A fiber-optic fluorescence microscope using a consumer-grade digital camera for in vivo cellular imaging.

PubMed

Shin, Dongsuk; Pierce, Mark C; Gillenwater, Ann M; Williams, Michelle D; Richards-Kortum, Rebecca R

2010-06-23

Early detection is an essential component of cancer management. Unfortunately, visual examination can often be unreliable, and many settings lack the financial capital and infrastructure to operate PET, CT, and MRI systems. Moreover, the infrastructure and expense associated with surgical biopsy and microscopy are a challenge to establishing cancer screening/early detection programs in low-resource settings. Improvements in performance and declining costs have led to the availability of optoelectronic components, which can be used to develop low-cost diagnostic imaging devices for use at the point-of-care. Here, we demonstrate a fiber-optic fluorescence microscope using a consumer-grade camera for in vivo cellular imaging. The fiber-optic fluorescence microscope includes an LED light, an objective lens, a fiber-optic bundle, and a consumer-grade digital camera. The system was used to image an oral cancer cell line labeled with 0.01% proflavine. A human tissue specimen was imaged following surgical resection, enabling dysplastic and cancerous regions to be evaluated. The oral mucosa of a healthy human subject was imaged in vivo, following topical application of 0.01% proflavine. The fiber-optic microscope resolved individual nuclei in all specimens and tissues imaged. This capability allowed qualitative and quantitative differences between normal and precancerous or cancerous tissues to be identified. The optical efficiency of the system permitted imaging of the human oral mucosa in real time. Our results indicate this device as a useful tool to assist in the identification of early neoplastic changes in epithelial tissues. This portable, inexpensive unit may be particularly appropriate for use at the point-of-care in low-resource settings.

Application of Tablet PCs to Lecture Demonstrations on Optical Mineralogy

ERIC Educational Resources Information Center

Hoisch, Thomas D.; Austin, Barbara A.; Newell, Shawn L.; Manone, Mark F.

2010-01-01

Learning optical mineralogy requires students to integrate a complex theory with microscope manipulations and image interpretation. To assist student learning, we performed lecture demonstrations during which digital photomicrographs were taken and delivered to students using Tablet PCs, whereupon they were imported into note-taking software and…

Fast and Accurate Cell Tracking by a Novel Optical-Digital Hybrid Method

NASA Astrophysics Data System (ADS)

Torres-Cisneros, M.; Aviña-Cervantes, J. G.; Pérez-Careta, E.; Ambriz-Colín, F.; Tinoco, Verónica; Ibarra-Manzano, O. G.; Plascencia-Mora, H.; Aguilera-Gómez, E.; Ibarra-Manzano, M. A.; Guzman-Cabrera, R.; Debeir, Olivier; Sánchez-Mondragón, J. J.

2013-09-01

An innovative methodology to detect and track cells using microscope images enhanced by optical cross-correlation techniques is proposed in this paper. In order to increase the tracking sensibility, image pre-processing has been implemented as a morphological operator on the microscope image. Results show that the pre-processing process allows for additional frames of cell tracking, therefore increasing its robustness. The proposed methodology can be used in analyzing different problems such as mitosis, cell collisions, and cell overlapping, ultimately designed to identify and treat illnesses and malignancies.

Three-dimensional motion-picture imaging of dynamic object by parallel-phase-shifting digital holographic microscopy using an inverted magnification optical system

NASA Astrophysics Data System (ADS)

Fukuda, Takahito; Shinomura, Masato; Xia, Peng; Awatsuji, Yasuhiro; Nishio, Kenzo; Matoba, Osamu

2017-04-01

We constructed a parallel-phase-shifting digital holographic microscopy (PPSDHM) system using an inverted magnification optical system, and succeeded in three-dimensional (3D) motion-picture imaging for 3D displacement of a microscopic object. In the PPSDHM system, the inverted and afocal magnification optical system consisted of a microscope objective (16.56 mm focal length and 0.25 numerical aperture) and a convex lens (300 mm focal length and 82 mm aperture diameter). A polarization-imaging camera was used to record multiple phase-shifted holograms with a single-shot exposure. We recorded an alum crystal, sinking down in aqueous solution of alum, by the constructed PPSDHM system at 60 frames/s for about 20 s and reconstructed high-quality 3D motion-picture image of the crystal. Then, we calculated amounts of displacement of the crystal from the amounts in the focus plane and the magnifications of the magnification optical system, and obtained the 3D trajectory of the crystal by that amounts.

Compact, light-weight and cost-effective microscope based on lensless incoherent holography for telemedicine applications.

PubMed

Mudanyali, Onur; Tseng, Derek; Oh, Chulwoo; Isikman, Serhan O; Sencan, Ikbal; Bishara, Waheb; Oztoprak, Cetin; Seo, Sungkyu; Khademhosseini, Bahar; Ozcan, Aydogan

2010-06-07

Despite the rapid progress in optical imaging, most of the advanced microscopy modalities still require complex and costly set-ups that unfortunately limit their use beyond well equipped laboratories. In the meantime, microscopy in resource-limited settings has requirements significantly different from those encountered in advanced laboratories, and such imaging devices should be cost-effective, compact, light-weight and appropriately accurate and simple to be usable by minimally trained personnel. Furthermore, these portable microscopes should ideally be digitally integrated as part of a telemedicine network that connects various mobile health-care providers to a central laboratory or hospital. Toward this end, here we demonstrate a lensless on-chip microscope weighing approximately 46 grams with dimensions smaller than 4.2 cm x 4.2 cm x 5.8 cm that achieves sub-cellular resolution over a large field of view of approximately 24 mm(2). This compact and light-weight microscope is based on digital in-line holography and does not need any lenses, bulky optical/mechanical components or coherent sources such as lasers. Instead, it utilizes a simple light-emitting-diode (LED) and a compact opto-electronic sensor-array to record lensless holograms of the objects, which then permits rapid digital reconstruction of regular transmission or differential interference contrast (DIC) images of the objects. Because this lensless incoherent holographic microscope has orders-of-magnitude improved light collection efficiency and is very robust to mechanical misalignments it may offer a cost-effective tool especially for telemedicine applications involving various global health problems in resource limited settings.

Perspective: Electronic systems of knowledge in the world of virtual microscopy.

PubMed

Maybury, Terrence; Farah, Camile S

2009-09-01

Across a broad range of medical disciplines, learning how to use an optical or light microscope has been a mandatory inclusion in the undergraduate curriculum. The development of virtual microscopy (VM) technology during the past 10 years has called into question the use of the optical microscope in educational contexts. VM allows slide specimens to be digitized, which, in turn, allows the computer to mimic the workings of the light microscope. This move from analog technology (the light microscope) to digital technology (the computer as microscope) is part of the many significant changes going on in education, a singular manifestation of the broader move from print-literate traditions of knowledge (requiring literacy) to an electronics-literate, or "electrate," mode (requiring "electracy"). VM is here used as an exemplar of this broad transition from literacy to electracy, some components of which include data deluge, a multimodal structure, and modularity. Understandably, this transition is important to clarify educationally, especially in a global context mediated via digital means. A related aspect of these educational changes is the move from teacher-directed learning to student-centered learning, or "user-led education," which points to a redefinition of "pedagogy" as "andragogy." The dissemination of the specific value of VM, then, is critical to both learners and teachers and to a more coherent understanding of electracy. A practical consequence of this clarity might be a better application of this knowledge in the evolving fields of computer simulation and telemedicine, areas in which today's medical students will need future expertise.

Digital holographic microscope with low-frequency attenuation filter for position measurement of a nanoparticle.

PubMed

Pham, Quang Duc; Kusumi, Yuichi; Hasegawa, Satoshi; Hayasaki, Yoshio

2012-10-01

We propose a new method for three-dimensional (3D) position measurement of nanoparticles using an in-line digital holographic microscope. The method improves the signal-to-noise ratio of the amplitude of the interference fringes to achieve higher accuracy in the position measurement by increasing weak scattered light from a nanoparticle relative to the reference light by using a low spatial frequency attenuation filter. We demonstrated the improvements of signal-to-noise ratio of the optical system and contrast of the interference fringes, allowing the 3D positions of nanoparticles to be determined more precisely.

Optical fabrication and testing; Proceedings of the Meeting, Singapore, Oct. 22-27, 1990

NASA Astrophysics Data System (ADS)

Lorenzen, Manfred; Campbell, Duncan R.; Johnson, Craig W.

1991-03-01

Various papers on optical fabrication and testing are presented. Individual topics addressed include: interferometry with laser diodes, new methods for economic production of prisms and lenses, interferometer accuracy and precision, optical testing with wavelength scanning interferometer, digital Talbot interferometer, high-sensitivity interferometric technique for strain measurements, absolute interferometric testing of spherical surfaces, contouring using gratings created on an LCD panel, three-dimensional inspection using laser-based dynamic fringe projection, noncontact optical microtopography, laser scan microscope and infrared laser scan microscope, photon scanning tunneling microscopy. Also discussed are: combination-matching problems in the layout design of minilaser rangefinder, design and testing of a cube-corner array for laser ranging, mode and far-field pattern of diode laser-phased arrays, new glasses for optics and optoelectronics, optical properties of Li-doped ZnO films, application and machining of Zerodur for optical purposes, finish machining of optical components in mass production.

Optical fabrication and testing; Proceedings of the Meeting, Singapore, Oct. 22-27, 1990

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

Lorenzen, M.; Campbell, D.R.; Johnson, C.W.

1991-01-01

Various papers on optical fabrication and testing are presented. Individual topics addressed include: interferometry with laser diodes, new methods for economic production of prisms and lenses, interferometer accuracy and precision, optical testing with wavelength scanning interferometer, digital Talbot interferometer, high-sensitivity interferometric technique for strain measurements, absolute interferometric testing of spherical surfaces, contouring using gratings created on an LCD panel, three-dimensional inspection using laser-based dynamic fringe projection, noncontact optical microtopography, laser scan microscope and infrared laser scan microscope, photon scanning tunneling microscopy. Also discussed are: combination-matching problems in the layout design of minilaser rangefinder, design and testing of a cube-corner arraymore » for laser ranging, mode and far-field pattern of diode laser-phased arrays, new glasses for optics and optoelectronics, optical properties of Li-doped ZnO films, application and machining of Zerodur for optical purposes, finish machining of optical components in mass production.« less

Digital optical imaging of green fluorescent proteins for tracking vascular gene expression: feasibility study in rabbit and human cell models.

PubMed

Yang, X; Liu, H; Li, D; Zhou, X; Jung, W C; Deans, A E; Cui, Y; Cheng, L

2001-04-01

To investigate the feasibility of using a sensitive digital optical imaging technique to detect green fluorescent protein (GFP) expressed in rabbit vasculature and human arterial smooth muscle cells. A GFP plasmid was transfected into human arterial smooth muscle cells to obtain a GFP-smooth muscle cell solution. This solution was imaged in cell phantoms by using a prototype digital optical imaging system. For in vivo validation, a GFP-lentivirus vector was transfected during surgery into the carotid arteries of two rabbits, and GFP-targeted vessels were harvested for digital optical imaging ex vivo. Optical imaging of cell phantoms resulted in a spatial resolution of 25 microm/pixel. Fluorescent signals were detected as diffusely distributed bright spots. At ex vivo optical imaging of arterial tissues, the average fluorescent signal was significantly higher (P <.05) in GFP-targeted tissues (mean +/- SD, 9,357.3 absolute units of density +/- 1,001.3) than in control tissues (5,633.7 absolute units of density +/- 985.2). Both fluorescence microscopic and immunohistochemical findings confirmed these differences between GFP-targeted and control vessels. The digital optical imaging system was sensitive to GFPs and may potentially provide an in vivo imaging tool to monitor and track vascular gene transfer and expression in experimental investigations.

Lensless digital holographic microscopy and its applications in biomedicine and environmental monitoring.

PubMed

Wu, Yichen; Ozcan, Aydogan

2018-03-01

Optical compound microscope has been a major tool in biomedical imaging for centuries. Its performance relies on relatively complicated, bulky and expensive lenses and alignment mechanics. In contrast, the lensless microscope digitally reconstructs microscopic images of specimens without using any lenses, as a result of which it can be made much smaller, lighter and lower-cost. Furthermore, the limited space-bandwidth product of objective lenses in a conventional microscope can be significantly surpassed by a lensless microscope. Such lensless imaging designs have enabled high-resolution and high-throughput imaging of specimens using compact, portable and cost-effective devices to potentially address various point-of-care, global-health and telemedicine related challenges. In this review, we discuss the operation principles and the methods behind lensless digital holographic on-chip microscopy. We also go over various applications that are enabled by cost-effective and compact implementations of lensless microscopy, including some recent work on air quality monitoring, which utilized machine learning for high-throughput and accurate quantification of particulate matter in air. Finally, we conclude with a brief future outlook of this computational imaging technology. Copyright © 2017 Elsevier Inc. All rights reserved.

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.

Suspension and simple optical characterization of two-dimensional membranes

NASA Astrophysics Data System (ADS)

Northeast, David B.; Knobel, Robert G.

2018-03-01

We report on a method for suspending two-dimensional crystal materials in an electronic circuit using an only photoresists and solvents. Graphene and NbSe2 are suspended tens of nanometers above metal electrodes with clamping diameters of several microns. The optical cavity formed from the membrane/air/metal structures enables a quick method to measure the number of layers and the gap separation using comparisons between the expected colour and optical microscope images. This characterization technique can be used with just an illuminated microscope with a digital camera which makes it adaptable to environments where other means of characterization are not possible, such as inside nitrogen glove boxes used in handling oxygen-sensitive materials.

Digital micromirror devices: principles and applications in imaging.

PubMed

Bansal, Vivek; Saggau, Peter

2013-05-01

A digital micromirror device (DMD) is an array of individually switchable mirrors that can be used in many advanced optical systems as a rapid spatial light modulator. With a DMD, several implementations of confocal microscopy, hyperspectral imaging, and fluorescence lifetime imaging can be realized. The DMD can also be used as a real-time optical processor for applications such as the programmable array microscope and compressive sensing. Advantages and disadvantages of the DMD for these applications as well as methods to overcome some of the limitations will be discussed in this article. Practical considerations when designing with the DMD and sample optical layouts of a completely DMD-based imaging system and one in which acousto-optic deflectors (AODs) are used in the illumination pathway are also provided.

Particle detection, number estimation, and feature measurement in gene transfer studies: optical fractionator stereology integrated with digital image processing and analysis.

PubMed

King, Michael A; Scotty, Nicole; Klein, Ronald L; Meyer, Edwin M

2002-10-01

Assessing the efficacy of in vivo gene transfer often requires a quantitative determination of the number, size, shape, or histological visualization characteristics of biological objects. The optical fractionator has become a choice stereological method for estimating the number of objects, such as neurons, in a structure, such as a brain subregion. Digital image processing and analytic methods can increase detection sensitivity and quantify structural and/or spectral features located in histological specimens. We describe a hardware and software system that we have developed for conducting the optical fractionator process. A microscope equipped with a video camera and motorized stage and focus controls is interfaced with a desktop computer. The computer contains a combination live video/computer graphics adapter with a video frame grabber and controls the stage, focus, and video via a commercial imaging software package. Specialized macro programs have been constructed with this software to execute command sequences requisite to the optical fractionator method: defining regions of interest, positioning specimens in a systematic uniform random manner, and stepping through known volumes of tissue for interactive object identification (optical dissectors). The system affords the flexibility to work with count regions that exceed the microscope image field size at low magnifications and to adjust the parameters of the fractionator sampling to best match the demands of particular specimens and object types. Digital image processing can be used to facilitate object detection and identification, and objects that meet criteria for counting can be analyzed for a variety of morphometric and optical properties. Copyright 2002 Elsevier Science (USA)

Field-portable lensfree tomographic microscope.

PubMed

Isikman, Serhan O; Bishara, Waheb; Sikora, Uzair; Yaglidere, Oguzhan; Yeah, John; Ozcan, Aydogan

2011-07-07

We present a field-portable lensfree tomographic microscope, which can achieve sectional imaging of a large volume (∼20 mm(3)) on a chip with an axial resolution of <7 μm. In this compact tomographic imaging platform (weighing only ∼110 grams), 24 light-emitting diodes (LEDs) that are each butt-coupled to a fibre-optic waveguide are controlled through a cost-effective micro-processor to sequentially illuminate the sample from different angles to record lensfree holograms of the sample that is placed on the top of a digital sensor array. In order to generate pixel super-resolved (SR) lensfree holograms and hence digitally improve the achievable lateral resolution, multiple sub-pixel shifted holograms are recorded at each illumination angle by electromagnetically actuating the fibre-optic waveguides using compact coils and magnets. These SR projection holograms obtained over an angular range of ±50° are rapidly reconstructed to yield projection images of the sample, which can then be back-projected to compute tomograms of the objects on the sensor-chip. The performance of this compact and light-weight lensfree tomographic microscope is validated by imaging micro-beads of different dimensions as well as a Hymenolepis nana egg, which is an infectious parasitic flatworm. Achieving a decent three-dimensional spatial resolution, this field-portable on-chip optical tomographic microscope might provide a useful toolset for telemedicine and high-throughput imaging applications in resource-poor settings. This journal is © The Royal Society of Chemistry 2011

A Low-Cost Digital Microscope with Real-Time Fluorescent Imaging Capability.

PubMed

Hasan, Md Mehedi; Alam, Mohammad Wajih; Wahid, Khan A; Miah, Sayem; Lukong, Kiven Erique

2016-01-01

This paper describes the development of a prototype of a low-cost digital fluorescent microscope built from commercial off-the-shelf (COTS) components. The prototype was tested to detect malignant tumor cells taken from a living organism in a preclinical setting. This experiment was accomplished by using Alexa Fluor 488 conjugate dye attached to the cancer cells. Our prototype utilizes a torch along with an excitation filter as a light source for fluorophore excitation, a dichroic mirror to reflect the excitation and pass the emitted green light from the sample under test and a barrier filter to permit only appropriate wavelength. The system is designed out of a microscope using its optical zooming property and an assembly of exciter filter, dichroic mirror and transmitter filter. The microscope is connected to a computer or laptop through universal serial bus (USB) that allows real-time transmission of captured florescence images; this also offers real-time control of the microscope. The designed system has comparable features of high-end commercial fluorescent microscopes while reducing cost, power, weight and size.

A Low-Cost Digital Microscope with Real-Time Fluorescent Imaging Capability

PubMed Central

Hasan, Md. Mehedi; Wahid, Khan A.; Miah, Sayem; Lukong, Kiven Erique

2016-01-01

This paper describes the development of a prototype of a low-cost digital fluorescent microscope built from commercial off-the-shelf (COTS) components. The prototype was tested to detect malignant tumor cells taken from a living organism in a preclinical setting. This experiment was accomplished by using Alexa Fluor 488 conjugate dye attached to the cancer cells. Our prototype utilizes a torch along with an excitation filter as a light source for fluorophore excitation, a dichroic mirror to reflect the excitation and pass the emitted green light from the sample under test and a barrier filter to permit only appropriate wavelength. The system is designed out of a microscope using its optical zooming property and an assembly of exciter filter, dichroic mirror and transmitter filter. The microscope is connected to a computer or laptop through universal serial bus (USB) that allows real-time transmission of captured florescence images; this also offers real-time control of the microscope. The designed system has comparable features of high-end commercial fluorescent microscopes while reducing cost, power, weight and size. PMID:27977709

A fast low-power optical memory based on coupled micro-ring lasers

NASA Astrophysics Data System (ADS)

Hill, Martin T.; Dorren, Harmen J. S.; de Vries, Tjibbe; Leijtens, Xaveer J. M.; den Besten, Jan Hendrik; Smalbrugge, Barry; Oei, Yok-Siang; Binsma, Hans; Khoe, Giok-Djan; Smit, Meint K.

2004-11-01

The increasing speed of fibre-optic-based telecommunications has focused attention on high-speed optical processing of digital information. Complex optical processing requires a high-density, high-speed, low-power optical memory that can be integrated with planar semiconductor technology for buffering of decisions and telecommunication data. Recently, ring lasers with extremely small size and low operating power have been made, and we demonstrate here a memory element constructed by interconnecting these microscopic lasers. Our device occupies an area of 18 × 40µm2 on an InP/InGaAsP photonic integrated circuit, and switches within 20ps with 5.5fJ optical switching energy. Simulations show that the element has the potential for much smaller dimensions and switching times. Large numbers of such memory elements can be densely integrated and interconnected on a photonic integrated circuit: fast digital optical information processing systems employing large-scale integration should now be viable.

Real-time processing of interferograms for monitoring protein crystal growth on the Space Station

NASA Technical Reports Server (NTRS)

Choudry, A.; Dupuis, N.

1988-01-01

The possibility of using microscopic interferometric techniques to monitor the growth of protein crystals on the Space Station is studied. Digital image processing techniques are used to develop a system for the real-time analysis of microscopic interferograms of nucleation sites during protein crystal growth. Features of the optical setup and the image processing system are discussed and experimental results are presented.

Standardization of whole slide image morphologic assessment with definition of a new application: Digital slide dynamic morphometry.

PubMed

Puppa, Giacomo; Risio, Mauro; Sheahan, Kieran; Vieth, Michael; Zlobec, Inti; Lugli, Alessandro; Pecori, Sara; Wang, Lai Mun; Langner, Cord; Mitomi, Hiroyuki; Nakamura, Takatoshi; Watanabe, Masahiko; Ueno, Hideki; Chasle, Jacques; Senore, Carlo; Conley, Stephen A; Herlin, Paulette; Lauwers, Gregory Y

2011-01-01

In histopathology, the quantitative assessment of various morphologic features is based on methods originally conceived on specific areas observed through the microscope used. Failure to reproduce the same reference field of view using a different microscope will change the score assessed. Visualization of a digital slide on a screen through a dedicated viewer allows selection of the magnification. However, the field of view is rectangular, unlike the circular field of optical microscopy. In addition, the size of the selected area is not evident, and must be calculated. A digital slide morphometric system was conceived to reproduce the various methods published for assessing tumor budding in colorectal cancer. Eighteen international experts in colorectal cancer were invited to participate in a web-based study by assessing tumor budding with five different methods in 100 digital slides. The specific areas to be tested by each method were marked by colored circles. The areas were grouped in a target-like pattern and then saved as an .xml file. When a digital slide was opened, the .xml file was imported in order to perform the measurements. Since the morphometric tool is composed of layers that can be freely moved on top of the digital slide, the technique was named digital slide dynamic morphometry. Twelve investigators completed the task, the majority of them performing the multiple evaluations of each of the cases in less than 12 minutes. Digital slide dynamic morphometry has various potential applications and might be a useful tool for the assessment of histologic parameters originally conceived for optical microscopy that need to be quantified.

Wide-field computational imaging of pathology slides using lens-free on-chip microscopy.

PubMed

Greenbaum, Alon; Zhang, Yibo; Feizi, Alborz; Chung, Ping-Luen; Luo, Wei; Kandukuri, Shivani R; Ozcan, Aydogan

2014-12-17

Optical examination of microscale features in pathology slides is one of the gold standards to diagnose disease. However, the use of conventional light microscopes is partially limited owing to their relatively high cost, bulkiness of lens-based optics, small field of view (FOV), and requirements for lateral scanning and three-dimensional (3D) focus adjustment. We illustrate the performance of a computational lens-free, holographic on-chip microscope that uses the transport-of-intensity equation, multi-height iterative phase retrieval, and rotational field transformations to perform wide-FOV imaging of pathology samples with comparable image quality to a traditional transmission lens-based microscope. The holographically reconstructed image can be digitally focused at any depth within the object FOV (after image capture) without the need for mechanical focus adjustment and is also digitally corrected for artifacts arising from uncontrolled tilting and height variations between the sample and sensor planes. Using this lens-free on-chip microscope, we successfully imaged invasive carcinoma cells within human breast sections, Papanicolaou smears revealing a high-grade squamous intraepithelial lesion, and sickle cell anemia blood smears over a FOV of 20.5 mm(2). The resulting wide-field lens-free images had sufficient image resolution and contrast for clinical evaluation, as demonstrated by a pathologist's blinded diagnosis of breast cancer tissue samples, achieving an overall accuracy of ~99%. By providing high-resolution images of large-area pathology samples with 3D digital focus adjustment, lens-free on-chip microscopy can be useful in resource-limited and point-of-care settings. Copyright © 2014, American Association for the Advancement of Science.

Structured illumination 3D microscopy using adaptive lenses and multimode fibers

NASA Astrophysics Data System (ADS)

Czarske, Jürgen; Philipp, Katrin; Koukourakis, Nektarios

2017-06-01

Microscopic techniques with high spatial and temporal resolution are required for in vivo studying biological cells and tissues. Adaptive lenses exhibit strong potential for fast motion-free axial scanning. However, they also lead to a degradation of the achievable resolution because of aberrations. This hurdle can be overcome by digital optical technologies. We present a novel High-and-Low-frequency (HiLo) 3D-microscope using structured illumination and an adaptive lens. Uniform illumination is used to obtain optical sectioning for the high-frequency (Hi) components of the image, and nonuniform illumination is needed to obtain optical sectioning for the low-frequency (Lo) components of the image. Nonuniform illumination is provided by a multimode fiber. It ensures robustness against optical aberrations of the adaptive lens. The depth-of-field of our microscope can be adjusted a-posteriori by computational optics. It enables to create flexible scans, which compensate for irregular axial measurement positions. The adaptive HiLo 3D-microscope provides an axial scanning range of 1 mm with an axial resolution of about 4 microns and sub-micron lateral resolution over the full scanning range. In result, volumetric measurements with high temporal and spatial resolution are provided. Demonstration measurements of zebrafish embryos with reporter gene-driven fluorescence in the thyroid gland are presented.

Simulation study on compressive laminar optical tomography for cardiac action potential propagation

PubMed Central

Harada, Takumi; Tomii, Naoki; Manago, Shota; Kobayashi, Etsuko; Sakuma, Ichiro

2017-01-01

To measure the activity of tissue at the microscopic level, laminar optical tomography (LOT), which is a microscopic form of diffuse optical tomography, has been developed. However, obtaining sufficient recording speed to determine rapidly changing dynamic activity remains major challenges. For a high frame rate of the reconstructed data, we here propose a new LOT method using compressed sensing theory, called compressive laminar optical tomography (CLOT), in which novel digital micromirror device-based illumination and data reduction in a single reconstruction are applied. In the simulation experiments, the reconstructed volumetric images of the action potentials that were acquired from 5 measured images with random pattern featured a wave border at least to a depth of 2.5 mm. Consequently, it was shown that CLOT has potential for over 200 fps required for the cardiac electrophysiological phenomena. PMID:28736675

Optical digital microscopy for cyto- and hematological studies in vitro

NASA Astrophysics Data System (ADS)

Ganilova, Yu. A.; Dolmashkin, A. A.; Doubrovski, V. A.; Yanina, I. Yu.; Tuchin, V. V.

2013-08-01

The dependence of the spatial resolution and field of view of an optical microscope equipped with a CCD camera on the objective magnification has been experimentally investigated. Measurement of these characteristics has shown that a spatial resolution of 20-25 px/μm at a field of view of about 110 μm is quite realistic; this resolution is acceptable for a detailed study of the processes occurring in cell. It is proposed to expand the dynamic range of digital camera by measuring and approximating its light characteristics with subsequent plotting of the corresponding calibration curve. The biological objects of study were human adipose tissue cells, as well as erythrocytes and their immune complexes in human blood; both objects have been investigated in vitro. Application of optical digital microscopy for solving specific problems of cytology and hematology can be useful in both biomedical studies in experiments with objects of nonbiological origin.

Field-portable lensfree tomographic microscope†

PubMed Central

Isikman, Serhan O.; Bishara, Waheb; Sikora, Uzair; Yaglidere, Oguzhan; Yeah, John; Ozcan, Aydogan

2011-01-01

We present a field-portable lensfree tomographic microscope, which can achieve sectional imaging of a large volume (~20 mm3) on a chip with an axial resolution of <7 μm. In this compact tomographic imaging platform (weighing only ~110 grams), 24 light-emitting diodes (LEDs) that are each butt-coupled to a fibre-optic waveguide are controlled through a cost-effective micro-processor to sequentially illuminate the sample from different angles to record lensfree holograms of the sample that is placed on the top of a digital sensor array. In order to generate pixel super-resolved (SR) lensfree holograms and hence digitally improve the achievable lateral resolution, multiple sub-pixel shifted holograms are recorded at each illumination angle by electromagnetically actuating the fibre-optic waveguides using compact coils and magnets. These SR projection holograms obtained over an angular range of ~50° are rapidly reconstructed to yield projection images of the sample, which can then be back-projected to compute tomograms of the objects on the sensor-chip. The performance of this compact and light-weight lensfree tomographic microscope is validated by imaging micro-beads of different dimensions as well as a Hymenolepis nana egg, which is an infectious parasitic flatworm. Achieving a decent three-dimensional spatial resolution, this field-portable on-chip optical tomographic microscope might provide a useful toolset for telemedicine and high-throughput imaging applications in resource-poor settings. PMID:21573311

HOMER: the Holographic Optical Microscope for Education and Research

NASA Astrophysics Data System (ADS)

Luviano, Anali

Holography was invented in 1948 by Dennis Gabor and has undergone major advancements since the 2000s leading to the development of commercial digital holographic microscopes (DHM). This noninvasive form of microscopy produces a three-dimensional (3-D) digital model of a sample without altering or destroying the sample, thus allowing the same sample to be studied multiple times. HOMER-the Holographic Optical Microscope for Education and Research-produces a 3-D image from a two-dimensional (2-D) interference pattern captured by a camera that is then put through reconstruction software. This 2-D pattern is created when a reference wave interacts with the sample to produce a secondary wave that interferes with the unaltered part of the reference wave. I constructed HOMER to be an efficient, portable in-line DHM using inexpensive material and free reconstruction software. HOMER uses three different-colored LEDs as light sources. I am testing the performance of HOMER with the goal of producing tri-color images of samples. I'm using small basic biological samples to test the effectiveness of HOMER and plan to transition to complex cellular and biological specimens as I pursue my interest in biophysics. Norwich University.

High contrast imaging and flexible photomanipulation for quantitative in vivo multiphoton imaging with polygon scanning microscope.

PubMed

Li, Yongxiao; Montague, Samantha J; Brüstle, Anne; He, Xuefei; Gillespie, Cathy; Gaus, Katharina; Gardiner, Elizabeth E; Lee, Woei Ming

2018-02-28

In this study, we introduce two key improvements that overcome limitations of existing polygon scanning microscopes while maintaining high spatial and temporal imaging resolution over large field of view (FOV). First, we proposed a simple and straightforward means to control the scanning angle of the polygon mirror to carry out photomanipulation without resorting to high speed optical modulators. Second, we devised a flexible data sampling method directly leading to higher image contrast by over 2-fold and digital images with 100 megapixels (10 240 × 10 240) per frame at 0.25 Hz. This generates sub-diffraction limited pixels (60 nm per pixels over the FOV of 512 μm) which increases the degrees of freedom to extract signals computationally. The unique combined optical and digital control recorded fine fluorescence recovery after localized photobleaching (r ~10 μm) within fluorescent giant unilamellar vesicles and micro-vascular dynamics after laser-induced injury during thrombus formation in vivo. These new improvements expand the quantitative biological-imaging capacity of any polygon scanning microscope system. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Comparison study of five different display modalities for whole slide images in surgical pathology and cytopathology in Europe

NASA Astrophysics Data System (ADS)

D'Haene, Nicky; Maris, Calliope; Rorive, Sandrine; Moles Lopez, Xavier; Rostang, Johan; Marchessoux, Cédric; Pantanowitz, Liron; Parwani, Anil V.; Salmon, Isabelle

2013-03-01

User experience with viewing images in pathology is crucial for accurate interpretation and diagnosis. With digital pathology, images are being read on a display system, and this poses new types of questions: such as what is the difference in terms of pixelation, refresh lag or obscured features compared to an optical microscope. Is there a resultant change in user performance in terms of speed of slide review, perception of adequacy and quality or in diagnostic confidence? A prior psychophysical study was carried out comparing various display modalities on whole slide imaging (WSI) in pathology at the University of Pittsburgh Medical Center (UPMC) in the USA. This prior study compared professional and non-professional grade display modalities and highlighted the importance of using a medical grade display to view pathological digital images. This study was duplicated in Europe at the Department of Pathology in Erasme Hospital (Université Libre de Bruxelles (ULB)) in an attempt to corroborate these findings. Digital WSI with corresponding glass slides of 58 cases including surgical pathology and cytopathology slides of varying difficulty were employed. Similar non-professional and professional grade display modalities were compared to an optical microscope (Olympus BX51). Displays ranged from a laptop (DELL Latitude D620), to a consumer grade display (DELL E248WFPb), to two professional grade monitors (Eizo CG245W and Barco MDCC-6130). Three pathologists were selected from the Department of Pathology in Erasme Hospital (ULB) in Belgium to view and interpret the pathological images on these different displays. The results show that non-professional grade displays (laptop and consumer) have inferior user experience compared to professional grade monitors and the optical microscope.

Holographic fluorescence microscopy with incoherent digital holographic adaptive optics

NASA Astrophysics Data System (ADS)

Jang, Changwon; Kim, Jonghyun; Clark, David C.; Lee, Byoungho; Kim, Myung K.

2015-03-01

Introduction of adaptive optics technology into astronomy and ophthalmology has made great contributions in these fields, allowing one to recover images blurred by atmospheric turbulence or aberrations of the eye. Similar adaptive optics improvement in microscopic imaging is also of interest to researchers using various techniques. Current technology of adaptive optics typically contains three key elements: wavefront sensor, wavefront corrector and controller. These hardware elements tend to be bulky, expensive, and limited in resolution, involving, e.g., lenslet arrays for sensing or multi-acuator deformable mirrors for correcting. We have previously introduced an alternate approach to adaptive optics based on unique capabilities of digital holography, namely direct access to the phase profile of an optical field and the ability to numerically manipulate the phase profile. We have also demonstrated that direct access and compensation of the phase profile is possible not only with the conventional coherent type of digital holography, but also with a new type of digital holography using incoherent light: self-interference incoherent digital holography (SIDH). The SIDH generates complex - i.e. amplitude plus phase - hologram from one or several interferograms acquired with incoherent light, such as LEDs, lamps, sunlight, or fluorescence. The complex point spread function can be measured using a guide star illumination and it allows deterministic deconvolution of the full-field image. We present experimental demonstration of aberration compensation in holographic fluorescence microscopy using SIDH. The adaptive optics by SIDH provides new tools for improved cellular fluorescence microscopy through intact tissue layers or other types of aberrant media.

Volumetric Light-field Encryption at the Microscopic Scale

PubMed Central

Li, Haoyu; Guo, Changliang; Muniraj, Inbarasan; Schroeder, Bryce C.; Sheridan, John T.; Jia, Shu

2017-01-01

We report a light-field based method that allows the optical encryption of three-dimensional (3D) volumetric information at the microscopic scale in a single 2D light-field image. The system consists of a microlens array and an array of random phase/amplitude masks. The method utilizes a wave optics model to account for the dominant diffraction effect at this new scale, and the system point-spread function (PSF) serves as the key for encryption and decryption. We successfully developed and demonstrated a deconvolution algorithm to retrieve both spatially multiplexed discrete data and continuous volumetric data from 2D light-field images. Showing that the method is practical for data transmission and storage, we obtained a faithful reconstruction of the 3D volumetric information from a digital copy of the encrypted light-field image. The method represents a new level of optical encryption, paving the way for broad industrial and biomedical applications in processing and securing 3D data at the microscopic scale. PMID:28059149

Volumetric Light-field Encryption at the Microscopic Scale

NASA Astrophysics Data System (ADS)

Li, Haoyu; Guo, Changliang; Muniraj, Inbarasan; Schroeder, Bryce C.; Sheridan, John T.; Jia, Shu

2017-01-01

We report a light-field based method that allows the optical encryption of three-dimensional (3D) volumetric information at the microscopic scale in a single 2D light-field image. The system consists of a microlens array and an array of random phase/amplitude masks. The method utilizes a wave optics model to account for the dominant diffraction effect at this new scale, and the system point-spread function (PSF) serves as the key for encryption and decryption. We successfully developed and demonstrated a deconvolution algorithm to retrieve both spatially multiplexed discrete data and continuous volumetric data from 2D light-field images. Showing that the method is practical for data transmission and storage, we obtained a faithful reconstruction of the 3D volumetric information from a digital copy of the encrypted light-field image. The method represents a new level of optical encryption, paving the way for broad industrial and biomedical applications in processing and securing 3D data at the microscopic scale.

Multimodal full-field optical coherence tomography on biological tissue: toward all optical digital pathology

NASA Astrophysics Data System (ADS)

Harms, F.; Dalimier, E.; Vermeulen, P.; Fragola, A.; Boccara, A. C.

2012-03-01

Optical Coherence Tomography (OCT) is an efficient technique for in-depth optical biopsy of biological tissues, relying on interferometric selection of ballistic photons. Full-Field Optical Coherence Tomography (FF-OCT) is an alternative approach to Fourier-domain OCT (spectral or swept-source), allowing parallel acquisition of en-face optical sections. Using medium numerical aperture objective, it is possible to reach an isotropic resolution of about 1x1x1 ìm. After stitching a grid of acquired images, FF-OCT gives access to the architecture of the tissue, for both macroscopic and microscopic structures, in a non-invasive process, which makes the technique particularly suitable for applications in pathology. Here we report a multimodal approach to FF-OCT, combining two Full-Field techniques for collecting a backscattered endogeneous OCT image and a fluorescence exogeneous image in parallel. Considering pathological diagnosis of cancer, visualization of cell nuclei is of paramount importance. OCT images, even for the highest resolution, usually fail to identify individual nuclei due to the nature of the optical contrast used. We have built a multimodal optical microscope based on the combination of FF-OCT and Structured Illumination Microscopy (SIM). We used x30 immersion objectives, with a numerical aperture of 1.05, allowing for sub-micron transverse resolution. Fluorescent staining of nuclei was obtained using specific fluorescent dyes such as acridine orange. We present multimodal images of healthy and pathological skin tissue at various scales. This instrumental development paves the way for improvements of standard pathology procedures, as a faster, non sacrificial, operator independent digital optical method compared to frozen sections.

Image Montaging for Creating a Virtual Pathology Slide: An Innovative and Economical Tool to Obtain a Whole Slide Image

PubMed Central

Pandurangappa, Rohit; Annavajjula, Saileela; Rajashekaraiah, Premalatha Bidadi

2016-01-01

Background. Microscopes are omnipresent throughout the field of biological research. With microscopes one can see in detail what is going on at the cellular level in tissues. Though it is a ubiquitous tool, the limitation is that with high magnification there is a small field of view. It is often advantageous to see an entire sample at high magnification. Over the years technological advancements in optics have helped to provide solutions to this limitation of microscopes by creating the so-called dedicated “slide scanners” which can provide a “whole slide digital image.” These scanners can provide seamless, large-field-of-view, high resolution image of entire tissue section. The only disadvantage of such complete slide imaging system is its outrageous cost, thereby hindering their practical use by most laboratories, especially in developing and low resource countries. Methods. In a quest for their substitute, we tried commonly used image editing software Adobe Photoshop along with a basic image capturing device attached to a trinocular microscope to create a digital pathology slide. Results. The seamless image created using Adobe Photoshop maintained its diagnostic quality. Conclusion. With time and effort photomicrographs obtained from a basic camera-microscope set up can be combined and merged in Adobe Photoshop to create a whole slide digital image of practically usable quality at a negligible cost. PMID:27747147

Image Montaging for Creating a Virtual Pathology Slide: An Innovative and Economical Tool to Obtain a Whole Slide Image.

PubMed

Banavar, Spoorthi Ravi; Chippagiri, Prashanthi; Pandurangappa, Rohit; Annavajjula, Saileela; Rajashekaraiah, Premalatha Bidadi

2016-01-01

Background . Microscopes are omnipresent throughout the field of biological research. With microscopes one can see in detail what is going on at the cellular level in tissues. Though it is a ubiquitous tool, the limitation is that with high magnification there is a small field of view. It is often advantageous to see an entire sample at high magnification. Over the years technological advancements in optics have helped to provide solutions to this limitation of microscopes by creating the so-called dedicated "slide scanners" which can provide a "whole slide digital image." These scanners can provide seamless, large-field-of-view, high resolution image of entire tissue section. The only disadvantage of such complete slide imaging system is its outrageous cost, thereby hindering their practical use by most laboratories, especially in developing and low resource countries. Methods . In a quest for their substitute, we tried commonly used image editing software Adobe Photoshop along with a basic image capturing device attached to a trinocular microscope to create a digital pathology slide. Results . The seamless image created using Adobe Photoshop maintained its diagnostic quality. Conclusion . With time and effort photomicrographs obtained from a basic camera-microscope set up can be combined and merged in Adobe Photoshop to create a whole slide digital image of practically usable quality at a negligible cost.

Note: A three-dimensional calibration device for the confocal microscope.

PubMed

Jensen, K E; Weitz, D A; Spaepen, F

2013-01-01

Modern confocal microscopes enable high-precision measurement in three dimensions by collecting stacks of 2D (x-y) images that can be assembled digitally into a 3D image. It is difficult, however, to ensure position accuracy, particularly along the optical (z) axis where scanning is performed by a different physical mechanism than in x-y. We describe a simple device to calibrate simultaneously the x, y, and z pixel-to-micrometer conversion factors for a confocal microscope. By taking a known 2D pattern and positioning it at a precise angle with respect to the microscope axes, we created a 3D reference standard. The device is straightforward to construct and easy to use.

Holographic fluorescence microscopy with incoherent digital holographic adaptive optics

NASA Astrophysics Data System (ADS)

Jang, Changwon; Kim, Jonghyun; Clark, David C.; Lee, Seungjae; Lee, Byoungho; Kim, Myung K.

2015-11-01

Introduction of adaptive optics technology into astronomy and ophthalmology has made great contributions in these fields, allowing one to recover images blurred by atmospheric turbulence or aberrations of the eye. Similar adaptive optics improvement in microscopic imaging is also of interest to researchers using various techniques. Current technology of adaptive optics typically contains three key elements: a wavefront sensor, wavefront corrector, and controller. These hardware elements tend to be bulky, expensive, and limited in resolution, involving, for example, lenslet arrays for sensing or multiactuator deformable mirrors for correcting. We have previously introduced an alternate approach based on unique capabilities of digital holography, namely direct access to the phase profile of an optical field and the ability to numerically manipulate the phase profile. We have also demonstrated that direct access and compensation of the phase profile are possible not only with conventional coherent digital holography, but also with a new type of digital holography using incoherent light: self­interference incoherent digital holography (SIDH). The SIDH generates a complex-i.e., amplitude plus phase-hologram from one or several interferograms acquired with incoherent light, such as LEDs, lamps, sunlight, or fluorescence. The complex point spread function can be measured using guide star illumination and it allows deterministic deconvolution of the full-field image. We present experimental demonstration of aberration compensation in holographic fluorescence microscopy using SIDH. Adaptive optics by SIDH provides new tools for improved cellular fluorescence microscopy through intact tissue layers or other types of aberrant media.

Holographic fluorescence microscopy with incoherent digital holographic adaptive optics.

PubMed

Jang, Changwon; Kim, Jonghyun; Clark, David C; Lee, Seungjae; Lee, Byoungho; Kim, Myung K

2015-01-01

Introduction of adaptive optics technology into astronomy and ophthalmology has made great contributions in these fields, allowing one to recover images blurred by atmospheric turbulence or aberrations of the eye. Similar adaptive optics improvement in microscopic imaging is also of interest to researchers using various techniques. Current technology of adaptive optics typically contains three key elements: a wavefront sensor, wavefront corrector, and controller. These hardware elements tend to be bulky, expensive, and limited in resolution, involving, for example, lenslet arrays for sensing or multiactuator deformable mirrors for correcting. We have previously introduced an alternate approach based on unique capabilities of digital holography, namely direct access to the phase profile of an optical field and the ability to numerically manipulate the phase profile. We have also demonstrated that direct access and compensation of the phase profile are possible not only with conventional coherent digital holography, but also with a new type of digital holography using incoherent light: selfinterference incoherent digital holography (SIDH). The SIDH generates a complex—i.e., amplitude plus phase—hologram from one or several interferograms acquired with incoherent light, such as LEDs, lamps, sunlight, or fluorescence. The complex point spread function can be measured using guide star illumination and it allows deterministic deconvolution of the full-field image. We present experimental demonstration of aberration compensation in holographic fluorescence microscopy using SIDH. Adaptive optics by SIDH provides new tools for improved cellular fluorescence microscopy through intact tissue layers or other types of aberrant media.

Temporal focusing-based multiphoton excitation microscopy via digital micromirror device.

PubMed

Yih, Jenq-Nan; Hu, Yvonne Yuling; Sie, Yong Da; Cheng, Li-Chung; Lien, Chi-Hsiang; Chen, Shean-Jen

2014-06-01

This Letter presents an enhanced temporal focusing-based multiphoton excitation (MPE) microscope in which the conventional diffraction grating is replaced by a digital micromirror device (DMD). Experimental results from imaging a thin fluorescence film show that the 4.0 μm axial resolution of the microscope is comparable with that of a setup incorporating a 600  lines/mm grating; hence, the optical sectioning ability of the proposed setup is demonstrated. Similar to a grating, the DMD diffracts illuminating light frequencies for temporal focusing; additionally, it generates arbitrary patterns. Since the DMD is placed on the image-conjugate plane of the objective lens' focal plane, the MPE pattern can be projected on the focal plane precisely.

Continuous stacking computational approach based automated microscope slide scanner

NASA Astrophysics Data System (ADS)

Murali, Swetha; Adhikari, Jayesh Vasudeva; Jagannadh, Veerendra Kalyan; Gorthi, Sai Siva

2018-02-01

Cost-effective and automated acquisition of whole slide images is a bottleneck for wide-scale deployment of digital pathology. In this article, a computation augmented approach for the development of an automated microscope slide scanner is presented. The realization of a prototype device built using inexpensive off-the-shelf optical components and motors is detailed. The applicability of the developed prototype to clinical diagnostic testing is demonstrated by generating good quality digital images of malaria-infected blood smears. Further, the acquired slide images have been processed to identify and count the number of malaria-infected red blood cells and thereby perform quantitative parasitemia level estimation. The presented prototype would enable cost-effective deployment of slide-based cyto-diagnostic testing in endemic areas.

Dual-mode optical microscope based on single-pixel imaging

NASA Astrophysics Data System (ADS)

Rodríguez, A. D.; Clemente, P.; Tajahuerce, E.; Lancis, J.

2016-07-01

We demonstrate an inverted microscope that can image specimens in both reflection and transmission modes simultaneously with a single light source. The microscope utilizes a digital micromirror device (DMD) for patterned illumination altogether with two single-pixel photosensors for efficient light detection. The system, a scan-less device with no moving parts, works by sequential projection of a set of binary intensity patterns onto the sample that are codified onto a modified commercial DMD. Data to be displayed are geometrically transformed before written into a memory cell to cancel optical artifacts coming from the diamond-like shaped structure of the micromirror array. The 24-bit color depth of the display is fully exploited to increase the frame rate by a factor of 24, which makes the technique practicable for real samples. Our commercial DMD-based LED-illumination is cost effective and can be easily coupled as an add-on module for already existing inverted microscopes. The reflection and transmission information provided by our dual microscope complement each other and can be useful for imaging non-uniform samples and to prevent self-shadowing effects.

Review of advanced imaging techniques

PubMed Central

Chen, Yu; Liang, Chia-Pin; Liu, Yang; Fischer, Andrew H.; Parwani, Anil V.; Pantanowitz, Liron

2012-01-01

Pathology informatics encompasses digital imaging and related applications. Several specialized microscopy techniques have emerged which permit the acquisition of digital images (“optical biopsies”) at high resolution. Coupled with fiber-optic and micro-optic components, some of these imaging techniques (e.g., optical coherence tomography) are now integrated with a wide range of imaging devices such as endoscopes, laparoscopes, catheters, and needles that enable imaging inside the body. These advanced imaging modalities have exciting diagnostic potential and introduce new opportunities in pathology. Therefore, it is important that pathology informaticists understand these advanced imaging techniques and the impact they have on pathology. This paper reviews several recently developed microscopic techniques, including diffraction-limited methods (e.g., confocal microscopy, 2-photon microscopy, 4Pi microscopy, and spatially modulated illumination microscopy) and subdiffraction techniques (e.g., photoactivated localization microscopy, stochastic optical reconstruction microscopy, and stimulated emission depletion microscopy). This article serves as a primer for pathology informaticists, highlighting the fundamentals and applications of advanced optical imaging techniques. PMID:22754737

Three-dimensional microscopic tomographic imagings of the cataract in a human lens in vivo

NASA Astrophysics Data System (ADS)

Masters, Barry R.

1998-10-01

The problem of three-dimensional visualization of a human lens in vivo has been solved by a technique of volume rendering a transformed series of 60 rotated Scheimpflug (a dual slit reflected light microscope) digital images. The data set was obtained by rotating the Scheimpflug camera about the optic axis of the lens in 3 degree increments. The transformed set of optical sections were first aligned to correct for small eye movements, and then rendered into a volume reconstruction with volume rendering computer graphics techniques. To help visualize the distribution of lens opacities (cataracts) in the living, human lens the intensity of light scattering was pseudocolor coded and the cataract opacities were displayed as a movie.

eeDAP: An Evaluation Environment for Digital and Analog Pathology.

