Quantitative shear wave imaging optical coherence tomography for noncontact mechanical characterization of myocardium

NASA Astrophysics Data System (ADS)

Wang, Shang; Lopez, Andrew L.; Morikawa, Yuka; Tao, Ge; Li, Jiasong; Larina, Irina V.; Martin, James F.; Larin, Kirill V.

2015-03-01

Optical coherence elastography (OCE) is an emerging low-coherence imaging technique that provides noninvasive assessment of tissue biomechanics with high spatial resolution. Among various OCE methods, the capability of quantitative measurement of tissue elasticity is of great importance for tissue characterization and pathology detection across different samples. Here we report a quantitative OCE technique, termed quantitative shear wave imaging optical coherence tomography (Q-SWI-OCT), which enables noncontact measurement of tissue Young's modulus based on the ultra-fast imaging of the shear wave propagation inside the sample. A focused air-puff device is used to interrogate the tissue with a low-pressure short-duration air stream that stimulates a localized displacement with the scale at micron level. The propagation of this tissue deformation in the form of shear wave is captured by a phase-sensitive OCT system running with the scan of the M-mode imaging over the path of the wave propagation. The temporal characteristics of the shear wave is quantified based on the cross-correlation of the tissue deformation profiles at all the measurement locations, and linear regression is utilized to fit the data plotted in the domain of time delay versus wave propagation distance. The wave group velocity is thus calculated, which results in the quantitative measurement of the Young's modulus. As the feasibility demonstration, experiments are performed on tissuemimicking phantoms with different agar concentrations and the quantified elasticity values with Q-SWI-OCT agree well with the uniaxial compression tests. For functional characterization of myocardium with this OCE technique, we perform our pilot experiments on ex vivo mouse cardiac muscle tissues with two studies, including 1) elasticity difference of cardiac muscle under relaxation and contract conditions and 2) mechanical heterogeneity of the heart introduced by the muscle fiber orientation. Our results suggest the potential of using Q-SWI-OCT as an essential tool for nondestructive biomechanical evaluation of myocardium.

Errors in quantitative backscattered electron analysis of bone standardized by energy-dispersive x-ray spectrometry.

PubMed

Vajda, E G; Skedros, J G; Bloebaum, R D

1998-10-01

Backscattered electron (BSE) imaging has proven to be a useful method for analyzing the mineral distribution in microscopic regions of bone. However, an accepted method of standardization has not been developed, limiting the utility of BSE imaging for truly quantitative analysis. Previous work has suggested that BSE images can be standardized by energy-dispersive x-ray spectrometry (EDX). Unfortunately, EDX-standardized BSE images tend to underestimate the mineral content of bone when compared with traditional ash measurements. The goal of this study is to investigate the nature of the deficit between EDX-standardized BSE images and ash measurements. A series of analytical standards, ashed bone specimens, and unembedded bone specimens were investigated to determine the source of the deficit previously reported. The primary source of error was found to be inaccurate ZAF corrections to account for the organic phase of the bone matrix. Conductive coatings, methylmethacrylate embedding media, and minor elemental constituents in bone mineral introduced negligible errors. It is suggested that the errors would remain constant and an empirical correction could be used to account for the deficit. However, extensive preliminary testing of the analysis equipment is essential.

New Trends in Radionuclide Myocardial Perfusion Imaging

PubMed Central

Hung, Guang-Uei; Wang, Yuh-Feng; Su, Hung-Yi; Hsieh, Te-Chun; Ko, Chi-Lun; Yen, Ruoh-Fang

2016-01-01

Radionuclide myocardial perfusion imaging (MPI) with single photon emission computed tomography (SPECT) has been widely used clinically as one of the major functional imaging modalities for patients with coronary artery disease (CAD) for decades. Ample evidence has supported the use of MPI as a useful and important tool in the diagnosis, risk stratification and treatment planning for CAD. Although popular in the United States, MPI has become the most frequently used imaging modality among all nuclear medicine tests in Taiwan. However, it should be acknowledged that MPI SPECT does have its limitations. These include false-positive results due to certain artifacts, false-negative due to balanced ischemia, complexity and adverse reaction arising from current pharmacological stressors, time consuming nature of the imaging procedure, no blood flow quantitation and relatively high radiation exposure. The purpose of this article was to review the recent trends in nuclear cardiology, including the utilization of positron emission tomography (PET) for MPI, new stressor, new SPECT camera with higher resolution and higher sensitivity, dynamic SPECT protocol for blood flow quantitation, new software of phase analysis for evaluation of LV dyssynchrony, and measures utilized for reducing radiation exposure of MPI. PMID:27122946

Magnetic particle imaging: from proof of principle to preclinical applications

NASA Astrophysics Data System (ADS)

Knopp, T.; Gdaniec, N.; Möddel, M.

2017-07-01

Tomographic imaging has become a mandatory tool for the diagnosis of a majority of diseases in clinical routine. Since each method has its pros and cons, a variety of them is regularly used in clinics to satisfy all application needs. Magnetic particle imaging (MPI) is a relatively new tomographic imaging technique that images magnetic nanoparticles with a high spatiotemporal resolution in a quantitative way, and in turn is highly suited for vascular and targeted imaging. MPI was introduced in 2005 and now enters the preclinical research phase, where medical researchers get access to this new technology and exploit its potential under physiological conditions. Within this paper, we review the development of MPI since its introduction in 2005. Besides an in-depth description of the basic principles, we provide detailed discussions on imaging sequences, reconstruction algorithms, scanner instrumentation and potential medical applications.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stassi, D.; Ma, H.; Schmidt, T. G., E-mail: taly.gilat-schmidt@marquette.edu

Purpose: Reconstructing a low-motion cardiac phase is expected to improve coronary artery visualization in coronary computed tomography angiography (CCTA) exams. This study developed an automated algorithm for selecting the optimal cardiac phase for CCTA reconstruction. The algorithm uses prospectively gated, single-beat, multiphase data made possible by wide cone-beam imaging. The proposed algorithm differs from previous approaches because the optimal phase is identified based on vessel image quality (IQ) directly, compared to previous approaches that included motion estimation and interphase processing. Because there is no processing of interphase information, the algorithm can be applied to any sampling of image phases, makingmore » it suited for prospectively gated studies where only a subset of phases are available. Methods: An automated algorithm was developed to select the optimal phase based on quantitative IQ metrics. For each reconstructed slice at each reconstructed phase, an image quality metric was calculated based on measures of circularity and edge strength of through-plane vessels. The image quality metric was aggregated across slices, while a metric of vessel-location consistency was used to ignore slices that did not contain through-plane vessels. The algorithm performance was evaluated using two observer studies. Fourteen single-beat cardiac CT exams (Revolution CT, GE Healthcare, Chalfont St. Giles, UK) reconstructed at 2% intervals were evaluated for best systolic (1), diastolic (6), or systolic and diastolic phases (7) by three readers and the algorithm. Pairwise inter-reader and reader-algorithm agreement was evaluated using the mean absolute difference (MAD) and concordance correlation coefficient (CCC) between the reader and algorithm-selected phases. A reader-consensus best phase was determined and compared to the algorithm selected phase. In cases where the algorithm and consensus best phases differed by more than 2%, IQ was scored by three readers using a five point Likert scale. Results: There was no statistically significant difference between inter-reader and reader-algorithm agreement for either MAD or CCC metrics (p > 0.1). The algorithm phase was within 2% of the consensus phase in 15/21 of cases. The average absolute difference between consensus and algorithm best phases was 2.29% ± 2.47%, with a maximum difference of 8%. Average image quality scores for the algorithm chosen best phase were 4.01 ± 0.65 overall, 3.33 ± 1.27 for right coronary artery (RCA), 4.50 ± 0.35 for left anterior descending (LAD) artery, and 4.50 ± 0.35 for left circumflex artery (LCX). Average image quality scores for the consensus best phase were 4.11 ± 0.54 overall, 3.44 ± 1.03 for RCA, 4.39 ± 0.39 for LAD, and 4.50 ± 0.18 for LCX. There was no statistically significant difference (p > 0.1) between the image quality scores of the algorithm phase and the consensus phase. Conclusions: The proposed algorithm was statistically equivalent to a reader in selecting an optimal cardiac phase for CCTA exams. When reader and algorithm phases differed by >2%, image quality as rated by blinded readers was statistically equivalent. By detecting the optimal phase for CCTA reconstruction, the proposed algorithm is expected to improve coronary artery visualization in CCTA exams.« less

Nuclear medicine and imaging research (instrumentation and quantitative methods of evaluation): Comprehensive 3-year progress report for the period January 15, 1986-January 14, 1989

DOE Office of Scientific and Technical Information (OSTI.GOV)

Beck, R.N.; Cooper, M.D.

1988-06-01

This program addresses the problems involving the basic science and technology underlying the physical and conceptual tools of radioactive tracer methodology as they relate to the measurement of structural and functional parameters of physiologic importance in health and disease. The principal tool is quantitative radionuclide imaging. The overall objective of this program is to further the development and transfer of radiotracer methodology from basic theory to routine clinical practice in order that individual patients and society as a whole will receive the maximum net benefit from the new knowledge gained. The focus of the research is on the development ofmore » new instruments and radiopharmaceuticals, and the evaluation of these through the phase of clinical feasibility. 58 refs., 15 figs., 4 tabs.« less

A novel dual gating approach using joint inertial sensors: implications for cardiac PET imaging

NASA Astrophysics Data System (ADS)

Jafari Tadi, Mojtaba; Teuho, Jarmo; Lehtonen, Eero; Saraste, Antti; Pänkäälä, Mikko; Koivisto, Tero; Teräs, Mika

2017-10-01

Positron emission tomography (PET) is a non-invasive imaging technique which may be considered as the state of art for the examination of cardiac inflammation due to atherosclerosis. A fundamental limitation of PET is that cardiac and respiratory motions reduce the quality of the achieved images. Current approaches for motion compensation involve gating the PET data based on the timing of quiescent periods of cardiac and respiratory cycles. In this study, we present a novel gating method called microelectromechanical (MEMS) dual gating which relies on joint non-electrical sensors, i.e. tri-axial accelerometer and gyroscope. This approach can be used for optimized selection of quiescent phases of cardiac and respiratory cycles. Cardiomechanical activity according to echocardiography observations was investigated to confirm whether this dual sensor solution can provide accurate trigger timings for cardiac gating. Additionally, longitudinal chest motions originating from breathing were measured by accelerometric- and gyroscopic-derived respiratory (ADR and GDR) tracking. The ADR and GDR signals were evaluated against Varian real-time position management (RPM) signals in terms of amplitude and phase. Accordingly, high linear correlation and agreement were achieved between the reference electrocardiography, RPM, and measured MEMS signals. We also performed a Ge-68 phantom study to evaluate possible metal artifacts caused by the integrated read-out electronics including mechanical sensors and semiconductors. The reconstructed phantom images did not reveal any image artifacts. Thus, it was concluded that MEMS-driven dual gating can be used in PET studies without an effect on the quantitative or visual accuracy of the PET images. Finally, the applicability of MEMS dual gating for cardiac PET imaging was investigated with two atherosclerosis patients. Dual gated PET images were successfully reconstructed using only MEMS signals and both qualitative and quantitative assessments revealed encouraging results that warrant further investigation of this method.