PubMed

Gallas, Brandon D; Cheng, Wei-Chung; Gavrielides, Marios A; Ivansky, Adam; Keay, Tyler; Wunderlich, Adam; Hipp, Jason; Hewitt, Stephen M

2014-01-01

The purpose of this work is to present a platform for designing and executing studies that compare pathologists interpreting histopathology of whole slide images (WSI) on a computer display to pathologists interpreting glass slides on an optical microscope. Here we present eeDAP, an evaluation environment for digital and analog pathology. The key element in eeDAP is the registration of the WSI to the glass slide. Registration is accomplished through computer control of the microscope stage and a camera mounted on the microscope that acquires images of the real time microscope view. Registration allows for the evaluation of the same regions of interest (ROIs) in both domains. This can reduce or eliminate disagreements that arise from pathologists interpreting different areas and focuses the comparison on image quality. We reduced the pathologist interpretation area from an entire glass slide (≈10-30 mm) 2 to small ROIs <(50 um) 2 . We also made possible the evaluation of individual cells. We summarize eeDAP's software and hardware and provide calculations and corresponding images of the microscope field of view and the ROIs extracted from the WSIs. These calculations help provide a sense of eeDAP's functionality and operating principles, while the images provide a sense of the look and feel of studies that can be conducted in the digital and analog domains. The eeDAP software can be downloaded from code.google.com (project: eeDAP) as Matlab source or as a precompiled stand-alone license-free application.

Development of a combined portable x-ray fluorescence and Raman spectrometer for in situ analysis.

PubMed

Guerra, M; Longelin, S; Pessanha, S; Manso, M; Carvalho, M L

2014-06-01

In this work, we have built a portable X-ray fluorescence (XRF) spectrometer in a planar configuration coupled to a Raman head and a digital optical microscope, for in situ analysis. Several geometries for the XRF apparatus and digital microscope are possible in order to overcome spatial constraints and provide better measurement conditions. With this combined spectrometer, we are now able to perform XRF and Raman measurements in the same point without the need for sample collection, which can be crucial when dealing with cultural heritage objects, as well as forensic analysis. We show the capabilities of the spectrometer by measuring several standard reference materials, as well as other samples usually encountered in cultural heritage, geological, as well as biomedical studies.

Optical Scatter Imaging with a digital micromirror device.

PubMed

Zheng, Jing-Yi; Pasternack, Robert M; Boustany, Nada N

2009-10-26

We had developed Optical Scatter Imaging (OSI) as a method which combines light scattering spectroscopy with microscopic imaging to probe local particle size in situ. Using a variable diameter iris as a Fourier spatial filter, the technique consisted of collecting images that encoded the intensity ratio of wide-to-narrow angle scatter at each pixel in the full field of view. In this paper, we replace the variable diameter Fourier filter with a digital micromirror device (DMD) to extend our assessment of morphology to the characterization of particle shape and orientation. We describe our setup in detail and demonstrate how to eliminate aberrations associated with the placement of the DMD in a conjugate Fourier plane of our microscopic imaging system. Using bacteria and polystyrene spheres, we show how this system can be used to assess particle aspect ratio even when imaged at low resolution. We also show the feasibility of detecting alterations in organelle aspect ratio in situ within living cells. This improved OSI system could be further developed to automate morphological quantification and sorting of non-spherical particles in situ.

Lensfree microscopy on a cellphone

PubMed Central

Tseng, Derek; Mudanyali, Onur; Oztoprak, Cetin; Isikman, Serhan O.; Sencan, Ikbal; Yaglidere, Oguzhan; Ozcan, Aydogan

2010-01-01

We demonstrate lensfree digital microscopy on a cellphone. This compact and light-weight holographic microscope installed on a cellphone does not utilize any lenses, lasers or other bulky optical components and it may offer a cost-effective tool for telemedicine applications to address various global health challenges. Weighing ~38 grams (<1.4 ounces), this lensfree imaging platform can be mechanically attached to the camera unit of a cellphone where the samples are loaded from the side, and are vertically illuminated by a simple light-emitting diode (LED). This incoherent LED light is then scattered from each micro-object to coherently interfere with the background light, creating the lensfree hologram of each object on the detector array of the cellphone. These holographic signatures captured by the cellphone permit reconstruction of microscopic images of the objects through rapid digital processing. We report the performance of this lensfree cellphone microscope by imaging various sized micro-particles, as well as red blood cells, white blood cells, platelets and a waterborne parasite (Giardia lamblia). PMID:20445943

Comparison of marginal fit of cemented zirconia copings manufactured after digital impression with lava™ C.O.S and conventional impression technique.

PubMed

Dauti, Rinet; Cvikl, Barbara; Franz, Alexander; Schwarze, Uwe Yacine; Lilaj, Bledar; Rybaczek, Tina; Moritz, Andreas

2016-12-08

Evaluation of the marginal fit of cemented zirconia copings manufactured after digital impression with Lava™ Chairside Oral Scanner in comparison to that of zirconia copings manufactured after conventional impressions with polyvinyl siloxane. A prepared typodont tooth #36, was replicated 40 times with a vinyl silicone and precise model resin. The dies were randomly divided into two groups according to the impression taking technique. Digital impressions with Lava™ C.O.S. and conventional impressions were taken according to the group. Subsequently zirconia copings were manufactured and cemented on their respective dies with zinc oxide phosphate cement. After embedding in resin, mesio-distal section of each coping was performed with a diamond saw in order to obtain two slices. One half of the specimen was used for evaluation with an optical microscope (OM) and the other half for evaluation with a scanning electron microscope (SEM). Marginal gap (MG) and absolute marginal discrepancy (AMD) were measured mesial and distal on each slice. No significant difference of the marginal parameters between the digital and the conventional group was found. The mean values for MG in the digital group were 96.28 μm (+/-43.21 μm) measured with the OM and 99.26 μm (+/-48.73 μm) measured with the SEM, respectively. AMD mean values were 191.54 μm (+/-85.42 μm) measured with the optical microscope and 211.6 μm (+/-96.55 μm) with the SEM. For the conventional group the mean MG values were 94.84 μm (+/-50.77 μm) measured with the OM and 83.37 μm (+/-44.38 μm) measured with the SEM, respectively. AMD mean values were 158.60 μm (+/-69.14 μm) for the OM and 152.72 μm (+/-72.36) for the SEM. Copings manufactured after digital impression with Lava™ C.O.S. show comparable marginal parameters with the copings manufactured after conventional impression with polyvinyl syloxane. The mean MG values of both groups fit in the clinically acceptable range.

Optical method for high magnification imaging and video recording of live cells at sub-micron resolution

NASA Astrophysics Data System (ADS)

Romo, Jaime E., Jr.

Optical microscopy, the most common technique for viewing living microorganisms, is limited in resolution by Abbe's criterion. Recent microscopy techniques focus on circumnavigating the light diffraction limit by using different methods to obtain the topography of the sample. Systems like the AFM and SEM provide images with fields of view in the nanometer range with high resolvable detail, however these techniques are expensive, and limited in their ability to document live cells. The Dino-Lite digital microscope coupled with the Zeiss Axiovert 25 CFL microscope delivers a cost-effective method for recording live cells. Fields of view ranging from 8 microns to 300 microns with fair resolution provide a reliable method for discovering native cell structures at the nanoscale. In this report, cultured HeLa cells are recorded using different optical configurations resulting in documentation of cell dynamics at high magnification and resolution.

Studies of mechanisms of decay and recovery in organic dye-doped polymers using spatially resolved white light interferometry

NASA Astrophysics Data System (ADS)

Anderson, Benjamin; Bernhardt, Elizabeth; Kuzyk, Mark

2012-10-01

Several organic dyes have been shown to self heal when doped in a polymer matrix. Most measurements to date use optical absorbance, amplified spontaneous emission, or digital imaging as a probe. Each method determines a subset of the relevant parameters. We have constructed a white light interferometric microscope, which measures the absorption spectrum and change in refractive index during decay and recovery simultaneously at multiple points in the material. We report on preliminary measurements and results concerning the microscopes spatial resolution.

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.

Diagnostic Efficiency in Digital Pathology: A Comparison of Optical Versus Digital Assessment in 510 Surgical Pathology Cases.

PubMed

Mills, Anne M; Gradecki, Sarah E; Horton, Bethany J; Blackwell, Rebecca; Moskaluk, Christopher A; Mandell, James W; Mills, Stacey E; Cathro, Helen P

2018-01-01

Prior work has shown that digital images and microscopic slides can be interpreted with comparable diagnostic accuracy. Although accuracy has been well-validated, the interpretative time for digital images has scarcely been studied and concerns about efficiency remain a major barrier to adoption. We investigated the efficiency of digital pathology when compared with glass slide interpretation in the diagnosis of surgical pathology biopsy and resection specimens. Slides were pulled from 510 surgical pathology cases from 5 organ systems (gastrointestinal, gynecologic, liver, bladder, and brain). Original diagnoses were independently confirmed by 2 validating pathologists. Diagnostic slides were scanned using the Philips IntelliSite Pathology Solution. Each case was assessed independently on digital and optical by 3 reading pathologists, with a ≥6 week washout period between modalities. Reading pathologists recorded assessment times for each modality; digital times included time to load the case. Diagnostic accuracy was determined based on whether a rendered diagnosis differed significantly from the original diagnosis. Statistical analysis was performed to assess for differences in interpretative times across modalities. All 3 reading pathologists showed comparable diagnostic accuracy across optical and digital modalities (mean major discordance rates with original diagnosis: 4.8% vs. 4.4%, respectively). Mean assessment times ranged from 1.2 to 9.1 seconds slower on digital versus optical. The slowest reader showed a significant learning effect during the course of the study so that digital assessment times decreased over time and were comparable with optical times by the end of the series. Organ site and specimen type did not significantly influence differences in interpretative times. In summary, digital image reading times compare favorably relative to glass slides across a variety of organ systems and specimen types. Mean increase in assessment time is 4 seconds/case. This time can be minimized with experience and may be further balanced by the improved ease of electronic chart access allowed by digital slide viewing, as well as quantitative assessments which can be expedited on digital images.

Digital holographic microscopy applied to measurement of a flow in a T-shaped micromixer

NASA Astrophysics Data System (ADS)

Ooms, T. A.; Lindken, R.; Westerweel, J.

2009-12-01

In this paper, we describe measurements of a three-dimensional (3D) flow in a T-shaped micromixer by means of digital holographic microscopy. Imaging tracer particles in a microscopic flow with conventional microscopy is accompanied by a small depth-of-field, which hinders true volumetric flow measurements. In holographic microscopy, the depth of the measurement domain does not have this limitation because any desired image plane can be reconstructed after recording. Our digital holographic microscope (DHM) consists of a conventional in-line recording system with an added magnifying optical element. The measured flow velocity and the calculated vorticity illustrate four streamwise vortices in the micromixer outflow channel. Because the investigated flow is stationary and strongly 3D, the DHM performance (i.e. accuracy and resolution) can be precisely investigated. The obtained Dynamic spatial range and Dynamic velocity range are larger than 20 and 30, respectively. High-speed multiple-frame measurements illustrate the capability to simultaneously track about 80 particles in a volumetric measurement domain.

Scanning digital lithography providing high speed large area patterning with diffraction limited sub-micron resolution

NASA Astrophysics Data System (ADS)

Wen, Sy-Bor; Bhaskar, Arun; Zhang, Hongjie

2018-07-01

A scanning digital lithography system using computer controlled digital spatial light modulator, spatial filter, infinity correct optical microscope and high precision translation stage is proposed and examined. Through utilizing the spatial filter to limit orders of diffraction modes for light delivered from the spatial light modulator, we are able to achieve diffraction limited deep submicron spatial resolution with the scanning digital lithography system by using standard one inch level optical components with reasonable prices. Raster scanning of this scanning digital lithography system using a high speed high precision x-y translation stage and piezo mount to real time adjust the focal position of objective lens allows us to achieve large area sub-micron resolved patterning with high speed (compared with e-beam lithography). It is determined in this study that to achieve high quality stitching of lithography patterns with raster scanning, a high-resolution rotation stage will be required to ensure the x and y directions of the projected pattern are in the same x and y translation directions of the nanometer precision x-y translation stage.

3D refractive index measurements of special optical fibers

NASA Astrophysics Data System (ADS)

Yan, Cheng; Huang, Su-Juan; Miao, Zhuang; Chang, Zheng; Zeng, Jun-Zhang; Wang, Ting-Yun

2016-09-01

A digital holographic microscopic chromatography-based approach with considerably improved accuracy, simplified configuration and performance stability is proposed to measure three dimensional refractive index of special optical fibers. Based on the approach, a measurement system is established incorporating a modified Mach-Zehnder interferometer and lab-developed supporting software for data processing. In the system, a phase projection distribution of an optical fiber is utilized to obtain an optimal digital hologram recorded by a CCD, and then an angular spectrum theory-based algorithm is adopted to extract the phase distribution information of an object wave. The rotation of the optic fiber enables the experimental measurements of multi-angle phase information. Based on the filtered back projection algorithm, a 3D refraction index of the optical fiber is thus obtained at high accuracy. To evaluate the proposed approach, both PANDA fibers and special elliptical optical fiber are considered in the system. The results measured in PANDA fibers agree well with those measured using S14 Refractive Index Profiler, which is, however, not suitable for measuring the property of a special elliptical fiber.

Evaluation environment for digital and analog pathology: a platform for validation studies

PubMed Central

Gallas, Brandon D.; Gavrielides, Marios A.; Conway, Catherine M.; Ivansky, Adam; Keay, Tyler C.; Cheng, Wei-Chung; Hipp, Jason; Hewitt, Stephen M.

2014-01-01

Abstract. We present a platform for designing and executing studies that compare pathologists interpreting histopathology of whole slide images (WSIs) on a computer display to pathologists interpreting glass slides on an optical microscope. eeDAP is an evaluation environment for digital and analog pathology. The key element in eeDAP is the registration of the WSI to the glass slide. Registration is accomplished through computer control of the microscope stage and a camera mounted on the microscope that acquires real-time images of the microscope field of view (FOV). Registration allows for the evaluation of the same regions of interest (ROIs) in both domains. This can reduce or eliminate disagreements that arise from pathologists interpreting different areas and focuses on the comparison of image quality. We reduced the pathologist interpretation area from an entire glass slide (10 to 30  mm2) to small ROIs (<50  μm2). We also made possible the evaluation of individual cells. We summarize eeDAP’s software and hardware and provide calculations and corresponding images of the microscope FOV and the ROIs extracted from the WSIs. The eeDAP software can be downloaded from the Google code website (project: eeDAP) as a MATLAB source or as a precompiled stand-alone license-free application. PMID:26158076

Evaluation environment for digital and analog pathology: a platform for validation studies.

PubMed

Gallas, Brandon D; Gavrielides, Marios A; Conway, Catherine M; Ivansky, Adam; Keay, Tyler C; Cheng, Wei-Chung; Hipp, Jason; Hewitt, Stephen M

2014-10-01

We present a platform for designing and executing studies that compare pathologists interpreting histopathology of whole slide images (WSIs) on a computer display to pathologists interpreting glass slides on an optical microscope. eeDAP is an evaluation environment for digital and analog pathology. The key element in eeDAP is the registration of the WSI to the glass slide. Registration is accomplished through computer control of the microscope stage and a camera mounted on the microscope that acquires real-time images of the microscope field of view (FOV). Registration allows for the evaluation of the same regions of interest (ROIs) in both domains. This can reduce or eliminate disagreements that arise from pathologists interpreting different areas and focuses on the comparison of image quality. We reduced the pathologist interpretation area from an entire glass slide (10 to [Formula: see text]) to small ROIs ([Formula: see text]). We also made possible the evaluation of individual cells. We summarize eeDAP's software and hardware and provide calculations and corresponding images of the microscope FOV and the ROIs extracted from the WSIs. The eeDAP software can be downloaded from the Google code website (project: eeDAP) as a MATLAB source or as a precompiled stand-alone license-free application.

Quantitative luminescence imaging system

DOEpatents

Erwin, D.N.; Kiel, J.L.; Batishko, C.R.; Stahl, K.A.

1990-08-14

The QLIS images and quantifies low-level chemiluminescent reactions in an electromagnetic field. It is capable of real time nonperturbing measurement and simultaneous recording of many biochemical and chemical reactions such as luminescent immunoassays or enzyme assays. The system comprises image transfer optics, a low-light level digitizing camera with image intensifying microchannel plates, an image process or, and a control computer. The image transfer optics may be a fiber image guide with a bend, or a microscope, to take the light outside of the RF field. Output of the camera is transformed into a localized rate of cumulative digitalized data or enhanced video display or hard-copy images. The system may be used as a luminescent microdosimetry device for radiofrequency or microwave radiation, as a thermal dosimeter, or in the dosimetry of ultra-sound (sonoluminescence) or ionizing radiation. It provides a near-real-time system capable of measuring the extremely low light levels from luminescent reactions in electromagnetic fields in the areas of chemiluminescence assays and thermal microdosimetry, and is capable of near-real-time imaging of the sample to allow spatial distribution analysis of the reaction. It can be used to instrument three distinctly different irradiation configurations, comprising (1) RF waveguide irradiation of a small Petri-dish-shaped sample cell, (2) RF irradiation of samples in a microscope for the microscopic imaging and measurement, and (3) RF irradiation of small to human body-sized samples in an anechoic chamber. 22 figs.

Common path in-line holography using enhanced joint object reference digital interferometers

PubMed Central

Kelner, Roy; Katz, Barak; Rosen, Joseph

2014-01-01

Joint object reference digital interferometer (JORDI) is a recently developed system capable of recording holograms of various types [Opt. Lett. 38(22), 4719 (2013)24322115]. Presented here is a new enhanced system design that is based on the previous JORDI. While the previous JORDI has been based purely on diffractive optical elements, displayed on spatial light modulators, the present design incorporates an additional refractive objective lens, thus enabling hologram recording with improved resolution and increased system applicability. Experimental results demonstrate successful hologram recording for various types of objects, including transmissive, reflective, three-dimensional, phase and highly scattering objects. The resolution limit of the system is analyzed and experimentally validated. Finally, the suitability of JORDI for microscopic applications is verified as a microscope objective based configuration of the system is demonstrated. PMID:24663838

Sub-micrometer resolution proximity X-ray microscope with digital image registration

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

Chkhalo, N. I.; Salashchenko, N. N.; Sherbakov, A. V., E-mail: SherbakovAV@ipm.sci-nnov.ru

A compact laboratory proximity soft X-ray microscope providing submicrometer spatial resolution and digital image registration is described. The microscope consists of a laser-plasma soft X-ray radiation source, a Schwarzschild objective to illuminate the test sample, and a two-coordinate detector for image registration. Radiation, which passes through the sample under study, generates an absorption image on the front surface of the detector. Optical ceramic YAG:Ce was used to convert the X-rays into visible light. An image was transferred from the scintillator to a charge-coupled device camera with a Mitutoyo Plan Apo series lens. The detector’s design allows the use of lensesmore » with numerical apertures of NA = 0.14, 0.28, and 0.55 without changing the dimensions and arrangement of the elements of the device. This design allows one to change the magnification, spatial resolution, and field of view of the X-ray microscope. A spatial resolution better than 0.7 μm and an energy conversion efficiency of the X-ray radiation with a wavelength of 13.5 nm into visible light collected by the detector of 7.2% were achieved with the largest aperture lens.« less

eeDAP: An Evaluation Environment for Digital and Analog Pathology

PubMed Central

Gallas, Brandon D.; Cheng, Wei-Chung; Gavrielides, Marios A.; Ivansky, Adam; Keay, Tyler; Wunderlich, Adam; Hipp, Jason; Hewitt, Stephen M.

2017-01-01

Purpose The purpose of this work is to present a platform for designing and executing studies that compare pathologists interpreting histopathology of whole slide images (WSI) on a computer display to pathologists interpreting glass slides on an optical microscope. Methods Here we present eeDAP, an evaluation environment for digital and analog pathology. The key element in eeDAP is the registration of the WSI to the glass slide. Registration is accomplished through computer control of the microscope stage and a camera mounted on the microscope that acquires images of the real time microscope view. Registration allows for the evaluation of the same regions of interest (ROIs) in both domains. This can reduce or eliminate disagreements that arise from pathologists interpreting different areas and focuses the comparison on image quality. Results We reduced the pathologist interpretation area from an entire glass slide (≈10–30 mm)2 to small ROIs <(50 um)2. We also made possible the evaluation of individual cells. Conclusions We summarize eeDAP’s software and hardware and provide calculations and corresponding images of the microscope field of view and the ROIs extracted from the WSIs. These calculations help provide a sense of eeDAP’s functionality and operating principles, while the images provide a sense of the look and feel of studies that can be conducted in the digital and analog domains. The eeDAP software can be downloaded from code.google.com (project: eeDAP) as Matlab source or as a precompiled stand-alone license-free application. PMID:28845079

Single-shot measurement of phase and amplitude by using a heterodyne time-lens system and ultrafast digital time-holography

NASA Astrophysics Data System (ADS)

Tikan, Alexey; Bielawski, Serge; Szwaj, Christophe; Randoux, Stéphane; Suret, Pierre

2018-04-01

Temporal imaging systems are outstanding tools for single-shot observation of optical signals that have irregular and ultrafast dynamics. They allow long time windows to be recorded with femtosecond resolution, and do not rely on complex algorithms. However, simultaneous recording of amplitude and phase remains an open challenge for these systems. Here, we present a new heterodyne time-lens arrangement that efficiently records both the amplitude and phase of complex and random signals over large temporal windows (tens of picoseconds). Phase and time are encoded onto the two spatial dimensions of a camera. We implement this phase-sensitive time-lens system in two configurations: a time microscope and a digital temporal-holography device that enables single-shot measurement with a temporal resolution of 80 fs. We demonstrate direct application of our heterodyne time-lens to turbulent-like optical fields and optical rogue waves generated from nonlinear propagation of partially coherent waves inside optical fibres.

Optical monitoring of neuronal activity at high frame rate with a digital random-access multiphoton (RAMP) microscope.

PubMed

Otsu, Yo; Bormuth, Volker; Wong, Jerome; Mathieu, Benjamin; Dugué, Guillaume P; Feltz, Anne; Dieudonné, Stéphane

2008-08-30

Two-photon microscopy offers the promise of monitoring brain activity at multiple locations within intact tissue. However, serial sampling of voxels has been difficult to reconcile with millisecond timescales characteristic of neuronal activity. This is due to the conflicting constraints of scanning speed and signal amplitude. The recent use of acousto-optic deflector scanning to implement random-access multiphoton microscopy (RAMP) potentially allows to preserve long illumination dwell times while sampling multiple points-of-interest at high rates. However, the real-life abilities of RAMP microscopy regarding sensitivity and phototoxicity issues, which have so far impeded prolonged optical recordings at high frame rates, have not been assessed. Here, we describe the design, implementation and characterisation of an optimised RAMP microscope. We demonstrate the application of the microscope by monitoring calcium transients in Purkinje cells and cortical pyramidal cell dendrites and spines. We quantify the illumination constraints imposed by phototoxicity and show that stable continuous high-rate recordings can be obtained. During these recordings the fluorescence signal is large enough to detect spikes with a temporal resolution limited only by the calcium dye dynamics, improving upon previous techniques by at least an order of magnitude.

Time-resolved wide-field optically sectioned fluorescence microscopy

NASA Astrophysics Data System (ADS)

Dupuis, Guillaume; Benabdallah, Nadia; Chopinaud, Aurélien; Mayet, Céline; Lévêque-Fort, Sandrine

2013-02-01

We present the implementation of a fast wide-field optical sectioning technique called HiLo microscopy on a fluorescence lifetime imaging microscope. HiLo microscopy is based on the fusion of two images, one with structured illumination and another with uniform illumination. Optically sectioned images are then digitally generated thanks to a fusion algorithm. HiLo images are comparable in quality with confocal images but they can be acquired faster over larger fields of view. We obtain 4D imaging by combining HiLo optical sectioning, time-gated detection, and z-displacement. We characterize the performances of this set-up in terms of 3D spatial resolution and time-resolved capabilities in both fixed- and live-cell imaging modes.

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.

Ion photon emission microscope

DOEpatents

Doyle, Barney L.

2003-04-22

An ion beam analysis system that creates microscopic multidimensional image maps of the effects of high energy ions from an unfocussed source upon a sample by correlating the exact entry point of an ion into a sample by projection imaging of the ion-induced photons emitted at that point with a signal from a detector that measures the interaction of that ion within the sample. The emitted photons are collected in the lens system of a conventional optical microscope, and projected on the image plane of a high resolution single photon position sensitive detector. Position signals from this photon detector are then correlated in time with electrical effects, including the malfunction of digital circuits, detected within the sample that were caused by the individual ion that created these photons initially.

Stretching of red blood cells by optical tweezers quantified by digital holographic microscopy

NASA Astrophysics Data System (ADS)

Cardenas, Nelson; Yu, Lingfeng; Mohanty, Samarendra K.

2011-03-01

Red blood cells (RBC) possess unique viscoelastic characteristics which allow them to pass through capillaries narrower than their size. Measurement of viscoelastic property of cells (e.g. RBC) in low-force regime is of high significance as it represents conditions of membrane fluctuation in response to physiological conditions. Estimation of visco-elastic properties of RBC requires measurement of extent of deformation in RBC subjected to known force. Optical tweezers, being gentle and absolutely sterile, are emerging as the tool of choice for application of localized force on cells. However, stretching of RBC in very low force regime has not been quantified. Further, though deformations in transverse directions have been measured, vertical deformations due to stretching of cells cannot be quantified by classical microscopic images. Here, we report realization of offaxis digital holographic microscopy (DHM) for highly sensitive axial changes in RBC shape due to stretching by optical tweezers without attaching microscopic beads. The RBC was stretched in axial direction with nanometer precision by change of divergence of the trapping beam. The obtained deformation patterns were compared with the axial position of the tweezers focus. Since the pathophysiology of progression of diseases like malaria and cancer is reflected in the biophysical (both mechanical and material) properties of the cells, it is possible to identify the changes by simultaneous measurement of refractive index and elasticity using this approach.

Chromatic confocal microscope using hybrid aspheric diffractive lenses

NASA Astrophysics Data System (ADS)

Rayer, Mathieu; Mansfield, Daniel

2014-05-01

A chromatic confocal microscope is a single point non-contact distance measurement sensor. For three decades the vast majority of the chromatic confocal microscope use refractive-based lenses to code the measurement axis chromatically. However, such an approach is limiting the range of applications. In this paper the performance of refractive, diffractive and Hybrid aspheric diffractive are compared. Hybrid aspheric diffractive lenses combine the low geometric aberration of a diffractive lens with the high optical power of an aspheric lens. Hybrid aspheric diffractive lenses can reduce the number of elements in an imaging system significantly or create large hyper- chromatic lenses for sensing applications. In addition, diffractive lenses can improve the resolution and the dynamic range of a chromatic confocal microscope. However, to be suitable for commercial applications, the diffractive optical power must be significant. Therefore, manufacturing such lenses is a challenge. We show in this paper how a theoretical manufacturing model can demonstrate that the hybrid aspheric diffractive configuration with the best performances is achieved by step diffractive surface. The high optical quality of step diffractive surface is then demonstrated experimentally. Publisher's Note: This paper, originally published on 5/10/14, was replaced with a corrected/revised version on 5/19/14. If you downloaded the original PDF but are unable to access the revision, please contact SPIE Digital Library Customer Service for assistance.

A simple optical tweezers for trapping polystyrene particles

NASA Astrophysics Data System (ADS)

Shiddiq, Minarni; Nasir, Zulfa; Yogasari, Dwiyana

2013-09-01

Optical tweezers is an optical trap. For decades, it has become an optical tool that can trap and manipulate any particle from the very small size like DNA to the big one like bacteria. The trapping force comes from the radiation pressure of laser light which is focused to a group of particles. Optical tweezers has been used in many research areas such as atomic physics, medical physics, biophysics, and chemistry. Here, a simple optical tweezers has been constructed using a modified Leybold laboratory optical microscope. The ocular lens of the microscope has been removed for laser light and digital camera accesses. A laser light from a Coherent diode laser with wavelength λ = 830 nm and power 50 mW is sent through an immersion oil objective lens with magnification 100 × and NA 1.25 to a cell made from microscope slides containing polystyrene particles. Polystyrene particles with size 3 μm and 10 μm are used. A CMOS Thorlabs camera type DCC1545M with USB Interface and Thorlabs camera lens 35 mm are connected to a desktop and used to monitor the trapping and measure the stiffness of the trap. The camera is accompanied by camera software which makes able for the user to capture and save images. The images are analyzed using ImageJ and Scion macro. The polystyrene particles have been trapped successfully. The stiffness of the trap depends on the size of the particles and the power of the laser. The stiffness increases linearly with power and decreases as the particle size larger.

Experimental research of digital holographic microscopic measuring

NASA Astrophysics Data System (ADS)

Zhu, Xueliang; Chen, Feifei; Li, Jicheng

2013-06-01

Digital holography is a new imaging technique, which is developed on the base of optical holography, Digital processing, and Computer techniques. It is using CCD instead of the conventional silver to record hologram, and then reproducing the 3D contour of the object by the way of computer simulation. Compared with the traditional optical holographic, the whole process is of simple measuring, lower production cost, faster the imaging speed, and with the advantages of non-contact real-time measurement. At present, it can be used in the fields of the morphology detection of tiny objects, micro deformation analysis, and biological cells shape measurement. It is one of the research hot spot at home and abroad. This paper introduced the basic principles and relevant theories about the optical holography and Digital holography, and researched the basic questions which influence the reproduce images in the process of recording and reconstructing of the digital holographic microcopy. In order to get a clear digital hologram, by analyzing the optical system structure, we discussed the recording distance and of the hologram. On the base of the theoretical studies, we established a measurement and analyzed the experimental conditions, then adjusted them to the system. To achieve a precise measurement of tiny object in three-dimension, we measured MEMS micro device for example, and obtained the reproduction three-dimensional contour, realized the three dimensional profile measurement of tiny object. According to the experiment results consider: analysis the reference factors between the zero-order term and a pair of twin-images by the choice of the object light and the reference light and the distance of the recording and reconstructing and the characteristics of reconstruction light on the measurement, the measurement errors were analyzed. The research result shows that the device owns certain reliability.

Optical diffraction tomography with fully and partially coherent illumination in high numerical aperture label-free microscopy [Invited].

PubMed

Soto, Juan M; Rodrigo, José A; Alieva, Tatiana

2018-01-01

Quantitative label-free imaging is an important tool for the study of living microorganisms that, during the last decade, has attracted wide attention from the optical community. Optical diffraction tomography (ODT) is probably the most relevant technique for quantitative label-free 3D imaging applied in wide-field microscopy in the visible range. The ODT is usually performed using spatially coherent light illumination and specially designed holographic microscopes. Nevertheless, the ODT is also compatible with partially coherent illumination and can be realized in conventional wide-field microscopes by applying refocusing techniques, as it has been recently demonstrated. Here, we compare these two ODT modalities, underlining their pros and cons and discussing the optical setups for their implementation. In particular, we pay special attention to a system that is compatible with a conventional wide-field microscope that can be used for both ODT modalities. It consists of two easily attachable modules: the first for sample illumination engineering based on digital light processing technology; the other for focus scanning by using an electrically driven tunable lens. This hardware allows for a programmable selection of the wavelength and the illumination design, and provides fast data acquisition as well. Its performance is experimentally demonstrated in the case of ODT with partially coherent illumination providing speckle-free 3D quantitative imaging.

Optical sectioning using a digital Fresnel incoherent-holography-based confocal imaging system

PubMed Central

Kelner, Roy; Katz, Barak; Rosen, Joseph

2015-01-01

We propose a new type of confocal microscope using Fresnel incoherent correlation holography (FINCH). Presented here is a confocal configuration of FINCH using a phase pinhole and point illumination that is able to suppress out-of-focus information from the recorded hologram and hence combine the super-resolution capabilities of FINCH with the sectioning capabilities of confocal microscopy. PMID:26413560

Preparing and probing many-body correlated systems in a Quantum Gas Microscope by engineering arbitrary landscape potentials

NASA Astrophysics Data System (ADS)

Rispoli, Matthew; Lukin, Alexander; Ma, Ruichao; Preiss, Philipp; Tai, M. Eric; Islam, Rajibul; Greiner, Markus

2015-05-01

Ultracold atoms in optical lattices provide a versatile tool box for observing the emergence of strongly correlated physics in quantum systems. Dynamic control of optical potentials on the single-site level allows us to prepare and probe many-body quantum states through local Hamiltonian engineering. We achieve these high precision levels of optical control through spatial light modulation with a DMD (digital micro-mirror device). This allows for both arbitrary beam shaping and aberration compensation in our imaging system to produce high fidelity optical potentials. We use these techniques to control state initialization, Hamiltonian dynamics, and measurement in experiments investigating low-dimensional many-body physics - from one-dimensional correlated quantum walks to characterizing entanglement.

Use of digital micromirror devices as dynamic pinhole arrays for adaptive confocal fluorescence microscopy

NASA Astrophysics Data System (ADS)

Pozzi, Paolo; Wilding, Dean; Soloviev, Oleg; Vdovin, Gleb; Verhaegen, Michel

2018-02-01

In this work, we present a new confocal laser scanning microscope capable to perform sensorless wavefront optimization in real time. The device is a parallelized laser scanning microscope in which the excitation light is structured in a lattice of spots by a spatial light modulator, while a deformable mirror provides aberration correction and scanning. A binary DMD is positioned in an image plane of the detection optical path, acting as a dynamic array of reflective confocal pinholes, images by a high performance cmos camera. A second camera detects images of the light rejected by the pinholes for sensorless aberration correction.

Web-based virtual microscopy at the RWTH Aachen University: didactic concept, methods and analysis of acceptance by the students.

PubMed

Merk, Magdalene; Knuechel, Ruth; Perez-Bouza, Alberto

2010-12-20

Fundamental knowledge of microscopic anatomy and pathology has always been an essential part in medical education. The traditional didactic concept comprises theoretical and practical lessons using a light microscope and glass slides. High-speed Internet connections and technical improvement in whole-slide digital microscopy (commonly termed "virtual microscopy") provide a new and attractive approach for both teachers and students. High picture quality and unlimited temporal and spatial availability of histology samples from different fields are key advantages of web-based digital microscopy. In this report we discuss the technical requirements, system efficiency, optical resolution and didactic concept. Furthermore, we present a review of the experience gained in the course of one year based on an analysis of student acceptance. Three groups with a total of 192 students between the 3rd and 5th year of medical studies attending the practical courses of general and advanced histopathology had access to both glass-mounted and digitalized slides. Prior to exams, students were asked to answer an anonymous questionnaire. The results of the study reflect the high acceptance and intensive use of the web-based digital histology by students, thus encouraging the development of further Web-based learning strategies for the teaching of histology and pathology. 2010 Elsevier GmbH. All rights reserved.

Panning artifacts in digital pathology images

NASA Astrophysics Data System (ADS)

Avanaki, Ali R. N.; Lanciault, Christian; Espig, Kathryn S.; Xthona, Albert; Kimpe, Tom R. L.

2017-03-01

In making a pathologic diagnosis, a pathologist uses cognitive processes: perception, attention, memory, and search (Pena and Andrade-Filho, 2009). Typically, this involves focus while panning from one region of a slide to another, using either a microscope in a traditional workflow or software program and display in a digital pathology workflow (DICOM Standard Committee, 2010). We theorize that during panning operation, the pathologist receives information important to diagnosis efficiency and/or correctness. As compared to an optical microscope, panning in a digital pathology image involves some visual artifacts due to the following: (i) the frame rate is finite; (ii) time varying visual signals are reconstructed using imperfect zero-order hold. Specifically, after pixel's digital drive is changed, it takes time for a pixel to emit the expected amount of light. Previous work suggests that 49% of navigation is conducted in low-power/overview with digital pathology (Molin et al., 2015), but the influence of display factors has not been measured. We conducted a reader study to establish a relationship between display frame rate, panel response time, and threshold panning speed (above which the artifacts become noticeable). Our results suggest visual tasks that involve tissue structure are more impacted by the simulated panning artifacts than those that only involve color (e.g., staining intensity estimation), and that the panning artifacts versus normalized panning speed has a peak behavior which is surprising and may change for a diagnostic task. This is work in progress and our final findings should be considered in designing future digital pathology systems.

Three-dimensional imaging of micro-specimen by optical scanning holography

NASA Astrophysics Data System (ADS)

Liu, Jung-Ping; Tsou, Cheng-Hao

2017-04-01

Optical scanning holography (OSH) is a scanning-type digital holographic technique. In OSH, a heterodyne interference pattern is generated to raster scan the object. OSH can be operated in the incoherent mode and thus is able to record a fluorescence hologram. In addition, resolution of the OSH is proportional to the density of the interference pattern. Here we use a high-NA microscope objective to generate a dynamic Fresnel zone plate to record a hologram of micro-specimen. The achieved transverse resolution and longitudinal resolution are 0.78μm and 3.1μm, respectively.

Fabrication and characterization of optical super-smooth surfaces

NASA Astrophysics Data System (ADS)

Schmitt, Dirk-Roger; Kratz, Frank; Ringel, Gabriele A.; Mangelsdorf, Juergen; Creuzet, Francois; Garratt, John D.

1995-08-01

Intercomparison roughness measurements have been carried out at supersmooth artefacts fabricated from BK7, fused silica, and Zerodur. The surface parameters were determined using a special prototype of the mechanical profiler Nanostep (Rank Taylor Hobson), the Optical Heterodyne Profiler Z5500 (Zygo), and an Atomic Force Microscope (Park Scientific) with an improved acquisition technique. The intercomparison was performed after the range of collected spatial wavelength for each instrument was adjusted using digital filtering techniques. It is demonstrated for different roughness ranges that are applied superpolishing techniques yield supersmooth artefacts which can be used for more intercomparisons.

Realisation of a holographic microlaser scalpel using a digital micromirror device

NASA Astrophysics Data System (ADS)

Zwick, Susanne; Warber, Michael; Haist, Tobias; Osten, Wolfgang

2007-06-01

Modern spatial light modulators (SLM) enable the generation of more or less arbitrary light fields in three dimensions. Such light fields can be used for different future applications in the field of biomedical optics. One example is the processing/cutting of biological material on a microscopic scale. By displaying computer generated holograms by suitable SLMs it is possible to ablate complex structures into three-dimensional objects without scanning with very high accuracy on a microscopic scale. To effectively cut biological materials by light, pulsed ultraviolet light is preferable. We will present a combined setup of a holographic laser scalpel using a digital micromirror device (DMD) and holographic optical tweezers using a liquid crystal display (LCD). The setup enables to move and cut or process micro-scaled objects like biological cells or tissue in three dimensions with high accuracy and without any mechanical movements just by changing the hologram displayed by the SLMs. We will show that holograms can be used to compensate aberrations implemented by the DMD or other optical components of the setup. Also we can generate arbitrary light fields like stripes, circles or arbitrary curves. Additionally we will present results for the fast optimization of holograms for the system. In particular we will show results obtained by implementing iterative Fourier transform based algorithms on a standard consumer graphics board (Nvidia 8800GLX). By this approach we are able to compute more than 360 complex 2D FFTs (512 × 512 pixels) per second with floating point precision.

Digital holography applications in ophthalmology, biometry, and optical trapping characterization

NASA Astrophysics Data System (ADS)

Potcoava, Mariana Camelia

This dissertation combines various holographic techniques with application on the two- and three-dimensional imaging of ophthalmic tissue, fingerprints, and microsphere samples with micrometer resolution. Digital interference holography (DIH) uses scanned wavelengths to synthesize short-coherence interference tomographic images. We used DIH for in vitro imaging of human optic nerve head and retina. Tomographic images were produced by superposition of holograms. Holograms were obtained with a signal-to-noise ratio of approximately 50 dB. Optic nerve head characteristics (shape, diameter, cup depth, and cup width) were quantified with a few micron resolution (4.06--4.8mum). Multiple layers were distinguishable in cross-sectional images of the macula. To our knowledge, this is the first report of DIH use to image human macular and optic nerve tissue. Holographic phase microscopy is used to produce images of thin film patterns left by latent fingerprints. Two or more holographic phase images with different wavelengths are combined for optical phase unwrapping of images of patent prints. We demonstrated digital interference holography images of a plastic print, and latent prints. These demonstrations point to significant contributions to biometry by using digital interference holography to identify and quantify Level 1 (pattern), Level 2 (minutia points), and Level 3 (pores and ridge contours). Quantitative studies of physical and biological processes and precise non-contact manipulation of nanometer/micrometer trapped objects can be effectuated with nanometer accuracy due to the development of optical tweezers. A three-dimensional gradient trap is produced at the focus position of a high NA microscope objective. Particles are trapped axially and laterally due to the gradient force. The particle is confined in a potential well and the trap acts as a harmonic spring. The elastic constant or the stiffness along any axis is determined from the particle displacements in time along each specific axis. Thus, we report the sensing of small particles using optical trapping in combination with the digital Gabor holography to calibrate the optical force and the position and of the copolymer microsphere in the x, y, z direction with nm precision.

Array microscopy technology and its application to digital detection of Mycobacterium tuberculosis

NASA Astrophysics Data System (ADS)

McCall, Brian P.

Tuberculosis causes more deaths worldwide than any other curable infectious disease. This is the case despite tuberculosis appearing to be on the verge of eradication midway through the last century. Efforts at reversing the spread of tuberculosis have intensified since the early 1990s. Since then, microscopy has been the primary frontline diagnostic. In this dissertation, advances in clinical microscopy towards array microscopy for digital detection of Mycobacterium tuberculosis are presented. Digital array microscopy separates the tasks of microscope operation and pathogen detection and will reduce the specialization needed in order to operate the microscope. Distributing the work and reducing specialization will allow this technology to be deployed at the point of care, taking the front-line diagnostic for tuberculosis from the microscopy center to the community health center. By improving access to microscopy centers, hundreds of thousands of lives can be saved. For this dissertation, a lens was designed that can be manufactured as 4x6 array of microscopes. This lens design is diffraction limited, having less than 0.071 waves of aberration (root mean square) over the entire field of view. A total area imaged onto a full-frame digital image sensor is expected to be 3.94 mm2, which according to tuberculosis microscopy guidelines is more than sufficient for a sensitive diagnosis. The design is tolerant to single point diamond turning manufacturing errors, as found by tolerance analysis and by fabricating a prototype. Diamond micro-milling, a fabrication technique for lens array molds, was applied to plastic plano-concave and plano-convex lens arrays, and found to produce high quality optical surfaces. The micro-milling technique did not prove robust enough to produce bi-convex and meniscus lens arrays in a variety of lens shapes, however, and it required lengthy fabrication times. In order to rapidly prototype new lenses, a new diamond machining technique was developed called 4-axis single point diamond machining. This technique is 2-10x faster than micro-milling, depending on how advanced the micro-milling equipment is. With array microscope fabrication still in development, a single prototype of the lens designed for an array microscope was fabricated using single point diamond turning. The prototype microscope objective was validated in a pre-clinical trial. The prototype was compared with a standard clinical microscope objective in diagnostic tests. High concordance, a Fleiss's kappa of 0.88, was found between diagnoses made using the prototype and standard microscope objectives and a reference test. With the lens designed and validated and an advanced fabrication process developed, array microscopy technology is advanced to the point where it is feasible to rapidly prototype an array microscope for detection of tuberculosis and translate array microscope from an innovative concept to a device that can save lives.

Characterization of the strain-life fatigue properties of thin sheet metal using an optical extensometer

NASA Astrophysics Data System (ADS)

Zhang, Shuiqiang; Mao, Shuangshuang; Arola, Dwayne; Zhang, Dongsheng

2014-09-01

Characterizing the strain-life fatigue behavior of thin sheet metals is often challenging since the required specimens have short gauge lengths to avoid buckling, thereby preventing the use of conventional mechanical extensometers. To overcome this obstacle a microscopic optical imaging system has been developed to measure the strain amplitude during fatigue testing using Digital Image Correlation (DIC). A strategy for rapidly recording images is utilized to enable sequential image sampling rates of at least 10 frames per second (fps) using a general digital camera. An example of a complete strain-life fatigue test for thin sheet steel under constant displacement control is presented in which the corresponding strain within the gage section of the specimen is measured using the proposed imaging system. The precision in strain measurement is assessed and methods for improving the image sampling rates in dynamic testing are discussed.

Construction of a Quantum Matter Synthesizer

NASA Astrophysics Data System (ADS)

Trisnadi, Jonathan; McDonald, Mickey; Chin, Cheng

2017-04-01

We report progress on the construction of a new platform to manipulate ultracold atoms. The ``Quantum Matter Synthesizer (QMS)'' will have the capability of deterministically preparing large 2D arrays of atoms with single site addressability. Cesium atoms are first transferred into a science cell (specially textured to reduce reflectance to 0.1% across a wide range of wavelengths and incident angles) via a moving 1D lattice, where they are loaded into a magic-wavelength, far-detuned 2D optical lattice. Two NA=0.8 microscope objectives surround the science cell from above and below. The lower objective will be used to project an array of optical tweezers created via a digital micromirror device (DMD) onto the atom-trapping plane, which will be used to rearrange atoms into a desired configuration after first taking a site-resolved fluorescence image of the initial atomic distribution with the upper objective. We provide updates on our magnetic-optical trap and Raman-sideband cooling performance, characterization of the resolution of our microscope objectives, and stability tests for the objective mounting structure.

A targeted illumination optical fiber probe for high resolution fluorescence imaging and optical switching

NASA Astrophysics Data System (ADS)

Shinde, Anant; Perinchery, Sandeep Menon; Murukeshan, Vadakke Matham

2017-04-01

An optical imaging probe with targeted multispectral and spatiotemporal illumination features has applications in many diagnostic biomedical studies. However, these systems are mostly adapted in conventional microscopes, limiting their use for in vitro applications. We present a variable resolution imaging probe using a digital micromirror device (DMD) with an achievable maximum lateral resolution of 2.7 μm and an axial resolution of 5.5 μm, along with precise shape selective targeted illumination ability. We have demonstrated switching of different wavelengths to image multiple regions in the field of view. Moreover, the targeted illumination feature allows enhanced image contrast by time averaged imaging of selected regions with different optical exposure. The region specific multidirectional scanning feature of this probe has facilitated high speed targeted confocal imaging.

Digital Hilbert transformation for separation measurement of thicknesses and refractive indices of layered objects by use of a wavelength-scanning heterodyne interference confocal microscope.

PubMed

Watanabe, Yuuki; Yamaguchi, Ichirou

2002-08-01

A wavelength-scanning heterodyne interference confocal microscope quickly accomplishes the simultaneous measurement of the thickness and the refractive index of a sample by detection of the amplitude and the phase of the interference signal during a sample scan. However, the measurement range of the optical path difference (OPD) that is obtained from the phase changes is limited by the time response of the phase-locked loop circuit in the FM demodulator. To overcome this limitation and to improve the accuracy of the separation measurement, we propose an OPD detection using digital signal processing with a Hilbert transform. The measurement range is extended approximately five times, and the resolution of the OPD is improved to 5.5 from 9 microm without the electrical noise of the FM demodulator circuit. By applying this method for simultaneous measurement of thickness and the refractive index, we can measure samples 20-30-microm thick with refractive indices between 1 and 1.5.

Digital Hilbert transformation for separation measurement of thicknesses and refractive indices of layered objects by use of a wavelength-scanning heterodyne interference confocal microscope

NASA Astrophysics Data System (ADS)

Watanabe, Yuuki; Yamaguchi, Ichirou

2002-08-01

A wavelength-scanning heterodyne interference confocal microscope quickly accomplishes the simultaneous measurement of the thickness and the refractive index of a sample by detection of the amplitude and the phase of the interference signal during a sample scan. However, the measurement range of the optical path difference (OPD) that is obtained from the phase changes is limited by the time response of the phase-locked loop circuit in the FM demodulator. To overcome this limitation and to improve the accuracy of the separation measurement, we propose an OPD detection using digital signal processing with a Hilbert transform. The measurement range is extended approximately five times, and the resolution of the OPD is improved to 5.5 from 9 mum without the electrical noise of the FM demodulator circuit. By applying this method for simultaneous measurement of thickness and the refractive index, we can measure samples 20-30-mum thick with refractive indices between 1 and 1.5.

Noise removal in extended depth of field microscope images through nonlinear signal processing.

PubMed

Zahreddine, Ramzi N; Cormack, Robert H; Cogswell, Carol J

2013-04-01

Extended depth of field (EDF) microscopy, achieved through computational optics, allows for real-time 3D imaging of live cell dynamics. EDF is achieved through a combination of point spread function engineering and digital image processing. A linear Wiener filter has been conventionally used to deconvolve the image, but it suffers from high frequency noise amplification and processing artifacts. A nonlinear processing scheme is proposed which extends the depth of field while minimizing background noise. The nonlinear filter is generated via a training algorithm and an iterative optimizer. Biological microscope images processed with the nonlinear filter show a significant improvement in image quality and signal-to-noise ratio over the conventional linear filter.

Operative record using intraoperative digital data in neurosurgery.

PubMed

Houkin, K; Kuroda, S; Abe, H

2000-01-01

The purpose of this study was to develop a new method for more efficient and accurate operative records using intra-operative digital data in neurosurgery, including macroscopic procedures and microscopic procedures under an operating microscope. Macroscopic procedures were recorded using a digital camera and microscopic procedures were also recorded using a microdigital camera attached to an operating microscope. Operative records were then recorded digitally and filed in a computer using image retouch software and database base software. The time necessary for editing of the digital data and completing the record was less than 30 minutes. Once these operative records are digitally filed, they are easily transferred and used as database. Using digital operative records along with digital photography, neurosurgeons can document their procedures more accurately and efficiently than by the conventional method (handwriting). A complete digital operative record is not only accurate but also time saving. Construction of a database, data transfer and desktop publishing can be achieved using the intra-operative data, including intra-operative photographs.

Smartphone adapters for digital photomicrography.