Feasibility Study of Using Gemstone Spectral Imaging (GSI) and Adaptive Statistical Iterative Reconstruction (ASIR) for Reducing Radiation and Iodine Contrast Dose in Abdominal CT Patients with High BMI Values.

PubMed

Zhu, Zheng; Zhao, Xin-ming; Zhao, Yan-feng; Wang, Xiao-yi; Zhou, Chun-wu

2015-01-01

To prospectively investigate the effect of using Gemstone Spectral Imaging (GSI) and adaptive statistical iterative reconstruction (ASIR) for reducing radiation and iodine contrast dose in abdominal CT patients with high BMI values. 26 patients (weight > 65kg and BMI ≥ 22) underwent abdominal CT using GSI mode with 300mgI/kg contrast material as study group (group A). Another 21 patients (weight ≤ 65kg and BMI ≥ 22) were scanned with a conventional 120 kVp tube voltage for noise index (NI) of 11 with 450mgI/kg contrast material as control group (group B). GSI images were reconstructed at 60keV with 50%ASIR and the conventional 120kVp images were reconstructed with FBP reconstruction. The CT values, standard deviation (SD), signal-noise-ratio (SNR), contrast-noise-ratio (CNR) of 26 landmarks were quantitatively measured and image quality qualitatively assessed using statistical analysis. As for the quantitative analysis, the difference of CNR between groups A and B was all significant except for the mesenteric vein. The SNR in group A was higher than B except the mesenteric artery and splenic artery. As for the qualitative analysis, all images had diagnostic quality and the agreement for image quality assessment between the reviewers was substantial (kappa = 0.684). CT dose index (CTDI) values for non-enhanced, arterial phase and portal phase in group A were decreased by 49.04%, 40.51% and 40.54% compared with group B (P = 0.000), respectively. The total dose and the injection rate for the contrast material were reduced by 14.40% and 14.95% in A compared with B. The use of GSI and ASIR provides similar enhancement in vessels and image quality with reduced radiation dose and contrast dose, compared with the use of conventional scan protocol.

Controlling the near-field excitation of nano-antennas with phase-change materials.

PubMed

Kao, Tsung Sheng; Chen, Yi Guo; Hong, Ming Hui

2013-01-01

By utilizing the strongly induced plasmon coupling between discrete nano-antennas and quantitatively controlling the crystalline proportions of an underlying Ge2Sb2Te5 (GST) phase-change thin layer, we show that nanoscale light localizations in the immediate proximity of plasmonic nano-antennas can be spatially positioned. Isolated energy hot-spots at a subwavelength scale can be created and adjusted across the landscape of the plasmonic system at a step resolution of λ/20. These findings introduce a new approach for nano-circuitry, bio-assay addressing and imaging applications.

Spin echo SPI methods for quantitative analysis of fluids in porous media.

PubMed

Li, Linqing; Han, Hui; Balcom, Bruce J

2009-06-01

Fluid density imaging is highly desirable in a wide variety of porous media measurements. The SPRITE class of MRI methods has proven to be robust and general in their ability to generate density images in porous media, however the short encoding times required, with correspondingly high magnetic field gradient strengths and filter widths, and low flip angle RF pulses, yield sub-optimal S/N images, especially at low static field strength. This paper explores two implementations of pure phase encode spin echo 1D imaging, with application to a proposed new petroleum reservoir core analysis measurement. In the first implementation of the pulse sequence, we modify the spin echo single point imaging (SE-SPI) technique to acquire the k-space origin data point, with a near zero evolution time, from the free induction decay (FID) following a 90 degrees excitation pulse. Subsequent k-space data points are acquired by separately phase encoding individual echoes in a multi-echo acquisition. T(2) attenuation of the echo train yields an image convolution which causes blurring. The T(2) blur effect is moderate for porous media with T(2) lifetime distributions longer than 5 ms. As a robust, high S/N, and fast 1D imaging method, this method will be highly complementary to SPRITE techniques for the quantitative analysis of fluid content in porous media. In the second implementation of the SE-SPI pulse sequence, modification of the basic measurement permits fast determination of spatially resolved T(2) distributions in porous media through separately phase encoding each echo in a multi-echo CPMG pulse train. An individual T(2) weighted image may be acquired from each echo. The echo time (TE) of each T(2) weighted image may be reduced to 500 micros or less. These profiles can be fit to extract a T(2) distribution from each pixel employing a variety of standard inverse Laplace transform methods. Fluid content 1D images are produced as an essential by product of determining the spatially resolved T(2) distribution. These 1D images do not suffer from a T(2) related blurring. The above SE-SPI measurements are combined to generate 1D images of the local saturation and T(2) distribution as a function of saturation, upon centrifugation of petroleum reservoir core samples. The logarithm mean T(2) is observed to shift linearly with water saturation. This new reservoir core analysis measurement may provide a valuable calibration of the Coates equation for irreducible water saturation, which has been widely implemented in NMR well logging measurements.

Dual-energy-X-ray imaging to measure phase volume fractions in a transient multiphase flow

NASA Astrophysics Data System (ADS)

Loewen, Eric Paul

1999-12-01

The objective of this research was to visualize the pre-mixing phase of a fuel-coolant interaction (FCI) by using combinations of high-speed cinematography and dual energy X-ray imaging to identify and quantify the spatial and temporal characteristics of the three FCI phases---metal (fuel), liquid (coolant water), and voids (generated steam). (1) The high-speed cinematography imaging subsystem and the low-energy X-ray imaging subsystem provided visual photographs and distinguished generated voids from water. (2) The high-energy X-Ray imaging subsystem provided additional discernment of metal from water and vapor. This is the first time that dynamic dual X-ray images have been provided with quantitative results. The data provide new information concerning the melt fractions, melt jet configuration, melt jet velocity, and qualitative spatial and temporal quantification of the pre-mixing event. This information provides new insight into the FCI phenomenon that could not have been deduced from visible-light imaging or other instrumentation such as thermocouples, void sensors, or pressure transmitters. Significant findings include: (1) the fuel column (molten Pb jet) penetrated deeply (<7 cm) into the coolant (water) while maintaining its columnar shape. (2) Energetic FCIs occurred (and were imaged) below the melt-coolant interface temperature equal to the homogenous nucleation temperature (310°C). (3) The molten jet breakup was observed to be caused by hydrodynamic forces. (4) The Pb/water thermal interaction zone was imaged over melt temperatures from 330°C to 640°C and coolant subcooling of 4°C to 80°C. (5) The interface regions between the molten Pb and coolant was observed to grow with decreasing coolant subcooling. This imaging process can be applied to further study of the FCI phenomena at other test facilities. It can also be applied for observation of other two- or three-phase flow phenomena previously opaque to conventional imaging systems.

Susceptibility Tensor Imaging (STI) of the Brain

PubMed Central

Li, Wei; Liu, Chunlei; Duong, Timothy Q.; van Zijl, Peter C.M.; Li, Xu

2016-01-01

Susceptibility tensor imaging (STI) is a recently developed MRI technique that allows quantitative determination of orientation-independent magnetic susceptibility parameters from the dependence of gradient echo signal phase on the orientation of biological tissues with respect to the main magnetic field. By modeling the magnetic susceptibility of each voxel as a symmetric rank-2 tensor, individual magnetic susceptibility tensor elements as well as the mean magnetic susceptibility (MMS) and magnetic susceptibility anisotropy (MSA) can be determined for brain tissues that would still show orientation dependence after conventional scalar-based quantitative susceptibility mapping (QSM) to remove such dependence. Similar to diffusion tensor imaging (DTI), STI allows mapping of brain white matter fiber orientations and reconstruction of 3D white matter pathways using the principal eigenvectors of the susceptibility tensor. In contrast to diffusion anisotropy, the main determinant factor of susceptibility anisotropy in brain white matter is myelin. Another unique feature of susceptibility anisotropy of white matter is its sensitivity to gadolinium-based contrast agents. Mechanistically, MRI-observed susceptibility anisotropy is mainly attributed to the highly ordered lipid molecules in myelin sheath. STI provides a consistent interpretation of the dependence of phase and susceptibility on orientation at multiple scales. This article reviews the key experimental findings and physical theories that led to the development of STI, its practical implementations, and its applications for brain research. PMID:27120169

Fracture mechanics by three-dimensional crack-tip synchrotron X-ray microscopy

PubMed Central

Withers, P. J.

2015-01-01

To better understand the relationship between the nucleation and growth of defects and the local stresses and phase changes that cause them, we need both imaging and stress mapping. Here, we explore how this can be achieved by bringing together synchrotron X-ray diffraction and tomographic imaging. Conventionally, these are undertaken on separate synchrotron beamlines; however, instruments capable of both imaging and diffraction are beginning to emerge, such as ID15 at the European Synchrotron Radiation Facility and JEEP at the Diamond Light Source. This review explores the concept of three-dimensional crack-tip X-ray microscopy, bringing them together to probe the crack-tip behaviour under realistic environmental and loading conditions and to extract quantitative fracture mechanics information about the local crack-tip environment. X-ray diffraction provides information about the crack-tip stress field, phase transformations, plastic zone and crack-face tractions and forces. Time-lapse CT, besides providing information about the three-dimensional nature of the crack and its local growth rate, can also provide information as to the activation of extrinsic toughening mechanisms such as crack deflection, crack-tip zone shielding, crack bridging and crack closure. It is shown how crack-tip microscopy allows a quantitative measure of the crack-tip driving force via the stress intensity factor or the crack-tip opening displacement. Finally, further opportunities for synchrotron X-ray microscopy are explored. PMID:25624521

Colorectal carcinoma: Ex vivo evaluation using 3-T high-spatial-resolution quantitative T2 mapping and its correlation with histopathologic findings.