PubMed

Roy, Somak; Pantanowitz, Liron; Amin, Milon; Seethala, Raja R; Ishtiaque, Ahmed; Yousem, Samuel A; Parwani, Anil V; Cucoranu, Ioan; Hartman, Douglas J

2014-01-01

Photomicrographs in Anatomic Pathology provide a means of quickly sharing information from a glass slide for consultation, education, documentation and publication. While static image acquisition historically involved the use of a permanently mounted camera unit on a microscope, such cameras may be expensive, need to be connected to a computer, and often require proprietary software to acquire and process images. Another novel approach for capturing digital microscopic images is to use smartphones coupled with the eyepiece of a microscope. Recently, several smartphone adapters have emerged that allow users to attach mobile phones to the microscope. The aim of this study was to test the utility of these various smartphone adapters. We surveyed the market for adapters to attach smartphones to the ocular lens of a conventional light microscope. Three adapters (Magnifi, Skylight and Snapzoom) were tested. We assessed the designs of these adapters and their effectiveness at acquiring static microscopic digital images. All adapters facilitated the acquisition of digital microscopic images with a smartphone. The optimal adapter was dependent on the type of phone used. The Magnifi adapters for iPhone were incompatible when using a protective case. The Snapzoom adapter was easiest to use with iPhones and other smartphones even with protective cases. Smartphone adapters are inexpensive and easy to use for acquiring digital microscopic images. However, they require some adjustment by the user in order to optimize focus and obtain good quality images. Smartphone microscope adapters provide an economically feasible method of acquiring and sharing digital pathology photomicrographs.

Zoom microscope objective using electrowetting lenses.

PubMed

Li, Lei; Wang, Di; Liu, Chao; Wang, Qiong-Hua

2016-02-08

We report a zoom microscope objective which can achieve continuous zoom change and correct the aberrations dynamically. The objective consists of three electrowetting liquid lenses and two glass lenses. The magnification is changed by applying voltages on the three electrowetting lenses. Besides, the three electrowetting liquid lenses can play a role to correct the aberrations. A digital microscope based on the proposed objective is demonstrated. We analyzed the properties of the proposed objective. In contrast to the conventional objectives, the proposed objective can be tuned from ~7.8 × to ~13.2 × continuously. For our objective, the working distance is fixed, which means no movement parts are needed to refocus or change its magnification. Moreover, the zoom objective can be dynamically optimized for a wide range of wavelength. Using such an objective, the fabrication tolerance of the optical system is larger than that of a conventional system, which can decrease the fabrication cost. The proposed zoom microscope objective cannot only take place of the conventional objective, but also has potential application in the 3D microscopy.

Quantitative DIC microscopy using an off-axis self-interference approach.

PubMed

Fu, Dan; Oh, Seungeun; Choi, Wonshik; Yamauchi, Toyohiko; Dorn, August; Yaqoob, Zahid; Dasari, Ramachandra R; Feld, Michael S

2010-07-15

Traditional Normarski differential interference contrast (DIC) microscopy is a very powerful method for imaging nonstained biological samples. However, one of its major limitations is the nonquantitative nature of the imaging. To overcome this problem, we developed a quantitative DIC microscopy method based on off-axis sample self-interference. The digital holography algorithm is applied to obtain quantitative phase gradients in orthogonal directions, which leads to a quantitative phase image through a spiral integration of the phase gradients. This method is practically simple to implement on any standard microscope without stringent requirements on polarization optics. Optical sectioning can be obtained through enlarged illumination NA.

Digital holographic microtomography of fusion spliced optical fibers

NASA Astrophysics Data System (ADS)

Deng, Yating; Xiao, Wen; Ma, Xichao; Pan, Feng

2017-03-01

In this paper, we report three-dimensional(3D) measurement results of structural parameters of fusion spliced optical fibers using digital holographic microtomography. A holographic setup in microscopy configuration with the sample-fixed and setup-rotating scheme is established. A series of holograms is recorded from various incident angles. Then the filtered backprojection algorithm is applied to reconstruct the 3D refractive index (RI) distributions of the fusion spliced optical fibers inserted in the index-matching liquid. Experimental results exhibit the internal and external shapes of three kinds of fusion splices between different fibers, including a single-mode fiber(SMF) and a multimode fiber, an SMF and a panda polarization maintaining fiber (Panda PMF), and an SMF and a bow-tie polarization maintaining fiber (Bow-Tie PMF). With 3D maps of RI, it is intuitive to observe internal structural details of fused fibers and evaluate the splicing quality. This paper describes a powerful method for non-invasive microscopic measurement of fiber splicing. Furthermore, it provides the possibility of detecting fiber splicing loss by 3D structures.

Compact whole-body fluorescent imaging of nude mice bearing EGFP expressing tumor

NASA Astrophysics Data System (ADS)

Chen, Yanping; Xiong, Tao; Chu, Jun; Yu, Li; Zeng, Shaoqun; Luo, Qingming

2005-01-01

Issue of tumor has been a hotspot of current medicine. It is important for tumor research to detect tumors bearing in animal models easily, fast, repetitively and noninvasivly. Many researchers have paid their increasing interests on the detecting. Some contrast agents, such as green fluorescent protein (GFP) and Discosoma red fluorescent protein (Dsred) were applied to enhance image quality. Three main kinds of imaging scheme were adopted to visualize fluorescent protein expressing tumors in vivo. These schemes based on fluorescence stereo microscope, cooled charge-coupled-device (CCD) or camera as imaging set, and laser or mercury lamp as excitation light source. Fluorescence stereo microscope, laser and cooled CCD are expensive to many institutes. The authors set up an inexpensive compact whole-body fluorescent imaging tool, which consisted of a Kodak digital camera (model DC290), fluorescence filters(B and G2;HB Optical, Shenyang, Liaoning, P.R. China) and a mercury 50-W lamp power supply (U-LH50HG;Olympus Optical, Japan) as excitation light source. The EGFP was excited directly by mercury lamp with D455/70 nm band-pass filter and fluorescence was recorded by digital camera with 520nm long-pass filter. By this easy operation tool, the authors imaged, in real time, fluorescent tumors growing in live mice. The imaging system is external and noninvasive. For half a year our experiments suggested the imaging scheme was feasible. Whole-body fluorescence optical imaging for fluorescent expressing tumors in nude mouse is an ideal tool for antitumor, antimetastatic, and antiangiogenesis drug screening.

Full field study of strain distribution near the crack tip in the fracture of solid propellants via large strain digital image correlation and optical microscopy

NASA Astrophysics Data System (ADS)

Gonzalez, Javier

A full field method for visualizing deformation around the crack tip in a fracture process with large strains is developed. A digital image correlation program (DIC) is used to incrementally compute strains and displacements between two consecutive images of a deformation process. Values of strain and displacements for consecutive deformations are added, this way solving convergence problems in the DIC algorithm when large deformations are investigated. The method developed is used to investigate the strain distribution within 1 mm of the crack tip in a particulate composite solid (propellant) using microscopic visualization of the deformation process.

TRIIG - Time-lapse reproduction of images through interactive graphics. [digital processing of quality hard copy

NASA Technical Reports Server (NTRS)

Buckner, J. D.; Council, H. W.; Edwards, T. R.

1974-01-01

Description of the hardware and software implementing the system of time-lapse reproduction of images through interactive graphics (TRIIG). The system produces a quality hard copy of processed images in a fast and inexpensive manner. This capability allows for optimal development of processing software through the rapid viewing of many image frames in an interactive mode. Three critical optical devices are used to reproduce an image: an Optronics photo reader/writer, the Adage Graphics Terminal, and Polaroid Type 57 high speed film. Typical sources of digitized images are observation satellites, such as ERTS or Mariner, computer coupled electron microscopes for high-magnification studies, or computer coupled X-ray devices for medical research.

On the influence of lipid-induced optical anisotropy for the bioimaging of exo- or endocytosis with interference microscopic imaging.

PubMed

Marques, D; Miranda, A; Silva, A G; Munro, P R T; DE Beule, P A A

2018-05-01

Some implementations of interference microscopy imaging use digital holographic measurements of complex scattered fields to reconstruct three-dimensional refractive index maps of weakly scattering, semi-transparent objects, frequently encountered in biological investigations. Reconstruction occurs through application of the object scattering potential which assumes an isotropic refractive index throughout the object. Here, we demonstrate that this assumption can in some circumstances be invalid for biological imaging due to the presence of lipid-induced optical anisotropy. We show that the nanoscale organization of lipids in the observation of cellular endocytosis with polarized light induces a significant change in far-field scattering. We obtain this result by presenting a general solution to Maxwell's equations describing light scattering of core-shell particles near an isotropic substrate covered with an anisotropic thin film. This solution is based on an extension of the Bobbert-Vlieger solution for particle scattering near a substrate delivering an exact solution to the scattering problem in the near field as well as far field. By applying this solution to study light scattering by a lipid vesicle near a lipid bilayer, whereby the lipids are represented through a biaxial optical model, we conclude through ellipsometry concepts that effective amounts of lipid-induced optical anisotropy significantly alter far-field optical scattering in respect to an equivalent optical model that neglects the presence of optical anisotropy. © 2018 The Authors Journal of Microscopy © 2018 Royal Microscopical Society.

Smartphone adapters for digital photomicrography

PubMed Central

Roy, Somak; Pantanowitz, Liron; Amin, Milon; Seethala, Raja R.; Ishtiaque, Ahmed; Yousem, Samuel A.; Parwani, Anil V.; Cucoranu, Ioan; Hartman, Douglas J.

2014-01-01

Background: Photomicrographs in Anatomic Pathology provide a means of quickly sharing information from a glass slide for consultation, education, documentation and publication. While static image acquisition historically involved the use of a permanently mounted camera unit on a microscope, such cameras may be expensive, need to be connected to a computer, and often require proprietary software to acquire and process images. Another novel approach for capturing digital microscopic images is to use smartphones coupled with the eyepiece of a microscope. Recently, several smartphone adapters have emerged that allow users to attach mobile phones to the microscope. The aim of this study was to test the utility of these various smartphone adapters. Materials and Methods: We surveyed the market for adapters to attach smartphones to the ocular lens of a conventional light microscope. Three adapters (Magnifi, Skylight and Snapzoom) were tested. We assessed the designs of these adapters and their effectiveness at acquiring static microscopic digital images. Results: All adapters facilitated the acquisition of digital microscopic images with a smartphone. The optimal adapter was dependent on the type of phone used. The Magnifi adapters for iPhone were incompatible when using a protective case. The Snapzoom adapter was easiest to use with iPhones and other smartphones even with protective cases. Conclusions: Smartphone adapters are inexpensive and easy to use for acquiring digital microscopic images. However, they require some adjustment by the user in order to optimize focus and obtain good quality images. Smartphone microscope adapters provide an economically feasible method of acquiring and sharing digital pathology photomicrographs. PMID:25191623

3D imaging of optically cleared tissue using a simplified CLARITY method and on-chip microscopy

PubMed Central

Zhang, Yibo; Shin, Yoonjung; Sung, Kevin; Yang, Sam; Chen, Harrison; Wang, Hongda; Teng, Da; Rivenson, Yair; Kulkarni, Rajan P.; Ozcan, Aydogan

2017-01-01

High-throughput sectioning and optical imaging of tissue samples using traditional immunohistochemical techniques can be costly and inaccessible in resource-limited areas. We demonstrate three-dimensional (3D) imaging and phenotyping in optically transparent tissue using lens-free holographic on-chip microscopy as a low-cost, simple, and high-throughput alternative to conventional approaches. The tissue sample is passively cleared using a simplified CLARITY method and stained using 3,3′-diaminobenzidine to target cells of interest, enabling bright-field optical imaging and 3D sectioning of thick samples. The lens-free computational microscope uses pixel super-resolution and multi-height phase recovery algorithms to digitally refocus throughout the cleared tissue and obtain a 3D stack of complex-valued images of the sample, containing both phase and amplitude information. We optimized the tissue-clearing and imaging system by finding the optimal illumination wavelength, tissue thickness, sample preparation parameters, and the number of heights of the lens-free image acquisition and implemented a sparsity-based denoising algorithm to maximize the imaging volume and minimize the amount of the acquired data while also preserving the contrast-to-noise ratio of the reconstructed images. As a proof of concept, we achieved 3D imaging of neurons in a 200-μm-thick cleared mouse brain tissue over a wide field of view of 20.5 mm2. The lens-free microscope also achieved more than an order-of-magnitude reduction in raw data compared to a conventional scanning optical microscope imaging the same sample volume. Being low cost, simple, high-throughput, and data-efficient, we believe that this CLARITY-enabled computational tissue imaging technique could find numerous applications in biomedical diagnosis and research in low-resource settings. PMID:28819645

Quantitative luminescence imaging system

DOEpatents

Erwin, David N.; Kiel, Johnathan L.; Batishko, Charles R.; Stahl, Kurt A.

1990-01-01

The QLIS images and quantifies low-level chemiluminescent reactions in an electromagnetic field. It is capable of real time nonperturbing measurement and simultaneous recording of many biochemical and chemical reactions such as luminescent immunoassays or enzyme assays. The system comprises image transfer optics, a low-light level digitizing camera with image intensifying microchannel plates, an image process or, and a control computer. The image transfer optics may be a fiber image guide with a bend, or a microscope, to take the light outside of the RF field. Output of the camera is transformed into a localized rate of cumulative digitalized data or enhanced video display or hard-copy images. The system may be used as a luminescent microdosimetry device for radiofrequency or microwave radiation, as a thermal dosimeter, or in the dosimetry of ultra-sound (sonoluminescence) or ionizing radiation. It provides a near-real-time system capable of measuring the extremely low light levels from luminescent reactions in electromagnetic fields in the areas of chemiluminescence assays and thermal microdosimetry, and is capable of near-real-time imaging of the sample to allow spatial distribution analysis of the reaction. It can be used to instrument three distinctly different irradiation configurations, comprising (1) RF waveguide irradiation of a small Petri-dish-shaped sample cell, (2) RF irradiation of samples in a microscope for the microscopie imaging and measurement, and (3) RF irradiation of small to human body-sized samples in an anechoic chamber.

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.

Study on mechanical properties and damage behaviors of Kevlar fiber reinforced epoxy composites by digital image correlation technique under optical microscope

NASA Astrophysics Data System (ADS)

Gao, Xiang; Shao, Wenquan; Ji, Hongwei

2010-10-01

Kevlar fiber-reinforced epoxy (KFRE) composites are widely used in the fields of aerospace, weapon, shipping, and civil industry, due to their outstanding capabilities. In this paper, mechanical properties and damage behaviors of KFRE laminate (02/902) were tested and studied under tension condition. To precisely measure the tensile mechanical properties of the material and investigate its micro-scale damage evolution, a micro-image measuring system with in-situ tensile device was designed. The measuring system, by which the in-situ tensile test can be carried out and surface morphology evolution of the tensile specimen can be visually monitored and recorded during the process of loading, includes an ultra-long working distance zoom microscope and a in-situ tensile loading device. In this study, a digital image correlation method (DICM) was used to calculate the deformation of the tensile specimen under different load levels according to the temporal series images captured by an optical microscope and CCD camera. Then, the elastic modulus and Poisson's ratio of the KFRE was obtained accordingly. The damage progresses of the KFRE laminates were analyzed. Experimental results indicated that: (1) the KFRE laminate (02/902) is almost elastic, its failure mode is brittle tensile fracture.(2) Mechanical properties parameters of the material are as follows: elastic modulus is 14- 16GPa, and tensile ultimate stress is 450-480 Mpa respectively. (3) The damage evolution of the material is that cracks appear in epoxy matrix firstly, then, with the increasing of the tensile loading, matrix cracks add up and extend along a 45° angle direction with tensile load. Furthermore, decohesion between matrix and fibers as well as delamination occurs. Eventually, fibers break and the material is damaged.

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.

Optical analysis of electro-optical systems by MTF calculus

NASA Astrophysics Data System (ADS)

Barbarini, Elisa Signoreto; Dos Santos, Daniel, Jr.; Stefani, Mário Antonio; Yasuoka, Fátima Maria Mitsue; Castro Neto, Jarbas C.; Rodrigues, Evandro Luís Linhari

2011-08-01

One of the widely used methods for performance analysis of an optical system is the determination of the Modulation Transfer Function (MTF). The MTF represents a quantitative and direct measure of image quality, and, besides being an objective test, it can be used on concatenated optical system. This paper presents the application of software called SMTF (software modulation transfer function), built in C++ and Open CV platforms for MTF calculation on electro-optical system. Through this technique, it is possible to develop specific method to measure the real time performance of a digital fundus camera, an infrared sensor and an ophthalmological surgery microscope. Each optical instrument mentioned has a particular device to measure the MTF response, which is being developed. Then the MTF information assists the analysis of the optical system alignment, and also defines its resolution limit by the MTF graphic. The result obtained from the implemented software is compared with the theoretical MTF curve from the analyzed systems.

Development of an imaging method for quantifying a large digital PCR droplet

NASA Astrophysics Data System (ADS)

Huang, Jen-Yu; Lee, Shu-Sheng; Hsu, Yu-Hsiang

2017-02-01

Portable devices have been recognized as the future linkage between end-users and lab-on-a-chip devices. It has a user friendly interface and provides apps to interface headphones, cameras, and communication duct, etc. In particular, the digital resolution of cameras installed in smartphones or pads already has a high imaging resolution with a high number of pixels. This unique feature has triggered researches to integrate optical fixtures with smartphone to provide microscopic imaging capabilities. In this paper, we report our study on developing a portable diagnostic tool based on the imaging system of a smartphone and a digital PCR biochip. A computational algorithm is developed to processing optical images taken from a digital PCR biochip with a smartphone in a black box. Each reaction droplet is recorded in pixels and is analyzed in a sRGB (red, green, and blue) color space. Multistep filtering algorithm and auto-threshold algorithm are adopted to minimize background noise contributed from ccd cameras and rule out false positive droplets, respectively. Finally, a size-filtering method is applied to identify the number of positive droplets to quantify target's concentration. Statistical analysis is then performed for diagnostic purpose. This process can be integrated in an app and can provide a user friendly interface without professional training.

Algorithms and applications of aberration correction and American standard-based digital evaluation in surface defects evaluating system

NASA Astrophysics Data System (ADS)

Wu, Fan; Cao, Pin; Yang, Yongying; Li, Chen; Chai, Huiting; Zhang, Yihui; Xiong, Haoliang; Xu, Wenlin; Yan, Kai; Zhou, Lin; Liu, Dong; Bai, Jian; Shen, Yibing

2016-11-01

The inspection of surface defects is one of significant sections of optical surface quality evaluation. Based on microscopic scattering dark-field imaging, sub-aperture scanning and stitching, the Surface Defects Evaluating System (SDES) can acquire full-aperture image of defects on optical elements surface and then extract geometric size and position information of defects with image processing such as feature recognization. However, optical distortion existing in the SDES badly affects the inspection precision of surface defects. In this paper, a distortion correction algorithm based on standard lattice pattern is proposed. Feature extraction, polynomial fitting and bilinear interpolation techniques in combination with adjacent sub-aperture stitching are employed to correct the optical distortion of the SDES automatically in high accuracy. Subsequently, in order to digitally evaluate surface defects with American standard by using American military standards MIL-PRF-13830B to judge the surface defects information obtained from the SDES, an American standard-based digital evaluation algorithm is proposed, which mainly includes a judgment method of surface defects concentration. The judgment method establishes weight region for each defect and adopts the method of overlap of weight region to calculate defects concentration. This algorithm takes full advantage of convenience of matrix operations and has merits of low complexity and fast in running, which makes itself suitable very well for highefficiency inspection of surface defects. Finally, various experiments are conducted and the correctness of these algorithms are verified. At present, these algorithms have been used in SDES.

The different ways to obtain digital images of urine microscopy findings: Their advantages and limitations.

PubMed

Fogazzi, G B; Garigali, G

2017-03-01

We describe three ways to take digital images of urine sediment findings. Way 1 encompasses a digital camera permanently mounted on the microscope and connected with a computer equipped with a proprietary software to acquire, process and store the images. Way 2 is based on the use of inexpensive compact digital cameras, held by hands - or mounted on a tripod - close to one eyepiece of the microscope. Way 3 is based on the use of smartphones, held by hands close to one eyepiece of the microscope or connected to the microscope by an adapter. The procedures, advantages and limitations of each way are reported. Copyright © 2017. Published by Elsevier B.V.

Surface defects evaluation system based on electromagnetic model simulation and inverse-recognition calibration method

NASA Astrophysics Data System (ADS)

Yang, Yongying; Chai, Huiting; Li, Chen; Zhang, Yihui; Wu, Fan; Bai, Jian; Shen, Yibing

2017-05-01

Digitized evaluation of micro sparse defects on large fine optical surfaces is one of the challenges in the field of optical manufacturing and inspection. The surface defects evaluation system (SDES) for large fine optical surfaces is developed based on our previously reported work. In this paper, the electromagnetic simulation model based on Finite-Difference Time-Domain (FDTD) for vector diffraction theory is firstly established to study the law of microscopic scattering dark-field imaging. Given the aberration in actual optical systems, point spread function (PSF) approximated by a Gaussian function is introduced in the extrapolation from the near field to the far field and the scatter intensity distribution in the image plane is deduced. Analysis shows that both diffraction-broadening imaging and geometrical imaging should be considered in precise size evaluation of defects. Thus, a novel inverse-recognition calibration method is put forward to avoid confusion caused by diffraction-broadening effect. The evaluation method is applied to quantitative evaluation of defects information. The evaluation results of samples of many materials by SDES are compared with those by OLYMPUS microscope to verify the micron-scale resolution and precision. The established system has been applied to inspect defects on large fine optical surfaces and can achieve defects inspection of surfaces as large as 850 mm×500 mm with the resolution of 0.5 μm.

A Comparative Study of Microscopic Images Captured by a Box Type Digital Camera Versus a Standard Microscopic Photography Camera Unit

PubMed Central

Desai, Nandini J.; Gupta, B. D.; Patel, Pratik Narendrabhai

2014-01-01

Introduction: Obtaining images of slides viewed by a microscope can be invaluable for both diagnosis and teaching.They can be transferred among technologically-advanced hospitals for further consultation and evaluation. But a standard microscopic photography camera unit (MPCU)(MIPS-Microscopic Image projection System) is costly and not available in resource poor settings. The aim of our endeavour was to find a comparable and cheaper alternative method for photomicrography. Materials and Methods: We used a NIKON Coolpix S6150 camera (box type digital camera) with Olympus CH20i microscope and a fluorescent microscope for the purpose of this study. Results: We got comparable results for capturing images of light microscopy, but the results were not as satisfactory for fluorescent microscopy. Conclusion: A box type digital camera is a comparable, less expensive and convenient alternative to microscopic photography camera unit. PMID:25478350

Functional imaging of glucose-evoked rat islet activities using transient intrinsic optical signals

NASA Astrophysics Data System (ADS)

Yao, Xin-Cheng; Cui, Wan-Xing; Li, Yi-Chao; Zhang, Wei; Lu, Rong-Wen; Thompson, Anthony; Amthor, Franklin; Wang, Xu-Jing

2012-05-01

We demonstrate intrinsic optical signal (IOS) imaging of intact rat islet, which consists of many endocrine cells working together. A near-infrared digital microscope was employed for optical monitoring of islet activities evoked by glucose stimulation. Dynamic NIR images revealed transient IOS responses in the islet activated by low-dose (2.75 mM) and high-dose (5.5 mM) glucose stimuli. Comparative experiments and quantitative analysis indicated that both glucose metabolism and calcium/insulin dynamics might contribute to the observed IOS responses. Further investigation of the IOS imaging technology may provide a high resolution method for ex vivo functional examination of the islet, which is important for advanced study of diabetes associated islet dysfunctions and for improved quality control of donor islets for transplantation.

A Minimal Optical Trapping and Imaging Microscopy System

PubMed Central

Hernández Candia, Carmen Noemí; Tafoya Martínez, Sara; Gutiérrez-Medina, Braulio

2013-01-01

We report the construction and testing of a simple and versatile optical trapping apparatus, suitable for visualizing individual microtubules (∼25 nm in diameter) and performing single-molecule studies, using a minimal set of components. This design is based on a conventional, inverted microscope, operating under plain bright field illumination. A single laser beam enables standard optical trapping and the measurement of molecular displacements and forces, whereas digital image processing affords real-time sample visualization with reduced noise and enhanced contrast. We have tested our trapping and imaging instrument by measuring the persistence length of individual double-stranded DNA molecules, and by following the stepping of single kinesin motor proteins along clearly imaged microtubules. The approach presented here provides a straightforward alternative for studies of biomaterials and individual biomolecules. PMID:23451216

A phased antenna array for surface plasmons

PubMed Central

Dikken, Dirk Jan W.; Korterik, Jeroen P.; Segerink, Frans B.; Herek, Jennifer L.; Prangsma, Jord C.

2016-01-01

Surface plasmon polaritons are electromagnetic waves that propagate tightly bound to metal surfaces. The concentration of the electromagnetic field at the surface as well as the short wavelength of surface plasmons enable sensitive detection methods and miniaturization of optics. We present an optical frequency plasmonic analog to the phased antenna array as it is well known in radar technology and radio astronomy. Individual holes in a thick gold film act as dipolar emitters of surface plasmon polaritons whose phase is controlled individually using a digital spatial light modulator. We show experimentally, using a phase sensitive near-field microscope, that this optical system allows accurate directional emission of surface waves. This compact and flexible method allows for dynamically shaping the propagation of plasmons and holds promise for nanophotonic applications employing propagating surface plasmons. PMID:27121099

Simple and fast spectral domain algorithm for quantitative phase imaging of living cells with digital holographic microscopy

NASA Astrophysics Data System (ADS)

Min, Junwei; Yao, Baoli; Ketelhut, Steffi; Kemper, Björn

2017-02-01

The modular combination of optical microscopes with digital holographic microscopy (DHM) has been proven to be a powerful tool for quantitative live cell imaging. The introduction of condenser and different microscope objectives (MO) simplifies the usage of the technique and makes it easier to measure different kinds of specimens with different magnifications. However, the high flexibility of illumination and imaging also causes variable phase aberrations that need to be eliminated for high resolution quantitative phase imaging. The existent phase aberrations compensation methods either require add additional elements into the reference arm or need specimen free reference areas or separate reference holograms to build up suitable digital phase masks. These inherent requirements make them unpractical for usage with highly variable illumination and imaging systems and prevent on-line monitoring of living cells. In this paper, we present a simple numerical method for phase aberration compensation based on the analysis of holograms in spatial frequency domain with capabilities for on-line quantitative phase imaging. From a single shot off-axis hologram, the whole phase aberration can be eliminated automatically without numerical fitting or pre-knowledge of the setup. The capabilities and robustness for quantitative phase imaging of living cancer cells are demonstrated.

Structured light optical microscopy for three-dimensional reconstruction of technical surfaces

NASA Astrophysics Data System (ADS)

Kettel, Johannes; Reinecke, Holger; Müller, Claas

2016-04-01

In microsystems technology quality control of micro structured surfaces with different surface properties is playing an ever more important role. The process of quality control incorporates three-dimensional (3D) reconstruction of specularand diffusive reflecting technical surfaces. Due to the demand on high measurement accuracy and data acquisition rates, structured light optical microscopy has become a valuable solution to solve this problem providing high vertical and lateral resolution. However, 3D reconstruction of specular reflecting technical surfaces still remains a challenge to optical measurement principles. In this paper we present a measurement principle based on structured light optical microscopy which enables 3D reconstruction of specular- and diffusive reflecting technical surfaces. It is realized using two light paths of a stereo microscope equipped with different magnification levels. The right optical path of the stereo microscope is used to project structured light onto the object surface. The left optical path is used to capture the structured illuminated object surface with a camera. Structured light patterns are generated by a Digital Light Processing (DLP) device in combination with a high power Light Emitting Diode (LED). Structured light patterns are realized as a matrix of discrete light spots to illuminate defined areas on the object surface. The introduced measurement principle is based on multiple and parallel processed point measurements. Analysis of the measured Point Spread Function (PSF) by pattern recognition and model fitting algorithms enables the precise calculation of 3D coordinates. Using exemplary technical surfaces we demonstrate the successful application of our measurement principle.

High-resolution optical control of spatiotemporal neuronal activity patterns in zebrafish using a digital micromirror device.

PubMed

Zhu, Peixin; Fajardo, Otto; Shum, Jennifer; Zhang Schärer, Yan-Ping; Friedrich, Rainer W

2012-06-28

Optogenetic approaches allow the manipulation of neuronal activity patterns in space and time by light, particularly in small animals such as zebrafish. However, most techniques cannot control neuronal activity independently at different locations. Here we describe equipment and provide a protocol for single-photon patterned optical stimulation of neurons using a digital micromirror device (DMD). This method can create arbitrary spatiotemporal light patterns with spatial and temporal resolutions in the micrometer and submillisecond range, respectively. Different options to integrate a DMD into a multiphoton microscope are presented and compared. We also describe an ex vivo preparation of the adult zebrafish head that greatly facilitates optogenetic and other experiments. After assembly, the initial alignment takes about one day and the zebrafish preparation takes <30 min. The method has previously been used to activate channelrhodopsin-2 and manipulate oscillatory synchrony among spatially distributed neurons in the zebrafish olfactory bulb. It can be adapted easily to a wide range of other species, optogenetic probes and scientific applications.

Manufacture and calibration of optical supersmooth roughness artifacts for intercomparisons

NASA Astrophysics Data System (ADS)

Ringel, Gabriele A.; Kratz, Frank; Schmitt, Dirk-Roger; Mangelsdorf, Juergen; Creuzet, Francois; Garratt, John D.

1995-09-01

Intercomparison roughness measurements have been carried out on supersmooth artifacts fabricated from BK7, fused silica, and Zerodur. The surface parameters were determined using the optical heterodyne profiler Z5500 (Zygo), a special prototype of the mechanical profiler Nanostep (Rank Taylor Hobson), and an Atomic Force Microscope (Park Scientific Instruments) with an improved acquisition technique. The intercomparison was performed after the range of collected spatial wavelengths for each instrument was adjusted using digital filtering techniques. It is demonstrated for different roughness ranges that the applied superpolishing techniques yield supersmooth artifacts which can be used for more intercomparisons. More than 100 samples were investigated. Criteria were developed to select artifacts from the sample stock.

Production and characterization of pure cryogenic inertial fusion targets

NASA Astrophysics Data System (ADS)

Boyd, B. A.; Kamerman, G. W.

An experimental cryogenic inertial fusion target generator and two optical techniques for automated target inspection are described. The generator produces 100 microns diameter solid hydrogen spheres at a rate compatible with fueling requirements of conceptual inertial fusion power plants. A jet of liquified hydrogen is disrupted into droplets by an ultrasonically excited nozzle. The droplets solidify into microspheres while falling through a chamber maintained below the hydrogen triple point pressure. Stable operation of the generator has been demonstrated for up to three hours. The optical inspection techniques are computer aided photomicrography and coarse diffraction pattern analysis (CDPA). The photomicrography system uses a conventional microscope coupled to a computer by a solid state camera and digital image memory. The computer enhances the stored image and performs feature extraction to determine pellet parameters. The CDPA technique uses Fourier transform optics and a special detector array to perform optical processing of a target image.

Use of a Digital Camera To Document Student Observations in a Microbiology Laboratory Class.

ERIC Educational Resources Information Center

Mills, David A.; Kelley, Kevin; Jones, Michael

2001-01-01

Points out the lack of microscopic images of wine-related microbes. Uses a digital camera during a wine microbiology laboratory to capture student-generated microscope images. Discusses the advantages of using a digital camera in a teaching lab. (YDS)

Miniature objective lens for array digital pathology: design improvement based on clinical evaluation

NASA Astrophysics Data System (ADS)

McCall, Brian; Pierce, Mark; Graviss, Edward A.; Richards-Kortum, Rebecca R.; Tkaczyk, Tomasz S.

2016-03-01

A miniature objective designed for digital detection of Mycobacterium tuberculosis (MTB) was evaluated for diagnostic accuracy. The objective was designed for array microscopy, but fabricated and evaluated at this stage of development as a single objective. The counts and diagnoses of patient samples were directly compared for digital detection and standard microscopy. The results were found to be correlated and highly concordant. The evaluation of this lens by direct comparison to standard fluorescence sputum smear microscopy presented unique challenges and led to some new insights in the role played by the system parameters of the microscope. The design parameters and how they were developed are reviewed in light of these results. New system parameters are proposed with the goal of easing the challenges of evaluating the miniature objective and maintaining the optical performance that produced the agreeable results presented without over-optimizing. A new design is presented that meets and exceeds these criteria.

The Scanning Optical Microscope: An Overview

NASA Astrophysics Data System (ADS)

Kino, G. S.; Corte, T. R.; Xiao, G. Q.

1988-07-01

In the last few years there has been a resurgence in research on optical microscopes. One reason stems from the invention of the acoustic microscope by Quate and Lemons,1 and the realization that some of the same principles could be applied to the optical microscope. The acoustic microscope has better transverse definition for the same wavelength than the standard optical microscope and at the same time has far better range definition. Consequently, Kompfner, who was involved with the work on the early acoustic microscope, decided to try out similar scanning microscope principles with optics, and started a group with Wilson and Sheppard to carry out such research at Oxford.2 Sometime earlier, Petran et a13 had invented the tandem scanning microscope which used many of the same principles. Now, in our laboratory at Stanford, these ideas on the tandem scanning microscope and the scanning optical microscope are converging. Another aspect of this work, which stems from the earlier experience with the acoustic microscope, involves measurement of both phase and amplitude of the optical beam. It is also possible to use scanned optical microscopy for other purposes. For instance, an optical beam can be used to excite electrons and holes in semiconductors, and the generated current can be measured. By scanning the optical beam over the semiconductor, an image can be obtained of the regions where there is strong or weak electron hole generation. This type of microscope is called OBIC (Optical Beam Induced Current). A second application involves fluorescent imaging of biological materials. Here we have the excellent range definition of a scanning optical microscope which eliminates unwanted glare from regions of the material where the beam is unfocused.3 A third application is focused on the heating effect of the light beam. With such a system, images can be obtained which are associated with changes in the thermal properties of a material, changes in recombination rates in semiconductors, and differences in material properties associated with either acoustic or thermal effects.4,5 Thus, the range of scanning optical microscopy applications is very large. In the main, the most important applications have been to semiconductors and to biology.

Astigmatism compensation in digital holographic microscopy using complex-amplitude correlation

NASA Astrophysics Data System (ADS)

Tamrin, Khairul Fikri; Rahmatullah, Bahbibi; Samuri, Suzani Mohamad

2015-07-01

Digital holographic microscopy (DHM) is a promising tool for a three-dimensional imaging of microscopic particles. It offers the possibility of wavefront processing by manipulating amplitude and phase of the recorded digital holograms. With a view to compensate for aberration in the reconstructed particle images, this paper discusses a new approach of aberration compensation based on complex amplitude correlation and the use of a priori information. The approach is applied to holograms of microscopic particles flowing inside a cylindrical micro-channel recorded using an off-axis digital holographic microscope. The approach results in improvements in the image and signal qualities.

Near real-time digital holographic microscope based on GPU parallel computing

NASA Astrophysics Data System (ADS)

Zhu, Gang; Zhao, Zhixiong; Wang, Huarui; Yang, Yan

2018-01-01

A transmission near real-time digital holographic microscope with in-line and off-axis light path is presented, in which the parallel computing technology based on compute unified device architecture (CUDA) and digital holographic microscopy are combined. Compared to other holographic microscopes, which have to implement reconstruction in multiple focal planes and are time-consuming the reconstruction speed of the near real-time digital holographic microscope can be greatly improved with the parallel computing technology based on CUDA, so it is especially suitable for measurements of particle field in micrometer and nanometer scale. Simulations and experiments show that the proposed transmission digital holographic microscope can accurately measure and display the velocity of particle field in micrometer scale, and the average velocity error is lower than 10%.With the graphic processing units(GPU), the computing time of the 100 reconstruction planes(512×512 grids) is lower than 120ms, while it is 4.9s using traditional reconstruction method by CPU. The reconstruction speed has been raised by 40 times. In other words, it can handle holograms at 8.3 frames per second and the near real-time measurement and display of particle velocity field are realized. The real-time three-dimensional reconstruction of particle velocity field is expected to achieve by further optimization of software and hardware. Keywords: digital holographic microscope,

DMD-based LED-illumination super-resolution and optical sectioning microscopy.

PubMed

Dan, Dan; Lei, Ming; Yao, Baoli; Wang, Wen; Winterhalder, Martin; Zumbusch, Andreas; Qi, Yujiao; Xia, Liang; Yan, Shaohui; Yang, Yanlong; Gao, Peng; Ye, Tong; Zhao, Wei

2013-01-01

Super-resolution three-dimensional (3D) optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. We present a novel approach of structured illumination microscopy (SIM) by using a digital micromirror device (DMD) for fringe projection and a low-coherence LED light for illumination. The lateral resolution of 90 nm and the optical sectioning depth of 120 μm were achieved. The maximum acquisition speed for 3D imaging in the optical sectioning mode was 1.6×10(7) pixels/second, which was mainly limited by the sensitivity and speed of the CCD camera. In contrast to other SIM techniques, the DMD-based LED-illumination SIM is cost-effective, ease of multi-wavelength switchable and speckle-noise-free. The 2D super-resolution and 3D optical sectioning modalities can be easily switched and applied to either fluorescent or non-fluorescent specimens.

DMD-based LED-illumination Super-resolution and optical sectioning microscopy

PubMed Central

Dan, Dan; Lei, Ming; Yao, Baoli; Wang, Wen; Winterhalder, Martin; Zumbusch, Andreas; Qi, Yujiao; Xia, Liang; Yan, Shaohui; Yang, Yanlong; Gao, Peng; Ye, Tong; Zhao, Wei

2013-01-01

Super-resolution three-dimensional (3D) optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. We present a novel approach of structured illumination microscopy (SIM) by using a digital micromirror device (DMD) for fringe projection and a low-coherence LED light for illumination. The lateral resolution of 90 nm and the optical sectioning depth of 120 μm were achieved. The maximum acquisition speed for 3D imaging in the optical sectioning mode was 1.6×107 pixels/second, which was mainly limited by the sensitivity and speed of the CCD camera. In contrast to other SIM techniques, the DMD-based LED-illumination SIM is cost-effective, ease of multi-wavelength switchable and speckle-noise-free. The 2D super-resolution and 3D optical sectioning modalities can be easily switched and applied to either fluorescent or non-fluorescent specimens. PMID:23346373

The application of digital image plane holography technology to identify Chinese herbal medicine

NASA Astrophysics Data System (ADS)

Wang, Huaying; Guo, Zhongjia; Liao, Wei; Zhang, Zhihui

2012-03-01

In this paper, the imaging technology of digital image plane holography to identify the Chinese herbal medicine is studied. The optical experiment system of digital image plane holography which is the special case of pre-magnification digital holography was built. In the record system, one is an object light by using plane waves which illuminates the object, and the other one is recording hologram by using spherical light wave as reference light. There is a Micro objective lens behind the object. The second phase factor which caus ed by the Micro objective lens can be eliminated by choosing the proper position of the reference point source when digital image plane holography is recorded by spherical light. In this experiment, we use the Lygodium cells and Onion cells as the object. The experiment results with Lygodium cells and Onion cells show that digital image plane holography avoid the process of finding recording distance by using auto-focusing approach, and the phase information of the object can be reconstructed more accurately. The digital image plane holography is applied to the microscopic imaging of cells more effectively, and it is suit to apply for the identify of Chinese Herbal Medicine. And it promotes the application of digital holographic in practice.

Space-resolved diffusing wave spectroscopy measurements of the macroscopic deformation and the microscopic dynamics in tensile strain tests

NASA Astrophysics Data System (ADS)

Nagazi, Med-Yassine; Brambilla, Giovanni; Meunier, Gérard; Marguerès, Philippe; Périé, Jean-Noël; Cipelletti, Luca

2017-01-01

We couple a laser-based, space-resolved dynamic light scattering apparatus to a universal traction machine for mechanical extensional tests. We perform simultaneous optical and mechanical measurements on polyether ether ketone, a semi-crystalline polymer widely used in the industry. Due to the high turbidity of the sample, light is multiply scattered by the sample and the diffusing wave spectroscopy (DWS) formalism is used to interpret the data. Space-resolved DWS yields spatial maps of the sample strain and of the microscopic dynamics. An excellent agreement is found between the strain maps thus obtained and those measured by a conventional stereo-digital image correlation technique. The microscopic dynamics reveals both affine motion and plastic rearrangements. Thanks to the extreme sensitivity of DWS to displacements as small as 1 nm, plastic activity and its spatial localization can be detected at an early stage of the sample strain, making the technique presented here a valuable complement to existing material characterization methods.

Low-cost mobile phone microscopy with a reversed mobile phone camera lens.

PubMed

Switz, Neil A; D'Ambrosio, Michael V; Fletcher, Daniel A

2014-01-01

The increasing capabilities and ubiquity of mobile phones and their associated digital cameras offer the possibility of extending low-cost, portable diagnostic microscopy to underserved and low-resource areas. However, mobile phone microscopes created by adding magnifying optics to the phone's camera module have been unable to make use of the full image sensor due to the specialized design of the embedded camera lens, exacerbating the tradeoff between resolution and field of view inherent to optical systems. This tradeoff is acutely felt for diagnostic applications, where the speed and cost of image-based diagnosis is related to the area of the sample that can be viewed at sufficient resolution. Here we present a simple and low-cost approach to mobile phone microscopy that uses a reversed mobile phone camera lens added to an intact mobile phone to enable high quality imaging over a significantly larger field of view than standard microscopy. We demonstrate use of the reversed lens mobile phone microscope to identify red and white blood cells in blood smears and soil-transmitted helminth eggs in stool samples.

Low-Cost Mobile Phone Microscopy with a Reversed Mobile Phone Camera Lens

PubMed Central

Fletcher, Daniel A.

2014-01-01

The increasing capabilities and ubiquity of mobile phones and their associated digital cameras offer the possibility of extending low-cost, portable diagnostic microscopy to underserved and low-resource areas. However, mobile phone microscopes created by adding magnifying optics to the phone's camera module have been unable to make use of the full image sensor due to the specialized design of the embedded camera lens, exacerbating the tradeoff between resolution and field of view inherent to optical systems. This tradeoff is acutely felt for diagnostic applications, where the speed and cost of image-based diagnosis is related to the area of the sample that can be viewed at sufficient resolution. Here we present a simple and low-cost approach to mobile phone microscopy that uses a reversed mobile phone camera lens added to an intact mobile phone to enable high quality imaging over a significantly larger field of view than standard microscopy. We demonstrate use of the reversed lens mobile phone microscope to identify red and white blood cells in blood smears and soil-transmitted helminth eggs in stool samples. PMID:24854188

Small Wonders Close Encounters

ERIC Educational Resources Information Center

Kniseley, MacGregor; Capraro, Karen

2013-01-01

This article introduces students to the world of digital microscopy. Looking at small objects through a digital microscope is like traveling through a foreign country for the first time. The experience is new, engaging, and exciting. A handheld digital microscope is an essential tool in a 21st century teacher's toolkit and the perfect tool to…

Improved Scanners for Microscopic Hyperspectral Imaging

NASA Technical Reports Server (NTRS)

Mao, Chengye

2009-01-01

Improved scanners to be incorporated into hyperspectral microscope-based imaging systems have been invented. Heretofore, in microscopic imaging, including spectral imaging, it has been customary to either move the specimen relative to the optical assembly that includes the microscope or else move the entire assembly relative to the specimen. It becomes extremely difficult to control such scanning when submicron translation increments are required, because the high magnification of the microscope enlarges all movements in the specimen image on the focal plane. To overcome this difficulty, in a system based on this invention, no attempt would be made to move either the specimen or the optical assembly. Instead, an objective lens would be moved within the assembly so as to cause translation of the image at the focal plane: the effect would be equivalent to scanning in the focal plane. The upper part of the figure depicts a generic proposed microscope-based hyperspectral imaging system incorporating the invention. The optical assembly of this system would include an objective lens (normally, a microscope objective lens) and a charge-coupled-device (CCD) camera. The objective lens would be mounted on a servomotor-driven translation stage, which would be capable of moving the lens in precisely controlled increments, relative to the camera, parallel to the focal-plane scan axis. The output of the CCD camera would be digitized and fed to a frame grabber in a computer. The computer would store the frame-grabber output for subsequent viewing and/or processing of images. The computer would contain a position-control interface board, through which it would control the servomotor. There are several versions of the invention. An essential feature common to all versions is that the stationary optical subassembly containing the camera would also contain a spatial window, at the focal plane of the objective lens, that would pass only a selected portion of the image. In one version, the window would be a slit, the CCD would contain a one-dimensional array of pixels, and the objective lens would be moved along an axis perpendicular to the slit to spatially scan the image of the specimen in pushbroom fashion. The image built up by scanning in this case would be an ordinary (non-spectral) image. In another version, the optics of which are depicted in the lower part of the figure, the spatial window would be a slit, the CCD would contain a two-dimensional array of pixels, the slit image would be refocused onto the CCD by a relay-lens pair consisting of a collimating and a focusing lens, and a prism-gratingprism optical spectrometer would be placed between the collimating and focusing lenses. Consequently, the image on the CCD would be spatially resolved along the slit axis and spectrally resolved along the axis perpendicular to the slit. As in the first-mentioned version, the objective lens would be moved along an axis perpendicular to the slit to spatially scan the image of the specimen in pushbroom fashion.

Virtual reality microscope versus conventional microscope regarding time to diagnosis: an experimental study.

PubMed

Randell, Rebecca; Ruddle, Roy A; Mello-Thoms, Claudia; Thomas, Rhys G; Quirke, Phil; Treanor, Darren

2013-01-01

  To create and evaluate a virtual reality (VR) microscope that is as efficient as the conventional microscope, seeking to support the introduction of digital slides into routine practice.   A VR microscope was designed and implemented by combining ultra-high-resolution displays with VR technology, techniques for fast interaction, and high usability. It was evaluated using a mixed factorial experimental design with technology and task as within-participant variables and grade of histopathologist as a between-participant variable. Time to diagnosis was similar for the conventional and VR microscopes. However, there was a significant difference in the mean magnification used between the two technologies, with participants working at a higher level of magnification on the VR microscope.   The results suggest that, with the right technology, efficient use of digital pathology for routine practice is a realistic possibility. Further work is required to explore what magnification is required on the VR microscope for histopathologists to identify diagnostic features, and the effect on this of the digital slide production process. © 2012 Blackwell Publishing Limited.

High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.

PubMed

Bullen, A; Patel, S S; Saggau, P

1997-07-01

The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging.

High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.

PubMed Central

Bullen, A; Patel, S S; Saggau, P

1997-01-01

The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging. Images FIGURE 6 PMID:9199810

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

2010-06-29

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P.; Chernobrod, Boris M.

2009-11-10

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of impaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P.; Chernobrod, Boris M.

2007-12-11

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

2010-07-13

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Spin microscope based on optically detected magnetic resonance

DOEpatents

Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

2009-10-27

The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

Diabetes screening by telecentric digital holographic microscopy.

PubMed

Doblas, A; Roche, E; Ampudia-Blasco, F J; Martínez-Corral, M; Saavedra, G; Garcia-Sucerquia, J

2016-03-01

Diabetes is currently the world's fastest growing chronic disease and it is caused by deficient production of insulin by the endocrine pancreas or by abnormal insulin action in peripheral tissues. This results in persistent hyperglycaemia that over time may produce chronic diabetic complications. Determination of glycated haemoglobin level is currently the gold standard method to evaluate and control sustained hyperglycaemia in diabetic people. This measurement is currently made by high-performance liquid chromatography, which is a complex chemical process that requires the extraction of blood from the antecubital vein. To reduce the complexity of that measurement, we propose a fully-optical technique that is based in the fact that there are changes in the optical properties of erythrocytes due to the presence of glucose-derived adducts in the haemoglobin molecule. To evaluate these changes, we propose to perform quantitative phase maps of erythrocytes by using telecentric digital holographic microscopy. Our experiments show that telecentric digital holographic microscopy allows detecting, almost in real time and from a single drop of blood, significant differences between erythrocytes of diabetic patients and healthy patients. Besides, our phase measurements are well correlated with the values of glycated haemoglobin and the blood glucose values. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Sub-nanosecond time-resolved near-field scanning magneto-optical microscope.

PubMed

Rudge, J; Xu, H; Kolthammer, J; Hong, Y K; Choi, B C

2015-02-01

We report on the development of a new magnetic microscope, time-resolved near-field scanning magneto-optical microscope, which combines a near-field scanning optical microscope and magneto-optical contrast. By taking advantage of the high temporal resolution of time-resolved Kerr microscope and the sub-wavelength spatial resolution of a near-field microscope, we achieved a temporal resolution of ∼50 ps and a spatial resolution of <100 nm. In order to demonstrate the spatiotemporal magnetic imaging capability of this microscope, the magnetic field pulse induced gyrotropic vortex dynamics occurring in 1 μm diameter, 20 nm thick CoFeB circular disks has been investigated. The microscope provides sub-wavelength resolution magnetic images of the gyrotropic motion of the vortex core at a resonance frequency of ∼240 MHz.

The Scanning Optical Microscope.

ERIC Educational Resources Information Center

Sheppard, C. J. R.

1978-01-01

Describes the principle of the scanning optical microscope and explains its advantages over the conventional microscope in the improvement of resolution and contrast, as well as the possibility of producing a picture from optical harmonies generated within the specimen.

Digital photography for the light microscope: results with a gated, video-rate CCD camera and NIH-image software.