PubMed

Yamada, Ichiro; Yoshino, Norio; Hikishima, Keigo; Miyasaka, Naoyuki; Yamauchi, Shinichi; Uetake, Hiroyuki; Yasuno, Masamichi; Saida, Yukihisa; Tateishi, Ukihide; Kobayashi, Daisuke; Eishi, Yoshinobu

2017-05-01

In this study, we aimed to evaluate the feasibility of determining the mural invasion depths of colorectal carcinomas using high-spatial-resolution (HSR) quantitative T2 mapping on a 3-T magnetic resonance (MR) scanner. Twenty colorectal specimens containing adenocarcinomas were imaged on a 3-T MR system equipped with a 4-channel phased-array surface coil. HSR quantitative T2 maps were acquired using a spin-echo sequence with a repetition time/echo time of 7650/22.6-361.6ms (16 echoes), 87×43.5-mm field of view, 2-mm section thickness, 448×224 matrix, and average of 1. HSR fast-spin-echo T2-weighted images were also acquired. Differences between the T2 values (ms) of the tumor tissue, colorectal wall layers, and fibrosis were measured, and the MR images and histopathologic findings were compared. In all specimens (20/20, 100%), the HSR quantitative T2 maps clearly depicted an 8-layer normal colorectal wall in which the T2 values of each layer differed from those of the adjacent layer(s) (P<0.001). Using this technique, fibrosis (73.6±9.4ms) and tumor tissue (104.2±6.4ms) could also be clearly differentiated (P<0.001). In 19 samples (95%), the HSR quantitative T2 maps and histopathologic data yielded the same findings regarding the tumor invasion depth. Our results indicate that 3-T HSR quantitative T2 mapping is useful for distinguishing colorectal wall layers and differentiating tumor and fibrotic tissues. Accordingly, this technique could be used to determine mural invasion by colorectal carcinomas with a high level of accuracy. Copyright © 2017 Elsevier Inc. All rights reserved.

Comparison of amplitude-decorrelation, speckle-variance and phase-variance OCT angiography methods for imaging the human retina and choroid

PubMed Central

Gorczynska, Iwona; Migacz, Justin V.; Zawadzki, Robert J.; Capps, Arlie G.; Werner, John S.

2016-01-01

We compared the performance of three OCT angiography (OCTA) methods: speckle variance, amplitude decorrelation and phase variance for imaging of the human retina and choroid. Two averaging methods, split spectrum and volume averaging, were compared to assess the quality of the OCTA vascular images. All data were acquired using a swept-source OCT system at 1040 nm central wavelength, operating at 100,000 A-scans/s. We performed a quantitative comparison using a contrast-to-noise (CNR) metric to assess the capability of the three methods to visualize the choriocapillaris layer. For evaluation of the static tissue noise suppression in OCTA images we proposed to calculate CNR between the photoreceptor/RPE complex and the choriocapillaris layer. Finally, we demonstrated that implementation of intensity-based OCT imaging and OCT angiography methods allows for visualization of retinal and choroidal vascular layers known from anatomic studies in retinal preparations. OCT projection imaging of data flattened to selected retinal layers was implemented to visualize retinal and choroidal vasculature. User guided vessel tracing was applied to segment the retinal vasculature. The results were visualized in a form of a skeletonized 3D model. PMID:27231598

Atomic scale dynamics of a solid state chemical reaction directly determined by annular dark-field electron microscopy.

PubMed

Pennycook, Timothy J; Jones, Lewys; Pettersson, Henrik; Coelho, João; Canavan, Megan; Mendoza-Sanchez, Beatriz; Nicolosi, Valeria; Nellist, Peter D

2014-12-22

Dynamic processes, such as solid-state chemical reactions and phase changes, are ubiquitous in materials science, and developing a capability to observe the mechanisms of such processes on the atomic scale can offer new insights across a wide range of materials systems. Aberration correction in scanning transmission electron microscopy (STEM) has enabled atomic resolution imaging at significantly reduced beam energies and electron doses. It has also made possible the quantitative determination of the composition and occupancy of atomic columns using the atomic number (Z)-contrast annular dark-field (ADF) imaging available in STEM. Here we combine these benefits to record the motions and quantitative changes in the occupancy of individual atomic columns during a solid-state chemical reaction in manganese oxides. These oxides are of great interest for energy-storage applications such as for electrode materials in pseudocapacitors. We employ rapid scanning in STEM to both drive and directly observe the atomic scale dynamics behind the transformation of Mn3O4 into MnO. The results demonstrate we now have the experimental capability to understand the complex atomic mechanisms involved in phase changes and solid state chemical reactions.

Extracting morphologies from third harmonic generation images of structurally normal human brain tissue.

PubMed

Zhang, Zhiqing; Kuzmin, Nikolay V; Groot, Marie Louise; de Munck, Jan C

2017-06-01

The morphologies contained in 3D third harmonic generation (THG) images of human brain tissue can report on the pathological state of the tissue. However, the complexity of THG brain images makes the usage of modern image processing tools, especially those of image filtering, segmentation and validation, to extract this information challenging. We developed a salient edge-enhancing model of anisotropic diffusion for image filtering, based on higher order statistics. We split the intrinsic 3-phase segmentation problem into two 2-phase segmentation problems, each of which we solved with a dedicated model, active contour weighted by prior extreme. We applied the novel proposed algorithms to THG images of structurally normal ex-vivo human brain tissue, revealing key tissue components-brain cells, microvessels and neuropil, enabling statistical characterization of these components. Comprehensive comparison to manually delineated ground truth validated the proposed algorithms. Quantitative comparison to second harmonic generation/auto-fluorescence images, acquired simultaneously from the same tissue area, confirmed the correctness of the main THG features detected. The software and test datasets are available from the authors. z.zhang@vu.nl. Supplementary data are available at Bioinformatics online. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

High-resolution breast tomography at high energy: a feasibility study of phase contrast imaging on a whole breast

NASA Astrophysics Data System (ADS)

Sztrókay, A.; Diemoz, P. C.; Schlossbauer, T.; Brun, E.; Bamberg, F.; Mayr, D.; Reiser, M. F.; Bravin, A.; Coan, P.

2012-05-01

Previous studies on phase contrast imaging (PCI) mammography have demonstrated an enhancement of breast morphology and cancerous tissue visualization compared to conventional imaging. We show here the first results of the PCI analyser-based imaging (ABI) in computed tomography (CT) mode on whole and large (>12 cm) tumour-bearing breast tissues. We demonstrate in this work the capability of the technique of working at high x-ray energies and producing high-contrast images of large and complex specimens. One entire breast of an 80-year-old woman with invasive ductal cancer was imaged using ABI-CT with monochromatic 70 keV x-rays and an area detector of 92×92 µm2 pixel size. Sagittal slices were reconstructed from the acquired data, and compared to corresponding histological sections. Comparison with conventional absorption-based CT was also performed. Five blinded radiologists quantitatively evaluated the visual aspects of the ABI-CT images with respect to sharpness, soft tissue contrast, tissue boundaries and the discrimination of different structures/tissues. ABI-CT excellently depicted the entire 3D architecture of the breast volume by providing high-resolution and high-contrast images of the normal and cancerous breast tissues. These results are an important step in the evolution of PCI-CT towards its clinical implementation.

Using X-Ray In-Line Phase-Contrast Imaging for the Investigation of Nude Mouse Hepatic Tumors

PubMed Central

Zhang, Lu; Luo, Shuqian

2012-01-01

The purpose of this paper is to report the noninvasive imaging of hepatic tumors without contrast agents. Both normal tissues and tumor tissues can be detected, and tumor tissues in different stages can be classified quantitatively. We implanted BEL-7402 human hepatocellular carcinoma cells into the livers of nude mice and then imaged the livers using X-ray in-line phase-contrast imaging (ILPCI). The projection images' texture feature based on gray level co-occurrence matrix (GLCM) and dual-tree complex wavelet transforms (DTCWT) were extracted to discriminate normal tissues and tumor tissues. Different stages of hepatic tumors were classified using support vector machines (SVM). Images of livers from nude mice sacrificed 6 days after inoculation with cancer cells show diffuse distribution of the tumor tissue, but images of livers from nude mice sacrificed 9, 12, or 15 days after inoculation with cancer cells show necrotic lumps in the tumor tissue. The results of the principal component analysis (PCA) of the texture features based on GLCM of normal regions were positive, but those of tumor regions were negative. The results of PCA of the texture features based on DTCWT of normal regions were greater than those of tumor regions. The values of the texture features in low-frequency coefficient images increased monotonically with the growth of the tumors. Different stages of liver tumors can be classified using SVM, and the accuracy is 83.33%. Noninvasive and micron-scale imaging can be achieved by X-ray ILPCI. We can observe hepatic tumors and small vessels from the phase-contrast images. This new imaging approach for hepatic cancer is effective and has potential use in the early detection and classification of hepatic tumors. PMID:22761929

Multimodality hard-x-ray imaging of a chromosome with nanoscale spatial resolution

DOE PAGES

Yan, Hanfei; Nazaretski, Evgeny; Lauer, Kenneth R.; ...

2016-02-05

Here, we developed a scanning hard x-ray microscope using a new class of x-ray nano-focusing optic called a multilayer Laue lens and imaged a chromosome with nanoscale spatial resolution. The combination of the hard x-ray's superior penetration power, high sensitivity to elemental composition, high spatial-resolution and quantitative analysis creates a unique tool with capabilities that other microscopy techniques cannot provide. Using this microscope, we simultaneously obtained absorption-, phase-, and fluorescence-contrast images of Pt-stained human chromosome samples. The high spatial-resolution of the microscope and its multi-modality imaging capabilities enabled us to observe the internal ultra-structures of a thick chromosome without sectioningmore » it.« less

Multimodality hard-x-ray imaging of a chromosome with nanoscale spatial resolution

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yan, Hanfei; Nazaretski, Evgeny; Lauer, Kenneth R.

Here, we developed a scanning hard x-ray microscope using a new class of x-ray nano-focusing optic called a multilayer Laue lens and imaged a chromosome with nanoscale spatial resolution. The combination of the hard x-ray's superior penetration power, high sensitivity to elemental composition, high spatial-resolution and quantitative analysis creates a unique tool with capabilities that other microscopy techniques cannot provide. Using this microscope, we simultaneously obtained absorption-, phase-, and fluorescence-contrast images of Pt-stained human chromosome samples. The high spatial-resolution of the microscope and its multi-modality imaging capabilities enabled us to observe the internal ultra-structures of a thick chromosome without sectioningmore » it.« less

A quantitative experimental phantom study on MRI image uniformity.

PubMed

Felemban, Doaa; Verdonschot, Rinus G; Iwamoto, Yuri; Uchiyama, Yuka; Kakimoto, Naoya; Kreiborg, Sven; Murakami, Shumei

2018-05-23

Our goal was to assess MR image uniformity by investigating aspects influencing said uniformity via a method laid out by the National Electrical Manufacturers Association (NEMA). Six metallic materials embedded in a glass phantom were scanned (i.e. Au, Ag, Al, Au-Ag-Pd alloy, Ti and Co-Cr alloy) as well as a reference image. Sequences included spin echo (SE) and gradient echo (GRE) scanned in three planes (i.e. axial, coronal, and sagittal). Moreover, three surface coil types (i.e. head and neck, Brain, and temporomandibular joint coils) and two image correction methods (i.e. surface coil intensity correction or SCIC, phased array uniformity enhancement or PURE) were employed to evaluate their effectiveness on image uniformity. Image uniformity was assessed using the National Electrical Manufacturers Association peak-deviation non-uniformity method. Results showed that temporomandibular joint coils elicited the least uniform image and brain coils outperformed head and neck coils when metallic materials were present. Additionally, when metallic materials were present, spin echo outperformed gradient echo especially for Co-Cr (particularly in the axial plane). Furthermore, both SCIC and PURE improved image uniformity compared to uncorrected images, and SCIC slightly surpassed PURE when metallic metals were present. Lastly, Co-Cr elicited the least uniform image while other metallic materials generally showed similar patterns (i.e. no significant deviation from images without metallic metals). Overall, a quantitative understanding of the factors influencing MR image uniformity (e.g. coil type, imaging method, metal susceptibility, and post-hoc correction method) is advantageous to optimize image quality, assists clinical interpretation, and may result in improved medical and dental care.