PubMed

Shaw, S L; Salmon, E D; Quatrano, R S

1995-12-01

In this report, we describe a relatively inexpensive method for acquiring, storing and processing light microscope images that combines the advantages of video technology with the powerful medium now termed digital photography. Digital photography refers to the recording of images as digital files that are stored, manipulated and displayed using a computer. This report details the use of a gated video-rate charge-coupled device (CCD) camera and a frame grabber board for capturing 256 gray-level digital images from the light microscope. This camera gives high-resolution bright-field, phase contrast and differential interference contrast (DIC) images but, also, with gated on-chip integration, has the capability to record low-light level fluorescent images. The basic components of the digital photography system are described, and examples are presented of fluorescence and bright-field micrographs. Digital processing of images to remove noise, to enhance contrast and to prepare figures for printing is discussed.

Sub-micron materials characterization using near-field optics

NASA Astrophysics Data System (ADS)

Blodgett, David Wesley

1998-12-01

High-resolution sub-surface materials characterization and inspection are critical in the microelectronics and thin films industries. To this end, a technique is described that couples the bulk property measurement capabilities of high-frequency ultrasound with the high-resolution surface imaging capabilities of the near-field optical microscope. Sensing bulk microstructure variations in the material, such as grain boundaries, requires a detection footprint smaller than the variation itself. The near-field optical microscope, with the ability to exceed the diffraction limit in optical resolution, meets this requirement. Two apertureless near-field optical microscopes, on-axis and off-axis illumination, have been designed and built. Near-field and far-field approach curves for both microscopes are presented. The sensitivity of the near-field approach curve was 8.3 muV/nm. Resolution studies for the near-field microscope indicate optical resolutions on the order of 50 nm, which exceeds the diffraction limit. The near-field microscope has been adapted to detect both contact-transducer-generated and laser-generated ultrasound. The successful detection of high-frequency ultrasound with the near-field optical microscope demonstrates the potential of this technique.

Time-gated FLIM microscope for corneal metabolic imaging

NASA Astrophysics Data System (ADS)

Silva, Susana F.; Batista, Ana; Domingues, José Paulo; Quadrado, Maria João.; Morgado, António Miguel

2016-03-01

Detecting corneal cells metabolic alterations may prove a valuable tool in the early diagnosis of corneal diseases. Nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) are autofluorescent metabolic co-factors that allow the assessment of metabolic changes through non-invasive optical methods. These co-factors exhibit double-exponential fluorescence decays, with well-separated short and lifetime components, which are related to their protein-bound and free-states. Corneal metabolism can be assessed by measuring the relative contributions of these two components. For that purpose, we have developed a wide-field time-gated fluorescence lifetime microscope based on structured illumination and one-photon excitation to record FAD lifetime images from corneas. NADH imaging was not considered as its UV excitation peak is regarded as not safe for in vivo measurements. The microscope relies on a pulsed blue diode laser (λ=443 nm) as excitation source, an ultra-high speed gated image intensifier coupled to a CCD camera to acquire fluorescence signals and a Digital Micromirror Device (DMD) to implement the Structured Illumination technique. The system has a lateral resolution better than 2.4 μm, a field of view of 160 per 120 μm and an optical sectioning of 6.91 +/- 0.45 μm when used with a 40x, 0.75 NA, Water Immersion Objective. With this setup we were able to measure FAD contributions from ex-vivo chicken corneas collected from a local slaughterhouse..

Development of an add-on kit for scanning confocal microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Guo, Kaikai; Zheng, Guoan

2017-03-01

Scanning confocal microscopy is a standard choice for many fluorescence imaging applications in basic biomedical research. It is able to produce optically sectioned images and provide acquisition versatility to address many samples and application demands. However, scanning a focused point across the specimen limits the speed of image acquisition. As a result, scanning confocal microscope only works well with stationary samples. Researchers have performed parallel confocal scanning using digital-micromirror-device (DMD), which was used to project a scanning multi-point pattern across the sample. The DMD based parallel confocal systems increase the imaging speed while maintaining the optical sectioning ability. In this paper, we report the development of an add-on kit for high-speed and low-cost confocal microscopy. By adapting this add-on kit to an existing regular microscope, one can convert it into a confocal microscope without significant hardware modifications. Compared with current DMD-based implementations, the reported approach is able to recover multiple layers along the z axis simultaneously. It may find applications in wafer inspection and 3D metrology of semiconductor circuit. The dissemination of the proposed add-on kit under $1000 budget could also lead to new types of experimental designs for biological research labs, e.g., cytology analysis in cell culture experiments, genetic studies on multicellular organisms, pharmaceutical drug profiling, RNA interference studies, investigation of microbial communities in environmental systems, and etc.

Microscope collision protection apparatus

DOEpatents

DeNure, Charles R.

2001-10-23

A microscope collision protection apparatus for a remote control microscope which protects the optical and associated components from damage in the event of an uncontrolled collision with a specimen, regardless of the specimen size or shape. In a preferred embodiment, the apparatus includes a counterbalanced slide for mounting the microscope's optical components. This slide replaces the rigid mounts on conventional upright microscopes with a precision ball bearing slide. As the specimen contacts an optical component, the contacting force will move the slide and the optical components mounted thereon. This movement will protect the optical and associated components from damage as the movement causes a limit switch to be actuated, thereby stopping all motors responsible for the collision.

Examination of a high resolution laser optical plankton counter and FlowCAM for measuring plankton concentration and size

NASA Astrophysics Data System (ADS)

Kydd, Jocelyn; Rajakaruna, Harshana; Briski, Elizabeta; Bailey, Sarah

2018-03-01

Many commercial ships will soon begin to use treatment systems to manage their ballast water and reduce the global transfer of harmful aquatic organisms and pathogens in accordance with upcoming International Maritime Organization regulations. As a result, rapid and accurate automated methods will be needed to monitoring compliance of ships' ballast water. We examined two automated particle counters for monitoring organisms ≥ 50 μm in minimum dimension: a High Resolution Laser Optical Plankton Counter (HR-LOPC), and a Flow Cytometer with digital imaging Microscope (FlowCAM), in comparison to traditional (manual) microscopy considering plankton concentration, size frequency distributions and particle size measurements. The automated tools tended to underestimate particle concentration compared to standard microscopy, but gave similar results in terms of relative abundance of individual taxa. For most taxa, particle size measurements generated by FlowCAM ABD (Area Based Diameter) were more similar to microscope measurements than were those by FlowCAM ESD (Equivalent Spherical Diameter), though there was a mismatch in size estimates for some organisms between the FlowCAM ABD and microscope due to orientation and complex morphology. When a single problematic taxon is very abundant, the resulting size frequency distribution curves can become skewed, as was observed with Asterionella in this study. In particular, special consideration is needed when utilizing automated tools to analyse samples containing colonial species. Re-analysis of the size frequency distributions with the removal of Asterionella from FlowCAM and microscope data resulted in more similar curves across methods with FlowCAM ABD having the best fit compared to the microscope, although microscope concentration estimates were still significantly higher than estimates from the other methods. The results of our study indicate that both automated tools can generate frequency distributions of particles that might be particularly useful if correction factors can be developed for known differences in well-studied aquatic ecosystems.

Optical second harmonic images of 90 deg domain structure in BaTiO3 and periodically inverted antiparallel domains in LiTaO3

NASA Astrophysics Data System (ADS)

Uesu, Y.; Kurimura, S.; Yamamoto, Y.

1995-04-01

Applied is a microscope to observations of 90 deg ferroelectric domain structure in BaTiO3 and inverted periodically are ferroelectric domains in LiTaO3. It is founded that the second harmonic generation microscope gives information which cannot be obtained by ordinary optical microscopes. The developed nonlinear optical microscope builds two dimensional second harmonic image of a specimen with inhomogenous distribution of d(sub ijk) and applied the microscope to observations of inhomogeneity in some nonlinear-optical organic microcrystals.

Programmable Illumination and High-Speed, Multi-Wavelength, Confocal Microscopy Using a Digital Micromirror

PubMed Central

Martial, Franck P.; Hartell, Nicholas A.

2012-01-01

Confocal microscopy is routinely used for high-resolution fluorescence imaging of biological specimens. Most standard confocal systems scan a laser across a specimen and collect emitted light passing through a single pinhole to produce an optical section of the sample. Sequential scanning on a point-by-point basis limits the speed of image acquisition and even the fastest commercial instruments struggle to resolve the temporal dynamics of rapid cellular events such as calcium signals. Various approaches have been introduced that increase the speed of confocal imaging. Nipkov disk microscopes, for example, use arrays of pinholes or slits on a spinning disk to achieve parallel scanning which significantly increases the speed of acquisition. Here we report the development of a microscope module that utilises a digital micromirror device as a spatial light modulator to provide programmable confocal optical sectioning with a single camera, at high spatial and axial resolution at speeds limited by the frame rate of the camera. The digital micromirror acts as a solid state Nipkov disk but with the added ability to change the pinholes size and separation and to control the light intensity on a mirror-by-mirror basis. The use of an arrangement of concave and convex mirrors in the emission pathway instead of lenses overcomes the astigmatism inherent with DMD devices, increases light collection efficiency and ensures image collection is achromatic so that images are perfectly aligned at different wavelengths. Combined with non-laser light sources, this allows low cost, high-speed, multi-wavelength image acquisition without the need for complex wavelength-dependent image alignment. The micromirror can also be used for programmable illumination allowing spatially defined photoactivation of fluorescent proteins. We demonstrate the use of this system for high-speed calcium imaging using both a single wavelength calcium indicator and a genetically encoded, ratiometric, calcium sensor. PMID:22937130

Programmable illumination and high-speed, multi-wavelength, confocal microscopy using a digital micromirror.

PubMed

Martial, Franck P; Hartell, Nicholas A

2012-01-01

Confocal microscopy is routinely used for high-resolution fluorescence imaging of biological specimens. Most standard confocal systems scan a laser across a specimen and collect emitted light passing through a single pinhole to produce an optical section of the sample. Sequential scanning on a point-by-point basis limits the speed of image acquisition and even the fastest commercial instruments struggle to resolve the temporal dynamics of rapid cellular events such as calcium signals. Various approaches have been introduced that increase the speed of confocal imaging. Nipkov disk microscopes, for example, use arrays of pinholes or slits on a spinning disk to achieve parallel scanning which significantly increases the speed of acquisition. Here we report the development of a microscope module that utilises a digital micromirror device as a spatial light modulator to provide programmable confocal optical sectioning with a single camera, at high spatial and axial resolution at speeds limited by the frame rate of the camera. The digital micromirror acts as a solid state Nipkov disk but with the added ability to change the pinholes size and separation and to control the light intensity on a mirror-by-mirror basis. The use of an arrangement of concave and convex mirrors in the emission pathway instead of lenses overcomes the astigmatism inherent with DMD devices, increases light collection efficiency and ensures image collection is achromatic so that images are perfectly aligned at different wavelengths. Combined with non-laser light sources, this allows low cost, high-speed, multi-wavelength image acquisition without the need for complex wavelength-dependent image alignment. The micromirror can also be used for programmable illumination allowing spatially defined photoactivation of fluorescent proteins. We demonstrate the use of this system for high-speed calcium imaging using both a single wavelength calcium indicator and a genetically encoded, ratiometric, calcium sensor.

Autoradiographic method for quantitation of deposition and distribution of radiocalcium in bone

PubMed Central

Lawrence Riggs, B; Bassingthwaighte, James B.; Jowsey, Jenifer; Peter Pequegnat, E

2010-01-01

A method is described for quantitating autoradiographs of bone-seeking isotopes in microscopic sections of bone. Autoradiographs of bone sections containing 45Ca and internal calibration standards are automatically scanned with a microdensitometer. The digitized optical density output is stored on magnetic tape and is converted by computer to equivalent activity of 45Ca per gram of bone. The computer determines the total 45Ca uptake in the bone section and, on the basis of optical density and anatomic position, quantitatively divides the uptake into 4 components, each representing a separate physiologic process (bone formation, secondary mineralization, diffuse long-term exchange, and surface short-term exchange). The method is also applicable for quantitative analysis of microradiographs of bone sections for mineral content and density. PMID:5416906

Adaptive optics plug-and-play setup for high-resolution microscopes with multi-actuator adaptive lens

NASA Astrophysics Data System (ADS)

Quintavalla, M.; Pozzi, P.; Verhaegen, Michelle; Bijlsma, Hielke; Verstraete, Hans; Bonora, S.

2018-02-01

Adaptive Optics (AO) has revealed as a very promising technique for high-resolution microscopy, where the presence of optical aberrations can easily compromise the image quality. Typical AO systems however, are almost impossible to implement on commercial microscopes. We propose a simple approach by using a Multi-actuator Adaptive Lens (MAL) that can be inserted right after the objective and works in conjunction with an image optimization software allowing for a wavefront sensorless correction. We presented the results obtained on several commercial microscopes among which a confocal microscope, a fluorescence microscope, a light sheet microscope and a multiphoton microscope.

Development of an ultrasound microscope combined with optical microscope for multiparametric characterization of a single cell.

PubMed

Arakawa, Mototaka; Shikama, Joe; Yoshida, Koki; Nagaoka, Ryo; Kobayashi, Kazuto; Saijo, Yoshifumi

2015-09-01

Biomechanics of the cell has been gathering much attention because it affects the pathological status in atherosclerosis and cancer. In the present study, an ultrasound microscope system combined with optical microscope for characterization of a single cell with multiple ultrasound parameters was developed. The central frequency of the transducer was 375 MHz and the scan area was 80 × 80 μm with up to 200 × 200 sampling points. An inverted optical microscope was incorporated in the design of the system, allowing for simultaneous optical observations of cultured cells. Two-dimensional mapping of multiple ultrasound parameters, such as sound speed, attenuation, and acoustic impedance, as well as the thickness, density, and bulk modulus of specimen/cell under investigation, etc., was realized by the system. Sound speed and thickness of a 3T3-L1 fibroblast cell were successfully obtained by the system. The ultrasound microscope system combined with optical microscope further enhances our understanding of cellular biomechanics.

Optical detection of Trypanosoma cruzi in blood samples for diagnosis purpose

NASA Astrophysics Data System (ADS)

Alanis, Elvio; Romero, Graciela; Alvarez, Liliana; Martinez, Carlos C.; Basombrio, Miguel A.

2004-10-01

An optical method for detection of Trypanosoma Cruzi (T. cruzi) parasites in blood samples of mice infected with Chagas disease is presented. The method is intended for use in human blood, for diagnosis purposes. A thin layer of blood infected by T. cruzi parasites, in small concentrations, is examined in an interferometric microscope in which the images of the vision field are taken by a CCD camera and temporarily stored in the memory of a host computer. The whole sample is scanned displacing the microscope plate by means of step motors driven by the computer. Several consecutive images of the same field are taken and digitally processed by means of image temporal differentiation in order to detect if a parasite is eventually present in the field. Each field of view is processed in the same fashion, until the full area of the sample is covered or until a parasite is detected, in which case an acoustical warning is activated and the corresponding image is displayed permitting the technician to corroborate the result visually. A discussion of the reliability of the method as well as a comparison with other well established techniques are presented.

Shear Stress Sensing with Elastic Microfence Structures

NASA Technical Reports Server (NTRS)

Cisotto, Alexxandra; Palmieri, Frank L.; Saini, Aditya; Lin, Yi; Thurman, Christopher S; Kim, Jinwook; Kim, Taeyang; Connell, John W.; Zhu, Yong; Gopalarathnam, Ashok;

Potential Engineering of Fermi-Hubbard Systems using a Quantum Gas Microscope

NASA Astrophysics Data System (ADS)

Ji, Geoffrey; Mazurenko, Anton; Chiu, Christie; Parsons, Maxwell; Kanász-Nagy, Márton; Schmidt, Richard; Grusdt, Fabian; Demler, Eugene; Greif, Daniel; Greiner, Markus

2017-04-01

Arbitrary control of optical potentials has emerged as an important tool in manipulating ultracold atomic systems, especially when combined with the single-site addressing afforded by quantum gas microscopy. Already, experiments have used digital micromirror devices (DMDs) to initialize and control ultracold atomic systems in the context of studying quantum walks, quantum thermalization, and many-body localization. Here, we report on progress in using a DMD located in the image plane of a quantum gas microscope to explore static and dynamic properties of a 2D Fermi-Hubbard system. By projecting a large, ring-shaped anti-confining potential, we demonstrate entropy redistribution and controlled doping of the system. Moreover, we use the DMD to prepare localized holes, which upon release interact with and disrupt the surrounding spin environment. These techniques pave the way for controlled investigations of dynamics in the low-temperature phases of the Hubbard model.

Mechanical properties of the rust layer induced by impressed current method in reinforced mortar

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

Care, S.; Nguyen, Q.T.; L'Hostis, V.

This paper describes the mechanical effects of rust layer formed in reinforced mortar through accelerated tests of corrosion. The morphological and physico-chemical properties (composition, structures) of the corrosion system were characterized at different stages by using optical microscope and scanning electron microscope coupled with energy dispersive spectroscopy. The corrosion pattern was mainly characterized by a rust layer confined at the interface between the steel and the mortar. Expansion coefficient of rust products was determined from the rust thickness and the Faraday's law. Furthermore, in order to understand the mechanical effects of corrosion on the damage of mortar, displacement field measurementsmore » were obtained by using digital image correlation. An analytical model (hollow cylinder subjected to inner and outer pressures) was used with a set of experimental data to deduce the time of cracking and the order of magnitude of the mechanical properties of the rust layer.« less

Research in Optical Symbolic Tasks

DTIC Science & Technology

1989-11-29

November 1989. Specifically, we have concentrated on the following topics: complexity studies for optical neural and digital systems, architecture and...1989. Specifically, we hav, concentrated on the following topics: complexity studies for optical neural and digital systems, architecture and models for...Digital Systems 1.1 Digital Optical Parallel System Complexity Our study of digital optical system complexity has included a comparison of optical and

Digital pathology for the primary diagnosis of breast histopathological specimens: an innovative validation and concordance study on digital pathology validation and training.

PubMed

Williams, Bethany Jill; Hanby, Andrew; Millican-Slater, Rebecca; Nijhawan, Anju; Verghese, Eldo; Treanor, Darren

2018-03-01

To train and individually validate a group of breast pathologists in specialty-specific digital primary diagnosis by using a novel protocol endorsed by the Royal College of Pathologists' new guideline for digital pathology. The protocol allows early exposure to live digital reporting, in a risk-mitigated environment, and focuses on patient safety and professional development. Three specialty breast pathologists completed training in the use of a digital microscopy system, and were exposed to a training set of 20 challenging cases, designed to help them identify personal digital diagnostic pitfalls. Following this, the three pathologists viewed a total of 694 live, entire breast cases. All primary diagnoses were made on digital slides, with immediate glass slide review and reconciliation before final case sign-out. There was complete clinical concordance between the glass and digital impression of the case in 98.8% of cases. Only 1.2% of cases had a clinically significant difference in diagnosis/prognosis on glass and digital slide reads. All pathologists elected to continue using the digital microscope as the standard for breast histopathology specimens, with deferral to glass for a limited number of clinical/histological scenarios as a safety net. Individual training and validation for digital primary diagnosis allows pathologists to develop competence and confidence in their digital diagnostic skills, and aids safe and responsible transition from the light microscope to the digital microscope. © 2017 John Wiley & Sons Ltd.

Optical analog-to-digital converter

DOEpatents

Vawter, G Allen [Corrales, NM; Raring, James [Goleta, CA; Skogen, Erik J [Albuquerque, NM

2009-07-21

An optical analog-to-digital converter (ADC) is disclosed which converts an input optical analog signal to an output optical digital signal at a sampling rate defined by a sampling optical signal. Each bit of the digital representation is separately determined using an optical waveguide interferometer and an optical thresholding element. The interferometer uses the optical analog signal and the sampling optical signal to generate a sinusoidally-varying output signal using cross-phase-modulation (XPM) or a photocurrent generated from the optical analog signal. The sinusoidally-varying output signal is then digitized by the thresholding element, which includes a saturable absorber or at least one semiconductor optical amplifier, to form the optical digital signal which can be output either in parallel or serially.

Limits of agreement between the optical pachymeter and a noncontact specular microscope.

PubMed

Ogbuehi, Kelechi C; Almubrad, Turki M

2005-07-01

To determine the limits of agreement between central corneal thickness (CCT) measurements made with the slit lamp-attached optical pachymeter and the SP2000P noncontact specular microscope. Triplicate readings for CCT were obtained for each of 130 (right) eyes of 130 patients, using the slit lamp-attached optical pachymeter and then the SP2000P noncontact specular microscope. The average CCT measured by each method was compared. Subsequently, the mean difference between both sets of measurements was assessed, and the 95% confidence interval (limits of agreement) between both techniques was determined. The mean +/- SD CCT measured by the optical pachymeter was 543 +/- 34 microm and 532 +/- 34 microm for the specular microscope. We found a statistically significant (P < 0.001) mean bias of 10 mum between CCT values measured with both types of equipment, with the optical pachymeter returning the higher values. The coefficient of variation was 6.3% for the optical pachymeter and 6.4% for the specular microscope. The right eye CCT measurements made by the optical pachymeter are, on average, 10 mum thicker than those made with the SP2000P specular microscope, which suggests that both pieces of equipment cannot be used interchangeably to monitor CCT changes in patients. Excluding left eye measurements, the reliability of the optical pachymeter is identical to that of the noncontact specular microscope.

Imaging Schwarzschild multilayer X-ray microscope

NASA Technical Reports Server (NTRS)

Hoover, Richard B.; Baker, Phillip C.; Shealy, David L.; Core, David B.; Walker, Arthur B. C., Jr.; Barbee, Troy W., Jr.; Kerstetter, Ted

1993-01-01

We have designed, analyzed, fabricated, and tested Schwarzschild multilayer X-ray microscopes. These instruments use flow-polished Zerodur mirror substrates which have been coated with multilayers optimized for maximum reflectivity at normal incidence at 135 A. They are being developed as prototypes for the Water Window Imaging X-Ray Microscope. Ultrasmooth mirror sets of hemlite grade sapphire have been fabricated and they are now being coated with multilayers to reflect soft X-rays at 38 A, within the biologically important 'water window'. In this paper, we discuss the fabrication of the microscope optics and structural components as well as the mounting of the optics and assembly of the microscopes. We also describe the optical alignment, interferometric and visible light testing of the microscopes, present interferometrically measured performance data, and provide the first results of optical imaging tests.

Capturing latent fingerprints from metallic painted surfaces using UV-VIS spectroscope

NASA Astrophysics Data System (ADS)

Makrushin, Andrey; Scheidat, Tobias; Vielhauer, Claus

2015-03-01

In digital crime scene forensics, contactless non-destructive detection and acquisition of latent fingerprints by means of optical devices such as a high-resolution digital camera, confocal microscope, or chromatic white-light sensor is the initial step prior to destructive chemical development. The applicability of an optical sensor to digitalize latent fingerprints primarily depends on reflection properties of a substrate. Metallic painted surfaces, for instance, pose a problem for conventional sensors which make use of visible light. Since metallic paint is a semi-transparent layer on top of the surface, visible light penetrates it and is reflected off of the metallic flakes randomly disposed in the paint. Fingerprint residues do not impede light beams making ridges invisible. Latent fingerprints can be revealed, however, using ultraviolet light which does not penetrate the paint. We apply a UV-VIS spectroscope that is capable of capturing images within the range from 163 to 844 nm using 2048 discrete levels. We empirically show that latent fingerprints left behind on metallic painted surfaces become clearly visible within the range from 205 to 385 nm. Our proposed streakiness score feature determining the proportion of a ridge-valley pattern in an image is applied for automatic assessment of a fingerprint's visibility and distinguishing between fingerprint and empty regions. The experiments are carried out with 100 fingerprint and 100 non-fingerprint samples.

Science 101: How Does an Electron Microscope Work?

ERIC Educational Resources Information Center

Robertson, Bill

2013-01-01

Contrary to popular opinion, electron microscopes are not used to look at electrons. They are used to look for structure in things that are too small to observe with an optical microscope, or to obtain images that are magnified much more than is obtainable with an optical microscope. To understand how electron microscopes work, it will help to go…

Adaptive optical microscope for brain imaging in vivo

NASA Astrophysics Data System (ADS)

Wang, Kai

2017-04-01

The optical heterogeneity of biological tissue imposes a major limitation to acquire detailed structural and functional information deep in the biological specimens using conventional microscopes. To restore optimal imaging performance, we developed an adaptive optical microscope based on direct wavefront sensing technique. This microscope can reliably measure and correct biological samples induced aberration. We demonstrated its performance and application in structural and functional brain imaging in various animal models, including fruit fly, zebrafish and mouse.

Digital image processing of bone - Problems and potentials

NASA Technical Reports Server (NTRS)

Morey, E. R.; Wronski, T. J.

1980-01-01

The development of a digital image processing system for bone histomorphometry and fluorescent marker monitoring is discussed. The system in question is capable of making measurements of UV or light microscope features on a video screen with either video or computer-generated images, and comprises a microscope, low-light-level video camera, video digitizer and display terminal, color monitor, and PDP 11/34 computer. Capabilities demonstrated in the analysis of an undecalcified rat tibia include the measurement of perimeter and total bone area, and the generation of microscope images, false color images, digitized images and contoured images for further analysis. Software development will be based on an existing software library, specifically the mini-VICAR system developed at JPL. It is noted that the potentials of the system in terms of speed and reliability far exceed any problems associated with hardware and software development.

PC-based control unit for a head-mounted operating microscope for augmented-reality visualization in surgical navigation

NASA Astrophysics Data System (ADS)

Figl, Michael; Birkfellner, Wolfgang; Watzinger, Franz; Wanschitz, Felix; Hummel, Johann; Hanel, Rudolf A.; Ewers, Rolf; Bergmann, Helmar

2002-05-01

Two main concepts of Head Mounted Displays (HMD) for augmented reality (AR) visualization exist, the optical and video-see through type. Several research groups have pursued both approaches for utilizing HMDs for computer aided surgery. While the hardware requirements for a video see through HMD to achieve acceptable time delay and frame rate seem to be enormous the clinical acceptance of such a device is doubtful from a practical point of view. Starting from previous work in displaying additional computer-generated graphics in operating microscopes, we have adapted a miniature head mounted operating microscope for AR by integrating two very small computer displays. To calibrate the projection parameters of this so called Varioscope AR we have used Tsai's Algorithm for camera calibration. Connection to a surgical navigation system was performed by defining an open interface to the control unit of the Varioscope AR. The control unit consists of a standard PC with a dual head graphics adapter to render and display the desired augmentation of the scene. We connected this control unit to a computer aided surgery (CAS) system by the TCP/IP interface. In this paper we present the control unit for the HMD and its software design. We tested two different optical tracking systems, the Flashpoint (Image Guided Technologies, Boulder, CO), which provided about 10 frames per second, and the Polaris (Northern Digital, Ontario, Canada) which provided at least 30 frames per second, both with a time delay of one frame.

Technological requirements of teleneuropathological systems.

PubMed

Szymaś, J

2000-01-01

Teleneuropathology is the practice of conducting remote neuropathological examinations with the use of telecommunication links. Because of a limited number of expert neuropathologists, some, especially smaller departments have the equipment to conduct the examination but do not have a specialist who would be able to evaluate material from the central nervous system. In case of teleneuropathology, a neuropathologist examines tissue fragments taken during an operation by means of a telemicroscope connected with the computer through a telecommunications network. It enables the neuropathologist to operate the microscope and camera remotely. Two basic systems exist for performing remote neuropathological examination: static and dynamic. Both have different needs in medical, computing and telecommunication aspect. Depending on the type of service the public telephone network, the integrated services digital network, or optical fibre should be used. Conditionally Internet can be used as a link for teleneuropathological system. However, for the newest developments in teleneuropathology such as teleconference and remote operation on robotized microscope only transmission over the integrated service digital network, which guarantees high speed of transmission gives a possibility to communicate. Because images are basic information element in teleneuropathological systems the high capacity of acquisition, processing, storing, transmission, and visualization equipment is necessary. The farther development of telecommunication as well as standardization of recording and transmission procedures of pictorial data is necessary.

Acquisition of a Surface Plasmon Resonance Imager, Digital Microscope, and Peristaltic Pumps for Defense-Based Research

DTIC Science & Technology

2016-05-05

SECURITY CLASSIFICATION OF: The goal of this proposal is to purchase the GWC Technologies, Inc. Horizontal Surface Plasmon Resonance Imaging (SPRi...Unlimited UU UU UU UU 05-05-2016 1-Feb-2014 31-Jan-2016 Final Report: Acquisition of a Surface Plasmon Resonance Imager, Digital Microscope, and...S) AND ADDRESS (ES) U.S. Army Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 Surface Plasmon Resonance Imager, Digital

Fostering Student Engagement with Digital Microscopic Images Using ThingLink, an Image Annotation Program

ERIC Educational Resources Information Center

Appasamy, Pierette

2018-01-01

The teaching of histology has changed dramatically with virtual microscopy. Fewer students of histology spend significant time viewing slides on a microscope and instead study images available in digital slide sets, generally accessible via the internet. However, students must still be able to correctly identify the defining characteristics of…

Computational imaging of sperm locomotion.

PubMed

Daloglu, Mustafa Ugur; Ozcan, Aydogan

2017-08-01

Not only essential for scientific research, but also in the analysis of male fertility and for animal husbandry, sperm tracking and characterization techniques have been greatly benefiting from computational imaging. Digital image sensors, in combination with optical microscopy tools and powerful computers, have enabled the use of advanced detection and tracking algorithms that automatically map sperm trajectories and calculate various motility parameters across large data sets. Computational techniques are driving the field even further, facilitating the development of unconventional sperm imaging and tracking methods that do not rely on standard optical microscopes and objective lenses, which limit the field of view and volume of the semen sample that can be imaged. As an example, a holographic on-chip sperm imaging platform, only composed of a light-emitting diode and an opto-electronic image sensor, has emerged as a high-throughput, low-cost and portable alternative to lens-based traditional sperm imaging and tracking methods. In this approach, the sample is placed very close to the image sensor chip, which captures lensfree holograms generated by the interference of the background illumination with the light scattered from sperm cells. These holographic patterns are then digitally processed to extract both the amplitude and phase information of the spermatozoa, effectively replacing the microscope objective lens with computation. This platform has further enabled high-throughput 3D imaging of spermatozoa with submicron 3D positioning accuracy in large sample volumes, revealing various rare locomotion patterns. We believe that computational chip-scale sperm imaging and 3D tracking techniques will find numerous opportunities in both sperm related research and commercial applications. © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Breast cancer histopathology image analysis: a review.

PubMed

Veta, Mitko; Pluim, Josien P W; van Diest, Paul J; Viergever, Max A

2014-05-01

This paper presents an overview of methods that have been proposed for the analysis of breast cancer histopathology images. This research area has become particularly relevant with the advent of whole slide imaging (WSI) scanners, which can perform cost-effective and high-throughput histopathology slide digitization, and which aim at replacing the optical microscope as the primary tool used by pathologist. Breast cancer is the most prevalent form of cancers among women, and image analysis methods that target this disease have a huge potential to reduce the workload in a typical pathology lab and to improve the quality of the interpretation. This paper is meant as an introduction for nonexperts. It starts with an overview of the tissue preparation, staining and slide digitization processes followed by a discussion of the different image processing techniques and applications, ranging from analysis of tissue staining to computer-aided diagnosis, and prognosis of breast cancer patients.

Digital photocontrol of the network of live excitable cells

NASA Astrophysics Data System (ADS)

Erofeev, I. S.; Magome, N.; Agladze, K. I.

2011-11-01

Recent development of tissue engineering techniques allows creating and maintaining almost indefinitely networks of excitable cells with desired architecture. We coupled the network of live excitable cardiac cells with a common computer by sensitizing them to light, projecting a light pattern on the layer of cells, and monitoring excitation with the aid of fluorescent probes (optical mapping). As a sensitizing substance we used azobenzene trimethylammonium bromide (AzoTAB). This substance undergoes cis-trans-photoisomerization and trans-isomer of AzoTAB inhibits excitation in the cardiac cells, while cis-isomer does not. AzoTAB-mediated sensitization allows, thus, reversible and dynamic control of the excitation waves through the entire cardiomyocyte network either uniformly, or in a preferred spatial pattern. Technically, it was achieved by coupling a common digital projector with a macroview microscope and using computer graphic software for creating the projected pattern of conducting pathways. This approach allows real time interactive photocontrol of the heart tissue.

New design of a cryostat-mounted scanning near-field optical microscope for single molecule spectroscopy

NASA Astrophysics Data System (ADS)

Durand, Yannig; Woehl, Jörg C.; Viellerobe, Bertrand; Göhde, Wolfgang; Orrit, Michel

1999-02-01

Due to the weakness of the fluorescence signal from a single fluorophore, a scanning near-field optical microscope for single molecule spectroscopy requires a very efficient setup for the collection and detection of emitted photons. We have developed a home-built microscope for operation in a l-He cryostat which uses a solid parabolic mirror in order to optimize the fluorescence collection efficiency. This microscope works with Al-coated, tapered optical fibers in illumination mode. The tip-sample separation is probed by an optical shear-force detection. First results demonstrate the capability of the microscope to image single molecules and achieve a topographical resolution of a few nanometers vertically and better than 50 nm laterally.

A stereo-compound hybrid microscope for combined intracellular and optical recording of invertebrate neural network activity.

PubMed

Frost, William N; Wang, Jean; Brandon, Christopher J

2007-05-15

Optical recording studies of invertebrate neural networks with voltage-sensitive dyes seldom employ conventional intracellular electrodes. This may in part be due to the traditional reliance on compound microscopes for such work. While such microscopes have high light-gathering power, they do not provide depth of field, making working with sharp electrodes difficult. Here we describe a hybrid microscope design, with switchable compound and stereo objectives, that eases the use of conventional intracellular electrodes in optical recording experiments. We use it, in combination with a voltage-sensitive dye and photodiode array, to identify neurons participating in the swim motor program of the marine mollusk Tritonia. This microscope design should be applicable to optical recording studies in many preparations.

Microscopy with spatial filtering for sorting particles and monitoring subcellular morphology

NASA Astrophysics Data System (ADS)

Zheng, Jing-Yi; Qian, Zhen; Pasternack, Robert M.; Boustany, Nada N.

2009-02-01

Optical scatter imaging (OSI) was developed to non-invasively track real-time changes in particle morphology with submicron sensitivity in situ without exogenous labeling, cell fixing, or organelle isolation. For spherical particles, the intensity ratio of wide-to-narrow angle scatter (OSIR, Optical Scatter Image Ratio) was shown to decrease monotonically with diameter and agree with Mie theory. In living cells, we recently reported this technique is able to detect mitochondrial morphological alterations, which were mediated by the Bcl-xL transmembrane domain, and could not be observed by fluorescence or differential interference contrast images. Here we further extend the ability of morphology assessment by adopting a digital micromirror device (DMD) for Fourier filtering. When placed in the Fourier plane the DMD can be used to select scattering intensities at desired combination of scattering angles. We designed an optical filter bank consisting of Gabor-like filters with various scales and rotations based on Gabor filters, which have been widely used for localization of spatial and frequency information in digital images and texture analysis. Using a model system consisting of mixtures of polystyrene spheres and bacteria, we show how this system can be used to sort particles on a microscopic slide based on their size, orientation and aspect ratio. We are currently applying this technique to characterize the morphology of subcellular organelles to help understand fundamental biological processes.

Multiplexed phase-space imaging for 3D fluorescence microscopy.

PubMed

Liu, Hsiou-Yuan; Zhong, Jingshan; Waller, Laura

2017-06-26

Optical phase-space functions describe spatial and angular information simultaneously; examples of optical phase-space functions include light fields in ray optics and Wigner functions in wave optics. Measurement of phase-space enables digital refocusing, aberration removal and 3D reconstruction. High-resolution capture of 4D phase-space datasets is, however, challenging. Previous scanning approaches are slow, light inefficient and do not achieve diffraction-limited resolution. Here, we propose a multiplexed method that solves these problems. We use a spatial light modulator (SLM) in the pupil plane of a microscope in order to sequentially pattern multiplexed coded apertures while capturing images in real space. Then, we reconstruct the 3D fluorescence distribution of our sample by solving an inverse problem via regularized least squares with a proximal accelerated gradient descent solver. We experimentally reconstruct a 101 Megavoxel 3D volume (1010×510×500µm with NA 0.4), demonstrating improved acquisition time, light throughput and resolution compared to scanning aperture methods. Our flexible patterning scheme further allows sparsity in the sample to be exploited for reduced data capture.

Development of a networked four-million-pixel pathological and radiological digital image presentation system and its application to medical conferences

NASA Astrophysics Data System (ADS)

Sakano, Toshikazu; Furukawa, Isao; Okumura, Akira; Yamaguchi, Takahiro; Fujii, Tetsuro; Ono, Sadayasu; Suzuki, Junji; Matsuya, Shoji; Ishihara, Teruo

2001-08-01

The wide spread of digital technology in the medical field has led to a demand for the high-quality, high-speed, and user-friendly digital image presentation system in the daily medical conferences. To fulfill this demand, we developed a presentation system for radiological and pathological images. It is composed of a super-high-definition (SHD) imaging system, a radiological image database (R-DB), a pathological image database (P-DB), and the network interconnecting these three. The R-DB consists of a 270GB RAID, a database server workstation, and a film digitizer. The P-DB includes an optical microscope, a four-million-pixel digital camera, a 90GB RAID, and a database server workstation. A 100Mbps Ethernet LAN interconnects all the sub-systems. The Web-based system operation software was developed for easy operation. We installed the whole system in NTT East Kanto Hospital to evaluate it in the weekly case conferences. The SHD system could display digital full-color images of 2048 x 2048 pixels on a 28-inch CRT monitor. The doctors evaluated the image quality and size, and found them applicable to the actual medical diagnosis. They also appreciated short image switching time that contributed to smooth presentation. Thus, we confirmed that its characteristics met the requirements.

X-ray laser microscope apparatus

DOEpatents

Suckewer, Szymon; DiCicco, Darrell S.; Hirschberg, Joseph G.; Meixler, Lewis D.; Sathre, Robert; Skinner, Charles H.

1990-01-01

A microscope consisting of an x-ray contact microscope and an optical microscope. The optical, phase contrast, microscope is used to align a target with respect to a source of soft x-rays. The source of soft x-rays preferably comprises an x-ray laser but could comprise a synchrotron or other pulse source of x-rays. Transparent resist material is used to support the target. The optical microscope is located on the opposite side of the transparent resist material from the target and is employed to align the target with respect to the anticipated soft x-ray laser beam. After alignment with the use of the optical microscope, the target is exposed to the soft x-ray laser beam. The x-ray sensitive transparent resist material whose chemical bonds are altered by the x-ray beam passing through the target mater GOVERNMENT LICENSE RIGHTS This invention was made with government support under Contract No. De-FG02-86ER13609 awarded by the Department of Energy. The Government has certain rights in this invention.

Evaluation of a miniature microscope objective designed for fluorescence array microscopy detection of Mycobacterium tuberculosis.

PubMed

McCall, Brian; Olsen, Randall J; Nelles, Nicole J; Williams, Dawn L; Jackson, Kevin; Richards-Kortum, Rebecca; Graviss, Edward A; Tkaczyk, Tomasz S

2014-03-01

A prototype miniature objective that was designed for a point-of-care diagnostic array microscope for detection of Mycobacterium tuberculosis and previously fabricated and presented in a proof of concept is evaluated for its effectiveness in detecting acid-fast bacteria. To evaluate the ability of the microscope to resolve submicron features and details in the image of acid-fast microorganisms stained with a fluorescent dye, and to evaluate the accuracy of clinical diagnoses made with digital images acquired with the objective. The lens prescription data for the microscope design are presented. A test platform is built by combining parts of a standard microscope, a prototype objective, and a digital single-lens reflex camera. Counts of acid-fast bacteria made with the prototype objective are compared to counts obtained with a standard microscope over matched fields of view. Two sets of 20 smears, positive and negative, are diagnosed by 2 pathologists as sputum smear positive or sputum smear negative, using both a standard clinical microscope and the prototype objective under evaluation. The results are compared to a reference diagnosis of the same sample. More bacteria are counted in matched fields of view in digital images taken with the prototype objective than with the standard clinical microscope. All diagnostic results are found to be highly concordant. An array microscope built with this miniature lens design will be able to detect M tuberculosis with high sensitivity and specificity.

Smart-phone based computational microscopy using multi-frame contact imaging on a fiber-optic array.

PubMed

Navruz, Isa; Coskun, Ahmet F; Wong, Justin; Mohammad, Saqib; Tseng, Derek; Nagi, Richie; Phillips, Stephen; Ozcan, Aydogan

2013-10-21

We demonstrate a cellphone based contact microscopy platform, termed Contact Scope, which can image highly dense or connected samples in transmission mode. Weighing approximately 76 grams, this portable and compact microscope is installed on the existing camera unit of a cellphone using an opto-mechanical add-on, where planar samples of interest are placed in contact with the top facet of a tapered fiber-optic array. This glass-based tapered fiber array has ~9 fold higher density of fiber optic cables on its top facet compared to the bottom one and is illuminated by an incoherent light source, e.g., a simple light-emitting-diode (LED). The transmitted light pattern through the object is then sampled by this array of fiber optic cables, delivering a transmission image of the sample onto the other side of the taper, with ~3× magnification in each direction. This magnified image of the object, located at the bottom facet of the fiber array, is then projected onto the CMOS image sensor of the cellphone using two lenses. While keeping the sample and the cellphone camera at a fixed position, the fiber-optic array is then manually rotated with discrete angular increments of e.g., 1-2 degrees. At each angular position of the fiber-optic array, contact images are captured using the cellphone camera, creating a sequence of transmission images for the same sample. These multi-frame images are digitally fused together based on a shift-and-add algorithm through a custom-developed Android application running on the smart-phone, providing the final microscopic image of the sample, visualized through the screen of the phone. This final computation step improves the resolution and also removes spatial artefacts that arise due to non-uniform sampling of the transmission intensity at the fiber optic array surface. We validated the performance of this cellphone based Contact Scope by imaging resolution test charts and blood smears.

Smart-phone based computational microscopy using multi-frame contact imaging on a fiber-optic array

PubMed Central

Navruz, Isa; Coskun, Ahmet F.; Wong, Justin; Mohammad, Saqib; Tseng, Derek; Nagi, Richie; Phillips, Stephen; Ozcan, Aydogan

2013-01-01

We demonstrate a cellphone based contact microscopy platform, termed Contact Scope, which can image highly dense or connected samples in transmission mode. Weighing approximately 76 grams, this portable and compact microscope is installed on the existing camera unit of a cellphone using an opto-mechanical add-on, where planar samples of interest are placed in contact with the top facet of a tapered fiber-optic array. This glass-based tapered fiber array has ∼9 fold higher density of fiber optic cables on its top facet compared to the bottom one and is illuminated by an incoherent light source, e.g., a simple light-emitting-diode (LED). The transmitted light pattern through the object is then sampled by this array of fiber optic cables, delivering a transmission image of the sample onto the other side of the taper, with ∼3× magnification in each direction. This magnified image of the object, located at the bottom facet of the fiber array, is then projected onto the CMOS image sensor of the cellphone using two lenses. While keeping the sample and the cellphone camera at a fixed position, the fiber-optic array is then manually rotated with discrete angular increments of e.g., 1-2 degrees. At each angular position of the fiber-optic array, contact images are captured using the cellphone camera, creating a sequence of transmission images for the same sample. These multi-frame images are digitally fused together based on a shift-and-add algorithm through a custom-developed Android application running on the smart-phone, providing the final microscopic image of the sample, visualized through the screen of the phone. This final computation step improves the resolution and also gets rid of spatial artefacts that arise due to non-uniform sampling of the transmission intensity at the fiber optic array surface. We validated the performance of this cellphone based Contact Scope by imaging resolution test charts and blood smears. PMID:23939637

Emerging Themes in Image Informatics and Molecular Analysis for Digital Pathology.

PubMed

Bhargava, Rohit; Madabhushi, Anant

2016-07-11

Pathology is essential for research in disease and development, as well as for clinical decision making. For more than 100 years, pathology practice has involved analyzing images of stained, thin tissue sections by a trained human using an optical microscope. Technological advances are now driving major changes in this paradigm toward digital pathology (DP). The digital transformation of pathology goes beyond recording, archiving, and retrieving images, providing new computational tools to inform better decision making for precision medicine. First, we discuss some emerging innovations in both computational image analytics and imaging instrumentation in DP. Second, we discuss molecular contrast in pathology. Molecular DP has traditionally been an extension of pathology with molecularly specific dyes. Label-free, spectroscopic images are rapidly emerging as another important information source, and we describe the benefits and potential of this evolution. Third, we describe multimodal DP, which is enabled by computational algorithms and combines the best characteristics of structural and molecular pathology. Finally, we provide examples of application areas in telepathology, education, and precision medicine. We conclude by discussing challenges and emerging opportunities in this area.

Emerging Themes in Image Informatics and Molecular Analysis for Digital Pathology

PubMed Central

Bhargava, Rohit; Madabhushi, Anant

2017-01-01

Pathology is essential for research in disease and development, as well as for clinical decision making. For more than 100 years, pathology practice has involved analyzing images of stained, thin tissue sections by a trained human using an optical microscope. Technological advances are now driving major changes in this paradigm toward digital pathology (DP). The digital transformation of pathology goes beyond recording, archiving, and retrieving images, providing new computational tools to inform better decision making for precision medicine. First, we discuss some emerging innovations in both computational image analytics and imaging instrumentation in DP. Second, we discuss molecular contrast in pathology. Molecular DP has traditionally been an extension of pathology with molecularly specific dyes. Label-free, spectroscopic images are rapidly emerging as another important information source, and we describe the benefits and potential of this evolution. Third, we describe multimodal DP, which is enabled by computational algorithms and combines the best characteristics of structural and molecular pathology. Finally, we provide examples of application areas in telepathology, education, and precision medicine. We conclude by discussing challenges and emerging opportunities in this area. PMID:27420575

A stereo-compound hybrid microscope for combined intracellular and optical recording of invertebrate neural network activity

PubMed Central

Frost, William N.; Wang, Jean; Brandon, Christopher J.

2007-01-01

Optical recording studies of invertebrate neural networks with voltage-sensitive dyes seldom employ conventional intracellular electrodes. This may in part be due to the traditional reliance on compound microscopes for such work. While such microscopes have high light-gathering power, they do not provide depth of field, making working with sharp electrodes difficult. Here we describe a hybrid microscope design, with switchable compound and stereo objectives, that eases the use of conventional intracellular electrodes in optical recording experiments. We use it, in combination with a voltage-sensitive dye and photodiode array, to identify neurons participating in the swim motor program of the marine mollusk Tritonia. This microscope design should be applicable to optical recording studies in many preparations. PMID:17306887

Teaching optics concepts through an approach that emphasizes the colors of nature

NASA Astrophysics Data System (ADS)

Pompea, Stephen M.; Carsten-Conner, Laura D.

2015-10-01

A wide variety of optics concepts can be taught using the overall perspective of "colors of nature" as a guiding and unifying theme. This approach is attractive and interesting with a wide appeal to children, nature enthusiasts, photographers, and artists. This approach also encourages a deep understanding of the natural world and the role of coloration in biology, remote sensing, the aurora, mineralogy, meteorology, in human-made objects, and astronomy, to name a few. Third, using this theme promotes a close look at optical phenomena at all size scales-from the microscopic (e.g. silica spheres in opals) to the mid-scale (the aurora), to the largest scale (astronomical phenomena such as gaseous emission nebula). Fourth, the natural and human-constructed world provides accessible and beautiful examples of complex phenomena such as interference, diffraction, atomic and molecular emissions, Rayleigh and Mie scattering, illumination engineering, and fluorescence. These areas can be explored successfully in the context of "colors of nature". Finally, using the "colors of nature" also promotes an understanding of technology, from flashlights to streetlights, from telescopes and binoculars, to spectrometers and digital cameras. For examples something as simple as how to set the white balance on a digital camera to get a realistic looking photograph can lead to a lengthy exploration of spectrally selective surfaces and their reflectance, the nature of different illumination sources, the meaning of color temperature, and role of calibration in a digital image. We have used this approach of teaching using the colors of nature as an organizing theme in our NSF-funded project "Project STEAM: Integrating Art with Science to Build Science Identities Among Girls" (colorsofnature.org).

Multimodal biophotonic workstation for live cell analysis.

PubMed

Esseling, Michael; Kemper, Björn; Antkowiak, Maciej; Stevenson, David J; Chaudet, Lionel; Neil, Mark A A; French, Paul W; von Bally, Gert; Dholakia, Kishan; Denz, Cornelia

2012-01-01

A reliable description and quantification of the complex physiology and reactions of living cells requires a multimodal analysis with various measurement techniques. We have investigated the integration of different techniques into a biophotonic workstation that can provide biological researchers with these capabilities. The combination of a micromanipulation tool with three different imaging principles is accomplished in a single inverted microscope which makes the results from all the techniques directly comparable. Chinese Hamster Ovary (CHO) cells were manipulated by optical tweezers while the feedback was directly analyzed by fluorescence lifetime imaging, digital holographic microscopy and dynamic phase-contrast microscopy. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Three-dimensional registration of intravascular optical coherence tomography and cryo-image volumes for microscopic-resolution validation.

PubMed

Prabhu, David; Mehanna, Emile; Gargesha, Madhusudhana; Brandt, Eric; Wen, Di; van Ditzhuijzen, Nienke S; Chamie, Daniel; Yamamoto, Hirosada; Fujino, Yusuke; Alian, Ali; Patel, Jaymin; Costa, Marco; Bezerra, Hiram G; Wilson, David L

2016-04-01

Evidence suggests high-resolution, high-contrast, [Formula: see text] intravascular optical coherence tomography (IVOCT) can distinguish plaque types, but further validation is needed, especially for automated plaque characterization. We developed experimental and three-dimensional (3-D) registration methods to provide validation of IVOCT pullback volumes using microscopic, color, and fluorescent cryo-image volumes with optional registered cryo-histology. A specialized registration method matched IVOCT pullback images acquired in the catheter reference frame to a true 3-D cryo-image volume. Briefly, an 11-parameter registration model including a polynomial virtual catheter was initialized within the cryo-image volume, and perpendicular images were extracted, mimicking IVOCT image acquisition. Virtual catheter parameters were optimized to maximize cryo and IVOCT lumen overlap. Multiple assessments suggested that the registration error was better than the [Formula: see text] spacing between IVOCT image frames. Tests on a digital synthetic phantom gave a registration error of only [Formula: see text] (signed distance). Visual assessment of randomly presented nearby frames suggested registration accuracy within 1 IVOCT frame interval ([Formula: see text]). This would eliminate potential misinterpretations confronted by the typical histological approaches to validation, with estimated 1-mm errors. The method can be used to create annotated datasets and automated plaque classification methods and can be extended to other intravascular imaging modalities.

The optics inside an automated single molecule array analyzer

NASA Astrophysics Data System (ADS)

McGuigan, William; Fournier, David R.; Watson, Gary W.; Walling, Les; Gigante, Bill; Duffy, David C.; Rissin, David M.; Kan, Cheuk W.; Meyer, Raymond E.; Piech, Tomasz; Fishburn, Matthew W.

2014-02-01

Quanterix and Stratec Biomedical have developed an instrument that enables the automated measurement of multiple proteins at concentration ~1000 times lower than existing immunoassays. The instrument is based on Quanterix's proprietary Single Molecule Array technology (Simoa™ ) that facilitates the detection and quantification of biomarkers previously difficult to measure, thus opening up new applications in life science research and in-vitro diagnostics. Simoa is based on trapping individual beads in arrays of femtoliter-sized wells that, when imaged with sufficient resolution, allows for counting of single molecules associated with each bead. When used to capture and detect proteins, this approach is known as digital ELISA (Enzyme-linked immunosorbent assay). The platform developed is a merger of many science and engineering disciplines. This paper concentrates on the optical technologies that have enabled the development of a fully-automated single molecule analyzer. At the core of the system is a custom, wide field-of-view, fluorescence microscope that images arrays of microwells containing single molecules bound to magnetic beads. A consumable disc containing 24 microstructure arrays was developed previously in collaboration with Sony DADC. The system cadence requirements, array dimensions, and requirement to detect single molecules presented significant optical challenges. Specifically, the wide field-of-view needed to image the entire array resulted in the need for a custom objective lens. Additionally, cost considerations for the system required a custom solution that leveraged the image processing capabilities. This paper will discuss the design considerations and resultant optical architecture that has enabled the development of an automated digital ELISA platform.

Apparatus and Method for Effecting Data Transfer Between Data Systems

NASA Technical Reports Server (NTRS)

Kirkpatrick, Joey V. (Inventor); Grosz, Francis B., Jr. (Inventor); Lannes, Kenny (Inventor); Maniscalco, David G. (Inventor)

2001-01-01

An apparatus for effecting data transfer between data systems comprising a first transceiver and a second transceiver. The first transceiver has an input for receiving digital data from one of the data systems, an output for serially outputting digital data to one of the data systems, at least one transmitter for converting digital data received at the input into optical signals, and at least one receiver for receiving optical signals and serially converting the received optical signals to digital data for output to the data output. The second transceiver has an input for receiving digital data from another one of the data systems, an output for serially outputting digital data to the another one of the data systems, at least one transmitter for serially converting digital data received at the input of the second transceiver into optical signals, and at least one receiver for receiving optical signals and serially converting the received optical signals to digital data for output to the output of the second transceiver. The apparatus further comprises an optical link connecting the first and second transceivers. The optical link comprising a pair of optical fibers. One of the optical fibers optically links the transmitter of the first transceiver to the receiver of the second transceiver. The other optical fiber optically links the receiver of the first transceiver to the transmitter of the second transceiver.

Radiation pressure excitation of a low temperature atomic force/magnetic force microscope for imaging in 4-300 K temperature range

NASA Astrophysics Data System (ADS)

Ćelik, Ümit; Karcı, Özgür; Uysallı, Yiǧit; Özer, H. Özgür; Oral, Ahmet

2017-01-01

We describe a novel radiation pressure based cantilever excitation method for imaging in dynamic mode atomic force microscopy (AFM) for the first time. Piezo-excitation is the most common method for cantilever excitation, however it may cause spurious resonance peaks. Therefore, the direct excitation of the cantilever plays a crucial role in AFM imaging. A fiber optic interferometer with a 1310 nm laser was used both for the excitation of the cantilever at the resonance and the deflection measurement of the cantilever in a commercial low temperature atomic force microscope/magnetic force microscope (AFM/MFM) from NanoMagnetics Instruments. The laser power was modulated at the cantilever's resonance frequency by a digital Phase Locked Loop (PLL). The laser beam is typically modulated by ˜500 μW, and ˜141.8 nmpp oscillation amplitude is obtained in moderate vacuum levels between 4 and 300 K. We have demonstrated the performance of the radiation pressure excitation in AFM/MFM by imaging atomic steps in graphite, magnetic domains in CoPt multilayers between 4 and 300 K and Abrikosov vortex lattice in BSCCO(2212) single crystal at 4 K for the first time.

Radiation pressure excitation of a low temperature atomic force/magnetic force microscope for imaging in 4-300 K temperature range.

PubMed

Çelik, Ümit; Karcı, Özgür; Uysallı, Yiğit; Özer, H Özgür; Oral, Ahmet

2017-01-01

We describe a novel radiation pressure based cantilever excitation method for imaging in dynamic mode atomic force microscopy (AFM) for the first time. Piezo-excitation is the most common method for cantilever excitation, however it may cause spurious resonance peaks. Therefore, the direct excitation of the cantilever plays a crucial role in AFM imaging. A fiber optic interferometer with a 1310 nm laser was used both for the excitation of the cantilever at the resonance and the deflection measurement of the cantilever in a commercial low temperature atomic force microscope/magnetic force microscope (AFM/MFM) from NanoMagnetics Instruments. The laser power was modulated at the cantilever's resonance frequency by a digital Phase Locked Loop (PLL). The laser beam is typically modulated by ∼500 μW, and ∼141.8 nm pp oscillation amplitude is obtained in moderate vacuum levels between 4 and 300 K. We have demonstrated the performance of the radiation pressure excitation in AFM/MFM by imaging atomic steps in graphite, magnetic domains in CoPt multilayers between 4 and 300 K and Abrikosov vortex lattice in BSCCO(2212) single crystal at 4 K for the first time.

Compact and light-weight automated semen analysis platform using lensfree on-chip microscopy.

PubMed

Su, Ting-Wei; Erlinger, Anthony; Tseng, Derek; Ozcan, Aydogan

2010-10-01

We demonstrate a compact and lightweight platform to conduct automated semen analysis using a lensfree on-chip microscope. This holographic on-chip imaging platform weighs ∼46 g, measures ∼4.2 × 4.2 × 5.8 cm, and does not require any lenses, lasers or other bulky optical components to achieve phase and amplitude imaging of sperms over ∼24 mm(2) field-of-view with an effective numerical aperture of ∼0.2. Using this wide-field lensfree on-chip microscope, semen samples are imaged for ∼10 s, capturing a total of ∼20 holographic frames. Digital subtraction of these consecutive lensfree frames, followed by appropriate processing of the reconstructed images, enables automated quantification of the count, the speed and the dynamic trajectories of motile sperms, while summation of the same frames permits counting of immotile sperms. Such a compact and lightweight automated semen analysis platform running on a wide-field lensfree on-chip microscope could be especially important for fertility clinics, personal male fertility tests, as well as for field use in veterinary medicine such as in stud farming and animal breeding applications.

Focusing mirrors for enhanced neutron radiography with thermal neutrons and application for irradiated nuclear fuel

NASA Astrophysics Data System (ADS)

Rai, Durgesh K.; Abir, Muhammad; Wu, Huarui; Khaykovich, Boris; Moncton, David E.

2018-01-01

Neutron radiography is a powerful method of probing the structure of materials based on attenuation of neutrons. This method is most suitable for materials containing heavy metals, which are not transparent to X-rays, for example irradiated nuclear fuel and other nuclear materials. Neutron radiography is one of the first non-distractive post-irradiated examination methods, which is applied to gain an overview of the integrity of irradiated nuclear fuel and other nuclear materials. However, very powerful gamma radiation emitted by the samples is damaging to the electronics of digital imaging detectors and has so far precluded the use of modern detectors. Here we describe a design of a neutron microscope based on focusing mirrors suitable for thermal neutrons. As in optical microscopes, the sample is separated from the detector, decreasing the effect of gamma radiation. In addition, the application of mirrors would result in a thirty-fold gain in flux and a resolution of better than 40 μm for a field-of-view of about 2.5 cm. Such a thermal neutron microscope can be useful for other applications of neutron radiography, where thermal neutrons are advantageous.

Wafer defect detection by a polarization-insensitive external differential interference contrast module.

PubMed

Nativ, Amit; Feldman, Haim; Shaked, Natan T

2018-05-01

We present a system that is based on a new external, polarization-insensitive differential interference contrast (DIC) module specifically adapted for detecting defects in semiconductor wafers. We obtained defect signal enhancement relative to the surrounding wafer pattern when compared with bright-field imaging. The new DIC module proposed is based on a shearing interferometer that connects externally at the output port of an optical microscope and enables imaging thin samples, such as wafer defects. This module does not require polarization optics (such as Wollaston or Nomarski prisms) and is insensitive to polarization, unlike traditional DIC techniques. In addition, it provides full control of the DIC shear and orientation, which allows obtaining a differential phase image directly on the camera (with no further digital processing) while enhancing defect detection capabilities, even if the size of the defect is smaller than the resolution limit. Our technique has the potential of future integration into semiconductor production lines.

RGB digital lensless holographic microscopy

NASA Astrophysics Data System (ADS)

Garcia-Sucerquia, Jorge

2013-11-01

The recent introduction of color digital lensless holographic microscopy (CDLHM) has shown the possibility of imaging microscopic specimens at full color without the need of lenses. Owing to the simplicity, robustness, and compactness of the digital lensless holographic microscopes (DLHM), they have been presented as the ideal candidates to being developed into portable holographic microscopes. However, in the case of CDLHM the utilization of three independent lasers hinders the portability option for this microscope. In this contribution an alternative to reduce the complexity of CDLHM aimed to recover the portability of this microscopy technology is presented. A super-bright white-light light-emitting diode (LED) is spectrally and spatially filtered to produce the needed illumination by CDLHM to work. CDLHM with LED illumination is used to image at full color a section of the head of a drosophila melanogaster fly (fruit fly). The LED-CDLHM method shows the capability of imaging objects of 2μm size in comparison with the micrometer resolution reported for LASER-CDLHM.

Evaluation of a completely robotized neurosurgical operating microscope.

PubMed

Kantelhardt, Sven R; Finke, Markus; Schweikard, Achim; Giese, Alf

2013-01-01

Operating microscopes are essential for most neurosurgical procedures. Modern robot-assisted controls offer new possibilities, combining the advantages of conventional and automated systems. We evaluated the prototype of a completely robotized operating microscope with an integrated optical coherence tomography module. A standard operating microscope was fitted with motors and control instruments, with the manual control mode and balance preserved. In the robot mode, the microscope was steered by a remote control that could be fixed to a surgical instrument. External encoders and accelerometers tracked microscope movements. The microscope was additionally fitted with an optical coherence tomography-scanning module. The robotized microscope was tested on model systems. It could be freely positioned, without forcing the surgeon to take the hands from the instruments or avert the eyes from the oculars. Positioning error was about 1 mm, and vibration faded in 1 second. Tracking of microscope movements, combined with an autofocus function, allowed determination of the focus position within the 3-dimensional space. This constituted a second loop of navigation independent from conventional infrared reflector-based techniques. In the robot mode, automated optical coherence tomography scanning of large surface areas was feasible. The prototype of a robotized optical coherence tomography-integrated operating microscope combines the advantages of a conventional manually controlled operating microscope with a remote-controlled positioning aid and a self-navigating microscope system that performs automated positioning tasks such as surface scans. This demonstrates that, in the future, operating microscopes may be used to acquire intraoperative spatial data, volume changes, and structural data of brain or brain tumor tissue.

The influence of the microscope lamp filament colour temperature on the process of digital images of histological slides acquisition standardization.

PubMed

Korzynska, Anna; Roszkowiak, Lukasz; Pijanowska, Dorota; Kozlowski, Wojciech; Markiewicz, Tomasz

2014-01-01

The aim of this study is to compare the digital images of the tissue biopsy captured with optical microscope using bright field technique under various light conditions. The range of colour's variation in immunohistochemically stained with 3,3'-Diaminobenzidine and Haematoxylin tissue samples is immense and coming from various sources. One of them is inadequate setting of camera's white balance to microscope's light colour temperature. Although this type of error can be easily handled during the stage of image acquisition, it can be eliminated with use of colour adjustment algorithms. The examination of the dependence of colour variation from microscope's light temperature and settings of the camera is done as an introductory research to the process of automatic colour standardization. Six fields of view with empty space among the tissue samples have been selected for analysis. Each field of view has been acquired 225 times with various microscope light temperature and camera white balance settings. The fourteen randomly chosen images have been corrected and compared, with the reference image, by the following methods: Mean Square Error, Structural SIMilarity and visual assessment of viewer. For two types of backgrounds and two types of objects, the statistical image descriptors: range, median, mean and its standard deviation of chromaticity on a and b channels from CIELab colour space, and luminance L, and local colour variability for objects' specific area have been calculated. The results have been averaged for 6 images acquired in the same light conditions and camera settings for each sample. The analysis of the results leads to the following conclusions: (1) the images collected with white balance setting adjusted to light colour temperature clusters in certain area of chromatic space, (2) the process of white balance correction for images collected with white balance camera settings not matched to the light temperature moves image descriptors into proper chromatic space but simultaneously the value of luminance changes. So the process of the image unification in a sense of colour fidelity can be solved in separate introductory stage before the automatic image analysis.

Light-sheet microscopy by confocal line scanning of dual-Bessel beams

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

Zhang, Pengfei; Phipps, Mary Elizabeth; Goodwin, Peter Marvin

Here, we have developed a light-sheet microscope that uses confocal scanning of dual-Bessel beams for illumination. A digital micromirror device (DMD) is placed in the intermediate image plane of the objective used to collect fluorescence and is programmed with two lines of pixels in the “on” state such that the DMD functions as a spatial filter to reject the out-of-focus background generated by the side-lobes of the Bessel beams. The optical sectioning and out-of-focus background rejection capabilities of this microscope were demonstrated by imaging of fluorescently stained actin in human A431 cells. The dual-Bessel beam system enables twice as manymore » photons to be detected per imaging scan, which is useful for low light applications (e.g., single-molecule localization) or imaging at high speed with a superior signal to noise. While demonstrated for two Bessel beams, this approach is scalable to a larger number of beams.« less

Light-sheet microscopy by confocal line scanning of dual-Bessel beams

DOE PAGES

Zhang, Pengfei; Phipps, Mary Elizabeth; Goodwin, Peter Marvin; ...

2016-10-25

Here, we have developed a light-sheet microscope that uses confocal scanning of dual-Bessel beams for illumination. A digital micromirror device (DMD) is placed in the intermediate image plane of the objective used to collect fluorescence and is programmed with two lines of pixels in the “on” state such that the DMD functions as a spatial filter to reject the out-of-focus background generated by the side-lobes of the Bessel beams. The optical sectioning and out-of-focus background rejection capabilities of this microscope were demonstrated by imaging of fluorescently stained actin in human A431 cells. The dual-Bessel beam system enables twice as manymore » photons to be detected per imaging scan, which is useful for low light applications (e.g., single-molecule localization) or imaging at high speed with a superior signal to noise. While demonstrated for two Bessel beams, this approach is scalable to a larger number of beams.« less

Isometric multimodal photoacoustic microscopy based on optically transparent micro-ring ultrasonic detection.

PubMed

Dong, Biqin; Li, Hao; Zhang, Zhen; Zhang, Kevin; Chen, Siyu; Sun, Cheng; Zhang, Hao F

2015-01-01

Photoacoustic microscopy (PAM) is an attractive imaging tool complementary to established optical microscopic modalities by providing additional molecular specificities through imaging optical absorption contrast. While the development of optical resolution photoacoustic microscopy (ORPAM) offers high lateral resolution, the acoustically-determined axial resolution is limited due to the constraint in ultrasonic detection bandwidth. ORPAM with isometric spatial resolution along both axial and lateral direction is yet to be developed. Although recently developed sophisticated optical illumination and reconstruction methods offer improved axial resolution in ORPAM, the image acquisition procedures are rather complicated, limiting their capabilities for high-speed imaging and being easily integrated with established optical microscopic modalities. Here we report an isometric ORPAM based on an optically transparent micro-ring resonator ultrasonic detector and a commercial inverted microscope platform. Owing to the superior spatial resolution and the ease of integrating our ORPAM with established microscopic modalities, single cell imaging with extrinsic fluorescence staining, intrinsic autofluorescence, and optical absorption can be achieved simultaneously. This technique holds promise to greatly improve the accessibility of PAM to the broader biomedical researchers.

The Pixel Paradox and Transition-Metal Spectroscopy: One of Many Uses of the Handheld Digital Microscope in Chemical Demonstrations

ERIC Educational Resources Information Center

Vitz, Ed

2010-01-01

A handheld digital microscope (HDM) interfaced to a computer with a presentation projector is used to project an out-of-focus yellow patch on the screen, then the patch is brought into focus to show that, paradoxically, there are red and green but no yellow pixels. Chromaticity diagrams are used to discuss this observation and spectroscopic…

Thin-section ratiometric Ca2+ images obtained by optical sectioning of fura-2 loaded mast cells

PubMed Central

1992-01-01

The availability of the ratiometric Ca2+ indicator dyes, fura-2, and indo-1, and advances in digital imaging and computer technology have made it possible to detect Ca2+ changes in single cells with high temporal and spatial resolution. However, the optical properties of the conventional epifluorescence microscope do not produce a perfect image of the specimen. Instead, the observed image is a spatial low pass filtered version of the object and is contaminated with out of focus information. As a result, the image has reduced contrast and an increased depth of field. This problem is especially important for measurements of localized Ca2+ concentrations. One solution to this problem is to use a scanning confocal microscope which only detects in focus information, but this approach has several disadvantages for low light fluorescence measurements in living cells. An alternative approach is to use digital image processing and a deblurring algorithm to remove the out of focus information by using a knowledge of the point spread function of the microscope. All of these algorithms require a stack of two-dimensional images taken at different focal planes, although the "nearest neighbor deblurring" algorithm only requires one image above and below the image plane. We have used a modification of this scheme to construct a simple inverse filter, which extracts optical sections comparable to those of the nearest neighbors scheme, but without the need for adjacent image sections. We have used this "no neighbors" processing scheme to deblur images of fura-2-loaded mast cells from beige mice and generate high resolution ratiometric Ca2+ images of thin sections through the cell. The shallow depth of field of these images is demonstrated by taking pairs of images at different focal planes, 0.5-microns apart. The secretory granules, which exclude the fura-2, appear in focus in all sections and distinct changes in their size and shape can be seen in adjacent sections. In addition, we show, with the aid of model objects, how the combination of inverse filtering and ratiometric imaging corrects for some of the inherent limitations of using an inverse filter and can be used for quantitative measurements of localized Ca2+ gradients. With this technique, we can observe Ca2+ transients in narrow regions of cytosol between the secretory granules and plasma membrane that can be less than 0.5-microns wide. Moreover, these Ca2+ increases can be seen to coincide with the swelling of the secretory granules that follows exocytotic fusion. PMID:1730775

Comprehensive study of unexpected microscope condensers formed in sample arrangements commonly used in optical microscopy.

PubMed

Desai, Darshan B; Aldawsari, Mabkhoot Mudith S; Alharbi, Bandar Mohammed H; Sen, Sanchari; Grave de Peralta, Luis

2015-09-01

We show that various setups for optical microscopy which are commonly used in biomedical laboratories behave like efficient microscope condensers that are responsible for observed subwavelength resolution. We present a series of experiments and simulations that reveal how inclined illumination from such unexpected condensers occurs when the sample is perpendicularly illuminated by a microscope's built-in white-light source. In addition, we demonstrate an inexpensive add-on optical module that serves as an efficient and lightweight microscope condenser. Using such add-on optical module in combination with a low-numerical-aperture objective lens and Fourier plane imaging microscopy technique, we demonstrate detection of photonic crystals with a period nearly eight times smaller than the Rayleigh resolution limit.

Imaging the effect of hemoglobin on properties of RBCs using common-path digital holographic microscope

NASA Astrophysics Data System (ADS)

Joglekar, M.; Shah, H.; Trivedi, V.; Mahajan, S.; Chhaniwal, V.; Leitgeb, R.; Javidi, B.; Anand, A.

2017-07-01

Adequate supply of oxygen to the body is the most essential requirement. In vertebrate species this function is performed by Hemoglobin contained in red blood cells. The mass concentration of the Hb determines the oxygen carrying capacity of the blood. Thus it becomes necessary to determine its concentration in the blood, which helps in monitoring the health of a person. If the amount of Hb crosses certain range, then it is considered critical. As the Hb constitutes upto 96% of red blood cells dry content, it would be interesting to examine various physical and mechanical parameters of RBCs which depends upon its concentration. Various diseases bring about significant variation in the amount of hemoglobin which may alter certain parameters of the RBC such as surface area, volume, membrane fluctuation etc. The study of the variations of these parameters may be helpful in determining Hb content which will reflect the state of health of a human body leading to disease diagnosis. Any increase or decrease in the amount of Hb will change the density and hence the optical thickness of the RBCs, which affects the cell membrane and thereby changing its mechanical and physical properties. Here we describe the use of lateral shearing digital holographic microscope for quantifying the cell parameters for studying the change in biophysical properties of cells due to variation in hemoglobin concentration.

Hyperlens-array-implemented optical microscopy

NASA Astrophysics Data System (ADS)

Iwanaga, Masanobu

2014-08-01

Limit of resolution of conventional optical microscopes has never reached below 100 nm under visible light illumination. We show that numerically designed high-transmittance hyperlens array (HLA) is implemented in an optical microscope and works in practice for achieving one-shot-recording optical images of in-situ placed objects with sub 50 nm resolution in lateral direction. Direct resolution test employing well-defined nanopatterns proves that the HLA-implemented imaging is super-resolution optical microscopy, which works even under nW/mm2 visible illumination for objects. The HLA implementation makes the resolution of conventional microscopes one-scale higher, leading to the 1/10 illumination wavelength range, that is, mesoscopic range.

Remote histology learning from static versus dynamic microscopic images.

PubMed

Mione, Sylvia; Valcke, Martin; Cornelissen, Maria

2016-05-06

Histology is the study of microscopic structures in normal tissue sections. Curriculum redesign in medicine has led to a decrease in the use of optical microscopes during practical classes. Other imaging solutions have been implemented to facilitate remote learning. With advancements in imaging technologies, learning material can now be digitized. Digitized microscopy images can be presented in either a static or dynamic format. This study of remote histology education identifies whether dynamic pictures are superior to static images for the acquisition of histological knowledge. Test results of two cohorts of second-year Bachelor in Medicine students at Ghent University were analyzed in two consecutive academic years: Cohort 1 (n = 190) and Cohort 2 (n = 174). Students in Cohort 1 worked with static images whereas students in Cohort 2 were presented with dynamic images. ANCOVA was applied to study differences in microscopy performance scores between the two cohorts, taking into account any possible initial differences in prior knowledge. The results show that practical histology scores are significantly higher with dynamic images as compared to static images (F (1,361) = 15.14, P < 0.01), regardless of student's gender and performance level. Several reasons for this finding can be explained in accordance with cognitivist learning theory. Since the findings suggest that knowledge construction with dynamic pictures is stronger as compared to static images, dynamic images should be introduced in a remote setting for microscopy education. Further implementation within a larger electronic learning management system needs to be explored in future research. Anat Sci Educ 9: 222-230. © 2015 American Association of Anatomists. © 2015 American Association of Anatomists.

High-resolution Episcopic Microscopy (HREM) - Simple and Robust Protocols for Processing and Visualizing Organic Materials

PubMed Central

Geyer, Stefan H.; Maurer-Gesek, Barbara; Reissig, Lukas F.; Weninger, Wolfgang J.

2017-01-01

We provide simple protocols for generating digital volume data with the high-resolution episcopic microscopy (HREM) method. HREM is capable of imaging organic materials with volumes up to 5 x 5 x 7 mm3 in typical numeric resolutions between 1 x 1 x 1 and 5 x 5 x 5 µm3. Specimens are embedded in methacrylate resin and sectioned on a microtome. After each section an image of the block surface is captured with a digital video camera that sits on the phototube connected to the compound microscope head. The optical axis passes through a green fluorescent protein (GFP) filter cube and is aligned with a position, at which the bock holder arm comes to rest after each section. In this way, a series of inherently aligned digital images, displaying subsequent block surfaces are produced. Loading such an image series in three-dimensional (3D) visualization software facilitates the immediate conversion to digital volume data, which permit virtual sectioning in various orthogonal and oblique planes and the creation of volume and surface rendered computer models. We present three simple, tissue specific protocols for processing various groups of organic specimens, including mouse, chick, quail, frog and zebra fish embryos, human biopsy material, uncoated paper and skin replacement material. PMID:28715372

High-resolution Episcopic Microscopy (HREM) - Simple and Robust Protocols for Processing and Visualizing Organic Materials.

PubMed

Geyer, Stefan H; Maurer-Gesek, Barbara; Reissig, Lukas F; Weninger, Wolfgang J

2017-07-07

We provide simple protocols for generating digital volume data with the high-resolution episcopic microscopy (HREM) method. HREM is capable of imaging organic materials with volumes up to 5 x 5 x 7 mm 3 in typical numeric resolutions between 1 x 1 x 1 and 5 x 5 x 5 µm 3 . Specimens are embedded in methacrylate resin and sectioned on a microtome. After each section an image of the block surface is captured with a digital video camera that sits on the phototube connected to the compound microscope head. The optical axis passes through a green fluorescent protein (GFP) filter cube and is aligned with a position, at which the bock holder arm comes to rest after each section. In this way, a series of inherently aligned digital images, displaying subsequent block surfaces are produced. Loading such an image series in three-dimensional (3D) visualization software facilitates the immediate conversion to digital volume data, which permit virtual sectioning in various orthogonal and oblique planes and the creation of volume and surface rendered computer models. We present three simple, tissue specific protocols for processing various groups of organic specimens, including mouse, chick, quail, frog and zebra fish embryos, human biopsy material, uncoated paper and skin replacement material.

Optical domain analog to digital conversion methods and apparatus

DOEpatents

Vawter, Gregory A

2014-05-13

Methods and apparatus for optical analog to digital conversion are disclosed. An optical signal is converted by mapping the optical analog signal onto a wavelength modulated optical beam, passing the mapped beam through interferometers to generate analog bit representation signals, and converting the analog bit representation signals into an optical digital signal. A photodiode receives an optical analog signal, a wavelength modulated laser coupled to the photodiode maps the optical analog signal to a wavelength modulated optical beam, interferometers produce an analog bit representation signal from the mapped wavelength modulated optical beam, and sample and threshold circuits corresponding to the interferometers produce a digital bit signal from the analog bit representation signal.

Fusion of lens-free microscopy and mobile-phone microscopy images for high-color-accuracy and high-resolution pathology imaging

NASA Astrophysics Data System (ADS)

Zhang, Yibo; Wu, Yichen; Zhang, Yun; Ozcan, Aydogan

2017-03-01

Digital pathology and telepathology require imaging tools with high-throughput, high-resolution and accurate color reproduction. Lens-free on-chip microscopy based on digital in-line holography is a promising technique towards these needs, as it offers a wide field of view (FOV >20 mm2) and high resolution with a compact, low-cost and portable setup. Color imaging has been previously demonstrated by combining reconstructed images at three discrete wavelengths in the red, green and blue parts of the visible spectrum, i.e., the RGB combination method. However, this RGB combination method is subject to color distortions. To improve the color performance of lens-free microscopy for pathology imaging, here we present a wavelet-based color fusion imaging framework, termed "digital color fusion microscopy" (DCFM), which digitally fuses together a grayscale lens-free microscope image taken at a single wavelength and a low-resolution and low-magnification color-calibrated image taken by a lens-based microscope, which can simply be a mobile phone based cost-effective microscope. We show that the imaging results of an H&E stained breast cancer tissue slide with the DCFM technique come very close to a color-calibrated microscope using a 40x objective lens with 0.75 NA. Quantitative comparison showed 2-fold reduction in the mean color distance using the DCFM method compared to the RGB combination method, while also preserving the high-resolution features of the lens-free microscope. Due to the cost-effective and field-portable nature of both lens-free and mobile-phone microscopy techniques, their combination through the DCFM framework could be useful for digital pathology and telepathology applications, in low-resource and point-of-care settings.

Applications of virtual reality technology in pathology.

PubMed

Grimes, G J; McClellan, S A; Goldman, J; Vaughn, G L; Conner, D A; Kujawski, E; McDonald, J; Winokur, T; Fleming, W

1997-01-01

TelePath(SM) a telerobotic system utilizing virtual microscope concepts based on high quality still digital imaging and aimed at real-time support for surgery by remote diagnosis of frozen sections. Many hospitals and clinics have an application for the remote practice of pathology, particularly in the area of reading frozen sections in support of surgery, commonly called anatomic pathology. The goal is to project the expertise of the pathologist into the remote setting by giving the pathologist access to the microscope slides with an image quality and human interface comparable to what the pathologist would experience at a real rather than a virtual microscope. A working prototype of a virtual microscope has been defined and constructed which has the needed performance in both the image quality and human interface areas for a pathologist to work remotely. This is accomplished through the use of telerobotics and an image quality which provides the virtual microscope the same diagnostic capabilities as a real microscope. The examination of frozen sections is performed a two-dimensional world. The remote pathologist is in a virtual world with the same capabilities as a "real" microscope, but response times may be slower depending on the specific computing and telecommunication environments. The TelePath system has capabilities far beyond a normal biological microscope, such as the ability to create a low power image of the entire sample using multiple images digitally matched together; the ability to digitally retrace a viewing trajectory; and the ability to archive images using CD ROM and other mass storage devices.

Optical design and system characterization of an imaging microscope at 121.6 nm

NASA Astrophysics Data System (ADS)

Gao, Weichuan; Finan, Emily; Kim, Geon-Hee; Kim, Youngsik; Milster, Thomas D.

2018-03-01

We present the optical design and system characterization of an imaging microscope prototype at 121.6 nm. System engineering processes are demonstrated through the construction of a Schwarzschild microscope objective, including tolerance analysis, fabrication, alignment, and testing. Further improvements on the as-built system with a correction phase plate are proposed and analyzed. Finally, the microscope assembly and the imaging properties of the prototype are demonstrated.

Large area fabrication of plasmonic nanoparticle grating structure by conventional scanning electron microscope

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

Sudheer,, E-mail: sudheer@rrcat.gov.in; Tiwari, P.; Rai, V. N.

Plasmonic nanoparticle grating (PNG) structure of different periods has been fabricated by electron beam lithography using silver halide based transmission electron microscope film as a substrate. Conventional scanning electron microscope is used as a fabrication tool for electron beam lithography. Optical microscope and energy dispersive spectroscopy (EDS) have been used for its morphological and elemental characterization. Optical characterization is performed by UV-Vis absorption spectroscopic technique.

Signal digitizing system and method based on amplitude-to-time optical mapping

DOEpatents

Chou, Jason; Bennett, Corey V; Hernandez, Vince

2015-01-13

A signal digitizing system and method based on analog-to-time optical mapping, optically maps amplitude information of an analog signal of interest first into wavelength information using an amplitude tunable filter (ATF) to impress spectral changes induced by the amplitude of the analog signal onto a carrier signal, i.e. a train of optical pulses, and next from wavelength information to temporal information using a dispersive element so that temporal information representing the amplitude information is encoded in the time domain in the carrier signal. Optical-to-electrical conversion of the optical pulses into voltage waveforms and subsequently digitizing the voltage waveforms into a digital image enables the temporal information to be resolved and quantized in the time domain. The digital image may them be digital signal processed to digitally reconstruct the analog signal based on the temporal information with high fidelity.

Optical forces, torques, and force densities calculated at a microscopic level using a self-consistent hydrodynamics method

NASA Astrophysics Data System (ADS)

Ding, Kun; Chan, C. T.

2018-04-01

The calculation of optical force density distribution inside a material is challenging at the nanoscale, where quantum and nonlocal effects emerge and macroscopic parameters such as permittivity become ill-defined. We demonstrate that the microscopic optical force density of nanoplasmonic systems can be defined and calculated using the microscopic fields generated using a self-consistent hydrodynamics model that includes quantum, nonlocal, and retardation effects. We demonstrate this technique by calculating the microscopic optical force density distributions and the optical binding force induced by external light on nanoplasmonic dimers. This approach works even in the limit when the nanoparticles are close enough to each other so that electron tunneling occurs, a regime in which classical electromagnetic approach fails completely. We discover that an uneven distribution of optical force density can lead to a light-induced spinning torque acting on individual particles. The hydrodynamics method offers us an accurate and efficient approach to study optomechanical behavior for plasmonic systems at the nanoscale.

Point-of-care mobile digital microscopy and deep learning for the detection of soil-transmitted helminths and Schistosoma haematobium.

PubMed

Holmström, Oscar; Linder, Nina; Ngasala, Billy; Mårtensson, Andreas; Linder, Ewert; Lundin, Mikael; Moilanen, Hannu; Suutala, Antti; Diwan, Vinod; Lundin, Johan

2017-06-01

Microscopy remains the gold standard in the diagnosis of neglected tropical diseases. As resource limited, rural areas often lack laboratory equipment and trained personnel, new diagnostic techniques are needed. Low-cost, point-of-care imaging devices show potential in the diagnosis of these diseases. Novel, digital image analysis algorithms can be utilized to automate sample analysis. Evaluation of the imaging performance of a miniature digital microscopy scanner for the diagnosis of soil-transmitted helminths and Schistosoma haematobium, and training of a deep learning-based image analysis algorithm for automated detection of soil-transmitted helminths in the captured images. A total of 13 iodine-stained stool samples containing Ascaris lumbricoides, Trichuris trichiura and hookworm eggs and 4 urine samples containing Schistosoma haematobium were digitized using a reference whole slide-scanner and the mobile microscopy scanner. Parasites in the images were identified by visual examination and by analysis with a deep learning-based image analysis algorithm in the stool samples. Results were compared between the digital and visual analysis of the images showing helminth eggs. Parasite identification by visual analysis of digital slides captured with the mobile microscope was feasible for all analyzed parasites. Although the spatial resolution of the reference slide-scanner is higher, the resolution of the mobile microscope is sufficient for reliable identification and classification of all parasites studied. Digital image analysis of stool sample images captured with the mobile microscope showed high sensitivity for detection of all helminths studied (range of sensitivity = 83.3-100%) in the test set (n = 217) of manually labeled helminth eggs. In this proof-of-concept study, the imaging performance of a mobile, digital microscope was sufficient for visual detection of soil-transmitted helminths and Schistosoma haematobium. Furthermore, we show that deep learning-based image analysis can be utilized for the automated detection and classification of helminths in the captured images.

Point-of-care mobile digital microscopy and deep learning for the detection of soil-transmitted helminths and Schistosoma haematobium

PubMed Central

Holmström, Oscar; Linder, Nina; Ngasala, Billy; Mårtensson, Andreas; Linder, Ewert; Lundin, Mikael; Moilanen, Hannu; Suutala, Antti; Diwan, Vinod; Lundin, Johan

2017-01-01

ABSTRACT Background: Microscopy remains the gold standard in the diagnosis of neglected tropical diseases. As resource limited, rural areas often lack laboratory equipment and trained personnel, new diagnostic techniques are needed. Low-cost, point-of-care imaging devices show potential in the diagnosis of these diseases. Novel, digital image analysis algorithms can be utilized to automate sample analysis. Objective: Evaluation of the imaging performance of a miniature digital microscopy scanner for the diagnosis of soil-transmitted helminths and Schistosoma haematobium, and training of a deep learning-based image analysis algorithm for automated detection of soil-transmitted helminths in the captured images. Methods: A total of 13 iodine-stained stool samples containing Ascaris lumbricoides, Trichuris trichiura and hookworm eggs and 4 urine samples containing Schistosoma haematobium were digitized using a reference whole slide-scanner and the mobile microscopy scanner. Parasites in the images were identified by visual examination and by analysis with a deep learning-based image analysis algorithm in the stool samples. Results were compared between the digital and visual analysis of the images showing helminth eggs. Results: Parasite identification by visual analysis of digital slides captured with the mobile microscope was feasible for all analyzed parasites. Although the spatial resolution of the reference slide-scanner is higher, the resolution of the mobile microscope is sufficient for reliable identification and classification of all parasites studied. Digital image analysis of stool sample images captured with the mobile microscope showed high sensitivity for detection of all helminths studied (range of sensitivity = 83.3–100%) in the test set (n = 217) of manually labeled helminth eggs. Conclusions: In this proof-of-concept study, the imaging performance of a mobile, digital microscope was sufficient for visual detection of soil-transmitted helminths and Schistosoma haematobium. Furthermore, we show that deep learning-based image analysis can be utilized for the automated detection and classification of helminths in the captured images. PMID:28838305

Ultrawidefield microscope for high-speed fluorescence imaging and targeted optogenetic stimulation.

PubMed

Werley, Christopher A; Chien, Miao-Ping; Cohen, Adam E

2017-12-01

The rapid increase in the number and quality of fluorescent reporters and optogenetic actuators has yielded a powerful set of tools for recording and controlling cellular state and function. To achieve the full benefit of these tools requires improved optical systems with high light collection efficiency, high spatial and temporal resolution, and patterned optical stimulation, in a wide field of view (FOV). Here we describe our 'Firefly' microscope, which achieves these goals in a Ø6 mm FOV. The Firefly optical system is optimized for simultaneous photostimulation and fluorescence imaging in cultured cells. All but one of the optical elements are commercially available, yet the microscope achieves 10-fold higher light collection efficiency at its design magnification than the comparable commercially available microscope using the same objective. The Firefly microscope enables all-optical electrophysiology ('Optopatch') in cultured neurons with a throughput and information content unmatched by other neuronal phenotyping systems. This capability opens possibilities in disease modeling and phenotypic drug screening. We also demonstrate applications of the system to voltage and calcium recordings in human induced pluripotent stem cell derived cardiomyocytes.

Ultrawidefield microscope for high-speed fluorescence imaging and targeted optogenetic stimulation

PubMed Central

Werley, Christopher A.; Chien, Miao-Ping; Cohen, Adam E.

2017-01-01

The rapid increase in the number and quality of fluorescent reporters and optogenetic actuators has yielded a powerful set of tools for recording and controlling cellular state and function. To achieve the full benefit of these tools requires improved optical systems with high light collection efficiency, high spatial and temporal resolution, and patterned optical stimulation, in a wide field of view (FOV). Here we describe our ‘Firefly’ microscope, which achieves these goals in a Ø6 mm FOV. The Firefly optical system is optimized for simultaneous photostimulation and fluorescence imaging in cultured cells. All but one of the optical elements are commercially available, yet the microscope achieves 10-fold higher light collection efficiency at its design magnification than the comparable commercially available microscope using the same objective. The Firefly microscope enables all-optical electrophysiology (‘Optopatch’) in cultured neurons with a throughput and information content unmatched by other neuronal phenotyping systems. This capability opens possibilities in disease modeling and phenotypic drug screening. We also demonstrate applications of the system to voltage and calcium recordings in human induced pluripotent stem cell derived cardiomyocytes. PMID:29296505

A digital transducer and digital microphone using an optical technique

NASA Astrophysics Data System (ADS)

Ghelmansarai, F. A.

1996-09-01

A transducer is devised to measure pressure, displacements or angles by optical means. This transducer delivers a digital output without relying on interferometry techniques or analogue-to-digital converters. This device is based on an optical scanner and an optical detector. An inter-digital photoconductive detector (IDPC) is employed that delivers a series of pulses, whose number depends on the scan length. A pre-objective scanning configuration is used that allows for the possibility of a flat image plane. The optical scanner provides scanning of IDPC and the generated scan length is proportional to the measurand.

Diagnosis of major cancer resection specimens with virtual slides: impact of a novel digital pathology workstation.

PubMed

Randell, Rebecca; Ruddle, Roy A; Thomas, Rhys G; Mello-Thoms, Claudia; Treanor, Darren

2014-10-01

Digital pathology promises a number of benefits in efficiency in surgical pathology, yet the longer time required to review a virtual slide than a glass slide currently represents a significant barrier to the routine use of digital pathology. We aimed to create a novel workstation that enables pathologists to view a case as quickly as on the conventional microscope. The Leeds Virtual Microscope (LVM) was evaluated using a mixed factorial experimental design. Twelve consultant pathologists took part, each viewing one long cancer case (12-25 slides) on the LVM and one on a conventional microscope. Total time taken and diagnostic confidence were similar for the microscope and LVM, as was the mean slide viewing time. On the LVM, participants spent a significantly greater proportion of the total task time viewing slides and revisited slides more often. The unique design of the LVM, enabling real-time rendering of virtual slides while providing users with a quick and intuitive way to navigate within and between slides, makes use of digital pathology in routine practice a realistic possibility. With further practice with the system, diagnostic efficiency on the LVM is likely to increase yet more. Copyright © 2014 Elsevier Inc. All rights reserved.

Exploration of operator method digital optical computers for application to NASA

NASA Technical Reports Server (NTRS)

1990-01-01

Digital optical computer design has been focused primarily towards parallel (single point-to-point interconnection) implementation. This architecture is compared to currently developing VHSIC systems. Using demonstrated multichannel acousto-optic devices, a figure of merit can be formulated. The focus is on a figure of merit termed Gate Interconnect Bandwidth Product (GIBP). Conventional parallel optical digital computer architecture demonstrates only marginal competitiveness at best when compared to projected semiconductor implements. Global, analog global, quasi-digital, and full digital interconnects are briefly examined as alternative to parallel digital computer architecture. Digital optical computing is becoming a very tough competitor to semiconductor technology since it can support a very high degree of three dimensional interconnect density and high degrees of Fan-In without capacitive loading effects at very low power consumption levels.

[Clinical pathology on the verge of virtual microscopy].

PubMed

Tolonen, Teemu; Näpänkangas, Juha; Isola, Jorma

2015-01-01

For more than 100 years, examinations of pathology specimens have relied on the use of the light microscope. The technological progress of the last few years is enabling the digitizing of histologic specimen slides and application of the virtual microscope in diagnostics. Virtual microscopy will facilitate consultation possibilities, and digital image analysis serves to enhance the level of diagnostics. Organizing and monitoring clinicopathological meetings will become easier. Digital archive of histologic specimens and the virtual microscopy network are expected to benefit training and research as well, particularly what applies to the Finnish biobank network which is currently being established.

Quantification of Estrogen Receptor-Alpha Expression in Human Breast Carcinomas With a Miniaturized, Low-Cost Digital Microscope: A Comparison with a High-End Whole Slide-Scanner

PubMed Central

Holmström, Oscar; Linder, Nina; Lundin, Mikael; Moilanen, Hannu; Suutala, Antti; Turkki, Riku; Joensuu, Heikki; Isola, Jorma; Diwan, Vinod; Lundin, Johan

2015-01-01

Introduction A significant barrier to medical diagnostics in low-resource environments is the lack of medical care and equipment. Here we present a low-cost, cloud-connected digital microscope for applications at the point-of-care. We evaluate the performance of the device in the digital assessment of estrogen receptor-alpha (ER) expression in breast cancer samples. Studies suggest computer-assisted analysis of tumor samples digitized with whole slide-scanners may be comparable to manual scoring, here we study whether similar results can be obtained with the device presented. Materials and Methods A total of 170 samples of human breast carcinoma, immunostained for ER expression, were digitized with a high-end slide-scanner and the point-of-care microscope. Corresponding regions from the samples were extracted, and ER status was determined visually and digitally. Samples were classified as ER negative (<1% ER positivity) or positive, and further into weakly (1–10% positivity) and strongly positive. Interobserver agreement (Cohen’s kappa) was measured and correlation coefficients (Pearson’s product-momentum) were calculated for comparison of the methods. Results Correlation and interobserver agreement (r = 0.98, p < 0.001, kappa = 0.84, CI95% = 0.75–0.94) were strong in the results from both devices. Concordance of the point-of-care microscope and the manual scoring was good (r = 0.94, p < 0.001, kappa = 0.71, CI95% = 0.61–0.80), and comparable to the concordance between the slide scanner and manual scoring (r = 0.93, p < 0.001, kappa = 0.69, CI95% = 0.60–0.78). Fourteen (8%) discrepant cases between manual and device-based scoring were present with the slide scanner, and 16 (9%) with the point-of-care microscope, all representing samples of low ER expression. Conclusions Tumor ER status can be accurately quantified with a low-cost imaging device and digital image-analysis, with results comparable to conventional computer-assisted or manual scoring. This technology could potentially be expanded for other histopathological applications at the point-of-care. PMID:26659386

Optical path difference microscopy with a Shack-Hartmann wavefront sensor.

PubMed

Gong, Hai; Agbana, Temitope E; Pozzi, Paolo; Soloviev, Oleg; Verhaegen, Michel; Vdovin, Gleb

2017-06-01

In this Letter, we show that a Shack-Hartmann wavefront sensor can be used for the quantitative measurement of the specimen optical path difference (OPD) in an ordinary incoherent optical microscope, if the spatial coherence of the illumination light in the plane of the specimen is larger than the microscope resolution. To satisfy this condition, the illumination numerical aperture should be smaller than the numerical aperture of the imaging lens. This principle has been successfully applied to build a high-resolution reference-free instrument for the characterization of the OPD of micro-optical components and microscopic biological samples.

Design of small confocal endo-microscopic probe working under multiwavelength environment

NASA Astrophysics Data System (ADS)

Kim, Young-Duk; Ahn, MyoungKi; Gweon, Dae-Gab

2010-02-01

Recently, optical imaging system is widely used in medical purpose. By using optical imaging system specific diseases can be easily diagnosed at early stage because optical imaging system has high resolution performance and various imaging method. These methods are used to get high resolution image of human body and can be used to verify whether the cell is infected by virus. Confocal microscope is one of the famous imaging systems which is used for in-vivo imaging. Because most of diseases are accompanied with cellular level changes, doctors can diagnosis at early stage by observing the cellular image of human organ. Current research is focused in the development of endo-microscope that has great advantage in accessibility to human body. In this research, I designed small probe that is connected to confocal microscope through optical fiber bundle and work as endo-microscope. And this small probe is mainly designed to correct chromatic aberration to use various laser sources for both fluorescence type and reflection type confocal images. By using two kinds of laser sources at the same time we demonstrated multi-modality confocal endo-microscope.

Grid Gap Measurement for an NSTAR Ion Thruster

NASA Technical Reports Server (NTRS)

Diaz, Esther M.; Soulas, George C.

2006-01-01

The change in gap between the screen and accelerator grids of an engineering model NSTAR ion optics assembly was measured during thruster operation with beam extraction. The molybdenum ion optics assembly was mounted onto an engineering model NSTAR ion thruster. The measurement technique consisted of measuring the difference in height of an alumina pin relative to the downstream accelerator grid surface. The alumina pin was mechanically attached to the center aperture of the screen grid and protruded through the center aperture of the accelerator grid. The change in pin height was monitored using a long distance microscope coupled to a digital imaging system. Transient and steady-state hot grid gaps were measured at three power levels: 0.5, 1.5 and 2.3 kW. Also, the change in grid gap was measured during the transition between power levels, and during the startup with high voltage applied just prior to discharge ignition. Performance measurements, such as perveance, electron backstreaming limit and screen grid ion transparency, were also made to confirm that this ion optics assembly performed similarly to past testing. Results are compared to a prior test of 30 cm titanium ion optics.

Automatic and adaptive heterogeneous refractive index compensation for light-sheet microscopy.

PubMed

Ryan, Duncan P; Gould, Elizabeth A; Seedorf, Gregory J; Masihzadeh, Omid; Abman, Steven H; Vijayaraghavan, Sukumar; Macklin, Wendy B; Restrepo, Diego; Shepherd, Douglas P

2017-09-20

Optical tissue clearing has revolutionized researchers' ability to perform fluorescent measurements of molecules, cells, and structures within intact tissue. One common complication to all optically cleared tissue is a spatially heterogeneous refractive index, leading to light scattering and first-order defocus. We designed C-DSLM (cleared tissue digital scanned light-sheet microscopy) as a low-cost method intended to automatically generate in-focus images of cleared tissue. We demonstrate the flexibility and power of C-DSLM by quantifying fluorescent features in tissue from multiple animal models using refractive index matched and mismatched microscope objectives. This includes a unique measurement of myelin tracks within intact tissue using an endogenous fluorescent reporter where typical clearing approaches render such structures difficult to image. For all measurements, we provide independent verification using standard serial tissue sectioning and quantification methods. Paired with advancements in volumetric image processing, C-DSLM provides a robust methodology to quantify sub-micron features within large tissue sections.Optical clearing of tissue has enabled optical imaging deeper into tissue due to significantly reduced light scattering. Here, Ryan et al. tackle first-order defocus, an artefact of a non-uniform refractive index, extending light-sheet microscopy to partially cleared samples.