Two Phase Flow Measurements by Nuclear Magnetic Resonance (NMR)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Altobelli, Stephen A; Fukushima, Eiichi

In concentrated suspensions, there is a tendency for the solid phase to migrate from regions of high shear rate to regions of low shear (Leighton & Acrivos, 1987). In the early years that our effort was funded by the DOE Division of Basic Energy Science, quantitative measurement of this process in neutrally buoyant suspensions was a major focus (Abbott, et al., 1991; Altobelli, et al., 1991). Much of this work was used to improve multi-phase numerical models at Sandia National Laboratories. Later, our collaborators at Sandia and the University of New Mexico incorporated body forces into their numerical models ofmore » suspension flow (Rao, Mondy, Sun, et al., 2002). We developed experiments that allow us to study flows driven by buoyancy, to characterize these flows in well-known and useful engineering terms (Altobelli and Mondy, 2002) and to begin to explore the less well-understood area of flows with multiple solid phases (Beyea, Altobelli, et al., 2003). We also studied flows that combine the effects of shear and buoyancy, and flows of suspensions made from non-Newtonian liquids (Rao, Mondy, Baer, et al, 2002). We were able to demonstrate the usefulness of proton NMR imaging of liquid phase concentration and velocity and produced quantitative data not obtainable by other methods. Fluids flowing through porous solids are important in geophysics and in chemical processing. NMR techniques have been widely used to study liquid flow in porous media. We pioneered the extension of these studies to gas flows (Koptyug, et al, 2000, 2000, 2001, 2002). This extension allows us to investigate a wider range of Peclet numbers, and to gather data on problems of interest in catalysis. We devised two kinds of NMR experiments for three-phase systems. Both experiments employ two NMR visible phases and one phase that gives no NMR signal. The earlier method depends on the two visible phases differing in a NMR relaxation property. The second method (Beyea, Altobelli, et al., 2003) uses two different nuclei, protons and 19F. It also uses two different types of NMR image formation, a conventional spin-echo and a single-point method. The single-point method is notable for being useful for imaging materials which are much more rigid than can usually be studied by NMR imaging. We use it to image “low density” polyethylene (LDPE) plastic in this application. We have reduced the imaging time for this three-phase imaging method to less than 10 s per pair of profiles by using new hardware. Directly measuring the solid LDPE signal was a novel feature for multi-phase flow studies. We also used thermally polarized gas NMR (as opposed to hyper-polarized gas) which produces low signal to noise ratios because gas densities are on the order of 1000 times smaller than liquid densities. However since we used multi-atom molecules that have short T1's and operated at elevated pressures we could overcome some of the losses. Thermally polarized gases have advantages over hyperpolarized gases in the ease of preparation, and in maintaining a well-defined polarization. In these studies (Codd and Altobelli, 2003), we used stimulated echo sequences to successfully obtain propagators of gas in bead packs out to observation times of 300 ms. Zarraga, et al. (2000) used laser-sheet profilometry to investigate normal stress differences in concentrated suspensions. Recently we developed an NMR imaging analog for comparison with numerical work that is being performed by Rekha Rao at Sandia National Laboratories (Rao, Mondy, Sun, et al, 2002). A neutrally buoyant suspension of 100 mm PMMA spheres in a Newtonian liquid was sheared in a vertical Couette apparatus inside the magnet. The outer cylinder rotates and the inner cylinder is fixed. At these low rotation rates, the free-surface of the Newtonian liquid shows no measurable deformation, but the suspension clearly shows its non-Newtonian character.« less

Response to Comment on "Is the nuclear refractive index lower than cytoplasm? Validation of phase measurements and implications for light scattering technologies": A Comment on "How a phase image of a cell with nucleus refractive index smaller than that of the cytoplasm should look like?", e201800033.

PubMed

Steelman, Zachary A; Eldridge, Will J; Wax, Adam

2018-06-01

Recently, Maxim A. Yurkin commented on our paper "Is the nuclear refractive index lower than cytoplasm? Validation of phase measurements and implications for light scattering technologies" as well as on a complementary study "Cell nuclei have lower refractive index and mass density than cytoplasm" from Schürmann et al. In his comment, Yurkin concluded that quantitative phase images of cells with nuclei that are less optically dense than the cytoplasm must exhibit a characteristic concavity, the absence of which is evidence against our conclusion of a less-dense nucleus. In this response, we suggest that Yurkin's conclusion is reached through an oversimplification of the spatial refractive index distribution within cells, which does not account for high index inclusions such as the nucleolus. We further cite recent studies in 3-dimensional refractive index imaging, in which the preponderance of studies supports our conclusion. Finally, we comment on the current state of knowledge regarding subcellular refractive index distributions in living cells. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Investigation of BOLD fMRI Resonance Frequency Shifts and Quantitative Susceptibility Changes at 7 T

PubMed Central

Bianciardi, Marta; van Gelderen, Peter; Duyn, Jeff H.

2013-01-01

Although blood oxygenation level dependent (BOLD) functional magnetic resonance imaging (fMRI) experiments of brain activity generally rely on the magnitude of the signal, they also provide frequency information that can be derived from the phase of the signal. However, because of confounding effects of instrumental and physiological origin, BOLD related frequency information is difficult to extract and therefore rarely used. Here, we explored the use of high field (7 T) and dedicated signal processing methods to extract frequency information and use it to quantify and interpret blood oxygenation and blood volume changes. We found that optimized preprocessing improves detection of task-evoked and spontaneous changes in phase signals and resonance frequency shifts over large areas of the cortex with sensitivity comparable to that of magnitude signals. Moreover, our results suggest the feasibility of mapping BOLD quantitative susceptibility changes in at least part of the activated area and its largest draining veins. Comparison with magnitude data suggests that the observed susceptibility changes originate from neuronal activity through induced blood volume and oxygenation changes in pial and intracortical veins. Further, from frequency shifts and susceptibility values, we estimated that, relative to baseline, the fractional oxygen saturation in large vessels increased by 0.02–0.05 during stimulation, which is consistent to previously published estimates. Together, these findings demonstrate that valuable information can be derived from fMRI imaging of BOLD frequency shifts and quantitative susceptibility changes. PMID:23897623

Automated selection of the optimal cardiac phase for single-beat coronary CT angiography reconstruction.

PubMed

Stassi, D; Dutta, S; Ma, H; Soderman, A; Pazzani, D; Gros, E; Okerlund, D; Schmidt, T G

2016-01-01

Reconstructing a low-motion cardiac phase is expected to improve coronary artery visualization in coronary computed tomography angiography (CCTA) exams. This study developed an automated algorithm for selecting the optimal cardiac phase for CCTA reconstruction. The algorithm uses prospectively gated, single-beat, multiphase data made possible by wide cone-beam imaging. The proposed algorithm differs from previous approaches because the optimal phase is identified based on vessel image quality (IQ) directly, compared to previous approaches that included motion estimation and interphase processing. Because there is no processing of interphase information, the algorithm can be applied to any sampling of image phases, making it suited for prospectively gated studies where only a subset of phases are available. An automated algorithm was developed to select the optimal phase based on quantitative IQ metrics. For each reconstructed slice at each reconstructed phase, an image quality metric was calculated based on measures of circularity and edge strength of through-plane vessels. The image quality metric was aggregated across slices, while a metric of vessel-location consistency was used to ignore slices that did not contain through-plane vessels. The algorithm performance was evaluated using two observer studies. Fourteen single-beat cardiac CT exams (Revolution CT, GE Healthcare, Chalfont St. Giles, UK) reconstructed at 2% intervals were evaluated for best systolic (1), diastolic (6), or systolic and diastolic phases (7) by three readers and the algorithm. Pairwise inter-reader and reader-algorithm agreement was evaluated using the mean absolute difference (MAD) and concordance correlation coefficient (CCC) between the reader and algorithm-selected phases. A reader-consensus best phase was determined and compared to the algorithm selected phase. In cases where the algorithm and consensus best phases differed by more than 2%, IQ was scored by three readers using a five point Likert scale. There was no statistically significant difference between inter-reader and reader-algorithm agreement for either MAD or CCC metrics (p > 0.1). The algorithm phase was within 2% of the consensus phase in 15/21 of cases. The average absolute difference between consensus and algorithm best phases was 2.29% ± 2.47%, with a maximum difference of 8%. Average image quality scores for the algorithm chosen best phase were 4.01 ± 0.65 overall, 3.33 ± 1.27 for right coronary artery (RCA), 4.50 ± 0.35 for left anterior descending (LAD) artery, and 4.50 ± 0.35 for left circumflex artery (LCX). Average image quality scores for the consensus best phase were 4.11 ± 0.54 overall, 3.44 ± 1.03 for RCA, 4.39 ± 0.39 for LAD, and 4.50 ± 0.18 for LCX. There was no statistically significant difference (p > 0.1) between the image quality scores of the algorithm phase and the consensus phase. The proposed algorithm was statistically equivalent to a reader in selecting an optimal cardiac phase for CCTA exams. When reader and algorithm phases differed by >2%, image quality as rated by blinded readers was statistically equivalent. By detecting the optimal phase for CCTA reconstruction, the proposed algorithm is expected to improve coronary artery visualization in CCTA exams.

Imaging of blood antigen distribution on blood cells by thermal lens microscopy

NASA Astrophysics Data System (ADS)

Kimura, Hiroko; Sekiguchi, Kazuya; Nagao, Fumiko; Mukaida, Masahiro; Kitamori, Takehiko; Sawada, Tsuguo

2000-05-01

Blood group antigens on a cell were measured by a new microscopic method, i.e. thermal lens microscopy which involves spectrometry using a laser-induced thermal-lens effect. The blood group antigen was immunologically stained using antibody labeled with colloidal gold. Human leukocyte antigens (HLA) on lymphocytes and mononuclear leukocytes were observed by the thermal lens microscope, and Lewis blood group antigens on erythrocytes and polymorphonuclear leukocytes were also observed. The antigen distribution on each cell-surface was imaged using this technique. In spite of convex surface of living cells, colloidal gold was correctly quantified by adjusting the deviation of the focal point of the probe laser by the phase of the signal. In the measurement of leukocyte antigens, antigens of HLA-A, -B, -C loci on the lymphocytes were identified and quantitated by using a single cell. The image of HLA-A, -B, -C antigen distribution on a mononuclear leukocyte was obtained. In the measurement of erythrocyte antigens, a small quantity of Lewis antigens was detected on the cord erythrocytes. Localized small quantities of membrane antigens are better quantitated without extraction or cytolysis. Our thermal lens microscope is a powerful and highly sensitive analytical tool for detecting and quantitating localized antigens in single cells and/or cell-surface-associated molecules.

A ratiometric threshold for determining presence of cancer during fluorescence-guided surgery.

PubMed

Warram, Jason M; de Boer, Esther; Moore, Lindsay S; Schmalbach, Cecelia E; Withrow, Kirk P; Carroll, William R; Richman, Joshua S; Morlandt, Anthony B; Brandwein-Gensler, Margaret; Rosenthal, Eben L

2015-07-01

Fluorescence-guided imaging to assist in identification of malignant margins has the potential to dramatically improve oncologic surgery. However, a standardized method for quantitative assessment of disease-specific fluorescence has not been investigated. Introduced here is a ratiometric threshold derived from mean fluorescent tissue intensity that can be used to semi-quantitatively delineate tumor from normal tissue. Open-field and a closed-field imaging devices were used to quantify fluorescence in punch biopsy tissues sampled from primary tumors collected during a phase 1 trial evaluating the safety of cetuximab-IRDye800 in patients (n = 11) undergoing surgical intervention for head and neck cancer. Fluorescence ratios were calculated using mean fluorescence intensity (MFI) from punch biopsy normalized by MFI of patient-matched tissues. Ratios were compared to pathological assessment and a ratiometric threshold was established to predict presence of cancer. During open-field imaging using an intraoperative device, the threshold for muscle normalized tumor fluorescence was found to be 2.7, which produced a sensitivity of 90.5% and specificity of 78.6% for delineating disease tissue. The skin-normalized threshold generated greater sensitivity (92.9%) and specificity (81.0%). Successful implementation of a semi-quantitative threshold can provide a scientific methodology for delineating disease from normal tissue during fluorescence-guided resection of cancer. © 2015 Wiley Periodicals, Inc.

Atomic scale imaging of competing polar states in a Ruddlesden–Popper layered oxide

DOE PAGES

Stone, Greg; Ophus, Colin; Birol, Turan; ...

2016-08-31

Layered complex oxides offer an unusually rich materials platform for emergent phenomena through many built-in design knobs such as varied topologies, chemical ordering schemes and geometric tuning of the structure. A multitude of polar phases are predicted to compete in Ruddlesden-Popper (RP), A n+1 B n O 3n+1 , thin films by tuning layer dimension (n) and strain; however, direct atomic-scale evidence for such competing states is currently absent. Using aberration-corrected scanning transmission electron microscopy with sub-Ångstrom resolution in Sr n+1 Ti n O 3n+1 thin films, we demonstrate the coexistence of antiferroelectric, ferroelectric and new ordered and low-symmetry phases.more » We also directly image the atomic rumpling of the rock salt layer, a critical feature in RP structures that is responsible for the competing phases; exceptional quantitative agreement between electron microscopy and density functional theory is demonstrated. The study shows that layered topologies can enable multifunctionality through highly competitive phases exhibiting diverse phenomena in a single structure.« less

Label-free nanoscale characterization of red blood cell structure and dynamics using single-shot transport of intensity equation

NASA Astrophysics Data System (ADS)

Poola, Praveen Kumar; John, Renu

2017-10-01

We report the results of characterization of red blood cell (RBC) structure and its dynamics with nanometric sensitivity using transport of intensity equation microscopy (TIEM). Conventional transport of intensity technique requires three intensity images and hence is not suitable for studying real-time dynamics of live biological samples. However, assuming the sample to be homogeneous, phase retrieval using transport of intensity equation has been demonstrated with single defocused measurement with x-rays. We adopt this technique for quantitative phase light microscopy of homogenous cells like RBCs. The main merits of this technique are its simplicity, cost-effectiveness, and ease of implementation on a conventional microscope. The phase information can be easily merged with regular bright-field and fluorescence images to provide multidimensional (three-dimensional spatial and temporal) information without any extra complexity in the setup. The phase measurement from the TIEM has been characterized using polymeric microbeads and the noise stability of the system has been analyzed. We explore the structure and real-time dynamics of RBCs and the subdomain membrane fluctuations using this technique.

Nuclear medicine and imaging research (quantitative studies in radiopharmaceutical science)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cooper, M.D.; Beck, R.N.

1990-09-01

This is a report of progress in Year Two (January 1, 1990--December 31, 1990) of Grant FG02-86ER60438, Quantitative Studies in Radiopharmaceutical Science,'' awarded for the three-year period January 1, 1989--December 31, 1991 as a competitive renewal following site visit in the fall of 1988. This program addresses the problems involving the basic science and technology underlying the physical and conceptual tools of radioactive tracer methodology as they relate to the measurement of structural and functional parameters of physiologic importance in health and disease. The principal tool is quantitative radionuclide imaging. The overall objective of this program is to further themore » development and transfer of radiotracer methodology from basic theory to routine clinical practice in order that individual patients and society as a whole will receive the maximum net benefit from the new knowledge gained. The focus of the research is on the development of new instruments and radiopharmaceuticals, and the evaluation of these through the phase of clinical feasibility. 25 refs., 13 figs., 1 tab.« less

In vivo optical microscopy of peripheral nerve myelination with polarization sensitive-optical coherence tomography

PubMed Central

Henry, Francis P.; Wang, Yan; Rodriguez, Carissa L. R.; Randolph, Mark A.; Rust, Esther A. Z.; Winograd, Jonathan M.; de Boer, Johannes F.; Park, B. Hyle

2015-01-01

Abstract. Assessing nerve integrity and myelination after injury is necessary to provide insight for treatment strategies aimed at restoring neuromuscular function. Currently, this is largely done with electrical analysis, which lacks direct quantitative information. In vivo optical imaging with sufficient imaging depth and resolution could be used to assess the nerve microarchitecture. In this study, we examine the use of polarization sensitive-optical coherence tomography (PS-OCT) to quantitatively assess the sciatic nerve microenvironment through measurements of birefringence after applying a nerve crush injury in a rat model. Initial loss of function and subsequent recovery were demonstrated by calculating the sciatic function index (SFI). We found that the PS-OCT phase retardation slope, which is proportional to birefringence, increased monotonically with the SFI. Additionally, histomorphometric analysis of the myelin thickness and g-ratio shows that the PS-OCT slope is a good indicator of myelin health and recovery after injury. These results demonstrate that PS-OCT is capable of providing nondestructive and quantitative assessment of nerve health after injury and shows promise for continued use both clinically and experimentally in neuroscience. PMID:25858593

In vivo optical microscopy of peripheral nerve myelination with polarization sensitive-optical coherence tomography.

PubMed

Henry, Francis P; Wang, Yan; Rodriguez, Carissa L R; Randolph, Mark A; Rust, Esther A Z; Winograd, Jonathan M; de Boer, Johannes F; Park, B Hyle

2015-04-01

Assessing nerve integrity and myelination after injury is necessary to provide insight for treatment strategies aimed at restoring neuromuscular function. Currently, this is largely done with electrical analysis, which lacks direct quantitative information. In vivo optical imaging with sufficient imaging depth and resolution could be used to assess the nerve microarchitecture. In this study, we examine the use of polarization sensitive-optical coherence tomography (PS-OCT) to quantitatively assess the sciatic nerve microenvironment through measurements of birefringence after applying a nerve crush injury in a rat model. Initial loss of function and subsequent recovery were demonstrated by calculating the sciatic function index (SFI). We found that the PS-OCT phase retardation slope, which is proportional to birefringence, increased monotonically with the SFI. Additionally, histomorphometric analysis of the myelin thickness and g-ratio shows that the PS-OCT slope is a good indicator of myelin health and recovery after injury. These results demonstrate that PS-OCT is capable of providing nondestructive and quantitative assessment of nerve health after injury and shows promise for continued use both clinically and experimentally in neuroscience.

Superimpose signal processing method for micro-scale thermal imaging of solar salts at high temperature

NASA Astrophysics Data System (ADS)

Morikawa, Junko; Zamengo, Massimiliano; Kato, Yukitaka

2016-05-01

The global interest in energy applications activates the advanced study about the molten salts in the usage of fluids in the power cycle, such as for transport and heat storage in solar power facilities. However, the basic properties of molten salts show a general scattering in characterization especially in thermal properties. It is suggested that new studies are required on the measurement of thermal properties of solar salts using recent technologies. In this study, micro-scale heat transfer and phase change in molten salts are presented using our originally developed device: the micro-bolometer Infrared focal plane arrays (IR FPA) measuring system is a portable type instrument, which is re-designed to measure the thermal phenomena in high temperature up to 700 °C or higher. The superimpose system is newly setup adjusted to the signal processing in high temperature to realize the quantitative thermal imaging, simultaneously. The portable type apparatus for a quantitative micro-scale thermography using a micro-bolometer has been proposed based on an achromatic lens design to capture a micro-scale image in the long-wave infrared, a video signal superimposing for the real time emissivity correction, and a pseudo acceleration of a timeframe. Combined with the superimpose technique, the micro-scale thermal imaging in high temperature is achieved and the molten flows of the solar salts, sodium nitrate, and potassium nitrate are successfully observed. The solar salt, the mixture of sodium nitrate and potassium nitrate, shows a different shape of exothermic heat front morphology in the lower phase transition (solidification) temperature than the nitrates on cooling. The proposed measuring technique will be utilized to accelerate the screening step to determine the phase diagram and the eutectics of the multiple mixtures of candidate molten salts, which may be used as heat transport medium from the concentrated solar power to a processing plant for thermal energy storage.

Lens-free computational imaging of capillary morphogenesis within three-dimensional substrates

NASA Astrophysics Data System (ADS)

Weidling, John; Isikman, Serhan O.; Greenbaum, Alon; Ozcan, Aydogan; Botvinick, Elliot

2012-12-01

Endothelial cells cultured in three-dimensional (3-D) extracellular matrices spontaneously form microvessels in response to soluble and matrix-bound factors. Such cultures are common for the study of angiogenesis and may find widespread use in drug discovery. Vascular networks are imaged over weeks to measure the distribution of vessel morphogenic parameters. Measurements require micron-scale spatial resolution, which for light microscopy comes at the cost of limited field-of-view (FOV) and shallow depth-of-focus (DOF). Small FOVs and DOFs necessitate lateral and axial mechanical scanning, thus limiting imaging throughput. We present a lens-free holographic on-chip microscopy technique to rapidly image microvessels within a Petri dish over a large volume without any mechanical scanning. This on-chip method uses partially coherent illumination and a CMOS sensor to record in-line holographic images of the sample. For digital reconstruction of the measured holograms, we implement a multiheight phase recovery method to obtain phase images of capillary morphogenesis over a large FOV (24 mm2) with ˜1.5 μm spatial resolution. On average, measured capillary length in our method was within approximately 2% of lengths measured using a 10× microscope objective. These results suggest lens-free on-chip imaging is a useful toolset for high-throughput monitoring and quantitative analysis of microvascular 3-D networks.