Tomographic phase microscopy: principles and applications in bioimaging [Invited

PubMed Central

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

2017-01-01

Tomographic phase microscopy (TPM) is an emerging optical microscopic technique for bioimaging. TPM uses digital holographic measurements of complex scattered fields to reconstruct three-dimensional refractive index (RI) maps of cells with diffraction-limited resolution by solving inverse scattering problems. In this paper, we review the developments of TPM from the fundamental physics to its applications in bioimaging. We first provide a comprehensive description of the tomographic reconstruction physical models used in TPM. The RI map reconstruction algorithms and various regularization methods are discussed. Selected TPM applications for cellular imaging, particularly in hematology, are reviewed. Finally, we examine the limitations of current TPM systems, propose future solutions, and envision promising directions in biomedical research. PMID:29386746

Microscopy imaging device with advanced imaging properties

DOEpatents

Ghosh, Kunal; Burns, Laurie; El Gamal, Abbas; Schnitzer, Mark J.; Cocker, Eric; Ho, Tatt Wei

2015-11-24

Systems, methods and devices are implemented for microscope imaging solutions. One embodiment of the present disclosure is directed toward an epifluorescence microscope. The microscope includes an image capture circuit including an array of optical sensor. An optical arrangement is configured to direct excitation light of less than about 1 mW to a target object in a field of view of that is at least 0.5 mm.sup.2 and to direct epi-fluorescence emission caused by the excitation light to the array of optical sensors. The optical arrangement and array of optical sensors are each sufficiently close to the target object to provide at least 2.5 .mu.m resolution for an image of the field of view.

Microscopy imaging device with advanced imaging properties

DOEpatents

Ghosh, Kunal; Burns, Laurie; El Gamal, Abbas; Schnitzer, Mark J.; Cocker, Eric; Ho, Tatt Wei

2016-10-25

Systems, methods and devices are implemented for microscope imaging solutions. One embodiment of the present disclosure is directed toward an epifluorescence microscope. The microscope includes an image capture circuit including an array of optical sensor. An optical arrangement is configured to direct excitation light of less than about 1 mW to a target object in a field of view of that is at least 0.5 mm.sup.2 and to direct epi-fluorescence emission caused by the excitation light to the array of optical sensors. The optical arrangement and array of optical sensors are each sufficiently close to the target object to provide at least 2.5 .mu.m resolution for an image of the field of view.

Microscopy imaging device with advanced imaging properties

DOEpatents

Ghosh, Kunal; Burns, Laurie; El Gamal, Abbas; Schnitzer, Mark J.; Cocker, Eric; Ho, Tatt Wei

2016-11-22

Systems, methods and devices are implemented for microscope imaging solutions. One embodiment of the present disclosure is directed toward an epifluorescence microscope. The microscope includes an image capture circuit including an array of optical sensor. An optical arrangement is configured to direct excitation light of less than about 1 mW to a target object in a field of view of that is at least 0.5 mm.sup.2 and to direct epi-fluorescence emission caused by the excitation light to the array of optical sensors. The optical arrangement and array of optical sensors are each sufficiently close to the target object to provide at least 2.5 .mu.m resolution for an image of the field of view.

Microscopy imaging device with advanced imaging properties

DOEpatents

Ghosh, Kunal; Burns, Laurie; El Gamal, Abbas; Schnitzer, Mark J.; Cocker, Eric; Ho, Tatt Wei

2017-04-25

Systems, methods and devices are implemented for microscope imaging solutions. One embodiment of the present disclosure is directed toward an epifluorescence microscope. The microscope includes an image capture circuit including an array of optical sensor. An optical arrangement is configured to direct excitation light of less than about 1 mW to a target object in a field of view of that is at least 0.5 mm.sup.2 and to direct epi-fluorescence emission caused by the excitation light to the array of optical sensors. The optical arrangement and array of optical sensors are each sufficiently close to the target object to provide at least 2.5 .mu.m resolution for an image of the field of view.

Digital diffractive optics: Have diffractive optics entered mainstream industry yet?

NASA Astrophysics Data System (ADS)

Kress, Bernard; Hejmadi, Vic

2010-05-01

When a new technology is integrated into industry commodity products and consumer electronic devices, and sold worldwide in retail stores, it is usually understood that this technology has then entered the realm of mainstream technology and therefore mainstream industry. Such a leap however does not come cheap, as it has a double edge sword effect: first it becomes democratized and thus massively developed by numerous companies for various applications, but also it becomes a commodity, and thus gets under tremendous pressure to cut down its production and integration costs while not sacrificing to performance. We will show, based on numerous examples extracted from recent industry history, that the field of Diffractive Optics is about to undergo such a major transformation. Such a move has many impacts on all facets of digital diffractive optics technology, from the optical design houses to the micro-optics foundries (for both mastering and volume replication), to the final product integrators or contract manufacturers. The main causes of such a transformation are, as they have been for many other technologies in industry, successive technological bubbles which have carried and lifted up diffractive optics technology within the last decades. These various technological bubbles have been triggered either by real industry needs or by virtual investment hype. Both of these causes will be discussed in the paper. The adjective ""digital"" in "digital diffractive optics" does not refer only, as it is done in digital electronics, to the digital functionality of the element (digital signal processing), but rather to the digital way they are designed (by a digital computer) and fabricated (as wafer level optics using digital masking techniques). However, we can still trace a very strong similarity between the emergence of micro-electronics from analog electronics half a century ago, and the emergence of digital optics from conventional optics today.

Simultaneous imaging of cellular morphology and multiple biomarkers using an acousto-optic tunable filter-based bright field microscope.

PubMed

Wachman, Elliot S; Geyer, Stanley J; Recht, Joel M; Ward, Jon; Zhang, Bill; Reed, Murray; Pannell, Chris

2014-05-01

An acousto-optic tunable filter (AOTF)-based multispectral imaging microscope system allows the combination of cellular morphology and multiple biomarker stainings on a single microscope slide. We describe advances in AOTF technology that have greatly improved spectral purity, field uniformity, and image quality. A multispectral imaging bright field microscope using these advances demonstrates pathology results that have great potential for clinical use.

Multimodal optical workstation for simultaneous linear, nonlinear microscopy and nanomanipulation: upgrading a commercial confocal inverted microscope.

PubMed

Mathew, Manoj; Santos, Susana I C O; Zalvidea, Dobryna; Loza-Alvarez, Pablo

2009-07-01

In this work we propose and build a multimodal optical workstation that extends a commercially available confocal microscope (Nikon Confocal C1-Si) to include nonlinear/multiphoton microscopy and optical manipulation/stimulation tools such as nanosurgery. The setup allows both subsystems (confocal and nonlinear) to work independently and simultaneously. The workstation enables, for instance, nanosurgery along with simultaneous confocal and brightfield imaging. The nonlinear microscopy capabilities are added around the commercial confocal microscope by exploiting all the flexibility offered by this microscope and without need for any mechanical or electronic modification of the confocal microscope systems. As an example, the standard differential interference contrast condenser and diascopic detector in the confocal microscope are readily used as a forward detection mount for second harmonic generation imaging. The various capabilities of this workstation, as applied directly to biology, are demonstrated using the model organism Caenorhabditis elegans.

Apertureless near-field/far-field CW two-photon microscope for biological and material imaging and spectroscopic applications.

PubMed

Nowak, Derek B; Lawrence, A J; Sánchez, Erik J

2010-12-10

We present the development of a versatile spectroscopic imaging tool to allow for imaging with single-molecule sensitivity and high spatial resolution. The microscope allows for near-field and subdiffraction-limited far-field imaging by integrating a shear-force microscope on top of a custom inverted microscope design. The instrument has the ability to image in ambient conditions with optical resolutions on the order of tens of nanometers in the near field. A single low-cost computer controls the microscope with a field programmable gate array data acquisition card. High spatial resolution imaging is achieved with an inexpensive CW multiphoton excitation source, using an apertureless probe and simplified optical pathways. The high-resolution, combined with high collection efficiency and single-molecule sensitive optical capabilities of the microscope, are demonstrated with a low-cost CW laser source as well as a mode-locked laser source.

Images from Phoenix's MECA Instruments

NASA Technical Reports Server (NTRS)

2008-01-01

The image on the upper left is from NASA's Phoenix Mars Lander's Optical Microscope after a sample informally called 'Sorceress' was delivered to its silicon substrate on the 38th Martian day, or sol, of the mission (July 2, 2008).

A 3D representation of the same sample is on the right, as seen by Phoenix's Atomic Force Microscope. This is 100 times greater magnification than the view from the Optical Microscope, and the most highly magnified image ever seen from another world.

The Optical Microscope and the Atomic Force Microscope are part of Phoenix's Microscopy, Electrochemistry and Conductivity Analyzer instrument.

The Atomic Force Microscope was developed by a Swiss-led consortium in collaboration with Imperial College London.

The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

Experimental investigation on the caries characteristic of dental tissues by photothermal radiometry scanning imaging

NASA Astrophysics Data System (ADS)

Wang, Fei; Liu, Jun-yan; Wang, Xiao-chun; Wang, Yang

2018-03-01

In this paper, a one-dimensional (1D) thermal-wave model coupled diffuse-photon-density-wave for three-layer dental tissues using modulated laser stimulation was employed to illustrate the relationship between dental caries characteristic (i.e. caries layer thickness, optical absorption coefficient and optical scattering coefficient) and photothermal radiometry (PTR) signal. Experimental investigation of artificial caries was carried out using PTR scanning imaging. The PTR amplitude and phase delay were increased with dental demineralized treatment. The local caries characteristic parameters were obtained by the best-fitting method based on the 1D thermal-wave model. The PTR scanning imaging measurements illustrated that the optical absorption coefficient and scattering coefficient of caries region were much higher than those of the healthy enamel area. The demineralization thickness of caries region was measured by PTR scanning imaging and its average value shows in good agreement with the digital microscope. Experimental results show that PTR scanning imaging has the merits of high contrast for local inhomogeneity of dental caries; furthermore, this method is an allowance to provide a flexibility for non-contact quantitative evaluation of dental caries.

NeuroPG: open source software for optical pattern generation and data acquisition

PubMed Central

Avants, Benjamin W.; Murphy, Daniel B.; Dapello, Joel A.; Robinson, Jacob T.

2015-01-01

Patterned illumination using a digital micromirror device (DMD) is a powerful tool for optogenetics. Compared to a scanning laser, DMDs are inexpensive and can easily create complex illumination patterns. Combining these complex spatiotemporal illumination patterns with optogenetics allows DMD-equipped microscopes to probe neural circuits by selectively manipulating the activity of many individual cells or many subcellular regions at the same time. To use DMDs to study neural activity, scientists must develop specialized software to coordinate optical stimulation patterns with the acquisition of electrophysiological and fluorescence data. To meet this growing need we have developed an open source optical pattern generation software for neuroscience—NeuroPG—that combines, DMD control, sample visualization, and data acquisition in one application. Built on a MATLAB platform, NeuroPG can also process, analyze, and visualize data. The software is designed specifically for the Mightex Polygon400; however, as an open source package, NeuroPG can be modified to incorporate any data acquisition, imaging, or illumination equipment that is compatible with MATLAB’s Data Acquisition and Image Acquisition toolboxes. PMID:25784873

Advantages of microscope-integrated intraoperative online optical coherence tomography: usage in Boston keratoprosthesis type I surgery

NASA Astrophysics Data System (ADS)

Siebelmann, Sebastian; Steven, Philipp; Hos, Deniz; Hüttmann, Gereon; Lankenau, Eva; Bachmann, Björn; Cursiefen, Claus

2016-01-01

Boston keratoprosthesis (KPro) type I is a technique to treat patients with corneal diseases that are not amenable to conventional keratoplasty. Correct assembly and central implantation of the prosthesis are crucial for postoperative visual recovery. This study investigates the potential benefit of intraoperative optical coherence tomography (OCT) to monitor KPro surgery. Retrospective case series are presented for two patients who underwent Boston KPro type I implantation. The surgery in both patients was monitored intraoperatively using a commercially available intraoperative OCT (iOCT) device mounted on a surgical microscope. Microscope-integrated intraoperative OCT was able to evaluate the correct assembly and implantation of the KPro. All parts of the prosthesis were visible, and interfaces between the corneal graft and titanium backplate or anterior optics were clearly depictable. Moreover, iOCT visualized a gap between the backplate and graft in one case, and in the other case, a gap between the anterior optic and graft. Neither gap was visible with a conventional surgical microscope. The gap between the anterior optic and the graft could easily be corrected. Microscope-integrated iOCT delivers enhanced information, adding to the normal surgical microscope view during KPro surgery. Correct assembly can be controlled as well as the correct placement of the Boston KPro into the anterior chamber.

Development and Characterization of Embedded Sensory Particles Using Multi-Scale 3D Digital Image Correlation

NASA Technical Reports Server (NTRS)

Cornell, Stephen R.; Leser, William P.; Hochhalter, Jacob D.; Newman, John A.; Hartl, Darren J.

2014-01-01

A method for detecting fatigue cracks has been explored at NASA Langley Research Center. Microscopic NiTi shape memory alloy (sensory) particles were embedded in a 7050 aluminum alloy matrix to detect the presence of fatigue cracks. Cracks exhibit an elevated stress field near their tip inducing a martensitic phase transformation in nearby sensory particles. Detectable levels of acoustic energy are emitted upon particle phase transformation such that the existence and location of fatigue cracks can be detected. To test this concept, a fatigue crack was grown in a mode-I single-edge notch fatigue crack growth specimen containing sensory particles. As the crack approached the sensory particles, measurements of particle strain, matrix-particle debonding, and phase transformation behavior of the sensory particles were performed. Full-field deformation measurements were performed using a novel multi-scale optical 3D digital image correlation (DIC) system. This information will be used in a finite element-based study to determine optimal sensory material behavior and density.

Three-dimensional deformation of orthodontic brackets

PubMed Central

Melenka, Garrett W; Nobes, David S; Major, Paul W

2013-01-01

Braces are used by orthodontists to correct the misalignment of teeth in the mouth. Archwire rotation is a particular procedure used to correct tooth inclination. Wire rotation can result in deformation to the orthodontic brackets, and an orthodontic torque simulator has been designed to examine this wire–bracket interaction. An optical technique has been employed to measure the deformation due to size and geometric constraints of the orthodontic brackets. Images of orthodontic brackets are collected using a stereo microscope and two charge-coupled device cameras, and deformation of orthodontic brackets is measured using a three-dimensional digital image correlation technique. The three-dimensional deformation of orthodontic brackets will be evaluated. The repeatability of the three-dimensional digital image correlation measurement method was evaluated by performing 30 archwire rotation tests using the same bracket and archwire. Finally, five Damon 3MX and five In-Ovation R self-ligating brackets will be compared using this technique to demonstrate the effect of archwire rotation on bracket design. PMID:23762201

Three-dimensional deformation of orthodontic brackets.

PubMed

Melenka, Garrett W; Nobes, David S; Major, Paul W; Carey, Jason P

2013-01-01

Braces are used by orthodontists to correct the misalignment of teeth in the mouth. Archwire rotation is a particular procedure used to correct tooth inclination. Wire rotation can result in deformation to the orthodontic brackets, and an orthodontic torque simulator has been designed to examine this wire-bracket interaction. An optical technique has been employed to measure the deformation due to size and geometric constraints of the orthodontic brackets. Images of orthodontic brackets are collected using a stereo microscope and two charge-coupled device cameras, and deformation of orthodontic brackets is measured using a three-dimensional digital image correlation technique. The three-dimensional deformation of orthodontic brackets will be evaluated. The repeatability of the three-dimensional digital image correlation measurement method was evaluated by performing 30 archwire rotation tests using the same bracket and archwire. Finally, five Damon 3MX and five In-Ovation R self-ligating brackets will be compared using this technique to demonstrate the effect of archwire rotation on bracket design.

Design of a normal incidence multilayer imaging X-ray microscope

NASA Astrophysics Data System (ADS)

Shealy, David L.; Gabardi, David R.; Hoover, Richard B.; Walker, Arthur B. C., Jr.; Lindblom, Joakim F.

Normal incidence multilayer Cassegrain X-ray telescopes were flown on the Stanford/MSFC Rocket X-ray Spectroheliograph. These instruments produced high spatial resolution images of the sun and conclusively demonstrated that doubly reflecting multilayer X-ray optical systems are feasible. The images indicated that aplanatic imaging soft X-ray/EUV microscopes should be achievable using multilayer optics technology. A doubly reflecting normal incidence multilayer imaging X-ray microscope based on the Schwarzschild configuration has been designed. The design of the microscope and the results of the optical system ray trace analysis are discussed. High resolution aplanatic imaging X-ray microscopes using normal incidence multilayer X-ray mirrors should have many important applications in advanced X-ray astronomical instrumentation, X-ray lithography, biological, biomedical, metallurgical, and laser fusion research.

Color digital lensless holographic microscopy: laser versus LED illumination.

PubMed

Garcia-Sucerquia, Jorge

2016-08-20

A comparison of the performance of color digital lensless holographic microscopy (CDLHM) as utilized for illumination of RGB lasers or a super-bright white-light LED with a set of spectral filters is presented. As the use of lasers in CDLHM conceals the possibility of having a compact, lightweight, portable, and low cost microscope, and additionally the limited available laser radiation wavelengths limit a real multispectral imaging microscope, here we present the use of super-bright white-light LED and spectral filters for illuminating the sample. The performance of RGB laser-CDLHM and LED-CDLHM is evaluated on imaging a section of the head of a Drosophila melanogaster fly. This comparison shows that there is trade-off between the spatial resolution of the microscope and the light sources utilized, which can be understood with regard to the coherence properties of the illuminating light. Despite the smaller spatial coherence features of LED-CDLHM in comparison with laser-CDLHM, the former shows promise as a portable RGB digital lensless holographic microscope that could be extended to other wavelengths by the use of different spectral filters.

A versatile localization system for microscopic multiparametric analysis of cells.

PubMed

Thaw, H H; Rundquist, I; Johansson, U; Svensson, I; Collins, V P

1983-03-01

A new, simple and relatively inexpensive electronic digital position readout (DPRO) system which can be applied to the rapid localization and recovery of microscopic material is described. It is based upon a commercially available digital position readout system which is routinely utilized by industry for small machine tools and measuring equipment. This has been mounted onto the stage of various microscopic instrumentation to provide X and Y coordinates relative to an arbitrary reference point. The integration of small computers interfaced to scanning interferometric, microdensitometric and fluorescence microscopes were used to demonstrate the reliability, versatility and ease of application of this system to problems of multiparametric measurements and analysis of cultured cells. The system may be expanded and applied to clinical material to obtain automatized, multiparametric measurements of cells in haematology and clinical cytology.

SlideJ: An ImageJ plugin for automated processing of whole slide images.

PubMed

Della Mea, Vincenzo; Baroni, Giulia L; Pilutti, David; Di Loreto, Carla

2017-01-01

The digital slide, or Whole Slide Image, is a digital image, acquired with specific scanners, that represents a complete tissue sample or cytological specimen at microscopic level. While Whole Slide image analysis is recognized among the most interesting opportunities, the typical size of such images-up to Gpixels- can be very demanding in terms of memory requirements. Thus, while algorithms and tools for processing and analysis of single microscopic field images are available, Whole Slide images size makes the direct use of such tools prohibitive or impossible. In this work a plugin for ImageJ, named SlideJ, is proposed with the objective to seamlessly extend the application of image analysis algorithms implemented in ImageJ for single microscopic field images to a whole digital slide analysis. The plugin has been complemented by examples of macro in the ImageJ scripting language to demonstrate its use in concrete situations.

SlideJ: An ImageJ plugin for automated processing of whole slide images

PubMed Central

Baroni, Giulia L.; Pilutti, David; Di Loreto, Carla

2017-01-01

The digital slide, or Whole Slide Image, is a digital image, acquired with specific scanners, that represents a complete tissue sample or cytological specimen at microscopic level. While Whole Slide image analysis is recognized among the most interesting opportunities, the typical size of such images—up to Gpixels- can be very demanding in terms of memory requirements. Thus, while algorithms and tools for processing and analysis of single microscopic field images are available, Whole Slide images size makes the direct use of such tools prohibitive or impossible. In this work a plugin for ImageJ, named SlideJ, is proposed with the objective to seamlessly extend the application of image analysis algorithms implemented in ImageJ for single microscopic field images to a whole digital slide analysis. The plugin has been complemented by examples of macro in the ImageJ scripting language to demonstrate its use in concrete situations. PMID:28683129

All-optical negabinary adders using Mach-Zehnder interferometer

NASA Astrophysics Data System (ADS)

Cherri, A. K.

2011-02-01

In contrast to optoelectronics, all-optical adders are proposed where all-optical signals are used to represent the input numbers and the control signals. In addition, the all-optical adders use the negabinary modified signed-digit number representation (an extension of the negabinary number system) to represent the input digits. Further, the ultra-speed of the designed circuits is achieved due to the use of ultra-fast all-optical switching property of the semiconductor optical amplifier and Mach-Zehnder interferometer (SOA-MZI). Furthermore, two-bit per digit binary encoding scheme is employed to represent the trinary values of the negabinary modified signed-digits.

A scalable, self-analyzing digital locking system for use on quantum optics experiments.

PubMed

Sparkes, B M; Chrzanowski, H M; Parrain, D P; Buchler, B C; Lam, P K; Symul, T

2011-07-01

Digital control of optics experiments has many advantages over analog control systems, specifically in terms of the scalability, cost, flexibility, and the integration of system information into one location. We present a digital control system, freely available for download online, specifically designed for quantum optics experiments that allows for automatic and sequential re-locking of optical components. We show how the inbuilt locking analysis tools, including a white-noise network analyzer, can be used to help optimize individual locks, and verify the long term stability of the digital system. Finally, we present an example of the benefits of digital locking for quantum optics by applying the code to a specific experiment used to characterize optical Schrödinger cat states.

Time for Slime

ERIC Educational Resources Information Center

Tessmer, Michael; Cowlishaw, Richard

2011-01-01

An introduction to microscopy is common in the elementary curriculum, but microscope work with elementary school children can be a challenge. There is equipment maintenance to consider, as well as the difficulty of using the microscope for many children. These authors have found that using a digital microscope connected to a projector breaks down…

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NASA Astrophysics Data System (ADS)

2009-09-01

WE RECOMMEND Sustainable Energy—Without the Hot Air This excellent book makes sense of energy facts and figures Doppler Effect Unit Another simple, effective piece of kit from SEP Plastic Fantastic: How the Biggest Fraud in Physics Shook the Scientific World Intriguing and unique write-up of an intellectual fraud case Brunel Eyecam An affordable digital eyepiece for your microscope 200x Digital Microscope An adjustable digital flexcam for classroom use The Atom and the Apple: Twelve Tales from Contemporary Physics A fascinating round-up of the recent history of physics WORTH A LOOK The Physics of Rugby Book uses sport analogy and context to teach physics concepts Physics 2 for OCR Essential textbook for the course but otherwise pointless WEB WATCH Some free teaching materials are better than those you'd pay for

In Vivo Near Infrared Virtual Intraoperative Surgical Photoacoustic Optical Coherence Tomography

PubMed Central

Lee, Donghyun; Lee, Changho; Kim, Sehui; Zhou, Qifa; Kim, Jeehyun; Kim, Chulhong

2016-01-01

Since its first implementation in otolaryngological surgery nearly a century ago, the surgical microscope has improved the accuracy and the safety of microsurgeries. However, the microscope shows only a magnified surface view of the surgical region. To overcome this limitation, either optical coherence tomography (OCT) or photoacoustic microscopy (PAM) has been independently combined with conventional surgical microscope. Herein, we present a near-infrared virtual intraoperative photoacoustic optical coherence tomography (NIR-VISPAOCT) system that combines both PAM and OCT with a conventional surgical microscope. Using optical scattering and absorption, the NIR-VISPAOCT system simultaneously provides surgeons with real-time comprehensive biological information such as tumor margins, tissue structure, and a magnified view of the region of interest. Moreover, by utilizing a miniaturized beam projector, it can back-project 2D cross-sectional PAM and OCT images onto the microscopic view plane. In this way, both microscopic and cross-sectional PAM and OCT images are concurrently displayed on the ocular lens of the microscope. To verify the usability of the NIR-VISPAOCT system, we demonstrate simulated surgeries, including in vivo image-guided melanoma resection surgery and in vivo needle injection of carbon particles into a mouse thigh. The proposed NIR-VISPAOCT system has potential applications in neurosurgery, ophthalmological surgery, and other microsurgeries. PMID:27731390

In vivo fiber-optic confocal reflectance microscope with an injection-molded plastic miniature objective lens.

PubMed

Carlson, Kristen; Chidley, Matthew; Sung, Kung-Bin; Descour, Michael; Gillenwater, Ann; Follen, Michele; Richards-Kortum, Rebecca

2005-04-01

For in vivo optical diagnostic technologies to be distributed to the developed and developing worlds, optical imaging systems must be constructed of inexpensive components. We present a fiber-optic confocal reflectance microscope with a cost-effective injection-molded plastic miniature objective lens for in vivo imaging of human tissues in near real time. The measured lateral resolution is less than 2.2 microm, and the measured axial resolution is 10 microm. Confocal images of ex vivo cervical tissue biopsies and in vivo human lip taken at 15 frames/s demonstrate the microscope's capability of imaging cell morphology and tissue architecture.

An automated system for whole microscopic image acquisition and analysis.

PubMed

Bueno, Gloria; Déniz, Oscar; Fernández-Carrobles, María Del Milagro; Vállez, Noelia; Salido, Jesús

2014-09-01

The field of anatomic pathology has experienced major changes over the last decade. Virtual microscopy (VM) systems have allowed experts in pathology and other biomedical areas to work in a safer and more collaborative way. VMs are automated systems capable of digitizing microscopic samples that were traditionally examined one by one. The possibility of having digital copies reduces the risk of damaging original samples, and also makes it easier to distribute copies among other pathologists. This article describes the development of an automated high-resolution whole slide imaging (WSI) system tailored to the needs and problems encountered in digital imaging for pathology, from hardware control to the full digitization of samples. The system has been built with an additional digital monochromatic camera together with the color camera by default and LED transmitted illumination (RGB). Monochrome cameras are the preferred method of acquisition for fluorescence microscopy. The system is able to digitize correctly and form large high resolution microscope images for both brightfield and fluorescence. The quality of the digital images has been quantified using three metrics based on sharpness, contrast and focus. It has been proved on 150 tissue samples of brain autopsies, prostate biopsies and lung cytologies, at five magnifications: 2.5×, 10×, 20×, 40×, and 63×. The article is focused on the hardware set-up and the acquisition software, although results of the implemented image processing techniques included in the software and applied to the different tissue samples are also presented. © 2014 Wiley Periodicals, Inc.

Utility of Digital Stereo Images for Optic Disc Evaluation

PubMed Central

Ying, Gui-shuang; Pearson, Denise J.; Bansal, Mayank; Puri, Manika; Miller, Eydie; Alexander, Judith; Piltz-Seymour, Jody; Nyberg, William; Maguire, Maureen G.; Eledath, Jayan; Sawhney, Harpreet

2010-01-01

Purpose. To assess the suitability of digital stereo images for optic disc evaluations in glaucoma. Methods. Stereo color optic disc images in both digital and 35-mm slide film formats were acquired contemporaneously from 29 subjects with various cup-to-disc ratios (range, 0.26–0.76; median, 0.475). Using a grading scale designed to assess image quality, the ease of visualizing optic disc features important for glaucoma diagnosis, and the comparative diameters of the optic disc cup, experienced observers separately compared the primary digital stereo images to each subject's 35-mm slides, to scanned images of the same 35-mm slides, and to grayscale conversions of the digital images. Statistical analysis accounted for multiple gradings and comparisons and also assessed image formats under monoscopic viewing. Results. Overall, the quality of primary digital color images was judged superior to that of 35-mm slides (P < 0.001), including improved stereo (P < 0.001), but the primary digital color images were mostly equivalent to the scanned digitized images of the same slides. Color seemingly added little to grayscale optic disc images, except that peripapillary atrophy was best seen in color (P < 0.0001); both the nerve fiber layer (P < 0.0001) and the paths of blood vessels on the optic disc (P < 0.0001) were best seen in grayscale. The preference for digital over film images was maintained under monoscopic viewing conditions. Conclusions. Digital stereo optic disc images are useful for evaluating the optic disc in glaucoma and allow the application of advanced image processing applications. Grayscale images, by providing luminance distinct from color, may be informative for assessing certain features. PMID:20505199

Electron tomography simulator with realistic 3D phantom for evaluation of acquisition, alignment and reconstruction methods.

PubMed

Wan, Xiaohua; Katchalski, Tsvi; Churas, Christopher; Ghosh, Sreya; Phan, Sebastien; Lawrence, Albert; Hao, Yu; Zhou, Ziying; Chen, Ruijuan; Chen, Yu; Zhang, Fa; Ellisman, Mark H

2017-05-01

Because of the significance of electron microscope tomography in the investigation of biological structure at nanometer scales, ongoing improvement efforts have been continuous over recent years. This is particularly true in the case of software developments. Nevertheless, verification of improvements delivered by new algorithms and software remains difficult. Current analysis tools do not provide adaptable and consistent methods for quality assessment. This is particularly true with images of biological samples, due to image complexity, variability, low contrast and noise. We report an electron tomography (ET) simulator with accurate ray optics modeling of image formation that includes curvilinear trajectories through the sample, warping of the sample and noise. As a demonstration of the utility of our approach, we have concentrated on providing verification of the class of reconstruction methods applicable to wide field images of stained plastic-embedded samples. Accordingly, we have also constructed digital phantoms derived from serial block face scanning electron microscope images. These phantoms are also easily modified to include alignment features to test alignment algorithms. The combination of more realistic phantoms with more faithful simulations facilitates objective comparison of acquisition parameters, alignment and reconstruction algorithms and their range of applicability. With proper phantoms, this approach can also be modified to include more complex optical models, including distance-dependent blurring and phase contrast functions, such as may occur in cryotomography. Copyright © 2017 Elsevier Inc. All rights reserved.

X-ray microbeam stand-alone facility for cultured cells irradiation

NASA Astrophysics Data System (ADS)

Bożek, Sebastian; Bielecki, Jakub; Wiecheć, Anna; Lekki, Janusz; Stachura, Zbigniew; Pogoda, Katarzyna; Lipiec, Ewelina; Tkocz, Konrad; Kwiatek, Wojciech M.

2017-03-01

The article describes an X-ray microbeam standalone facility dedicated for irradiation of living cultured cells. The article can serve as an advice for such facilities construction, as it begins from engineering details, through mathematical modeling and experimental procedures, ending up with preliminary experimental results and conclusions. The presented system consists of an open type X-ray tube with microfocusing down to about 2 μm, an X-ray focusing system with optical elements arranged in the nested Kirckpatrick-Baez (or Montel) geometry, a sample stand and an optical microscope with a scientific digital CCD camera. For the beam visualisation an X-ray sensitive CCD camera and a spectral detector are used, as well as a scintillator screen combined with the microscope. A method of precise one by one irradiation of previously chosen cells is presented, as well as a fast method of uniform irradiation of a chosen sample area. Mathematical models of beam and cell with calculations of kerma and dose are presented. The experiments on dose-effect relationship, kinetics of DNA double strand breaks repair, as well as micronuclei observation were performed on PC-3 (Prostate Cancer) cultured cells. The cells were seeded and irradiated on Mylar foil, which covered a hole drilled in the Petri dish. DNA lesions were visualised with γ-H2AX marker combined with Alexa Fluor 488 fluorescent dye.

Analytical model of the optical vortex microscope.

PubMed

Płocinniczak, Łukasz; Popiołek-Masajada, Agnieszka; Masajada, Jan; Szatkowski, Mateusz

2016-04-20

This paper presents an analytical model of the optical vortex scanning microscope. In this microscope the Gaussian beam with an embedded optical vortex is focused into the sample plane. Additionally, the optical vortex can be moved inside the beam, which allows fine scanning of the sample. We provide an analytical solution of the whole path of the beam in the system (within paraxial approximation)-from the vortex lens to the observation plane situated on the CCD camera. The calculations are performed step by step from one optical element to the next. We show that at each step, the expression for light complex amplitude has the same form with only four coefficients modified. We also derive a simple expression for the vortex trajectory of small vortex displacements.

Fibre-optic nonlinear optical microscopy and endoscopy.

PubMed

Fu, L; Gu, M

2007-06-01

Nonlinear optical microscopy has been an indispensable laboratory tool of high-resolution imaging in thick tissue and live animals. Rapid developments of fibre-optic components in terms of growing functionality and decreasing size provide enormous opportunities for innovations in nonlinear optical microscopy. Fibre-based nonlinear optical endoscopy is the sole instrumentation to permit the cellular imaging within hollow tissue tracts or solid organs that are inaccessible to a conventional optical microscope. This article reviews the current development of fibre-optic nonlinear optical microscopy and endoscopy, which includes crucial technologies for miniaturized nonlinear optical microscopy and their embodiments of endoscopic systems. A particular attention is given to several classes of photonic crystal fibres that have been applied to nonlinear optical microscopy due to their unique properties for ultrashort pulse delivery and signal collection. Furthermore, fibre-optic nonlinear optical imaging systems can be classified into portable microscopes suitable for imaging behaving animals, rigid endoscopes that allow for deep tissue imaging with minimally invasive manners, and flexible endoscopes enabling imaging of internal organs. Fibre-optic nonlinear optical endoscopy is coming of age and a paradigm shift leading to optical microscope tools for early cancer detection and minimally invasive surgery.

Digital microscopy. Bringing new technology into focus.

PubMed

2010-06-01

Digital microscopy enables the scanning of microscope slides so that they can be viewed, analyzed, and archived on a computer. While the technology is not yet widely accepted by pathologists, a switch to digital microscopy systems seems to be inevitable in the near future.

Development and applications of optical interferometric micrometrology in the Angstrom and subangstrom range

NASA Technical Reports Server (NTRS)

Lauer, James L.; Abel, Phillip B.

1988-01-01

The characteristics of the scanning tunneling microscope and atomic force microscope (AFM) are briefly reviewed, and optical methods, mainly interferometry, of sufficient resolution to measure AFM deflections are discussed. The methods include optical resonators, laser interferometry, multiple-beam interferometry, and evanescent wave detection. Experimental results using AFM are reviewed.

The relationship between morphological changes of lens epithelial cells and intraocular lens optic material.

PubMed

Majima, K

1998-01-01

To examine the morphological changes of lens epithelial cells (LECs) occurring directly beneath and at regions contacting various intraocular lens (IOL) optic materials, human LECs were cultured on human anterior lens capsules and were further incubated upon placing above the cells lens optics made of polymethylmethacrylate, silicone, and soft acrylic material. Observations as to the morphological changes of LECs under phase-contrast microscope and scanning electron microscope were performed on the 14th day of incubation. Gatherings of LECs were observed at regions contacting the soft acrylic material under phase-contrast microscope, and gatherings of LECs were observed accurately at the same regions mentioned above under scanning electron microscope. On the other hand, LECs in contact with two other optic materials did not show morphological changes. The results suggest that LECs attached to and proliferated on not only the anterior lens capsules but also the soft acrylic IOL optics. The model used in this study may be useful in studying the relationship between cellular movement of LECs and IOL optic material.

Multiparallel Three-Dimensional Optical Microscopy

NASA Technical Reports Server (NTRS)

Nguyen, Lam K.; Price, Jeffrey H.; Kellner, Albert L.; Bravo-Zanoquera, Miguel

2010-01-01

Multiparallel three-dimensional optical microscopy is a method of forming an approximate three-dimensional image of a microscope sample as a collection of images from different depths through the sample. The imaging apparatus includes a single microscope plus an assembly of beam splitters and mirrors that divide the output of the microscope into multiple channels. An imaging array of photodetectors in each channel is located at a different distance along the optical path from the microscope, corresponding to a focal plane at a different depth within the sample. The optical path leading to each photodetector array also includes lenses to compensate for the variation of magnification with distance so that the images ultimately formed on all the photodetector arrays are of the same magnification. The use of optical components common to multiple channels in a simple geometry makes it possible to obtain high light-transmission efficiency with an optically and mechanically simple assembly. In addition, because images can be read out simultaneously from all the photodetector arrays, the apparatus can support three-dimensional imaging at a high scanning rate.

A high-accuracy optical linear algebra processor for finite element applications

NASA Technical Reports Server (NTRS)

Casasent, D.; Taylor, B. K.

1984-01-01

Optical linear processors are computationally efficient computers for solving matrix-matrix and matrix-vector oriented problems. Optical system errors limit their dynamic range to 30-40 dB, which limits their accuray to 9-12 bits. Large problems, such as the finite element problem in structural mechanics (with tens or hundreds of thousands of variables) which can exploit the speed of optical processors, require the 32 bit accuracy obtainable from digital machines. To obtain this required 32 bit accuracy with an optical processor, the data can be digitally encoded, thereby reducing the dynamic range requirements of the optical system (i.e., decreasing the effect of optical errors on the data) while providing increased accuracy. This report describes a new digitally encoded optical linear algebra processor architecture for solving finite element and banded matrix-vector problems. A linear static plate bending case study is described which quantities the processor requirements. Multiplication by digital convolution is explained, and the digitally encoded optical processor architecture is advanced.

Analysis of Polarizing Optical Systems for Digital Optical Computing with Symmetric Self Electrooptic Devices

DTIC Science & Technology

1991-03-31

I AD-A232 768 I Annual Report Analysis of Polarizing Optical Systems for Digital Optical Computing with I ’ Symmetric Self Electrooptic Devices I To...TTU AND SuSiIU S. PUNDIN mUMBERS Polarizing Optical Systems for Digital Optical Computing with Symmetric Self Electrooptic Devices AFOSR-89-0542 C...UTION COO$ UNLIMITED 13. ABSTRACT (MAxnum00woUw Two architectural approaches have dominated the field of optical computing . The first appAch uses

Acoustical nanometre-scale vibrations of live cells detected by a near-field optical setup

NASA Astrophysics Data System (ADS)

Piga, Rosaria; Micheletto, Ruggero; Kawakami, Yoichi

2007-04-01

The Scanning Near-field Optical Microscope (SNOM) is able to detect tiny vertical movement on the cell membrane in the range of only 1 nanometer or less, about 3 orders of magnitude better than conventional optical microscopes. Here we show intriguing data of cell membrane nanometer-scale dynamics associated to different phenomena of the cell’s The Scanning Near-field Optical Microscope (SNOM) is able to detect tiny vertical movement on the cell membrane in the range of only 1 nanometer or less, about 3 orders of magnitude better than conventional optical microscopes. Here we show intriguing data of cell membrane nanometer-scale dynamics associated to different phenomena of the cell’s life, such as cell cycle and cell death, on rat pheochromocytoma line PC12. Working in culture medium with alive and unperturbed samples, we could detect nanometer-sized movements; Fourier components revealed a clear distinct behavior associated to regulation of neurite outgrowth and changes on morphology after necrotic stimulus.

Antibacterial and tribological behavior of self-assembled monolayer on optical lens

NASA Astrophysics Data System (ADS)

Horng, J. H.; Jeng, Y. R.; Wei, C. C.; Tasi, Y. T.

2010-10-01

This paper studies the effects of the antibacterial and anti-adhesion properties of self-assembled monolayers (SAMs) on optical parts. Therefore, the experiments in this study prepared several kinds of SAMs, including alkyl and biphenyl spacer chains with different surface terminal groups (-CH3,-COOH) and head groups (-SH). This study reports the growth of eight self-assembled monolayers on optical parts: OTS, ODS, OTS with antibacterial solution, ODS with antibacterial solution, and pure antibacterial solution, with bio-compatibility. Experimental results regarding the contact angle of five self-assembled monolayers show that ODS with antibacterial illustrated the maximum contact angle 103° 12 hours after reaction. The solutions of OTS, ODS with antibacterial, OTS with antibacterial, and pure anti-bacterial showed contact angles of 102°, 99°, 101°, and 59° respectively. These results indicate that the antibacterial solution has negligible effects on anti-adhesion property of optical lenses. The results of digital optical microscope system analysis show that in the antibacterial experiment of eight kinds of selfassembled monolayers, the OTSanti50% effect cultured for 24 hours achieved the best results, with a growth rate of 12%. The descending order of antibacterial effect is antibacterial 10%>ODS>OTS> antibacterial 50%>ODSanti50%>OTSanti10%>ODSanti10%. In summary, the surface treatment of optical lenses involving OTSanti 50% is the most capable of effectively increasing antifouling and antibacterial functions.

Digital adaptive optics line-scanning confocal imaging system.

PubMed

Liu, Changgeng; Kim, Myung K

2015-01-01

A digital adaptive optics line-scanning confocal imaging (DAOLCI) system is proposed by applying digital holographic adaptive optics to a digital form of line-scanning confocal imaging system. In DAOLCI, each line scan is recorded by a digital hologram, which allows access to the complex optical field from one slice of the sample through digital holography. This complex optical field contains both the information of one slice of the sample and the optical aberration of the system, thus allowing us to compensate for the effect of the optical aberration, which can be sensed by a complex guide star hologram. After numerical aberration compensation, the corrected optical fields of a sequence of line scans are stitched into the final corrected confocal image. In DAOLCI, a numerical slit is applied to realize the confocality at the sensor end. The width of this slit can be adjusted to control the image contrast and speckle noise for scattering samples. DAOLCI dispenses with the hardware pieces, such as Shack–Hartmann wavefront sensor and deformable mirror, and the closed-loop feedbacks adopted in the conventional adaptive optics confocal imaging system, thus reducing the optomechanical complexity and cost. Numerical simulations and proof-of-principle experiments are presented that demonstrate the feasibility of this idea.

Soft control of scanning probe microscope with high flexibility.

PubMed

Liu, Zhenghui; Guo, Yuzheng; Zhang, Zhaohui; Zhu, Xing

2007-01-01

Most commercial scanning probe microscopes have multiple embedded digital microprocessors and utilize complex software for system control, which is not easily obtained or modified by researchers wishing to perform novel and special applications. In this paper, we present a simple and flexible control solution that just depends on software running on a single-processor personal computer with real-time Linux operating system to carry out all the control tasks including negative feedback, tip moving, data processing and user interface. In this way, we fully exploit the potential of a personal computer in calculating and programming, enabling us to manipulate the scanning probe as required without any special digital control circuits and related technical know-how. This solution has been successfully applied to a homemade ultrahigh vacuum scanning tunneling microscope and a multiprobe scanning tunneling microscope.

Measuring Roughnesses Of Optical Surfaces

NASA Technical Reports Server (NTRS)

Coulter, Daniel R.; Al-Jumaily, Gahnim A.; Raouf, Nasrat A.; Anderson, Mark S.

1994-01-01

Report discusses use of scanning tunneling microscopy and atomic force microscopy to measure roughnesses of optical surfaces. These techniques offer greater spatial resolution than other techniques. Report notes scanning tunneling microscopes and atomic force microscopes resolve down to 1 nm.

Hard X-Ray Scanning Microscope with Multilayer Laue Lens Nanofocusing Optics

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

Nazaretski, Evgeny

Evgeny Nazaretski, a physicist at Brookhaven Lab’s National Synchrotron Light Source II, spearheaded the development of a one-of-a-kind x-ray microscope with novel nanofocusing optics called multilayer Laue lenses.

Comparisons between conventional optical imaging and parametric indirect microscopic imaging on human skin detection

NASA Astrophysics Data System (ADS)

Liu, Guoyan; Gao, Kun; Liu, Xuefeng; Ni, Guoqiang

2016-10-01

We report a new method, polarization parameters indirect microscopic imaging with a high transmission infrared light source, to detect the morphology and component of human skin. A conventional reflection microscopic system is used as the basic optical system, into which a polarization-modulation mechanics is inserted and a high transmission infrared light source is utilized. The near-field structural characteristics of human skin can be delivered by infrared waves and material coupling. According to coupling and conduction physics, changes of the optical wave parameters can be calculated and curves of the intensity of the image can be obtained. By analyzing the near-field polarization parameters in nanoscale, we can finally get the inversion images of human skin. Compared with the conventional direct optical microscope, this method can break diffraction limit and achieve a super resolution of sub-100nm. Besides, the method is more sensitive to the edges, wrinkles, boundaries and impurity particles.

Stable and simple quantitative phase-contrast imaging by Fresnel biprism

NASA Astrophysics Data System (ADS)

Ebrahimi, Samira; Dashtdar, Masoomeh; Sánchez-Ortiga, Emilio; Martínez-Corral, Manuel; Javidi, Bahram

2018-03-01

Digital holographic (DH) microscopy has grown into a powerful nondestructive technique for the real-time study of living cells including dynamic membrane changes and cell fluctuations in nanometer and sub-nanometer scales. The conventional DH microscopy configurations require a separately generated coherent reference wave that results in a low phase stability and a necessity to precisely adjust the intensity ratio between two overlapping beams. In this work, we present a compact, simple, and very stable common-path DH microscope, employing a self-referencing configuration. The microscope is implemented by a diode laser as the source and a Fresnel biprism for splitting and recombining the beams simultaneously. In the overlapping area, linear interference fringes with high contrast are produced. The frequency of the interference pattern could be easily adjusted by displacement of the biprism along the optical axis without a decrease in fringe contrast. To evaluate the validity of the method, the spatial noise and temporal stability of the setup are compared with the common off-axis DH microscope based on a Mach-Zehnder interferometer. It is shown that the proposed technique has low mechanical noise as well as superb temporal stability with sub-nanometer precision without any external vibration isolation. The higher temporal stability improves the capabilities of the microscope for studying micro-object fluctuations, particularly in the case of biological specimens. Experimental results are presented using red blood cells and silica microspheres to demonstrate the system performance.

Mapping large extensions of flat dentin through digital microscopy: introduction to the method and possible applications.

PubMed

Reis, Claudia; De-Deus, Gustavo; Marins, Juliana; Fidel, Sandra; Fidel, Rivail; Paciornik, Sidnei

2012-08-01

To introduce a mapping method to characterize large dentin surfaces using digital microscopy and to discuss the advantages and possible applications of the method. Twenty unerupted third molars were sectioned transversally exposing coronal dentin surfaces. The microscopic mosaic method was used to generate a large field image with the resolution necessary to measure characteristics of dentin tubules. The AxioVision 4.7 software was used to control a motorized optical microscope and the process of acquiring approximately 400 small images to generate each dentin mosaic. An image analysis routine measured the number of tubules (NT) and the ratio between the total area of tubules and the area of the mosaic - the area fraction (AF) - of each mosaic. An automatic procedure transformed the mosaic image into a color map, providing a direct visual representation of tubule density through colors. The dentin maps were used for a comparative qualitative analysis of tubule density distribution of each sample. The results for NT (92450 to 196029 tubules/sample) and AF (4.12% to 11.10%) demonstrated a wide variation among dentin samples. The maps confirmed the microstructure variety, also revealing strong local variations in tubule density within each sample. The mapping method was able to perform dentin morphology characterization and is a valuable tool for producing a baseline for dentin adhesion studies. The method could be also useful in determining the real contribution of dentin structures to the final adhesion quality.

Modified-Signed-Digit Optical Computing Using Fan-Out

NASA Technical Reports Server (NTRS)

Liu, Hua-Kuang; Zhou, Shaomin; Yeh, Pochi

1996-01-01

Experimental optical computing system containing optical fan-out elements implements modified signed-digit (MSD) arithmetic and logic. In comparison with previous optical implementations of MSD arithmetic, this one characterized by larger throughput, greater flexibility, and simpler optics.

An optical/digital processor - Hardware and applications

NASA Technical Reports Server (NTRS)

Casasent, D.; Sterling, W. M.

1975-01-01

A real-time two-dimensional hybrid processor consisting of a coherent optical system, an optical/digital interface, and a PDP-11/15 control minicomputer is described. The input electrical-to-optical transducer is an electron-beam addressed potassium dideuterium phosphate (KD2PO4) light valve. The requirements and hardware for the output optical-to-digital interface, which is constructed from modular computer building blocks, are presented. Initial experimental results demonstrating the operation of this hybrid processor in phased-array radar data processing, synthetic-aperture image correlation, and text correlation are included. The applications chosen emphasize the role of the interface in the analysis of data from an optical processor and possible extensions to the digital feedback control of an optical processor.

Ultrafast superresolution fluorescence imaging with spinning disk confocal microscope optics.

PubMed

Hayashi, Shinichi; Okada, Yasushi

2015-05-01

Most current superresolution (SR) microscope techniques surpass the diffraction limit at the expense of temporal resolution, compromising their applications to live-cell imaging. Here we describe a new SR fluorescence microscope based on confocal microscope optics, which we name the spinning disk superresolution microscope (SDSRM). Theoretically, the SDSRM is equivalent to a structured illumination microscope (SIM) and achieves a spatial resolution of 120 nm, double that of the diffraction limit of wide-field fluorescence microscopy. However, the SDSRM is 10 times faster than a conventional SIM because SR signals are recovered by optical demodulation through the stripe pattern of the disk. Therefore a single SR image requires only a single averaged image through the rotating disk. On the basis of this theory, we modified a commercial spinning disk confocal microscope. The improved resolution around 120 nm was confirmed with biological samples. The rapid dynamics of micro-tubules, mitochondria, lysosomes, and endosomes were observed with temporal resolutions of 30-100 frames/s. Because our method requires only small optical modifications, it will enable an easy upgrade from an existing spinning disk confocal to a SR microscope for live-cell imaging. © 2015 Hayashi and Okada. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

HIGH TEMPERATURE MICROSCOPE AND FURNACE

DOEpatents

Olson, D.M.

1961-01-31

A high-temperature microscope is offered. It has a reflecting optic situated above a molten specimen in a furnace and reflecting the image of the same downward through an inert optic member in the floor of the furnace, a plurality of spaced reflecting plane mirrors defining a reflecting path around the furnace, a standard microscope supported in the path of and forming the end terminus of the light path.

Fiber optic accelerometer

NASA Technical Reports Server (NTRS)

August, R. R.

1981-01-01

Low-cost, rugged lightweight accelerometer has been developed that converts mechanical motion into digitized optical outputs and is immune to electromagnetic and electrostatic interferences. Instrument can be placed in hostile environment, such as engine under test, and output led out through miscellany of electrical fields, high temperatures, etc., by optic fiber cables to benign environment of test panel. There, digitized optical signals can be converted to electrical signals for use in standard electrical equipment or used directly in optical devices, such as optical digital computer.

Sedimentological Investigations of the Martian Surface using the Mars 2001 Robotic Arm Camera and MECA Optical Microscope

NASA Technical Reports Server (NTRS)

Rice, J. W., Jr.; Smith, P. H.; Marshall, J. R.

1999-01-01

The first microscopic sedimentological studies of the Martian surface will commence with the landing of the Mars Polar Lander (MPL) December 3, 1999. The Robotic Arm Camera (RAC) has a resolution of 25 um/p which will permit detailed micromorphological analysis of surface and subsurface materials. The Robotic Ann will be able to dig up to 50 cm below the surface. The walls of the trench will also be inspected by RAC to look for evidence of stratigraphic and / or sedimentological relationships. The 2001 Mars Lander will build upon and expand the sedimentological research begun by the RAC on MPL. This will be accomplished by: (1) Macroscopic (dm to cm): Descent Imager, Pancam, RAC; (2) Microscopic (mm to um RAC, MECA Optical Microscope (Figure 2), AFM This paper will focus on investigations that can be conducted by the RAC and MECA Optical Microscope.

Hard X-Ray Scanning Microscope with Multilayer Laue Lens Nanofocusing Optics

ScienceCinema

Nazaretski, Evgeny

2018-06-13

Evgeny Nazaretski, a physicist at Brookhaven Lab’s National Synchrotron Light Source II, spearheaded the development of a one-of-a-kind x-ray microscope with novel nanofocusing optics called multilayer Laue lenses.

The measurement of sperm motility by the fibre optic Doppler anemometer as a prediction of bovine fertility

NASA Astrophysics Data System (ADS)

Bullock, J. G.; Ross, D. A.

The fibre optic Doppler anemometer (FODA) has been used to develop an accurate quantitative method of routinely assessing bull fertility. This method is of importance to the artificial insemination industry because the present qualitative estimation, performed by viewing semen using a microscope, can only set broad limits of quality. Laser light from the FODA was directed into diluted semen samples and the back scattered light was measured. A digital correlator was used to calculate the signal correlation of the back scattered light. The resultant data curves were interpreted in terms of the collective motility and swimming speed of the spermatozoa using a microcomputer. These two parameters are accepted as being indicative of fertility. The accuracy of this method is demonstrated by examination of results obtained in an experiment where enzymes, thought to alter fertility, were added to semen. The effect of the enzymes on the swimming speed and motility was clearly demonstrated.

Digital optical conversion module

DOEpatents

Kotter, D.K.; Rankin, R.A.

1988-07-19

A digital optical conversion module used to convert an analog signal to a computer compatible digital signal including a voltage-to-frequency converter, frequency offset response circuitry, and an electrical-to-optical converter. Also used in conjunction with the digital optical conversion module is an optical link and an interface at the computer for converting the optical signal back to an electrical signal. Suitable for use in hostile environments having high levels of electromagnetic interference, the conversion module retains high resolution of the analog signal while eliminating the potential for errors due to noise and interference. The module can be used to link analog output scientific equipment such as an electrometer used with a mass spectrometer to a computer. 2 figs.

Digital optical conversion module

DOEpatents

Kotter, Dale K.; Rankin, Richard A.

1991-02-26

A digital optical conversion module used to convert an analog signal to a computer compatible digital signal including a voltage-to-frequency converter, frequency offset response circuitry, and an electrical-to-optical converter. Also used in conjunction with the digital optical conversion module is an optical link and an interface at the computer for converting the optical signal back to an electrical signal. Suitable for use in hostile environments having high levels of electromagnetic interference, the conversion module retains high resolution of the analog signal while eliminating the potential for errors due to noise and interference. The module can be used to link analog output scientific equipment such as an electrometer used with a mass spectrometer to a computer.

Virtual microscopy in a veterinary curriculum.

PubMed

Sims, Michael H; Mendis-Handagama, Chamindrani; Moore, Robert N

2007-01-01

Teaching faculty in the University of Tennessee College of Veterinary Medicine assist students in their professional education by providing a new way of viewing microscopic slides digitally. Faculty who teach classes in which glass slides are used participate in a program called Virtual Microscopy. Glass slides are digitized using a state-of-the-art integrated system, and a personal computer functions as the "microscope." Additionally, distribution of the interactive images is enhanced because they are available to students online. The digital slide offers equivalent quality and resolution to the original glass slide viewed on a microscope and has several additional advantages over microscopes. Students can choose to examine the entire slide at any of several objectives; they are able to access the slides (called WebSlides) from the college's server, using either Internet Explorer or a special browser developed by Bacus Laboratories, Inc.,(a) called the WebSlide browser, which lets the student simultaneously view a low-objective image and one or two high-objective images of the same slide. The student can "move the slide" by clicking and dragging the image to a new location. Easy archiving, annotation of images, and Web conferencing are additional features of the system.

The impact of the condenser on cytogenetic image quality in digital microscope system.

PubMed

Ren, Liqiang; Li, Zheng; Li, Yuhua; Zheng, Bin; Li, Shibo; Chen, Xiaodong; Liu, Hong

2013-01-01

Optimizing operational parameters of the digital microscope system is an important technique to acquire high quality cytogenetic images and facilitate the process of karyotyping so that the efficiency and accuracy of diagnosis can be improved. This study investigated the impact of the condenser on cytogenetic image quality and system working performance using a prototype digital microscope image scanning system. Both theoretical analysis and experimental validations through objectively evaluating a resolution test chart and subjectively observing large numbers of specimen were conducted. The results show that the optimal image quality and large depth of field (DOF) are simultaneously obtained when the numerical aperture of condenser is set as 60%-70% of the corresponding objective. Under this condition, more analyzable chromosomes and diagnostic information are obtained. As a result, the system shows higher working stability and less restriction for the implementation of algorithms such as autofocusing especially when the system is designed to achieve high throughput continuous image scanning. Although the above quantitative results were obtained using a specific prototype system under the experimental conditions reported in this paper, the presented evaluation methodologies can provide valuable guidelines for optimizing operational parameters in cytogenetic imaging using the high throughput continuous scanning microscopes in clinical practice.

A compact CCD-monitored atomic force microscope with optical vision and improved performances.

PubMed

Mingyue, Liu; Haijun, Zhang; Dongxian, Zhang

2013-09-01

A novel CCD-monitored atomic force microscope (AFM) with optical vision and improved performances has been developed. Compact optical paths are specifically devised for both tip-sample microscopic monitoring and cantilever's deflection detecting with minimized volume and optimal light-amplifying ratio. The ingeniously designed AFM probe with such optical paths enables quick and safe tip-sample approaching, convenient and effective tip-sample positioning, and high quality image scanning. An image stitching method is also developed to build a wider-range AFM image under monitoring. Experiments show that this AFM system can offer real-time optical vision for tip-sample monitoring with wide visual field and/or high lateral optical resolution by simply switching the objective; meanwhile, it has the elegant performances of nanometer resolution, high stability, and high scan speed. Furthermore, it is capable of conducting wider-range image measurement while keeping nanometer resolution. Copyright © 2013 Wiley Periodicals, Inc.

Optical anisotropy of the human cornea determined with a polarizing microscope.

PubMed

Bone, Richard A; Draper, Grenville

2007-12-01

We have investigated the optical anisotropy of the human cornea using a polarizing microscope normally used for optical mineralogy studies. The central part of the cornea was removed from 14 eyes (seven donors). With the sample placed on the microscope stage, we consistently observed hyperbolic isogyres characteristic of a negative biaxial material. The angle between the optic axes, generally similar in both eyes, ranged from 12 degrees to 40 degrees (mean+/-SD=31 degrees +/-8 degrees ). The optic axial plane always inclined downward in the nasal direction at 1 degrees -45 degrees below the horizontal (mean+/-SD=22+/-13 degrees ). The retardance produced by the corneas was estimated to be less than 200 nm. In conclusion, the human cornea possesses the anisotropy of a negative biaxial material. Both the angle between the optic axes and the retardance were fairly constant among the majority of samples, suggestive of uniformity in corneal structure.

Automated SEM Modal Analysis Applied to the Diogenites

NASA Technical Reports Server (NTRS)

Bowman, L. E.; Spilde, M. N.; Papike, James J.

1996-01-01

Analysis of volume proportions of minerals, or modal analysis, is routinely accomplished by point counting on an optical microscope, but the process, particularly on brecciated samples such as the diogenite meteorites, is tedious and prone to error by misidentification of very small fragments, which may make up a significant volume of the sample. Precise volume percentage data can be gathered on a scanning electron microscope (SEM) utilizing digital imaging and an energy dispersive spectrometer (EDS). This form of automated phase analysis reduces error, and at the same time provides more information than could be gathered using simple point counting alone, such as particle morphology statistics and chemical analyses. We have previously studied major, minor, and trace-element chemistry of orthopyroxene from a suite of diogenites. This abstract describes the method applied to determine the modes on this same suite of meteorites and the results of that research. The modal abundances thus determined add additional information on the petrogenesis of the diogenites. In addition, low-abundance phases such as spinels were located for further analysis by this method.

The long road to the use of microscope in clinical medicine in vivo: from early pioneering proposals to the modern perspectives of optical biopsy.

PubMed

Ponti, Giovanni; Muscatello, Umberto; Sgantzos, Markos

2015-01-01

For a long period the scientists did not recognized the potentialities of the compound microscope in medicine. Only few scientists recognized the potentialities of the microscope for the medicine; among them G. Campani who proposed the utilization of his microscope to investigate the skin lesions directly on the patient. The proposal was illustrated in a letter Acta Eruditorum of 1686. The recent development of optical techniques, capable of providing in-focus images of an object from different planes with high spatial resolution, significantly increased the diagnostic potential of the microscope directly on the patient.

3D pore-type digital rock modeling of natural gas hydrate for permafrost and numerical simulation of electrical properties

NASA Astrophysics Data System (ADS)

Dong, Huaimin; Sun, Jianmeng; Lin, Zhenzhou; Fang, Hui; Li, Yafen; Cui, Likai; Yan, Weichao

2018-02-01

Natural gas hydrate is being considered as an alternative energy source for sustainable development and has become a focus of research throughout the world. In this paper, based on CT scanning images of hydrate reservoir rocks, combined with the microscopic distribution of hydrate, a diffusion limited aggregation (DLA) model was used to construct 3D hydrate digital rocks of different distribution types, and the finite-element method was used to simulate their electrical characteristics in order to study the influence of different hydrate distribution types, hydrate saturation and formation of water salinity on electrical properties. The results show that the hydrate digital rocks constructed using the DLA model can be used to characterize the microscopic distribution of different types of hydrates. Under the same conditions, the resistivity of the adhesive hydrate digital rock is higher than the cemented and scattered type digital rocks, and the resistivity of the scattered hydrate digital rock is the smallest among the three types. Besides, the difference in the resistivity of the different types of hydrate digital rocks increases with an increase in hydrate saturation, especially when the saturation is larger than 55%, and the rate of increase of each of the hydrate types is quite different. Similarly, the resistivity of the three hydrate types decreases with an increase in the formation of water salinity. The single distribution hydrate digital rock constructed, combined with the law of microscopic distribution and influence of saturation on the electrical properties, can effectively improve the accuracy of logging identification of hydrate reservoirs and is of great significance for the estimation of hydrate reserves.

Design of an imaging microscope for soft X-ray applications

NASA Astrophysics Data System (ADS)

Hoover, Richard B.; Shealy, David L.; Gabardi, David R.; Walker, Arthur B. C., Jr.; Lindblom, Joakim F.

1988-01-01

An imaging soft X-ray microscope with a spatial resolution of 0.1 micron and normal incidence multilayer optics is discussed. The microscope has a Schwarzschild configuration, which consists of two concentric spherical mirrors with radii of curvature which minimize third-order spherical aberration, coma, and astigmatism. The performance of the Stanford/MSFC Cassegrain X-ray telescope and its relevance to the present microscope are addressed. A ray tracing analysis of the optical system indicates that diffraction-limited performance can be expected for an object height of 0.2 mm.

Sub-25-nm laboratory x-ray microscopy using a compound Fresnel zone plate.

PubMed

von Hofsten, Olov; Bertilson, Michael; Reinspach, Julia; Holmberg, Anders; Hertz, Hans M; Vogt, Ulrich

2009-09-01

Improving the resolution in x-ray microscopes is of high priority to enable future applications in nanoscience. However, high-resolution zone-plate optics often have low efficiency, which makes implementation in laboratory microscopes difficult. We present a laboratory x-ray microscope based on a compound zone plate. The compound zone plate utilizes multiple diffraction orders to achieve high resolution while maintaining reasonable efficiency. We analyze the illumination conditions necessary for this type of optics in order to suppress stray light and demonstrate microscopic imaging resolving 25 nm features.

Digital differential confocal microscopy based on spatial shift transformation.

PubMed

Liu, J; Wang, Y; Liu, C; Wilson, T; Wang, H; Tan, J

2014-11-01

Differential confocal microscopy is a particularly powerful surface profilometry technique in industrial metrology due to its high axial sensitivity and insensitivity to noise. However, the practical implementation of the technique requires the accurate positioning of point detectors in three-dimensions. We describe a simple alternative based on spatial transformation of a through-focus series of images obtained from a homemade beam scanning confocal microscope. This digital differential confocal microscopy approach is described and compared with the traditional Differential confocal microscopy approach. The ease of use of the digital differential confocal microscopy system is illustrated by performing measurements on a 3D standard specimen. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

A system for the automatic measurement and digital display of systolic and diastolic blood pressures

NASA Technical Reports Server (NTRS)

Schulze, A. E.

1971-01-01

Basic components of system are - occluding cuff with mounted cuff microscope, cuff pump deflator, pressure transducer, preamplifier unit, electrocardiograph machine, an analog to digital convertor unit, and digital display unit. System utilizes indirect auscultatory method, based on Korotkoff sounds, for measurement.

Classification of Salmonella serotypes with hyperspectral microscope imagery

USDA-ARS?s Scientific Manuscript database

Previous research has demonstrated an optical method with acousto-optic tunable filter (AOTF) based hyperspectral microscope imaging (HMI) had potential for classifying gram-negative from gram-positive foodborne pathogenic bacteria rapidly and nondestructively with a minimum sample preparation. In t...

Internal scanning method as unique imaging method of optical vortex scanning microscope

NASA Astrophysics Data System (ADS)

Popiołek-Masajada, Agnieszka; Masajada, Jan; Szatkowski, Mateusz

2018-06-01

The internal scanning method is specific for the optical vortex microscope. It allows to move the vortex point inside the focused vortex beam with nanometer resolution while the whole beam stays in place. Thus the sample illuminated by the focused vortex beam can be scanned just by the vortex point. We show that this method enables high resolution imaging. The paper presents the preliminary experimental results obtained with the first basic image recovery procedure. A prospect of developing more powerful tools for topography recovery with the optical vortex scanning microscope is discussed shortly.

High-resolution microscope for tip-enhanced optical processes in ultrahigh vacuum

NASA Astrophysics Data System (ADS)

Steidtner, Jens; Pettinger, Bruno

2007-10-01

An optical microscope based on tip-enhanced optical processes that can be used for studies on adsorbates as well as thin layers and nanostructures is presented. The microscope provides chemical and topographic informations with a resolution of a few nanometers and can be employed in ultrahigh vacuum as well as gas phase. The construction involves a number of improvements compared to conventional instruments. The central idea is to mount, within an UHV system, an optical platform with all necessary optical elements to a rigid frame that also carries the scanning tunneling microscope unit and to integrate a high numerical aperture parabolic mirror between the scanning probe microscope head and the sample. The parabolic mirror serves to focus the incident light and to collect a large fraction of the scattered light. The first experimental results of Raman measurements on silicon samples as well as brilliant cresyl blue layers on single crystalline gold and platinum surfaces in ultrahigh vacuum are presented. For dye adsorbates a Raman enhancement of ˜106 and a net signal gain of up to 4000 was observed. The focus diameter (˜λ/2) was measured by Raman imaging the focal region on a Si surface. The requirements of the parabolic mirror in terms of alignment accuracy were experimentally determined as well.

U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

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

Prabhakaran, Ramprashad; Joshi, Vineet V.; Rhodes, Mark A.

2016-10-01

The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

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

Prabhakaran, Ramprashad; Joshi, Vineet V.; Rhodes, Mark A.

2016-03-30

The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

Micropaleontological studies of lunar and terrestrial precambrian materials

NASA Technical Reports Server (NTRS)

Schope, J. W.

1974-01-01

Optical microscopic and scanning electron microscopic studies of rock chips and dust returned by Apollo 14, 15, 16, and 17 are analyzed along with optical microscopic studies of petrographic thin sections of breccias and basalts returned by Apollo 14, 15, and 16. Results show no evidence of modern or fossil lunar organisms. The lunar surface is now, and apparently has been throughout the geologic past, inimical to known biologic systems.

Increasing Student Understanding of Microscope Optics by Building and Testing the Limits of Simple, Hand-Made Model Microscopes†

PubMed Central

Drace, Kevin; Couch, Brett; Keeling, Patrick J.

2012-01-01

The ability to effectively use a microscope to observe microorganisms is a crucial skill required for many disciplines within biology, especially general microbiology and cell biology. A basic understanding of the optical properties of light microscopes is required for students to use microscopes effectively, but this subject can also be a challenge to make personally interesting to students. To explore basic optical principles of magnification and resolving power in a more engaging and hands-on fashion, students constructed handmade lenses and microscopes based on Antony van Leeuwenhoek’s design using simple materials—paper, staples, glass, and adhesive putty. Students determined the power of their lenses using a green laser pointer to magnify a copper grid of known size, which also allowed students to examine variables affecting the power and resolution of a lens such as diameter, working distance, and wavelength of light. To assess the effectiveness of the laboratory’s learning objectives, four sections of a general microbiology course were given a brief pre-activity assessment quiz to determine their background knowledge on the subject. One week after the laboratory activity, students were given the same quiz (unannounced) under similar conditions. Students showed significant gains in their understanding of microscope optics. PMID:23653781

A hybrid scanning force and light microscope for surface imaging and three-dimensional optical sectioning in differential interference contrast.

PubMed

Stemmer, A

1995-04-01

The design of a scanned-cantilever-type force microscope is presented which is fully integrated into an inverted high-resolution video-enhanced light microscope. This set-up allows us to acquire thin optical sections in differential interference contrast (DIC) or polarization while the force microscope is in place. Such a hybrid microscope provides a unique platform to study how cell surface properties determine, or are affected by, the three-dimensional dynamic organization inside the living cell. The hybrid microscope presented in this paper has proven reliable and versatile for biological applications. It is the only instrument that can image a specimen by force microscopy and high-power DIC without having either to translate the specimen or to remove the force microscope. Adaptation of the design features could greatly enhance the suitability of other force microscopes for biological work.

Manual stage acquisition and interactive display of digital slides in histopathology.

PubMed

Gherardi, Alessandro; Bevilacqua, Alessandro

2014-07-01

More powerful PC architectures, high-resolution cameras working at increasing frame rates, and more and more accurate motorized microscopes have boosted new applications in the field of biomedicine and medical imaging. In histopathology, the use of digital slides (DSs) imaging through dedicated hardware for digital pathology is increasing for several reasons: digital annotation of suspicious lesions, recorded clinical history, and telepathology as a collaborative environment. In this paper, we propose the first method known in the literature for real-time whole slide acquisition and displaying conceived for conventional nonautomated microscopes. Differently from DS scanner, our software enables biologists and histopathologists to build and view the DS in real time while inspecting the sample, as they are accustomed to. In addition, since our approach is compliant with existing common microscope positions, provided with camera and PC, this could contribute to disseminate the whole slide technology in the majority of small labs not endowed with DS hardware facilities. Experiments performed with different histologic specimens (referring to tumor tissues of different body parts as well as to tumor cells), acquired under different setup conditions and devices, prove the effectiveness of our approach both in terms of quality and speed performances.

High-resolution digital brain atlases: a Hubble telescope for the brain.

PubMed

Jones, Edward G; Stone, James M; Karten, Harvey J

2011-05-01

We describe implementation of a method for digitizing at microscopic resolution brain tissue sections containing normal and experimental data and for making the content readily accessible online. Web-accessible brain atlases and virtual microscopes for online examination can be developed using existing computer and internet technologies. Resulting databases, made up of hierarchically organized, multiresolution images, enable rapid, seamless navigation through the vast image datasets generated by high-resolution scanning. Tools for visualization and annotation of virtual microscope slides enable remote and universal data sharing. Interactive visualization of a complete series of brain sections digitized at subneuronal levels of resolution offers fine grain and large-scale localization and quantification of many aspects of neural organization and structure. The method is straightforward and replicable; it can increase accessibility and facilitate sharing of neuroanatomical data. It provides an opportunity for capturing and preserving irreplaceable, archival neurohistological collections and making them available to all scientists in perpetuity, if resources could be obtained from hitherto uninterested agencies of scientific support. © 2011 New York Academy of Sciences.

Nailfold capillaroscopy by digital microscope in an Indian population with systemic sclerosis.

PubMed

Bhakuni, Darshan S; Vasdev, Vivek; Garg, M K; Narayanan, Krishanan; Jain, Rahul; Mullick, Gautam

2012-02-01

Nailfold capillaroscopy (NFC) is a simple, non-invasive method with exceptional predictive value for the analysis of microvascular abnormalities, especially in systemic sclerosis (SSc) but remains underutilized due to cost factors of the nailfold videocapillaroscope, lack of expertise and availability issues. The aim of this study was to establish the utility of an inexpensive digital microscope to study NFC changes in SSc in correlation with disease subsets and extent of skin involvement. Twenty-two diffuse cutaneous SSc (DSS), 20 limited cutaneous SSc (LSS) patients and 42 controls were evaluated with NFC using a digital microscope at 30× and 100× magnification. Digital micrographs were used to study qualitative and quantitative changes in microvasculature. The capillary density was significantly less in all cases of SSc as compared to controls (5.3 ± 1.4 vs. 8.7 ± 1.2; P < 0.00001). Disorganized architecture was much more prevalent in DSS versus LSS (86.4%vs. 25%). The vascular deletion score (VDS) was significantly higher in DSS as compared to LSS (P < 0.0001). Scleroderma pattern (SP) was seen in 18 (81.9%) and 15 (75%) of patients with DSS and LSS, respectively. Only 4% of normal subjects showed non-specific pattern and none showed SP. The mean modified Rodnan skin score (MRSS) was positively correlated with vascular deletion score (r = 0.572; P < 0.001) and negatively with capillary density (r = -0.8; P < 0.001). Nailfold capillaroscopy changes in SSc are related to disease subset and MRSS. NFC with digital microscope is a simplified, inexpensive, outpatient procedure with results comparable to previous studies. © 2011 The Authors. International Journal of Rheumatic Diseases © 2011 Asia Pacific League of Associations for Rheumatology and Blackwell Publishing Asia Pty Ltd.

Coherent time-stretch transformation for real-time capture of wideband signals.

PubMed

Buckley, Brandon W; Madni, Asad M; Jalali, Bahram

2013-09-09

Time stretch transformation of wideband waveforms boosts the performance of analog-to-digital converters and digital signal processors by slowing down analog electrical signals before digitization. The transform is based on dispersive Fourier transformation implemented in the optical domain. A coherent receiver would be ideal for capturing the time-stretched optical signal. Coherent receivers offer improved sensitivity, allow for digital cancellation of dispersion-induced impairments and optical nonlinearities, and enable decoding of phase-modulated optical data formats. Because time-stretch uses a chirped broadband (>1 THz) optical carrier, a new coherent detection technique is required. In this paper, we introduce and demonstrate coherent time stretch transformation; a technique that combines dispersive Fourier transform with optically broadband coherent detection.

Wavefront coding for fast, high-resolution light-sheet microscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Olarte, Omar E.; Licea-Rodriguez, Jacob; Loza-Alvarez, Pablo

2017-02-01

Some biological experiments demand the observation of dynamics processes in 3D with high spatiotemporal resolution. The use of wavefront coding to extend the depth-of-field (DOF) of the collection arm of a light-sheet microscope is an interesting alternative for fast 3D imaging. Under this scheme, the 3D features of the sample are captured at high volumetric rates while the light sheet is swept rapidly within the extended DOF. The DOF is extended by coding the pupil function of the imaging lens by using a custom-designed phase mask. A posterior restoration step is required to decode the information of the captured images based on the applied phase mask [1]. This hybrid optical-digital approach is known as wavefront coding (WFC). Previously, we have demonstrated this method for performing fast 3D imaging of biological samples at medium resolution [2]. In this work, we present the extension of this approach for high-resolution microscopes. Under these conditions, the effective DOF of a standard high NA objective is of a few micrometers. Here we demonstrate that by the use of WFC, we can extend the DOF more than one order of magnitude keeping the high-resolution imaging. This is demonstrated for two designed phase masks using Zebrafish and C. elegans samples. [1] Olarte, O.E., Andilla, J., Artigas, D., and Loza-Alvarez, P., "Decoupled Illumination-Detection Microscopy. Selected Optics in Year 2105," in Optics and Photonics news 26, p. 41 (2015). [2] Olarte, O.E., Andilla, J., Artigas, D., and Loza-Alvarez, P., "Decoupled illumination detection in light sheet microscopy for fast volumetric imaging," Optica 2(8), 702 (2015).

Microscopic theory of linear light scattering from mesoscopic media and in near-field optics.

PubMed

Keller, Ole

2005-08-01

On the basis of quantum mechanical response theory a microscopic propagator theory of linear light scattering from mesoscopic systems is presented. The central integral equation problem is transferred to a matrix equation problem by discretization in transitions between pairs of (many-body) energy eigenstates. The local-field calculation which appears from this approach is valid down to the microscopic region. Previous theories based on the (macroscopic) dielectric constant concept make use of spatial (geometrical) discretization and cannot in general be trusted on the mesoscopic length scale. The present theory can be applied to light scattering studies in near-field optics. After a brief discussion of the macroscopic integral equation problem a microscopic potential description of the scattering process is established. In combination with the use of microscopic electromagnetic propagators the formalism allows one to make contact to the macroscopic theory of light scattering and to the spatial photon localization problem. The quantum structure of the microscopic conductivity response tensor enables one to establish a clear physical picture of the origin of local-field phenomena in mesoscopic and near-field optics. The Huygens scalar propagator formalism is revisited and its generality in microscopic physics pointed out.

Water window imaging x ray microscope

NASA Technical Reports Server (NTRS)

Hoover, Richard B. (Inventor)

1992-01-01

A high resolution x ray microscope for imaging microscopic structures within biological specimens has an optical system including a highly polished primary and secondary mirror coated with identical multilayer coatings, the mirrors acting at normal incidence. The coatings have a high reflectivity in the narrow wave bandpass between 23.3 and 43.7 angstroms and have low reflectivity outside of this range. The primary mirror has a spherical concave surface and the secondary mirror has a spherical convex surface. The radii of the mirrors are concentric about a common center of curvature on the optical axis of the microscope extending from the object focal plane to the image focal plane. The primary mirror has an annular configuration with a central aperture and the secondary mirror is positioned between the primary mirror and the center of curvature for reflecting radiation through the aperture to a detector. An x ray filter is mounted at the stage end of the microscope, and film sensitive to x rays in the desired band width is mounted in a camera at the image plane of the optical system. The microscope is mounted within a vacuum chamber for minimizing the absorption of x rays in air from a source through the microscope.

Microscopy with multimode fibers

NASA Astrophysics Data System (ADS)

Moser, Christophe; Papadopoulos, Ioannis; Farahi, Salma; Psaltis, Demetri

2013-04-01

Microscopes are usually thought of comprising imaging elements such as objectives and eye-piece lenses. A different type of microscope, used for endoscopy, consists of waveguiding elements such as fiber bundles, where each fiber in the bundle transports the light corresponding to one pixel in the image. Recently a new type of microscope has emerged that exploits the large number of propagating modes in a single multimode fiber. We have successfully produced fluorescence images of neural cells with sub-micrometer resolution via a 200 micrometer core multimode fiber. The method for achieving imaging consists of using digital phase conjugation to reproduce a focal spot at the tip of the multimode fiber. The image is formed by scanning the focal spot digitally and collecting the fluorescence point by point.

Quantitative optical imaging and sensing by joint design of point spread functions and estimation algorithms

NASA Astrophysics Data System (ADS)

Quirin, Sean Albert

The joint application of tailored optical Point Spread Functions (PSF) and estimation methods is an important tool for designing quantitative imaging and sensing solutions. By enhancing the information transfer encoded by the optical waves into an image, matched post-processing algorithms are able to complete tasks with improved performance relative to conventional designs. In this thesis, new engineered PSF solutions with image processing algorithms are introduced and demonstrated for quantitative imaging using information-efficient signal processing tools and/or optical-efficient experimental implementations. The use of a 3D engineered PSF, the Double-Helix (DH-PSF), is applied as one solution for three-dimensional, super-resolution fluorescence microscopy. The DH-PSF is a tailored PSF which was engineered to have enhanced information transfer for the task of localizing point sources in three dimensions. Both an information- and optical-efficient implementation of the DH-PSF microscope are demonstrated here for the first time. This microscope is applied to image single-molecules and micro-tubules located within a biological sample. A joint imaging/axial-ranging modality is demonstrated for application to quantifying sources of extended transverse and axial extent. The proposed implementation has improved optical-efficiency relative to prior designs due to the use of serialized cycling through select engineered PSFs. This system is demonstrated for passive-ranging, extended Depth-of-Field imaging and digital refocusing of random objects under broadband illumination. Although the serialized engineered PSF solution is an improvement over prior designs for the joint imaging/passive-ranging modality, it requires the use of multiple PSFs---a potentially significant constraint. Therefore an alternative design is proposed, the Single-Helix PSF, where only one engineered PSF is necessary and the chromatic behavior of objects under broadband illumination provides the necessary information transfer. The matched estimation algorithms are introduced along with an optically-efficient experimental system to image and passively estimate the distance to a test object. An engineered PSF solution is proposed for improving the sensitivity of optical wave-front sensing using a Shack-Hartmann Wave-front Sensor (SHWFS). The performance limits of the classical SHWFS design are evaluated and the engineered PSF system design is demonstrated to enhance performance. This system is fabricated and the mechanism for additional information transfer is identified.

An approach to the optical MSD adder

NASA Astrophysics Data System (ADS)

Takahashi, Hideya; Matsushita, Kenji; Shimizu, Eiji

1990-07-01

The intrinsic parallelism of optical elements for computation is presently taken fuller advantage of than heretofore possible through an optical implementation of the modified signed digit (MSD) number system, which yields carry-free addition and subtraction. In the present optical implementation of the MSD system, optical phase data are used to preclude negative value representation. Attention is given to an MSD adder array for addition operations on two n-digit trinary numbers; the output is composed of n + 1 trinary digits.

Fabrication and test of digital output interface devices for gas turbine electronic controls

NASA Technical Reports Server (NTRS)

Newirth, D. M.; Koenig, E. W.

1978-01-01

A program was conducted to develop an innovative digital output interface device, a digital effector with optical feedback of the fuel metering valve position, for future electronic controls for gas turbine engines. A digital effector (on-off solenoids driven directly by on-off signals from a digital electronic controller) with optical position feedback was fabricated, coupled with the fuel metering valve, and tested under simulated engine operating conditions. The testing indicated that a digital effector with optical position feedback is a suitable candidate, with proper development for future digital electronic gas turbine controls. The testing also identified several problem areas which would have to be overcome in a final production configuration.

Instant Grainification: Real-Time Grain-Size Analysis from Digital Images in the Field

NASA Astrophysics Data System (ADS)

Rubin, D. M.; Chezar, H.

2007-12-01

Over the past few years, digital cameras and underwater microscopes have been developed to collect in-situ images of sand-sized bed sediment, and software has been developed to measure grain size from those digital images (Chezar and Rubin, 2004; Rubin, 2004; Rubin et al., 2006). Until now, all image processing and grain- size analysis was done back in the office where images were uploaded from cameras and processed on desktop computers. Computer hardware has become small and rugged enough to process images in the field, which for the first time allows real-time grain-size analysis of sand-sized bed sediment. We present such a system consisting of weatherproof tablet computer, open source image-processing software (autocorrelation code of Rubin, 2004, running under Octave and Cygwin), and digital camera with macro lens. Chezar, H., and Rubin, D., 2004, Underwater microscope system: U.S. Patent and Trademark Office, patent number 6,680,795, January 20, 2004. Rubin, D.M., 2004, A simple autocorrelation algorithm for determining grain size from digital images of sediment: Journal of Sedimentary Research, v. 74, p. 160-165. Rubin, D.M., Chezar, H., Harney, J.N., Topping, D.J., Melis, T.S., and Sherwood, C.R., 2006, Underwater microscope for measuring spatial and temporal changes in bed-sediment grain size: USGS Open-File Report 2006-1360.

In vivo imaging of middle-ear and inner-ear microstructures of a mouse guided by SD-OCT combined with a surgical microscope

PubMed Central

Cho, Nam Hyun; Jang, Jeong Hun; Jung, Woonggyu; Kim, Jeehyun

2014-01-01

We developed an augmented-reality system that combines optical coherence tomography (OCT) with a surgical microscope. By sharing the common optical path in the microscope and OCT, we could simultaneously acquire OCT and microscope views. The system was tested to identify the middle-ear and inner-ear microstructures of a mouse. Considering the probability of clinical application including otorhinolaryngology, diseases such as middle-ear effusion were visualized using in vivo mouse and OCT images simultaneously acquired through the eyepiece of the surgical microscope during surgical manipulation using the proposed system. This system is expected to realize a new practical area of OCT application. PMID:24787787

(Gene sequencing by scanning molecular exciton microscopy)

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

Not Available

1991-01-01

This report details progress made in setting up a laboratory for optical microscopy of genes. The apparatus including a fluorescence microscope, a scanning optical microscope, various spectrometers, and supporting computers is described. Results in developing photon and exciton tips, and in preparing samples are presented. (GHH)

Design, assembly, and optical bench testing of a high-numerical-aperture miniature injection-molded objective for fiber-optic confocal reflectance microscopy.

PubMed

Chidley, Matthew D; Carlson, Kristen D; Richards-Kortum, Rebecca R; Descour, Michael R

2006-04-10

The design, analysis, assembly methods, and optical-bench test results for a miniature injection-molded plastic objective lens used in a fiber-optic confocal reflectance microscope are presented. The five-lens plastic objective was tested as a stand-alone optical system before its integration into a confocal microscope for in vivo imaging of cells and tissue. Changing the spacing and rotation of the individual optical elements can compensate for fabrication inaccuracies and improve performance. The system performance of the miniature objective lens is measured by use of an industry-accepted slanted-edge modulation transfer function (MTF) metric. An estimated Strehl ratio of 0.61 and a MTF value of 0.66 at the fiber-optic bundle Nyquist frequency have been obtained. The optical bench testing system is configured to permit interactive optical alignment during testing to optimize performance. These results are part of an effort to demonstrate the manufacturability of low-cost, high-performance biomedical optics for high-resolution in vivo imaging. Disposable endoscopic microscope objectives could help in vivo confocal microscopy technology mature to permit wide-scale clinical screening and detection of early cancers and precancerous lesions.

Miniaturized unified imaging system using bio-inspired fluidic lens

NASA Astrophysics Data System (ADS)

Tsai, Frank S.; Cho, Sung Hwan; Qiao, Wen; Kim, Nam-Hyong; Lo, Yu-Hwa

2008-08-01

Miniaturized imaging systems have become ubiquitous as they are found in an ever-increasing number of devices, such as cellular phones, personal digital assistants, and web cameras. Until now, the design and fabrication methodology of such systems have not been significantly different from conventional cameras. The only established method to achieve focusing is by varying the lens distance. On the other hand, the variable-shape crystalline lens found in animal eyes offers inspiration for a more natural way of achieving an optical system with high functionality. Learning from the working concepts of the optics in the animal kingdom, we developed bio-inspired fluidic lenses for a miniature universal imager with auto-focusing, macro, and super-macro capabilities. Because of the enormous dynamic range of fluidic lenses, the miniature camera can even function as a microscope. To compensate for the image quality difference between the central vision and peripheral vision and the shape difference between a solid-state image sensor and a curved retina, we adopted a hybrid design consisting of fluidic lenses for tunability and fixed lenses for aberration and color dispersion correction. A design of the world's smallest surgical camera with 3X optical zoom capabilities is also demonstrated using the approach of hybrid lenses.

Atmospheric scanning electron microscope for correlative microscopy.

PubMed

Morrison, Ian E G; Dennison, Clare L; Nishiyama, Hidetoshi; Suga, Mitsuo; Sato, Chikara; Yarwood, Andrew; O'Toole, Peter J

2012-01-01

The JEOL ClairScope is the first truly correlative scanning electron and optical microscope. An inverted scanning electron microscope (SEM) column allows electron images of wet samples to be obtained in ambient conditions in a biological culture dish, via a silicon nitride film window in the base. A standard inverted optical microscope positioned above the dish holder can be used to take reflected light and epifluorescence images of the same sample, under atmospheric conditions that permit biochemical modifications. For SEM, the open dish allows successive staining operations to be performed without moving the holder. The standard optical color camera used for fluorescence imaging can be exchanged for a high-sensitivity monochrome camera to detect low-intensity fluorescence signals, and also cathodoluminescence emission from nanophosphor particles. If these particles are applied to the sample at a suitable density, they can greatly assist the task of perfecting the correlation between the optical and electron images. Copyright © 2012 Elsevier Inc. All rights reserved.

Ultrasonographic analysis in vitro of parietal thickness of lower limb varicose veins.

PubMed

Bruschi, E; Como, G; Zuiani, C; Segatto, E; Rocco, M; Biasi, G; Bazzocchi, M

2006-09-01

The aim of this study was to evaluate the ability of ultrasound (US) to measure the parietal thickness of varicose veins. In a blind in vitro analysis, 28 great saphenous veins, obtained after stripping surgery from 28 patients with chronic venous insufficiency, were examined with a digital US scanner ATL-HDI5000, linear 5-1 to 2-MHz broadband probe, compound imaging technique and analogic-digital zooming. We obtained one to three progressive measurements for each vein wall (total 67 parietal thicknesses). The samples, fixed in formalin, were sent to the pathology laboratory: sections were obtained at the same level of the sonographic planes, and images were obtained by digital camera mounted on an optical microscope. Measurements obtained at histology were considered as the gold standard. K-statistic was applied to compare sonographic and histologic measurements. Considering only the hypoechoic wall portion, 29/29 (100%) diagnoses of hypotrophy (K=0.91), 19/22 (86%) diagnoses of normotrophy (K=0,47) and 12/16 (75%) diagnoses of hypertrophy (K=0.7) were obtained by sonography. In our preliminary experience, the in vitro study of varicose veins allows precise, at least morphological, detection of hypotrophic walls. If these preliminary data are confirmed in vivo, sonography could be used to discriminate patients eligible for conservative treatment instead of surgery.

Direct-to-digital holography and holovision

DOEpatents

Thomas, Clarence E.; Baylor, Larry R.; Hanson, Gregory R.; Rasmussen, David A.; Voelkl, Edgar; Castracane, James; Simkulet, Michelle; Clow, Lawrence

2000-01-01

Systems and methods for direct-to-digital holography are described. An apparatus includes a laser; a beamsplitter optically coupled to the laser; a reference beam mirror optically coupled to the beamsplitter; an object optically coupled to the beamsplitter, a focusing lens optically coupled to both the reference beam mirror and the object; and a digital recorder optically coupled to the focusing lens. A reference beam is incident upon the reference beam mirror at a non-normal angle, and the reference beam and an object beam are focused by the focusing lens at a focal plane of the digital recorder to form an image. The systems and methods provide advantages in that computer assisted holographic measurements can be made.

Virtual mask digital electron beam lithography

DOEpatents

Baylor, L.R.; Thomas, C.E.; Voelkl, E.; Moore, J.A.; Simpson, M.L.; Paulus, M.J.

1999-04-06

Systems and methods for direct-to-digital holography are described. An apparatus includes a laser; a beamsplitter optically coupled to the laser; a reference beam mirror optically coupled to the beamsplitter; an object optically coupled to the beamsplitter, a focusing lens optically coupled to both the reference beam mirror and the object; and a digital recorder optically coupled to the focusing lens. A reference beam is incident upon the reference beam mirror at a non-normal angle, and the reference beam and an object beam are focused by the focusing lens at a focal plane of the digital recorder to form an image. The systems and methods provide advantages in that computer assisted holographic measurements can be made. 5 figs.

Virtual mask digital electron beam lithography

DOEpatents

Baylor, Larry R.; Thomas, Clarence E.; Voelkl, Edgar; Moore, James A.; Simpson, Michael L.; Paulus, Michael J.

1999-01-01

Systems and methods for direct-to-digital holography are described. An apparatus includes a laser; a beamsplitter optically coupled to the laser; a reference beam mirror optically coupled to the beamsplitter; an object optically coupled to the beamsplitter, a focusing lens optically coupled to both the reference beam mirror and the object; and a digital recorder optically coupled to the focusing lens. A reference beam is incident upon the reference beam mirror at a non-normal angle, and the reference beam and an object beam are focused by the focusing lens at a focal plane of the digital recorder to form an image. The systems and methods provide advantages in that computer assisted holographic measurements can be made.

Optical digitizing and strategies to combine different views of an optical sensor

NASA Astrophysics Data System (ADS)

Duwe, Hans P.

1997-09-01

Non-contact digitization of objects and surfaces with optical sensors based on fringe or pattern projection in combination with a CCD-camera allows a representation of surfaces with pointclouds equals x, y, z data points. To digitize the total surface of an object, it is necessary to combine the different measurement data obtained by the optical sensor from different views. Depending on the size of the object and the required accuracy of the measured data, different sensor set-ups with handling system or a combination of linear and rotation axes are described. Furthermore, strategies to match the overlapping pointclouds of a digitized object are introduced. This is very important especially for the digitization of large objects like 1:1 car models, etc. With different sensor sizes, it is possible to digitize small objects like teeth, crowns, inlays, etc. with an overall accuracy of 20 micrometer as well as large objects like car models, with a total accuracy of 0.5 mm. The various applications in the field of optical digitization are described.

Design principles and applications of a cooled CCD camera for electron microscopy.

PubMed

Faruqi, A R

1998-01-01

Cooled CCD cameras offer a number of advantages in recording electron microscope images with CCDs rather than film which include: immediate availability of the image in a digital format suitable for further computer processing, high dynamic range, excellent linearity and a high detective quantum efficiency for recording electrons. In one important respect however, film has superior properties: the spatial resolution of CCD detectors tested so far (in terms of point spread function or modulation transfer function) are inferior to film and a great deal of our effort has been spent in designing detectors with improved spatial resolution. Various instrumental contributions to spatial resolution have been analysed and in this paper we discuss the contribution of the phosphor-fibre optics system in this measurement. We have evaluated the performance of a number of detector components and parameters, e.g. different phosphors (and a scintillator), optical coupling with lens or fibre optics with various demagnification factors, to improve the detector performance. The camera described in this paper, which is based on this analysis, uses a tapered fibre optics coupling between the phosphor and the CCD and is installed on a Philips CM12 electron microscope equipped to perform cryo-microscopy. The main use of the camera so far has been in recording electron diffraction patterns from two dimensional crystals of bacteriorhodopsin--from wild type and from different trapped states during the photocycle. As one example of the type of data obtained with the CCD camera a two dimensional Fourier projection map from the trapped O-state is also included. With faster computers, it will soon be possible to undertake this type of work on an on-line basis. Also, with improvements in detector size and resolution, CCD detectors, already ideal for diffraction, will be able to compete with film in the recording of high resolution images.

Wide spectral range confocal microscope based on endlessly single-mode fiber.

PubMed

Hubbard, R; Ovchinnikov, Yu B; Hayes, J; Richardson, D J; Fu, Y J; Lin, S D; See, P; Sinclair, A G

2010-08-30

We report an endlessly single mode, fiber-optic confocal microscope, based on a large mode area photonic crystal fiber. The microscope confines a very broad spectral range of excitation and emission wavelengths to a single spatial mode in the fiber. Single-mode operation over an optical octave is feasible. At a magnification of 10 and λ = 900 nm, its resolution was measured to be 1.0 μm (lateral) and 2.5 μm (axial). The microscope's use is demonstrated by imaging single photons emitted by individual InAs quantum dots in a pillar microcavity.

Structural properties of liposomes from digital holographic microscopy

NASA Astrophysics Data System (ADS)

Di Maio, Isabelle L.; Carl, Daniel; Langehanenberg, Patrik; Valenzuela, Stella M.; Battle, Andrew R.; Al Khazaaly, Sabah; Killingsworth, Murray; Kemper, Bjorn; von Bally, Gert; Martin, Donald K.

2006-01-01

We have constructed liposomes from L alpha Phosphatidylcholine (PC) lipids, which are biomimetic lipids similar to those present in the membranes of mammalian cells. We propose an advance in the use of liposomes, such as for drug delivery, to incorporate into the liposomal membranes transport proteins that have been extracted from the lipid membranes of mammalian cells. In this paper, we describe the usage of a novel optical microscope to characterize the nanomechanical properties of these liposomes. We have applied the technique of digital holographic microscopy, using an instrument recently developed at the University of Münster, Germany. This system enabled us to measure quantitatively the structural changes in liposomes. We have investigated the deformations of these biomimetic lipids comprising these liposomes by applying osmotic stresses, in order to gain insight into the membrane environment prior to incorporation of cloned membrane transport proteins. This control of the nanomechanical properties is important in the stresses transmitted to mechanosensitive ion channels that we have incorporated into the liposomal membranes. These liposomes provide transporting vesicles that respond to mechanical stresses, such as those that occur during implantation.

Digital learning programs - competition for the classical microscope?

PubMed

Schmidt, Peter

2013-01-01

The development of digital media has been impressive in recent years which is also among the reason for their increasing use in academic teaching. This is especially true for teaching Anatomy and Histology in the first two years in medical and dental curricula. Modern digital technologies allow for efficient, affordable and easily accessible distribution of histological images in high quality. Microscopy depends almost exclusively on such images. Since 20 years numerous digital teaching systems have been developed for this purpose. Respective developments have changed the ways students acquire knowledge and prepare for exams. Teaching staff should adapt lectures, seminars and labs accordingly. As a first step, a collection of high resolution digital microscopic slides was made available for students at the Friedrich-Schiller-University in Jena. The aim of the present study was to evaluate the importance of conventional light microscopy and related technologies in current and future medical and dental education aswell. A survey was done among 172 medical and dental students at the Friedrich-Schiller-University Jena. 51% of students use now frequently new digital media for learning histology in contrast to 5% in the year 2000 [1]. Digital media including Internet, CD- based learning combined with social networks successfully compete with classical light microscopy.

The Pathologist 2.0: An Update on Digital Pathology in Veterinary Medicine.

PubMed

Bertram, Christof A; Klopfleisch, Robert

2017-09-01

Using light microscopy to describe the microarchitecture of normal and diseased tissues has changed very little since the middle of the 19th century. While the premise of histologic analysis remains intact, our relationship with the microscope is changing dramatically. Digital pathology offers new forms of visualization, and delivery of images is facilitated in unprecedented ways. This new technology can untether us entirely from our light microscopes, with many pathologists already performing their jobs using virtual microscopy. Several veterinary colleges have integrated virtual microscopy in their curriculum, and some diagnostic histopathology labs are switching to virtual microscopy as their main tool for the assessment of histologic specimens. Considering recent technical advancements of slide scanner and viewing software, digital pathology should now be considered a serious alternative to traditional light microscopy. This review therefore intends to give an overview of the current digital pathology technologies and their potential in all fields of veterinary pathology (ie, research, diagnostic service, and education). A future integration of digital pathology in the veterinary pathologist's workflow seems to be inevitable, and therefore it is proposed that trainees should be taught in digital pathology to keep up with the unavoidable digitization of the profession.

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.

Thermal injury secondary to laparoscopic fiber-optic cables.

PubMed

Hindle, A Katharine; Brody, Fred; Hopkins, Vernon; Rosales, Greg; Gonzalez, Florencia; Schwartz, Arnold

2009-08-01

Laparoscopy requires a reliable light source to provide adequate visualization. However, thermal energy is produced as a by-product from the optical cable. This study attempts to quantify the degree of possible thermal damage secondary to the fiber-optic light source. Using a digital thermometer, temperature measurements were recorded at the tip of optical cables from five different light sources (Karl Storz, Inc., Tuttlingen, Germany). Temperature measurements were recorded with new and old bulbs. The tip of the cable was applied to surgical drapes and the time to charring was recorded. Subsequently, the tip of the optical cable was applied to a porcine model and tissue samples were obtained after varying amounts of time (5, 15, 30, 60, and 90 s). Sections of the damaged tissue were prepared for microscopic evaluation. Parameters for thermal injury included extent of epidermal, dermal, and subcutaneous fat damage and necrosis. The lateral extent and depth of injury were measured. The maximum temperature at the tip of the optical cable varied between 119.5 degrees C and 268.6 degrees C. When surgical drapes were exposed to the tip of the light source, the time to char was 3-6 s. The degree and volume of injury increased with longer exposure times, and significant injury was recorded with the optical cable 3 mm from the skin. This study demonstrates that the temperature at the tip of the optical light cord can induce extensive damage. The by-product of light, heat, can produce immediate superficial tissue necrosis that can extend into the subcutaneous fat even when the optical tip is not in direct contact with the skin. In addition, our study shows the variation in temperature that exists between light sources and bulb status. Overall, surgeons must realize and respect the potential complications associated with optical technology.

Electronic polarization-division demultiplexing based on digital signal processing in intensity-modulation direct-detection optical communication systems.