Investigation on microfluidic particles manipulation by holographic 3D tracking strategies

NASA Astrophysics Data System (ADS)

Cacace, Teresa; Paturzo, Melania; Memmolo, Pasquale; Vassalli, Massimo; Fraldi, Massimiliano; Mensitieri, Giuseppe; Ferraro, Pietro

2017-06-01

We demonstrate a 3D holographic tracking method to investigate particles motion in a microfluidic channel while unperturbed while inducing their migration through microfluidic manipulation. Digital holography (DH) in microscopy is a full-field, label-free imaging technique able to provide quantitative phase-contrast. The employed 3D tracking method is articulated in steps. First, the displacements along the optical axis are assessed by numerical refocusing criteria. In particular, an automatic refocusing method to recover the particles axial position is implemented employing a contrast-based refocusing criterion. Then, the transverse position of the in-focus object is evaluated through quantitative phase map segmentation methods and centroid-based 2D tracking strategy. The introduction of DH is thus suggested as a powerful approach for control of particles and biological samples manipulation, as well as a possible aid to precise design and implementation of advanced lab-on-chip microfluidic devices.

Visualization and automatic detection of defect distribution in GaN atomic structure from sampling Moiré phase.

PubMed

Wang, Qinghua; Ri, Shien; Tsuda, Hiroshi; Kodera, Masako; Suguro, Kyoichi; Miyashita, Naoto

2017-09-19

Quantitative detection of defects in atomic structures is of great significance to evaluating product quality and exploring quality improvement process. In this study, a Fourier transform filtered sampling Moire technique was proposed to visualize and detect defects in atomic arrays in a large field of view. Defect distributions, defect numbers and defect densities could be visually and quantitatively determined from a single atomic structure image at low cost. The effectiveness of the proposed technique was verified from numerical simulations. As an application, the dislocation distributions in a GaN/AlGaN atomic structure in two directions were magnified and displayed in Moire phase maps, and defect locations and densities were detected automatically. The proposed technique is able to provide valuable references to material scientists and engineers by checking the effect of various treatments for defect reduction. © 2017 IOP Publishing Ltd.

Multispectral Imaging Broadens Cellular Analysis

NASA Technical Reports Server (NTRS)

2007-01-01

Amnis Corporation, a Seattle-based biotechnology company, developed ImageStream to produce sensitive fluorescence images of cells in flow. The company responded to an SBIR solicitation from Ames Research Center, and proposed to evaluate several methods of extending the depth of field for its ImageStream system and implement the best as an upgrade to its commercial products. This would allow users to view whole cells at the same time, rather than just one section of each cell. Through Phase I and II SBIR contracts, Ames provided Amnis the funding the company needed to develop this extended functionality. For NASA, the resulting high-speed image flow cytometry process made its way into Medusa, a life-detection instrument built to collect, store, and analyze sample organisms from erupting hydrothermal vents, and has the potential to benefit space flight health monitoring. On the commercial end, Amnis has implemented the process in ImageStream, combining high-resolution microscopy and flow cytometry in a single instrument, giving researchers the power to conduct quantitative analyses of individual cells and cell populations at the same time, in the same experiment. ImageStream is also built for many other applications, including cell signaling and pathway analysis; classification and characterization of peripheral blood mononuclear cell populations; quantitative morphology; apoptosis (cell death) assays; gene expression analysis; analysis of cell conjugates; molecular distribution; and receptor mapping and distribution.

Quantitative evaluation of high-resolution features in images of negatively stained Tobacco Mosaic Virus.

PubMed

Chang, C F; Williams, R C; Grano, D A; Downing, K H; Glaeser, R M

1983-01-01

This study investigates the causes of the apparent differences between the optical diffraction pattern of a micrograph of a Tobacco Mosaic Virus (TMV) particle, the optical diffraction pattern of a ten-fold photographically averaged image, and the computed diffraction pattern of the original micrograph. Peak intensities along the layer lines in the transform of the averaged image appear to be quite unlike those in the diffraction pattern of the original micrograph, and the diffraction intensities for the averaged image extend to unexpectedly high resolution. A carefully controlled, quantitative comparison reveals, however, that the optical diffraction pattern of the original micrograph and that of the ten-fold averaged image are essentially equivalent. Using computer-based image processing, we discovered that the peak intensities on the 6th layer line have values very similar in magnitude to the neighboring noise, in contrast to what was expected from the optical diffraction pattern of the original micrograph. This discrepancy was resolved by recording a series of optical diffraction patterns when the original micrograph was immersed in oil. These patterns revealed the presence of a substantial phase grating effect, which exaggerated the peak intensities on the 6th layer line, causing an erroneous impression that the high resolution features possessed a good signal-to-noise ratio. This study thus reveals some pitfalls and misleading results that can be encountered when using optical diffraction patterns to evaluate image quality.

An Analysis and Classification of Dying AGB Stars Transitioning to Pre-Planetary Nebulae

NASA Technical Reports Server (NTRS)

Blake, Adam C.

2011-01-01

The principal objective of the project is to understand part of the life and death process of a star. During the end of a star's life, it expels its mass at a very rapid rate. We want to understand how these Asymptotic Giant Branch (AGB) stars begin forming asymmetric structures as they start evolving towards the planetary nebula phase and why planetary nebulae show a very large variety of non-round geometrical shapes. To do this, we analyzed images of just-forming pre-planetary nebula from Hubble surveys. These images were run through various image correction processes like saturation correction and cosmic ray removal using in-house software to bring out the circumstellar structure. We classified the visible structure based on qualitative data such as lobe, waist, halo, and other structures. Radial and azimuthal intensity cuts were extracted from the images to quantitatively examine the circumstellar structure and measure departures from the smooth spherical outflow expected during most of the AGB mass-loss phase. By understanding the asymmetrical structure, we hope to understand the mechanisms that drive this stellar evolution.

Contrast features of breast cancer in frequency-domain laser scanning mammography

NASA Astrophysics Data System (ADS)

Moesta, K. Thomas; Fantini, Sergio; Jess, Helge; Totkas, Susan; Franceschini, Maria-Angela; Kaschke, Michael; Schlag, Peter M.

1998-04-01

Frequency-domain optical mammography has been advocated to improve contrast and thus cancer detectability in breast transillumination. To the best of our knowledge, this report provides the first systematic clinical results of a frequency-domain laser scanning mammograph (FLM). The instrument provides monochromatic light at 690 and 810 nm, whose intensity is modulated at 110.0008 MHz, respectively. The breast is scanned by stepwise positioning of source and detector, and amplitude and phase for both wavelengths are measured by a photomultiplier tube using heterodyne detection. Images are formed representing amplitude or phase data on linear gray scales. Furthermore, various algorithms carrying on more than one signal were essayed. Twenty visible cancers out of 25 cancers in the first 59 investigations were analyzed for their quantitative contrast with respect to the whole breast or to defined reference areas. Contrast definitions refer to the signal itself, to the signal noise, or were based on nonparametric comparison. The amplitude signal provides better contrast than the phase signal. Ratio images between red and IR amplitudes gave variable results; in some cases the tumor contrast was canceled. The algorithms to determine (mu) a and (mu) sPRM from amplitude and phase data did not significantly improve upon objective contrast. The N algorithm, using the phase signal to flatten the amplitude signal did significantly improve upon contrast according to contrast definitions 1 and 2, however, did not improve upon nonparametric contrast. Thus, with the current instrumentation, the phase signal is helpful to correct for the complex and variable geometry of the breast. However, an independent informational content for tumor differentiation could not be determined. The flat field algorithm did greatly enhance optical contrast in comparison with amplitude or amplitude ratio images. Further evaluation of FLM will have to be based on the N-algorithm images.

Phase Shift Interferometer and Growth Set Up to Step Pattern Formation During Growth From Solutions. Influence of the Oscillatory solution Flow on Stability

NASA Technical Reports Server (NTRS)

Chernov, Alex A.; Booth, N. A.; Vekilov, P. G.; Murray, B. T.; McFadden, G. B.

2000-01-01

We have assembled an experimental setup based on Michelson interferometry with the growing crystal surface as one of the reflective surfaces. The crystallization part of the device allows optical monitoring of a face of a crystal growing at temperature stable within 0.05 C in a flow of solution of controlled direction and speed. The reference arm of the interferometer contains a liquid crystal element that allows controlled shifts of the phase of the interferograms. We employ an image-processing algorithm, which combines five images with a pi/2 phase difference between each pair of images. The images are transferred to a computer by a camera capable of capturing 60 frames per second. The device allows data collection on surface morphology and kinetics during the face layers growth over a relatively large area (approximately 4 sq. mm) in situ and in real time during growth. The estimated depth resolution of the phase shifting interferometry is approximately 50 Angstroms. The data will be analyzed in order to reveal and monitor step bunching during the growth process. The crystal chosen as a model for study in this work is KH2PO4 (KDP). This optically non-linear material is widely used in frequency doubling applications. There have been a number of studies of the kinetics of KDP crystallization that can serve as a benchmark for our investigations. However, so far, systematic quantitative characteristics of step interaction and bunching are missing. We intend to present our first quantitative results on the onset, initial stages and development of instabilities in moving step trains on vicinal crystal surfaces at varying supersaturation, flow rate, and flow direction. Behavior of a vicinal face growing from solution flowing normal to the steps and periodically changing its direction in time was considered theoretically. It was found that this oscillating flow reduces both stabilization and destabilization effects resulted from the unidirectional solution flow directed up the step stream and down the step stream. This reduction of stabilization and destabilization comes from effective mixing which entangles the phase shifts between the spatially periodic interface perturbation and the concentration wave induced by this perturbation. Numerical results and simplified mixing criterion will be discussed.

Validation of motion correction techniques for liver CT perfusion studies

PubMed Central

Chandler, A; Wei, W; Anderson, E F; Herron, D H; Ye, Z; Ng, C S

2012-01-01

Objectives Motion in images potentially compromises the evaluation of temporally acquired CT perfusion (CTp) data; image registration should mitigate this, but first requires validation. Our objective was to compare the relative performance of manual, rigid and non-rigid registration techniques to correct anatomical misalignment in acquired liver CTp data sets. Methods 17 data sets in patients with liver tumours who had undergone a CTp protocol were evaluated. Each data set consisted of a cine acquisition during a breath-hold (Phase 1), followed by six further sets of cine scans (each containing 11 images) acquired during free breathing (Phase 2). Phase 2 images were registered to a reference image from Phase 1 cine using two semi-automated intensity-based registration techniques (rigid and non-rigid) and a manual technique (the only option available in the relevant vendor CTp software). The performance of each technique to align liver anatomy was assessed by four observers, independently and blindly, on two separate occasions, using a semi-quantitative visual validation study (employing a six-point score). The registration techniques were statistically compared using an ordinal probit regression model. Results 306 registrations (2448 observer scores) were evaluated. The three registration techniques were significantly different from each other (p=0.03). On pairwise comparison, the semi-automated techniques were significantly superior to the manual technique, with non-rigid significantly superior to rigid (p<0.0001), which in turn was significantly superior to manual registration (p=0.04). Conclusion Semi-automated registration techniques achieved superior alignment of liver anatomy compared with the manual technique. We hope this will translate into more reliable CTp analyses. PMID:22374283

Visualization of small lesions in rat cartilage by means of laboratory-based x-ray phase contrast imaging

NASA Astrophysics Data System (ADS)

Marenzana, Massimo; Hagen, Charlotte K.; Das Neves Borges, Patricia; Endrizzi, Marco; Szafraniec, Magdalena B.; Ignatyev, Konstantin; Olivo, Alessandro

2012-12-01

Being able to quantitatively assess articular cartilage in three-dimensions (3D) in small rodent animal models, with a simple laboratory set-up, would prove extremely important for the development of pre-clinical research focusing on cartilage pathologies such as osteoarthritis (OA). These models are becoming essential tools for the development of new drugs for OA, a disease affecting up to 1/3 of the population older than 50 years for which there is no cure except prosthetic surgery. However, due to limitations in imaging technology, high-throughput 3D structural imaging has not been achievable in small rodent models, thereby limiting their translational potential and their efficiency as research tools. We show that a simple laboratory system based on coded-aperture x-ray phase contrast imaging (CAXPCi) can correctly visualize the cartilage layer in slices of an excised rat tibia imaged both in air and in saline solution. Moreover, we show that small, surgically induced lesions are also correctly detected by the CAXPCi system, and we support this finding with histopathology examination. Following these successful proof-of-concept results in rat cartilage, we expect that an upgrade of the system to higher resolutions (currently underway) will enable extending the method to the imaging of mouse cartilage as well. From a technological standpoint, by showing the capability of the system to detect cartilage also in water, we demonstrate phase sensitivity comparable to other lab-based phase methods (e.g. grating interferometry). In conclusion, CAXPCi holds a strong potential for being adopted as a routine laboratory tool for non-destructive, high throughput assessment of 3D structural changes in murine articular cartilage, with a possible impact in the field similar to the revolution that conventional microCT brought into bone research.

Diagnostic Accuracy of Dynamic Contrast Enhanced Magnetic Resonance Imaging in Characterizing Lung Masses

PubMed Central

Inan, Nagihan; Arslan, Arzu; Donmez, Muhammed; Sarisoy, Hasan Tahsin

2016-01-01

Background Imaging plays a critical role not only in the detection, but also in the characterization of lung masses as benign or malignant. Objectives To determine the diagnostic accuracy of dynamic magnetic resonance imaging (MRI) in the differential diagnosis of benign and malignant lung masses. Patients and Methods Ninety-four masses were included in this prospective study. Five dynamic series of T1-weighted spoiled gradient echo (FFE) images were obtained, followed by a T1-weighted FFE sequence in the late phase (5th minutes). Contrast enhancement patterns in the early (25th second) and late (5th minute) phase images were evaluated. For the quantitative evaluation, signal intensity (SI)-time curves were obtained and the maximum relative enhancement, wash-in rate, and time-to-peak enhancement of masses in both groups were calculated. Results The early phase contrast enhancement patterns were homogeneous in 78.2% of the benign masses, while heterogeneous in 74.4% of the malignant tumors. On the late phase images, 70.8% of the benign masses showed homogeneous enhancement, while most of the malignant masses showed heterogeneous enhancement (82.4%). During the first pass, the maximum relative enhancement and wash-in rate values of malignant masses were significantly higher than those of the benign masses (P = 0.03 and 0.04, respectively). The cutoff value at 15% yielded a sensitivity of 85.4%, specificity of 61.2%, and positive predictive value of 68.7% for the maximum relative enhancement. Conclusion Contrast enhancement patterns and SI-time curve analysis of MRI are helpful in the differential diagnosis of benign and malignant lung masses. PMID:27703654

Full field vertical scanning in short coherence digital holographic microscope.

PubMed

Monemahghdoust, Zahra; Montfort, Frederic; Cuche, Etienne; Emery, Yves; Depeursinge, Christian; Moser, Christophe

2013-05-20

In Digital holography Microscopes (DHM) implemented in the so-called "off axis" configuration, the object and reference wave fronts are not co-planar but form an angle of a few degrees. This results into two main drawbacks. First, the contrast of the interference is not uniform spatially when the light source has low coherence. The interference contrast is optimal along a line, but decreases when moving away from it, resulting in a lower image quality. Second, the non-coplanarity between the coherence plane of both wavefronts impacts the coherence vertical scanning measurement mode: when the optical path difference between the signal and the reference beam is changed, the region of maximum interference contrast shifts laterally in the plane of the objective. This results in more complex calculations to extract the topography of the sample and requires scanning over a much larger vertical range, leading to a longer measurement time. We have previously shown that by placing a volume diffractive optical element (VDOE) in the reference arm, the wavefront can be made coplanar with the object wavefront and the image plane of the microscope objective, resulting in a uniform and optimal interferogram. In this paper, we demonstrate a vertical scanning speed improvement by an order of magnitude. Noise in the phase and intensity images caused by scattering and non-uniform diffraction in the VDOE is analyzed quantitatively. Five VDOEs were fabricated with an identical procedure. We observe that VDOEs introduce a small intensity non-uniformity in the reference beam which results in a 20% noise increase in the extracted phase image as compared to the noise in extracted phase image when the VDOE is removed. However, the VDOE has no impact on the temporal noise measured from extracted phase images.

Near-infrared lymphatic imaging demonstrates the dynamics of lymph flow and lymphangiogenesis during the acute versus chronic phases of arthritis in mice.

PubMed

Zhou, Quan; Wood, Ronald; Schwarz, Edward M; Wang, Yong-Jun; Xing, Lianping

2010-07-01

To develop an in vivo imaging method to assess lymphatic draining function in the K/BxN mouse model of inflammatory arthritis. Indocyanine green, a near-infrared fluorescent dye, was injected intradermally into the footpads of wild-type mice, mouse limbs were illuminated with an 806-nm near-infrared laser, and the movement of indocyanine green from the injection site to the draining popliteal lymph node (LN) was recorded with a CCD camera. Indocyanine green near-infrared images were analyzed to obtain 5 measures of lymphatic function across time. Images of K/BxN arthritic mice and control nonarthritic littermates were obtained at 1 month of age, when acute joint inflammation commenced, and again at 3 months of age, when joint inflammation became chronic. Lymphangiogenesis in popliteal LNs was assessed by immunochemistry. Indocyanine green and its transport within lymphatic vessels were readily visualized, and quantitative measures were derived. During the acute phase of arthritis, the lymphatic vessels were dilated, with increased indocyanine green signal intensity and lymphatic pulses, and popliteal LNs became fluorescent quickly. During the chronic phase, new lymphatic vessels were present near the foot. However, the appearance of indocyanine green in lymphatic vessels was delayed. The size and area of popliteal LN lymphatic sinuses progressively increased in the K/BxN mice. Our findings indicate that indocyanine green near-infrared lymphatic imaging is a valuable method for assessing the lymphatic draining function in mice with inflammatory arthritis. Indocyanine green-near-infrared imaging of K/BxN mice identified 2 distinct lymphatic phenotypes during the acute and chronic phase of inflammation. This technique can be used to assess new therapies for lymphatic disorders.

Near infrared lymphatic imaging demonstrates the dynamics of lymph flow and lymphangiogenesis during the acute vs. chronic phases of arthritis in mice

PubMed Central

Zhou, Quan; Wood, Ronald; Schwarz, Edward M.; Wang, Yong-Jun; Xing, Lianping

2010-01-01

Objective Development of an in vivo imaging method to assess lymphatic draining function in the K/B×N mouse model of inflammatory arthritis. Methods Indocyanine green (ICG), a near-infrared (NIR) fluorescent dye, was injected intradermally into the footpad of wild-type mice, the limb was illuminated with an 806 nm NIR laser, and the movement of ICG from the injection site to the draining popliteal lymph node (PLN) was recorded with a CCD camera. ICG-NIR images were analyzed to obtain 5 measures of lymphatic function across time. K/B×N arthritic mice and control non-arthritic littermates were imaged at one-month of age when acute joint inflammation commenced, and repeated at 3 months when joint inflammation became chronic. Lymphangiogenesis in PLNs was assessed by immunochemistry. Results ICG and its transport within lymphatic vessels were readily visualized and quantitative measures derived. During the acute phase of arthritis, the lymphatic vessels were dilated with increased ICG signal intensity and lymphatic pulses, and PLNs became fluorescent quickly. During the chronic phase, new lymphatic vessels were present near the foot. However, ICG appearance in lymphatic vessels was delayed. The size and area of PLN lymphatic sinuses progressively increased in the K/B×N mice. Conclusion ICG-NIR lymphatic imaging is a valuable method to assess the lymphatic draining function in mice with inflammatory arthritis. ICG-NIR imaging of K/B×N mice identified two distinct lymphatic phenotypes during the acute and chronic phase of inflammation. This technique can be used to assess new therapies for lymphatic disorders. PMID:20309866

Probing the regional distribution of pulmonary gas exchange through single-breath gas- and dissolved-phase 129Xe MR imaging.

PubMed

Kaushik, S Sivaram; Freeman, Matthew S; Cleveland, Zackary I; Davies, John; Stiles, Jane; Virgincar, Rohan S; Robertson, Scott H; He, Mu; Kelly, Kevin T; Foster, W Michael; McAdams, H Page; Driehuys, Bastiaan

2013-09-01

Although some central aspects of pulmonary function (ventilation and perfusion) are known to be heterogeneous, the distribution of diffusive gas exchange remains poorly characterized. A solution is offered by hyperpolarized 129Xe magnetic resonance (MR) imaging, because this gas can be separately detected in the lung's air spaces and dissolved in its tissues. Early dissolved-phase 129Xe images exhibited intensity gradients that favored the dependent lung. To quantitatively corroborate this finding, we developed an interleaved, three-dimensional radial sequence to image the gaseous and dissolved 129Xe distributions in the same breath. These images were normalized and divided to calculate "129Xe gas-transfer" maps. We hypothesized that, for healthy volunteers, 129Xe gas-transfer maps would retain the previously observed posture-dependent gradients. This was tested in nine subjects: when the subjects were supine, 129Xe gas transfer exhibited a posterior-anterior gradient of -2.00 ± 0.74%/cm; when the subjects were prone, the gradient reversed to 1.94 ± 1.14%/cm (P < 0.001). The 129Xe gas-transfer maps also exhibited significant heterogeneity, as measured by the coefficient of variation, that correlated with subject total lung capacity (r = 0.77, P = 0.015). Gas-transfer intensity varied nonmonotonically with slice position and increased in slices proximal to the main pulmonary arteries. Despite substantial heterogeneity, the mean gas transfer for all subjects was 1.00 ± 0.01 while supine and 1.01 ± 0.01 while prone (P = 0.25), indicating good "matching" between gas- and dissolved-phase distributions. This study demonstrates that single-breath gas- and dissolved-phase 129Xe MR imaging yields 129Xe gas-transfer maps that are sensitive to altered gas exchange caused by differences in lung inflation and posture.

Illumination Modulation for Improved Propagation-Based Phase Imaging

NASA Astrophysics Data System (ADS)

Chakraborty, Tonmoy

Propagation-based phase imaging enables the quantitative reconstruction of a light beam's phase from measurements of its intensity. Because the intensity depends on the time-averaged square of the field the relationship between intensity and phase is, in general, nonlinear. The transport of intensity equation (TIE), is a linear equation relating phase and propagated intensity that arises from restricting the propagation distance to be small. However, the TIE limits the spatial frequencies that can be reliably reconstructed to those below some cutoff, which limits the accuracy of reconstruction of fine features in phase. On the other hand, the low frequency components suffer from poor signal to noise ratio (SNR) unless the propagation distance is sufficiently large, which leads to low frequency artifacts that obscure the reconstruction. In this research, I will consider the use of incoherent primary sources of illumination, in a Kohler illumination setup, to enhance the low-frequency performance of the TIE. The necessary steps required to design and build a table-top imaging setup which is capable of capturing intensity at any defocused position while modulating the source will be explained. In addition, it will be shown how by employing such illumination, the steps required for computationally recovering the phase, i.e. Fourier transforms and frequency-domain filtering, may be performed in the optical system. While these methods can address the low-frequency performance of the TIE, they do not extend its high-frequency cutoff. To avoid this cutoff, for objects with slowly varying phase, the contrast transfer function (CTF) model, an alternative to the TIE, can be used to recover phase. By allowing the combination of longer propagation distances and incoherent sources, it will be shown how CTF can improve performance at both high and low frequencies.

Correlative Light and Scanning X-Ray Scattering Microscopy of Healthy and Pathologic Human Bone Sections

PubMed Central

Giannini, C.; Siliqi, D.; Bunk, O.; Beraudi, A.; Ladisa, M.; Altamura, D.; Stea, S.; Baruffaldi, F.

2012-01-01

Scanning small and wide angle X-ray scattering (scanning SWAXS) experiments were performed on healthy and pathologic human bone sections. Via crystallographic tools the data were transformed into quantitative images and as such compared with circularly polarized light (CPL) microscopy images. SWAXS and CPL images allowed extracting information of the mineral nanocrystalline phase embedded, with and without preferred orientation, in the collagen fibrils, mapping local changes at sub-osteon resolution. This favorable combination has been applied for the first time to biopsies of dwarfism syndrome and Paget's disease to shed light onto the cortical structure of natural bone in healthy and pathologic sections. PMID:22666538

Multi-organ segmentation from multi-phase abdominal CT via 4D graphs using enhancement, shape and location optimization.

PubMed

Linguraru, Marius George; Pura, John A; Chowdhury, Ananda S; Summers, Ronald M

2010-01-01

The interpretation of medical images benefits from anatomical and physiological priors to optimize computer-aided diagnosis (CAD) applications. Diagnosis also relies on the comprehensive analysis of multiple organs and quantitative measures of soft tissue. An automated method optimized for medical image data is presented for the simultaneous segmentation of four abdominal organs from 4D CT data using graph cuts. Contrast-enhanced CT scans were obtained at two phases: non-contrast and portal venous. Intra-patient data were spatially normalized by non-linear registration. Then 4D erosion using population historic information of contrast-enhanced liver, spleen, and kidneys was applied to multi-phase data to initialize the 4D graph and adapt to patient specific data. CT enhancement information and constraints on shape, from Parzen windows, and location, from a probabilistic atlas, were input into a new formulation of a 4D graph. Comparative results demonstrate the effects of appearance and enhancement, and shape and location on organ segmentation.

Impact of CT attenuation correction method on quantitative respiratory-correlated (4D) PET/CT imaging

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nyflot, Matthew J., E-mail: nyflot@uw.edu; Lee, Tzu-Cheng; Alessio, Adam M.

Purpose: Respiratory-correlated positron emission tomography (PET/CT) 4D PET/CT is used to mitigate errors from respiratory motion; however, the optimal CT attenuation correction (CTAC) method for 4D PET/CT is unknown. The authors performed a phantom study to evaluate the quantitative performance of CTAC methods for 4D PET/CT in the ground truth setting. Methods: A programmable respiratory motion phantom with a custom movable insert designed to emulate a lung lesion and lung tissue was used for this study. The insert was driven by one of five waveforms: two sinusoidal waveforms or three patient-specific respiratory waveforms. 3DPET and 4DPET images of the phantommore » under motion were acquired and reconstructed with six CTAC methods: helical breath-hold (3DHEL), helical free-breathing (3DMOT), 4D phase-averaged (4DAVG), 4D maximum intensity projection (4DMIP), 4D phase-matched (4DMATCH), and 4D end-exhale (4DEXH) CTAC. Recovery of SUV{sub max}, SUV{sub mean}, SUV{sub peak}, and segmented tumor volume was evaluated as RC{sub max}, RC{sub mean}, RC{sub peak}, and RC{sub vol}, representing percent difference relative to the static ground truth case. Paired Wilcoxon tests and Kruskal–Wallis ANOVA were used to test for significant differences. Results: For 4DPET imaging, the maximum intensity projection CTAC produced significantly more accurate recovery coefficients than all other CTAC methods (p < 0.0001 over all metrics). Over all motion waveforms, ratios of 4DMIP CTAC recovery were 0.2 ± 5.4, −1.8 ± 6.5, −3.2 ± 5.0, and 3.0 ± 5.9 for RC{sub max}, RC{sub peak}, RC{sub mean}, and RC{sub vol}. In comparison, recovery coefficients for phase-matched CTAC were −8.4 ± 5.3, −10.5 ± 6.2, −7.6 ± 5.0, and −13.0 ± 7.7 for RC{sub max}, RC{sub peak}, RC{sub mean}, and RC{sub vol}. When testing differences between phases over all CTAC methods and waveforms, end-exhale phases were significantly more accurate (p = 0.005). However, these differences were driven by the patient-specific respiratory waveforms; when testing patient and sinusoidal waveforms separately, patient waveforms were significantly different between phases (p < 0.0001) while the sinusoidal waveforms were not significantly different (p = 0.98). When considering only the subset of 4DMATCH images that corresponded to the end-exhale image phase, 4DEXH, mean and interquartile range were similar to 4DMATCH but variability was considerably reduced. Conclusions: Comparative advantages in accuracy and precision of SUV metrics and segmented volumes were demonstrated with the use of the maximum intensity projection and end-exhale CT attenuation correction. While respiratory phase-matched CTAC should in theory provide optimal corrections, image artifacts and differences in implementation of 4DCT and 4DPET sorting can degrade the benefit of this approach. These results may be useful to guide the implementation, analysis, and development of respiratory-correlated thoracic PET/CT in the radiation oncology and diagnostic settings.« less

Digital holographic microscopy combined with optical tweezers

NASA Astrophysics Data System (ADS)

Cardenas, Nelson; Yu, Lingfeng; Mohanty, Samarendra K.

2011-02-01

While optical tweezers have been widely used for the manipulation and organization of microscopic objects in three dimensions, observing the manipulated objects along axial direction has been quite challenging. In order to visualize organization and orientation of objects along axial direction, we report development of a Digital holographic microscopy combined with optical tweezers. Digital holography is achieved by use of a modified Mach-Zehnder interferometer with digital recording of interference pattern of the reference and sample laser beams by use of a single CCD camera. In this method, quantitative phase information is retrieved dynamically with high temporal resolution, only limited by frame rate of the CCD. Digital focusing, phase-unwrapping as well as online analysis and display of the quantitative phase images was performed on a software developed on LabView platform. Since phase changes observed in DHOT is very sensitive to optical thickness of trapped volume, estimation of number of particles trapped in the axial direction as well as orientation of non-spherical objects could be achieved with high precision. Since in diseases such as malaria and diabetics, change in refractive index of red blood cells occurs, this system can be employed to map such disease-specific changes in biological samples upon immobilization with optical tweezers.

New techniques for positron emission tomography in the study of human neurological disorders

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kuhl, D.E.

1993-01-01

This progress report describes accomplishments of four programs. The four programs are entitled (1) Faster,simpler processing of positron-computing precursors: New physicochemical approaches, (2) Novel solid phase reagents and methods to improve radiosynthesis and isotope production, (3) Quantitative evaluation of the extraction of information from PET images, and (4) Optimization of tracer kinetic methods for radioligand studies in PET.

Enlightening intracellular complexity of living cells with quantitative phase microscopy

NASA Astrophysics Data System (ADS)

Martinez Torres, C.; Laperrousaz, B.; Berguiga, L.; Boyer Provera, E.; Elezgaray, J.; Nicolini, F. E.; Maguer-Satta, V.; Arneodo, A.; Argoul, F.

2016-03-01

The internal distribution of refractive indices (RIs) of a living cell is much more complex than usually admitted in multi-shell models. The reconstruction of RI maps from single phase images has rarely been achieved for several reasons: (i) we still have very little knowledge of the impact of internal macromolecular complexes on the local RI and (ii) phase changes produced by light propagation through the sample are mixed with diffraction effects by internal cell bodies. We propose the implementation a 2D wavelet-based contour chain detection method to distinguish internal boundaries thanks to their greatest optical path difference gradients. These contour chains correspond to the highest image phase contrast and follow the local RI inhomogeneities linked to the intracellular structural intricacy. Their statistics and spatial distribution are morphological indicators for distinguishing cells of different origins and to follow their transformation in pathologic situations. We use this method to compare non adherent blood cells from primary and laboratory culture origins, in healthy and pathological situations (chronic myelogenous leukaemia). In a second part of this presentation, we concentrate on the temporal dynamics of the phase contour chains and we discuss the spectral decomposition of their dynamics in both health and disease.