PubMed

Kikuchi, Kazuro

2014-01-27

We propose a novel configuration of optical receivers for intensity-modulation direct-detection (IM · DD) systems, which can cope with dual-polarization (DP) optical signals electrically. Using a Stokes analyzer and a newly-developed digital signal-processing (DSP) algorithm, we can achieve polarization tracking and demultiplexing in the digital domain after direct detection. Simulation results show that the power penalty stemming from digital polarization manipulations is negligibly small.

Assessment of a liquid lens enabled in vivo optical coherence microscope.

PubMed

Murali, Supraja; Meemon, Panomsak; Lee, Kye-Sung; Kuhn, William P; Thompson, Kevin P; Rolland, Jannick P

2010-06-01

The optical aberrations induced by imaging through skin can be predicted using formulas for Seidel aberrations of a plane-parallel plate. Knowledge of these aberrations helps to guide the choice of numerical aperture (NA) of the optics we can use in an implementation of Gabor domain optical coherence microscopy (GD-OCM), where the focus is the only aberration adjustment made through depth. On this basis, a custom-designed, liquid-lens enabled dynamic focusing optical coherence microscope operating at 0.2 NA is analyzed and validated experimentally. As part of the analysis, we show that the full width at half-maximum metric, as a characteristic descriptor for the point spread function, while commonly used, is not a useful metric for quantifying resolution in non-diffraction-limited systems. Modulation transfer function (MTF) measurements quantify that the liquid lens performance is as predicted by design, even when accounting for the effect of gravity. MTF measurements in a skinlike scattering medium also quantify the performance of the microscope in its potential applications. To guide the fusion of images across the various focus positions of the microscope, as required in GD-OCM, we present depth of focus measurements that can be used to determine the effective number of focusing zones required for a given goal resolution. Subcellular resolution in an onion sample, and high-definition in vivo imaging in human skin are demonstrated with the custom-designed and built microscope.

The virtual microscopy database-sharing digital microscope images for research and education.

PubMed

Lee, Lisa M J; Goldman, Haviva M; Hortsch, Michael

2018-02-14

Over the last 20 years, virtual microscopy has become the predominant modus of teaching the structural organization of cells, tissues, and organs, replacing the use of optical microscopes and glass slides in a traditional histology or pathology laboratory setting. Although virtual microscopy image files can easily be duplicated, creating them requires not only quality histological glass slides but also an expensive whole slide microscopic scanner and massive data storage devices. These resources are not available to all educators and researchers, especially at new institutions in developing countries. This leaves many schools without access to virtual microscopy resources. The Virtual Microscopy Database (VMD) is a new resource established to address this problem. It is a virtual image file-sharing website that allows researchers and educators easy access to a large repository of virtual histology and pathology image files. With the support from the American Association of Anatomists (Bethesda, MD) and MBF Bioscience Inc. (Williston, VT), registration and use of the VMD are currently free of charge. However, the VMD site is restricted to faculty and staff of research and educational institutions. Virtual Microscopy Database users can upload their own collection of virtual slide files, as well as view and download image files for their own non-profit educational and research purposes that have been deposited by other VMD clients. Anat Sci Educ. © 2018 American Association of Anatomists. © 2018 American Association of Anatomists.

Excitation-scanning hyperspectral imaging system for microscopic and endoscopic applications

NASA Astrophysics Data System (ADS)

Mayes, Sam A.; Leavesley, Silas J.; Rich, Thomas C.

2016-04-01

Current microscopic and endoscopic technologies for cancer screening utilize white-light illumination sources. Hyper-spectral imaging has been shown to improve sensitivity while retaining specificity when compared to white-light imaging in both microscopy and in vivo imaging. However, hyperspectral imaging methods have historically suffered from slow acquisition times due to the narrow bandwidth of spectral filters. Often minutes are required to gather a full image stack. We have developed a novel approach called excitation-scanning hyperspectral imaging that provides 2-3 orders of magnitude increased signal strength. This reduces acquisition times significantly, allowing for live video acquisition. Here, we describe a preliminary prototype excitation-scanning hyperspectral imaging system that can be coupled with endoscopes or microscopes for hyperspectral imaging of tissues and cells. Our system is comprised of three subsystems: illumination, transmission, and imaging. The illumination subsystem employs light-emitting diode arrays to illuminate at different wavelengths. The transmission subsystem utilizes a unique geometry of optics and a liquid light guide. Software controls allow us to interface with and control the subsystems and components. Digital and analog signals are used to coordinate wavelength intensity, cycling and camera triggering. Testing of the system shows it can cycle 16 wavelengths at as fast as 1 ms per cycle. Additionally, more than 18% of the light transmits through the system. Our setup should allow for hyperspectral imaging of tissue and cells in real time.

The Impact of the Condenser on Cytogenetic Image Quality in Digital Microscope System

PubMed Central

Ren, Liqiang; Li, Zheng; Li, Yuhua; Zheng, Bin; Li, Shibo; Chen, Xiaodong; Liu, Hong

2013-01-01

Background: Optimizing operational parameters of the digital microscope system is an important technique to acquire high quality cytogenetic images and facilitate the process of karyotyping so that the efficiency and accuracy of diagnosis can be improved. OBJECTIVE: This study investigated the impact of the condenser on cytogenetic image quality and system working performance using a prototype digital microscope image scanning system. Methods: Both theoretical analysis and experimental validations through objectively evaluating a resolution test chart and subjectively observing large numbers of specimen were conducted. Results: The results show that the optimal image quality and large depth of field (DOF) are simultaneously obtained when the numerical aperture of condenser is set as 60%–70% of the corresponding objective. Under this condition, more analyzable chromosomes and diagnostic information are obtained. As a result, the system shows higher working stability and less restriction for the implementation of algorithms such as autofocusing especially when the system is designed to achieve high throughput continuous image scanning. Conclusions: Although the above quantitative results were obtained using a specific prototype system under the experimental conditions reported in this paper, the presented evaluation methodologies can provide valuable guidelines for optimizing operational parameters in cytogenetic imaging using the high throughput continuous scanning microscopes in clinical practice. PMID:23676284

Nanoscale Optical Imaging and Spectroscopy from Visible to Mid-Infrared

DTIC Science & Technology

2015-11-13

field characterization of nanoscale materials, it also complements the near- field scanning optical microscope currently available in the PI’s lab...field scanning optical microscope currently available in the PI’s lab. This equipment will begin making major impacts on at least three current DoD...SECURITY CLASSIFICATION OF: 1. REPORT DATE (DD-MM-YYYY) 4. TITLE AND SUBTITLE 13. SUPPLEMENTARY NOTES 12. DISTRIBUTION AVAILIBILITY STATEMENT 6

Microscopic Engine Powered by Critical Demixing

NASA Astrophysics Data System (ADS)

Schmidt, Falko; Magazzù, Alessandro; Callegari, Agnese; Biancofiore, Luca; Cichos, Frank; Volpe, Giovanni

2018-02-01

We experimentally demonstrate a microscopic engine powered by the local reversible demixing of a critical mixture. We show that, when an absorbing microsphere is optically trapped by a focused laser beam in a subcritical mixture, it is set into rotation around the optical axis of the beam because of the emergence of diffusiophoretic propulsion. This behavior can be controlled by adjusting the optical power, the temperature, and the criticality of the mixture.

Tapered fiber optical tweezers for microscopic particle trapping: fabrication and application

NASA Astrophysics Data System (ADS)

Liu, Zhihai; Guo, Chengkai; Yang, Jun; Yuan, Libo

2006-12-01

A novel single tapered fiber optical tweezers is proposed and fabricated by heating and drawing technology. The microscopic particle tapping performance of this special designed tapered fiber probe is demonstrated and investigated. The distribution of the optical field emerging from the tapered fiber tip is numerically calculated based on the beam propagation method. The trapping force FDTD analysis results, both axial and transverse, are also given.

Eliminating Bias In Acousto-Optical Spectrum Analysis

NASA Technical Reports Server (NTRS)

Ansari, Homayoon; Lesh, James R.

1992-01-01

Scheme for digital processing of video signals in acousto-optical spectrum analyzer provides real-time correction for signal-dependent spectral bias. Spectrum analyzer described in "Two-Dimensional Acousto-Optical Spectrum Analyzer" (NPO-18092), related apparatus described in "Three-Dimensional Acousto-Optical Spectrum Analyzer" (NPO-18122). Essence of correction is to average over digitized outputs of pixels in each CCD row and to subtract this from the digitized output of each pixel in row. Signal processed electro-optically with reference-function signals to form two-dimensional spectral image in CCD camera.

Micro-electro-mechanical systems (MEMS) and agile lensing-based modules for communications, sensing and signal processing

NASA Astrophysics Data System (ADS)

Reza, Syed Azer

This dissertation proposes the use of the emerging Micro-Electro-Mechanical Systems (MEMS) and agile lensing optical device technologies to design novel and powerful signal conditioning and sensing modules for advanced applications in optical communications, physical parameter sensing and RF/optical signal processing. For example, these new module designs have experimentally demonstrated exceptional features such as stable loss broadband operations and high > 60 dB optical dynamic range signal filtering capabilities. The first part of the dissertation describes the design and demonstration of digital MEMS-based signal processing modules for communication systems and sensor networks using the TI DLP (Digital Light Processing) technology. Examples of such modules include optical power splitters, narrowband and broadband variable fiber optical attenuators, spectral shapers and filters. Compared to prior works, these all-digital designs have advantages of repeatability, accuracy, and reliability that are essential for advanced communications and sensor applications. The next part of the dissertation proposes, analyzes and demonstrates the use of analog opto-fluidic agile lensing technology for sensor networks and test and measurement systems. Novel optical module designs for distance sensing, liquid level sensing, three-dimensional object shape sensing and variable photonic delay lines are presented and experimentally demonstrated. Compared to prior art module designs, the proposed analog-mode modules have exceptional performances, particularly for extreme environments (e.g., caustic liquids) where the free-space agile beam-based sensor provide remote non-contact access for physical sensing operations. The dissertation also presents novel modules involving hybrid analog-digital photonic designs that make use of the different optical device technologies to deliver the best features of both analog and digital optical device operations and controls. Digital controls are achieved through the use of the digital MEMS technology and analog controls are realized by employing opto-fluidic agile lensing technology and acousto-optic technology. For example, variable fiber-optic attenuators and spectral filters are proposed using the hybrid design. Compared to prior art module designs, these hybrid designs provide a higher module dynamic range and increased resolution that are critical in various advanced system applications. In summary, the dissertation shows the added power of hybrid optical designs using both the digital and analog photonic signal processing versus just all-digital or all-analog module designs.

Confocal microscopic observation of structural changes in glass-ionomer cements and tooth interfaces.

PubMed

Watson, T F; Pagliari, D; Sidhu, S K; Naasan, M A

1998-03-01

This study aimed to develop techniques to allow dynamic imaging of a cavity before, during and after placement of glass-ionomer restorative materials. Cavities were cut in recently extracted third molars and the teeth longitudinally sectioned. Each hemisected tooth surface was placed in green modelling compound at 90 to the optical axis of the microscope. The cavity surface was imaged using a video rate confocal microscope in conjunction with an internally focusable microscope objective. The sample on the stage was pushed up to the objective lens which 'clamped' the cover glass onto it. Water, glycerine or oil was placed below the coverglass, with oil above. Internal tooth structures were imaged by changing the internal focus of the objective. The restorative material was then placed into the cavity. Video images were stored either onto video tape or digitally, using a frame grabber, computer and mass memory storage. Software controls produced time-lapse recordings of the interface over time. Preliminary experiments have examined the placement and early maturation of conventional glass-ionomer cements and a syringeable resin-modified glass-ionomer cement. Initial contact of the cement matrix and glass particles was visible as the plastic material rolled past the enamel and dentine, before making a bond. Evidence for water movement from the dentine into the cement has also been seen. After curing, the early dimensional changes in the cements due to water flux were apparent using the time-lapse facility. This new technique enables examination of developing tooth/restoration interfaces and the tracking of movement in materials.

Optical transmission modules for multi-channel superconducting quantum interference device readouts.

PubMed

Kim, Jin-Mok; Kwon, Hyukchan; Yu, Kwon-kyu; Lee, Yong-Ho; Kim, Kiwoong

2013-12-01

We developed an optical transmission module consisting of 16-channel analog-to-digital converter (ADC), digital-noise filter, and one-line serial transmitter, which transferred Superconducting Quantum Interference Device (SQUID) readout data to a computer by a single optical cable. A 16-channel ADC sent out SQUID readouts data with 32-bit serial data of 8-bit channel and 24-bit voltage data at a sample rate of 1.5 kSample/s. A digital-noise filter suppressed digital noises generated by digital clocks to obtain SQUID modulation as large as possible. One-line serial transmitter reformed 32-bit serial data to the modulated data that contained data and clock, and sent them through a single optical cable. When the optical transmission modules were applied to 152-channel SQUID magnetoencephalography system, this system maintained a field noise level of 3 fT/√Hz @ 100 Hz.

The potential for early and rapid pathogen detection within poultry processing through hyperspectral microscopy

USDA-ARS?s Scientific Manuscript database

The acquisition of hyperspectral microscopic images containing both spatial and spectral data has shown potential for the early and rapid optical classification of foodborne pathogens. A hyperspectral microscope with a metal halide light source and acousto-optical tunable filter (AOTF) collects 89 ...

Acousto-Optic Tunable Filter Hyperspectral Microscope Imaging Method for Characterizing Spectra from Foodborne Pathogens.

USDA-ARS?s Scientific Manuscript database

Hyperspectral microscope imaging (HMI) method, which provides both spatial and spectral characteristics of samples, can be effective for foodborne pathogen detection. The acousto-optic tunable filter (AOTF)-based HMI method can be used to characterize spectral properties of biofilms formed by Salmon...

Three-dimensional digital breast histopathology imaging

NASA Astrophysics Data System (ADS)

Clarke, G. M.; Peressotti, C.; Mawdsley, G. E.; Eidt, S.; Ge, M.; Morgan, T.; Zubovits, J. T.; Yaffe, M. J.

2005-04-01

We have developed a digital histology imaging system that has the potential to improve the accuracy of surgical margin assessment in the treatment of breast cancer by providing finer sampling and 3D visualization. The system is capable of producing a 3D representation of histopathology from an entire lumpectomy specimen. We acquire digital photomicrographs of a stack of large (120 x 170 mm) histology slides cut serially through the entire specimen. The images are then registered and displayed in 2D and 3D. This approach dramatically improves sampling and can improve visualization of tissue structures compared to current, small-format histology. The system consists of a brightfield microscope, adapted with a freeze-frame digital video camera and a large, motorized translation stage. The image of each slide is acquired as a mosaic of adjacent tiles, each tile representing one field-of-view of the microscope, and the mosaic is assembled into a seamless composite image. The assembly is done by a program developed to build image sets at six different levels within a multiresolution pyramid. A database-linked viewing program has been created to efficiently register and display the animated stack of images, which occupies about 80 GB of disk space per lumpectomy at full resolution, on a high-resolution (3840 x 2400 pixels) colour monitor. The scanning or tiling approach to digitization is inherently susceptible to two artefacts which disrupt the composite image, and which impose more stringent requirements on system performance. Although non-uniform illumination across any one isolated tile may not be discernible, the eye readily detects this non-uniformity when the entire assembly of tiles is viewed. The pattern is caused by deficiencies in optical alignment, spectrum of the light source, or camera corrections. The imaging task requires that features as small as 3.2 &mum in extent be seamlessly preserved. However, inadequate accuracy in positioning of the translation stage produces visible discontinuities between adjacent features. Both of these effects can distract the viewer from the perception of diagnostically important features. Here we describe the system design and discuss methods for the correction of these artefacts. In addition, we outline our approach to rendering the processing and display of these large images computationally feasible.

Simple fiber-optic confocal microscopy with nanoscale depth resolution beyond the diffraction barrier.

PubMed

Ilev, Ilko; Waynant, Ronald; Gannot, Israel; Gandjbakhche, Amir

2007-09-01

A novel fiber-optic confocal approach for ultrahigh depth-resolution (

Hybrid Microscopy: Enabling Inexpensive High-Performance Imaging through Combined Physical and Optical Magnifications.

PubMed

Zhang, Yu Shrike; Chang, Jae-Byum; Alvarez, Mario Moisés; Trujillo-de Santiago, Grissel; Aleman, Julio; Batzaya, Byambaa; Krishnadoss, Vaishali; Ramanujam, Aishwarya Aravamudhan; Kazemzadeh-Narbat, Mehdi; Chen, Fei; Tillberg, Paul W; Dokmeci, Mehmet Remzi; Boyden, Edward S; Khademhosseini, Ali

2016-03-15

To date, much effort has been expended on making high-performance microscopes through better instrumentation. Recently, it was discovered that physical magnification of specimens was possible, through a technique called expansion microscopy (ExM), raising the question of whether physical magnification, coupled to inexpensive optics, could together match the performance of high-end optical equipment, at a tiny fraction of the price. Here we show that such "hybrid microscopy" methods--combining physical and optical magnifications--can indeed achieve high performance at low cost. By physically magnifying objects, then imaging them on cheap miniature fluorescence microscopes ("mini-microscopes"), it is possible to image at a resolution comparable to that previously attainable only with benchtop microscopes that present costs orders of magnitude higher. We believe that this unprecedented hybrid technology that combines expansion microscopy, based on physical magnification, and mini-microscopy, relying on conventional optics--a process we refer to as Expansion Mini-Microscopy (ExMM)--is a highly promising alternative method for performing cost-effective, high-resolution imaging of biological samples. With further advancement of the technology, we believe that ExMM will find widespread applications for high-resolution imaging particularly in research and healthcare scenarios in undeveloped countries or remote places.

All optical coherent receiver for self-homodyne detection of digitally phase modulated optical signals

NASA Astrophysics Data System (ADS)

Kiasaleh, Kamran

1994-02-01

A novel optical phase-locked loop (OPLL) system for the self-homodyne detection of digitally phase modulated optical signals is introduced. A Mach-Zehnder type interferometer is used to self-homodyne binary phase-modulated optical signals with an external phase modulator inserted in the control arm of the interferometer.

Joint Applied Optics and Chinese Optics Letters feature introduction: digital holography and three-dimensional imaging.

PubMed

Poon, Ting-Chung

2011-12-01

This feature issue serves as a pilot issue promoting the joint issue of Applied Optics and Chinese Optics Letters. It focuses upon topics of current relevance to the community working in the area of digital holography and 3-D imaging. © 2011 Optical Society of America

Planning for optical disk technology with digital cartography.

USGS Publications Warehouse

Light, D.L.

1986-01-01

A major shortfall that still exists in digital systems is the need for very large mass storage capacity. The decade of the 1980s has introduced laser optical disk storage technology, which may be the breakthrough needed for mass storage. This paper addresses system concepts for digital cartography during the transition period. Emphasis will be placed on determining USGS mass storage requirements and introducing laser optical disk technology for handling storage problems for digital data in this decade.-from Author

A pragmatic guide to multiphoton microscope design

PubMed Central

Young, Michael D.; Field, Jeffrey J.; Sheetz, Kraig E.; Bartels, Randy A.; Squier, Jeff

2016-01-01

Multiphoton microscopy has emerged as a ubiquitous tool for studying microscopic structure and function across a broad range of disciplines. As such, the intent of this paper is to present a comprehensive resource for the construction and performance evaluation of a multiphoton microscope that will be understandable to the broad range of scientific fields that presently exploit, or wish to begin exploiting, this powerful technology. With this in mind, we have developed a guide to aid in the design of a multiphoton microscope. We discuss source selection, optical management of dispersion, image-relay systems with scan optics, objective-lens selection, single-element light-collection theory, photon-counting detection, image rendering, and finally, an illustrated guide for building an example microscope. PMID:27182429

Design of a normal incidence multilayer imaging x-ray microscope.

PubMed

Shealy, D L; Gabardi, D R; Hoover, R B; Walker, A B; Lindblom, J F; Barbee, T W

1989-01-01

Normal incidence multilayer Cassegrain x-ray telescopes were flown on the Stanford/MSFC Rocket X-Ray Spectroheliograph. These instruments produced high spatial resolution images of the Sun and conclusively demonstrated that doubly reflecting multilayer x-ray optical systems are feasible. The images indicated that aplanatic imaging soft x-ray /EUV microscopes should be achievable using multilayer optics technology. We have designed a doubly reflecting normal incidence multilayer imaging x-ray microscope based on the Schwarzschild configuration. The Schwarzschild microscope utilizes two spherical mirrors with concentric radii of curvature which are chosen such that the third-order spherical aberration and coma are minimized. We discuss the design of the microscope and the results of the optical system ray trace analysis which indicates that diffraction-limited performance with 600 Å spatial resolution should be obtainable over a 1 mm field of view at a wavelength of 100 Å. Fabrication of several imaging soft x-ray microscopes based upon these designs, for use in conjunction with x-ray telescopes and laser fusion research, is now in progress. High resolution aplanatic imaging x-ray microscopes using normal incidence multilayer x-ray mirrors should have many important applications in advanced x-ray astronomical instrumentation, x-ray lithography, biological, biomedical, metallurgical, and laser fusion research.

Multi-gigabit optical interconnects for next-generation on-board digital equipment

NASA Astrophysics Data System (ADS)

Venet, Norbert; Favaro, Henri; Sotom, Michel; Maignan, Michel; Berthon, Jacques

2017-11-01

Parallel optical interconnects are experimentally assessed as a technology that may offer the high-throughput data communication capabilities required to the next-generation on-board digital processing units. An optical backplane interconnect was breadboarded, on the basis of a digital transparent processor that provides flexible connectivity and variable bandwidth in telecom missions with multi-beam antenna coverage. The unit selected for the demonstration required that more than tens of Gbit/s be supported by the backplane. The demonstration made use of commercial parallel optical link modules at 850 nm wavelength, with 12 channels running at up to 2.5 Gbit/s. A flexible optical fibre circuit was developed so as to route board-to-board connections. It was plugged to the optical transmitter and receiver modules through 12-fibre MPO connectors. BER below 10-14 and optical link budgets in excess of 12 dB were measured, which would enable to integrate broadcasting. Integration of the optical backplane interconnect was successfully demonstrated by validating the overall digital processor functionality.

Multi-gigabit optical interconnects for next-generation on-board digital equipment

NASA Astrophysics Data System (ADS)

Venet, Norbert; Favaro, Henri; Sotom, Michel; Maignan, Michel; Berthon, Jacques

2004-06-01

Parallel optical interconnects are experimentally assessed as a technology that may offer the high-throughput data communication capabilities required to the next-generation on-board digital processing units. An optical backplane interconnect was breadboarded, on the basis of a digital transparent processor that provides flexible connectivity and variable bandwidth in telecom missions with multi-beam antenna coverage. The unit selected for the demonstration required that more than tens of Gbit/s be supported by the backplane. The demonstration made use of commercial parallel optical link modules at 850 nm wavelength, with 12 channels running at up to 2.5 Gbit/s. A flexible optical fibre circuit was developed so as to route board-to-board connections. It was plugged to the optical transmitter and receiver modules through 12-fibre MPO connectors. BER below 10-14 and optical link budgets in excess of 12 dB were measured, which would enable to integrate broadcasting. Integration of the optical backplane interconnect was successfully demonstrated by validating the overall digital processor functionality.

KM3NeT Digital Optical Module electronics

NASA Astrophysics Data System (ADS)

Real, Diego

2016-04-01

The KM3NeT collaboration is currently building of a neutrino telescope with a volume of several cubic kilometres at the bottom of the Mediterranean Sea. The telescope consists of a matrix of Digital Optical Modules that will detect the Cherenkov light originated by the interaction of the neutrinos in the proximity of the detector. This contribution describes the main components of the read-out electronics of the Digital Optical Module: the Power Board, which delivers all the power supply required by the Digital Optical Molule electronics; the Central Logic Board, the main core of the read-out system, hosting 31 Time to Digital Converters with 1 ns resolution and the White Rabbit protocol embedded in the Central Logic Board Field Programmable Gate Array; the Octopus boards, that transfer the Low Voltage Digital Signals from the PMT bases to the Central Logic Board and finally the PMT bases, in charge of converting the analogue signal produced in the 31 3" PMTs into a Low Voltage Digital Signal.

Project support of practical training in biophysics.

PubMed

Mornstein, V; Vlk, D; Forytkova, L

2006-01-01

The Department of Biophysics ensures practical training in biophysics and related subjects for students of medical and health study programmes. Demonstrations of medical technology are an important part of this training. Teaching for Faculty of Sciences in biophysical study programmes becomes also very important. Some lectures and demonstrations of technology are involved, but the practical trainig is missing. About 1 mil. CZK for additional laboratory equipment was obtained from the HEIDF project No. 1866/ 2005 "The demonstration and measuring technology for education in medical biophysics and radiological physics" for measuring system DEWETRON for high frequency signal analysis, Fluke Ti30 IR camera, PM 9000B patient monitor, ARSENAL AF 1 fluorescence microscope, and Nikon Coolpix 4500 digital camera with accessories for microphotography. At the present time, further financial resources are being provided by a development project of Ministry of Education "Inter-university co-operation in biomedical technology and engineering using top technologies" in total amount of almost 5 mil CZK, whereas over 2 mil CZK from this project are reserved for student laboratory equipment. The main goal of this project is to ensure the participation of Medical Faculty in educational co-operation in the biomedical technology and engineering, namely with the Faculty of Electrical Engineering and Communication (FEEC), Brno University of Technology. There will be taught those areas of biophysics which are not covered by FEEC, thus forming a separate subject "General Biophysics". The following instruments will be installed: UV-VIS spectrophotometers, rotation viscometers, tensiometers, microscopes with digital image processing, cooled centrifuge, optical benches, and some smaller instruments for practical measurements.

Optical information processing at NASA Ames Research Center

NASA Technical Reports Server (NTRS)

Reid, Max B.; Bualat, Maria G.; Cho, Young C.; Downie, John D.; Gary, Charles K.; Ma, Paul W.; Ozcan, Meric; Pryor, Anna H.; Spirkovska, Lilly

1993-01-01

The combination of analog optical processors with digital electronic systems offers the potential of tera-OPS computational performance, while often requiring less power and weight relative to all-digital systems. NASA is working to develop and demonstrate optical processing techniques for on-board, real time science and mission applications. Current research areas and applications under investigation include optical matrix processing for space structure vibration control and the analysis of Space Shuttle Main Engine plume spectra, optical correlation-based autonomous vision for robotic vehicles, analog computation for robotic path planning, free-space optical interconnections for information transfer within digital electronic computers, and multiplexed arrays of fiber optic interferometric sensors for acoustic and vibration measurements.

Document Examination: Applications of Image Processing Systems.

PubMed

Kopainsky, B

1989-12-01

Dealing with images is a familiar business for an expert in questioned documents: microscopic, photographic, infrared, and other optical techniques generate images containing the information he or she is looking for. A recent method for extracting most of this information is digital image processing, ranging from the simple contrast and contour enhancement to the advanced restoration of blurred texts. When combined with a sophisticated physical imaging system, an image pricessing system has proven to be a powerful and fast tool for routine non-destructive scanning of suspect documents. This article reviews frequent applications, comprising techniques to increase legibility, two-dimensional spectroscopy (ink discrimination, alterations, erased entries, etc.), comparison techniques (stamps, typescript letters, photo substitution), and densitometry. Computerized comparison of handwriting is not included. Copyright © 1989 Central Police University.

Augmented microscopy with near-infrared fluorescence detection

NASA Astrophysics Data System (ADS)

Watson, Jeffrey R.; Martirosyan, Nikolay; Skoch, Jesse; Lemole, G. Michael; Anton, Rein; Romanowski, Marek

2015-03-01

Near-infrared (NIR) fluorescence has become a frequently used intraoperative technique for image-guided surgical interventions. In procedures such as cerebral angiography, surgeons use the optical surgical microscope for the color view of the surgical field, and then switch to an electronic display for the NIR fluorescence images. However, the lack of stereoscopic, real-time, and on-site coregistration adds time and uncertainty to image-guided surgical procedures. To address these limitations, we developed the augmented microscope, whereby the electronically processed NIR fluorescence image is overlaid with the anatomical optical image in real-time within the optical path of the microscope. In vitro, the augmented microscope can detect and display indocyanine green (ICG) concentrations down to 94.5 nM, overlaid with the anatomical color image. We prepared polyacrylamide tissue phantoms with embedded polystyrene beads, yielding scattering properties similar to brain matter. In this model, 194 μM solution of ICG was detectable up to depths of 5 mm. ICG angiography was then performed in anesthetized rats. A dynamic process of ICG distribution in the vascular system overlaid with anatomical color images was observed and recorded. In summary, the augmented microscope demonstrates NIR fluorescence detection with superior real-time coregistration displayed within the ocular of the stereomicroscope. In comparison to other techniques, the augmented microscope retains full stereoscopic vision and optical controls including magnification and focus, camera capture, and multiuser access. Augmented microscopy may find application in surgeries where the use of traditional microscopes can be enhanced by contrast agents and image guided delivery of therapeutics, including oncology, neurosurgery, and ophthalmology.

Microscope basics.

PubMed

Sluder, Greenfield; Nordberg, Joshua J

2013-01-01

This chapter provides information on how microscopes work and discusses some of the microscope issues to be considered in using a video camera on the microscope. There are two types of microscopes in use today for research in cell biology-the older finite tube-length (typically 160mm mechanical tube length) microscopes and the infinity optics microscopes that are now produced. The objective lens forms a magnified, real image of the specimen at a specific distance from the objective known as the intermediate image plane. All objectives are designed to be used with the specimen at a defined distance from the front lens element of the objective (the working distance) so that the image formed is located at a specific location in the microscope. Infinity optics microscopes differ from the finite tube-length microscopes in that the objectives are designed to project the image of the specimen to infinity and do not, on their own, form a real image of the specimen. Three types of objectives are in common use today-plan achromats, plan apochromats, and plan fluorite lenses. The concept of mounting video cameras on the microscope is also presented in the chapter. Copyright © 2003 Elsevier Inc. All rights reserved.

Diamond Quantum Nanoemitters: Cross Discipline Research on Hyperbolic Optical Systems for Control of Quantum Nanoemitters

DTIC Science & Technology

2017-05-05

results of this project there are: (1) the investigation of the effect of phonons on the optical properties of solid state emitters. A microscopic ...In  what  follows  we  list  the  main  results  and  undergoing  research.   2. Results 2.1   Microscopic  modeling...fluorescent  markers   for   biological   measurements.   Here,   we   present   a   first-­‐principles   microscopic   description

Comparison of a virtual microscope laboratory to a regular microscope laboratory for teaching histology.

PubMed

Harris, T; Leaven, T; Heidger, P; Kreiter, C; Duncan, J; Dick, F

2001-02-01

Emerging technology now exists to digitize a gigabyte of information from a glass slide, save it in a highly compressed file format, and deliver it over the web. By accessing these images with a standard web browser and viewer plug-in, a computer can emulate a real microscope and glass slide. Using this new technology, the immediate aims of our project were to digitize the glass slides from urinary tract, male genital, and endocrine units and implement them in the Spring 2000 Histology course at the University of Iowa, and to carry out a formative evaluation of the virtual slides of these three units in a side-by-side comparison with the regular microscope laboratory. The methods and results of this paper will describe the technology employed to create the virtual slides, and the formative evaluation carried out in the course. Anat Rec (New Anat) 265:10-14, 2001. Copyright 2001 Wiley-Liss, Inc.

Design and construction of a modular low-cost epifluorescence upright microscope for neuron visualized recording and fluorescence detection.

PubMed

Beltran-Parrazal, Luis; Morgado-Valle, Consuelo; Serrano, Raul E; Manzo, Jorge; Vergara, Julio L

2014-03-30

One of the limitations when establishing an electrophysiology setup, particularly in low resource settings, is the high cost of microscopes. The average cost for a microscope equipped with the optics for infrared (IR) contrast or microfluorometry is $40,000. We hypothesized that optical elements and features included in commercial microscopes are not necessary to IR video-visualize neurons or for microfluorometry. We present instructions for building a low-cost epifluorescence upright microscope suitable for visualized patch-clamp recording and fluorescence detection using mostly catalog-available parts. This microscope supports applications such as visualized whole-cell recording using IR oblique illumination (IR-OI), or more complex applications such as microfluorometry using a photodiode. In both IR-OI and fluorescence, actual resolution measured with 2-μm latex beads is close to theoretical resolution. The lack of movable parts to switch configurations ensures stability when doing intracellular recording. The low cost is a significant advantage of this microscope compared to existent custom-built microscopes. The cost of the simplest configuration with IR-OI is ∼$2000, whereas the cost of the configuration with epifluorescence is ∼$5000. Since this design does not use pieces discarded from commercial microscopes, it is completely reproducible. We suggest that this microscope is a viable alternative for doing in vitro electrophysiology and microfluorometry in low-resource settings. Characteristics such as an open box design, easy assembly, and low-cost make this microscope a useful instrument for science education and teaching for topics such as optics, biology, neuroscience, and for scientific "hands-on" workshops. Copyright © 2014 Elsevier B.V. All rights reserved.

Apertureless near-field scanning optical microscope working with or without laser source.

PubMed

Formanek, F; De Wilde, Y; Aigouy, L; Chen, Y

2004-01-01

An apertureless near-field scanning optical microscope (ANSOM), used indifferent configurations, is presented. Our versatile home-made setup, based on a sharp tungsten tip glued onto a quartz tuning fork and working in tapping mode, allows to perform imaging over a broad spectral range. We have recorded optical images in the visible (wavelength, lambda = 655 nm) and in the infrared (lambda = 10.6 microm), proving that the setup routinely achieves an optical resolution of <50 nm regardless of the illumination wavelength. We have also shown optical images recorded in the visible (lambda = 655 nm) in an inverted configuration where the tip does not perturb the focused spot of the illumination laser. Approach curves as well as image profiles have revealed that on demodulating the optical signal at higher harmonics, we can obtain an effective probe sharpening which results in an improvement of the resolution. Finally, we have presented optical images recorded in the infrared without any illumination, that is, the usual laser source is replaced by a simple heating of the sample. This has shown that the ANSOM can be used as a near-field thermal optical microscope (NTOM) to probe the near field generated by the thermal emission of the sample.

Modified Linnik microscopic interferometry for quantitative depth evaluation of diffraction-limited microgroove

NASA Astrophysics Data System (ADS)

Ye, Shiwei; Takahashi, Satoru; Michihata, Masaki; Takamasu, Kiyoshi

2018-05-01

The quality control of microgrooves is extremely crucial to ensure the performance and stability of microstructures and improve their fabrication efficiency. This paper introduces a novel optical inspection method and a modified Linnik microscopic interferometer measurement system to detect the depth of microgrooves with a width less than the diffraction limit. Using this optical method, the depth of diffraction-limited microgrooves can be related to the near-field optical phase difference, which cannot be practically observed but can be computed from practical far-field observations. Thus, a modified Linnik microscopic interferometer system based on three identical objective lenses and an optical path reversibility principle were developed. In addition, experiments for standard grating microgrooves on the silicon surface were carried out to demonstrate the feasibility and repeatability of the proposed method and developed measurement system.

A Closed-Loop Proportional-Integral (PI) Control Software for Fully Mechanically Controlled Automated Electron Microscopic Tomography

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

REN, GANG; LIU, JINXIN; LI, HONGCHANG

A closed-loop proportional-integral (PI) control software is provided for fully mechanically controlled automated electron microscopic tomography. The software is developed based on Gatan DigitalMicrograph, and is compatible with Zeiss LIBRA 120 transmission electron microscope. However, it can be expanded to other TEM instrument with modification. The software consists of a graphical user interface, a digital PI controller, an image analyzing unit, and other drive units (i.e.: image acquire unit and goniometer drive unit). During a tomography data collection process, the image analyzing unit analyzes both the accumulated shift and defocus value of the latest acquired image, and provides the resultsmore » to the digital PI controller. The digital PI control compares the results with the preset values and determines the optimum adjustments of the goniometer. The goniometer drive unit adjusts the spatial position of the specimen according to the instructions given by the digital PI controller for the next tilt angle and image acquisition. The goniometer drive unit achieves high precision positioning by using a backlash elimination method. The major benefits of the software are: 1) the goniometer drive unit keeps pre-aligned/optimized beam conditions unchanged and achieves position tracking solely through mechanical control; 2) the image analyzing unit relies on only historical data and therefore does not require additional images/exposures; 3) the PI controller enables the system to dynamically track the imaging target with extremely low system error.« less

[Gene sequencing by scanning molecular exciton microscopy]. Progress report, October 1, 1990--September 30, 1991

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

Not Available

1991-12-31

This report details progress made in setting up a laboratory for optical microscopy of genes. The apparatus including a fluorescence microscope, a scanning optical microscope, various spectrometers, and supporting computers is described. Results in developing photon and exciton tips, and in preparing samples are presented. (GHH)

Working at the microscope: analysis of the activities involved in diagnostic pathology.

PubMed

Randell, Rebecca; Ruddle, Roy A; Quirke, Phil; Thomas, Rhys G; Treanor, Darren

2012-02-01

To study the current work practice of histopathologists to inform the design of digital microscopy systems. Four gastrointestinal histopathologists were video-recorded as they undertook their routine work. Analysis of the video data shows a range of activities beyond viewing slides involved in reporting a case. There is much overlapping of activities, supported by the 'eyes free' nature of the pathologists' interaction with the microscope. The order and timing of activities varies according to consultant. In order to support the work of pathologists adequately, digital microscopy systems need to provide support for a range of activities beyond viewing slides. Digital microscopy systems should support multitasking, while also providing flexibility so that pathologists can adapt their use of the technology to their own working patterns. © 2011 Blackwell Publishing Ltd.

A simple tool for stereological assessment of digital images: the STEPanizer.

PubMed

Tschanz, S A; Burri, P H; Weibel, E R

2011-07-01

STEPanizer is an easy-to-use computer-based software tool for the stereological assessment of digitally captured images from all kinds of microscopical (LM, TEM, LSM) and macroscopical (radiology, tomography) imaging modalities. The program design focuses on providing the user a defined workflow adapted to most basic stereological tasks. The software is compact, that is user friendly without being bulky. STEPanizer comprises the creation of test systems, the appropriate display of digital images with superimposed test systems, a scaling facility, a counting module and an export function for the transfer of results to spreadsheet programs. Here we describe the major workflow of the tool illustrating the application on two examples from transmission electron microscopy and light microscopy, respectively. © 2011 The Authors Journal of Microscopy © 2011 Royal Microscopical Society.

Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry.

PubMed

Schwertner, M; Booth, M J; Neil, M A A; Wilson, T

2004-01-01

Confocal or multiphoton microscopes, which deliver optical sections and three-dimensional (3D) images of thick specimens, are widely used in biology. These techniques, however, are sensitive to aberrations that may originate from the refractive index structure of the specimen itself. The aberrations cause reduced signal intensity and the 3D resolution of the instrument is compromised. It has been suggested to correct for aberrations in confocal microscopes using adaptive optics. In order to define the design specifications for such adaptive optics systems, one has to know the amount of aberrations present for typical applications such as with biological samples. We have built a phase stepping interferometer microscope that directly measures the aberration of the wavefront. The modal content of the wavefront is extracted by employing Zernike mode decomposition. Results for typical biological specimens are presented. It was found for all samples investigated that higher order Zernike modes give only a small contribution to the overall aberration. Therefore, these higher order modes can be neglected in future adaptive optics sensing and correction schemes implemented into confocal or multiphoton microscopes, leading to more efficient designs.

PLANNING FOR OPTICAL DISK TECHNOLOGY WITH DIGITAL CARTOGRAPHY.

USGS Publications Warehouse

Light, Donald L.

1984-01-01

Progress in the computer field continues to suggest that the transition from traditional analog mapping systems to digital systems has become a practical possibility. A major shortfall that still exists in digital systems is the need for very large mass storage capacity. The decade of the 1980's has introduced laser optical disk storage technology, which may be the breakthrough needed for mass storage. This paper addresses system concepts for digital cartography during the transition period. Emphasis is placed on determining U. S. Geological Survey mass storage requirements and introducing laser optical disk technology for handling storage problems for digital data in this decade.

Measurement methods to build up the digital optical twin

NASA Astrophysics Data System (ADS)

Prochnau, Marcel; Holzbrink, Michael; Wang, Wenxin; Holters, Martin; Stollenwerk, Jochen; Loosen, Peter

2018-02-01

The realization of the Digital Optical Twin (DOT), which is in short the digital representation of the physical state of an optical system, is particularly useful in the context of an automated assembly process of optical systems. During the assembly process, the physical system status of the optical system is continuously measured and compared with the digital model. In case of deviations between physical state and the digital model, the latter one is adapted to match the physical state. To reach the goal described above, in a first step measurement/characterization technologies concerning their suitability to generate a precise digital twin of an existing optical system have to be identified and evaluated. This paper gives an overview of possible characterization methods and, finally, shows first results of evaluated, compared methods (e.g. spot-radius, MTF, Zernike-polynomials), to create a DOT. The focus initially lies on the unequivocalness of the optimization results as well as on the computational time required for the optimization to reach the characterized system state. Possible sources of error are the measurement accuracy (to characterize the system) , execution time of the measurement, time needed to map the digital to the physical world (optimization step) as well as interface possibilities to integrate the measurement tool into an assembly cell. Moreover, it is to be discussed whether the used measurement methods are suitable for a `seamless' integration into an assembly cell.

VISUALIZATION FROM INTRAOPERATIVE SWEPT-SOURCE MICROSCOPE-INTEGRATED OPTICAL COHERENCE TOMOGRAPHY IN VITRECTOMY FOR COMPLICATIONS OF PROLIFERATIVE DIABETIC RETINOPATHY.

PubMed

Gabr, Hesham; Chen, Xi; Zevallos-Carrasco, Oscar M; Viehland, Christian; Dandrige, Alexandria; Sarin, Neeru; Mahmoud, Tamer H; Vajzovic, Lejla; Izatt, Joseph A; Toth, Cynthia A

2018-01-10

To evaluate the use of live volumetric (4D) intraoperative swept-source microscope-integrated optical coherence tomography in vitrectomy for proliferative diabetic retinopathy complications. In this prospective study, we analyzed a subgroup of patients with proliferative diabetic retinopathy complications who required vitrectomy and who were imaged by the research swept-source microscope-integrated optical coherence tomography system. In near real time, images were displayed in stereo heads-up display facilitating intraoperative surgeon feedback. Postoperative review included scoring image quality, identifying different diabetic retinopathy-associated pathologies and reviewing the intraoperatively documented surgeon feedback. Twenty eyes were included. Indications for vitrectomy were tractional retinal detachment (16 eyes), combined tractional-rhegmatogenous retinal detachment (2 eyes), and vitreous hemorrhage (2 eyes). Useful, good-quality 2D (B-scans) and 4D images were obtained in 16/20 eyes (80%). In these eyes, multiple diabetic retinopathy complications could be imaged. Swept-source microscope-integrated optical coherence tomography provided surgical guidance, e.g., in identifying dissection planes under fibrovascular membranes, and in determining residual membranes and traction that would benefit from additional peeling. In 4/20 eyes (20%), acceptable images were captured, but they were not useful due to high tractional retinal detachment elevation which was challenging for imaging. Swept-source microscope-integrated optical coherence tomography can provide important guidance during surgery for proliferative diabetic retinopathy complications through intraoperative identification of different complications and facilitation of intraoperative decision making.

Status of emerging standards for removable computer storage media and related contributions of NIST

NASA Technical Reports Server (NTRS)

Podio, Fernando L.

1992-01-01

Standards for removable computer storage media are needed so that users may reliably interchange data both within and among various computer installations. Furthermore, media interchange standards support competition in industry and prevent sole-source lock-in. NIST participates in magnetic tape and optical disk standards development through Technical Committees X3B5, Digital Magnetic Tapes, X3B11, Optical Digital Data Disk, and the Joint Technical Commission on Data Permanence. NIST also participates in other relevant national and international standards committees for removable computer storage media. Industry standards for digital magnetic tapes require the use of Standard Reference Materials (SRM's) developed and maintained by NIST. In addition, NIST has been studying care and handling procedures required for digital magnetic tapes. NIST has developed a methodology for determining the life expectancy of optical disks. NIST is developing care and handling procedures for optical digital data disks and is involved in a program to investigate error reporting capabilities of optical disk drives. This presentation reflects the status of emerging magnetic tape and optical disk standards, as well as NIST's contributions in support of these standards.

Digital-holographic analysis of femtosecond laser-induced photodisruption in ocular tissue

NASA Astrophysics Data System (ADS)

Saerchen, Emanuel; Biessy, Kevin; Kemper, Björn; Lubatschowski, Holger

2014-02-01

High repetition rated femtosecond laser oscillator systems with low pulse energy are more often applied for precise and safer eye surgery. Especially, the cutting procedure in the crystalline lens is of high important for presbyopia treatment. Nevertheless, the fundamental laser tissue interaction process is not completely understood, because apparently a self-induced process takes place, were one modified region changes the focusing behavior of following laser pulses. We used a MHz repetition rate femtosecond laser system with nJ-pulse energy which were focused inside an ocular-tissue-phantom (Hydroxy-ethylmethacrylat - HEMA) to induce photodisruption. The material change, caused by the fs-pulses was measured simultaneously with a compact digital-holographic microscope. To investigate the material manipulation at different time scales, we used a continuously illuminating light source. The holographic images provide quantitative values for optical path length difference (OPL), which is equivalent to a refractive index change. This change of the optical properties may cause following pulses to obtain different focusing conditions. Time lapse measurements during the laser application were performed, which show the temporal evolution of OPL. An increase of OPL during the laser application was measured, which was followed by a decrease in OPL after laser processing. Furthermore, similar experiments were performed in distilled water and in native porcine crystalline lenses. The fs-laser cutting effects in HEMA and crystalline lens were transferable. Simultaneous measurements of the material modification during the cutting process give rise to better knowledge of treatment modalities during ocular tissue processing.

Novel microscope-integrated stereoscopic heads-up display for intrasurgical optical coherence tomography

PubMed Central

Shen, Liangbo; Carrasco-Zevallos, Oscar; Keller, Brenton; Viehland, Christian; Waterman, Gar; Hahn, Paul S.; Kuo, Anthony N.; Toth, Cynthia A.; Izatt, Joseph A.

2016-01-01

Intra-operative optical coherence tomography (OCT) requires a display technology which allows surgeons to visualize OCT data without disrupting surgery. Previous research and commercial intrasurgical OCT systems have integrated heads-up display (HUD) systems into surgical microscopes to provide monoscopic viewing of OCT data through one microscope ocular. To take full advantage of our previously reported real-time volumetric microscope-integrated OCT (4D MIOCT) system, we describe a stereoscopic HUD which projects a stereo pair of OCT volume renderings into both oculars simultaneously. The stereoscopic HUD uses a novel optical design employing spatial multiplexing to project dual OCT volume renderings utilizing a single micro-display. The optical performance of the surgical microscope with the HUD was quantitatively characterized and the addition of the HUD was found not to substantially effect the resolution, field of view, or pincushion distortion of the operating microscope. In a pilot depth perception subject study, five ophthalmic surgeons completed a pre-set dexterity task with 50.0% (SD = 37.3%) higher success rate and in 35.0% (SD = 24.8%) less time on average with stereoscopic OCT vision compared to monoscopic OCT vision. Preliminary experience using the HUD in 40 vitreo-retinal human surgeries by five ophthalmic surgeons is reported, in which all surgeons reported that the HUD did not alter their normal view of surgery and that live surgical maneuvers were readily visible in displayed stereoscopic OCT volumes. PMID:27231616

Development of HiLo Microscope and its use in In-Vivo Applications

NASA Astrophysics Data System (ADS)

Patel, Shreyas J.

The functionality of achieving optical sectioning in biomedical research is invaluable as it allows for visualization of a biological sample at different depths while being free of background scattering. Most current microscopy techniques that offer optical sectioning, unfortunately, require complex instrumentation and thus are generally costly. HiLo microscopy, on the other hand, offers the same functionality and advantage at a relatively low cost. Hence, the work described in this thesis involves the design, build, and application of a HiLo microscope. More specifically, a standalone HiLo microscope was built in addition to implementing HiLo microscopy on a standard fluorescence microscope. In HiLo microscopy, optical sectioning is achieved by acquiring two different types of images per focal plane. One image is acquired under uniform illumination and the other is acquired under speckle illumination. These images are processed using an algorithm that extracts in-focus information and removes features and glare that occur as a result of background fluorescence. To show the benefits of the HiLo microscopy, several imaging experiments on various samples were performed under a HiLo microscope and compared against a traditional fluorescence microscope and a confocal microscope, which is considered the gold standard in optical imaging. In-vitro and ex-vivo imaging was performed on a set of pollen grains, and optically cleared mouse brain and heart slices. Each of these experiments showed great reduction in background scattering at different depths under HiLo microscopy. More importantly, HiLo imaging of optically cleared heart slice demonstrated emergence of different vasculature at different depths. Reduction of out-of-focus light increased the spatial resolution and allowed better visualization of capillary vessels. Furthermore, HiLo imaging was tested in an in-vivo model of a rodent dorsal window chamber model. When imaging the same sample under confocal microscope, the results were comparable between the two modalities. Additionally, a method of achieving blood flow maps at different depth using a combination of HiLo and LSI imaging is also discussed. The significance of this combined technique could help categorize blood flow to particular depths; this can help improve outcomes of medical treatments such pulse dye laser and photodynamic therapy treatments.

Hybrid photonic signal processing

NASA Astrophysics Data System (ADS)

Ghauri, Farzan Naseer

This thesis proposes research of novel hybrid photonic signal processing systems in the areas of optical communications, test and measurement, RF signal processing and extreme environment optical sensors. It will be shown that use of innovative hybrid techniques allows design of photonic signal processing systems with superior performance parameters and enhanced capabilities. These applications can be divided into domains of analog-digital hybrid signal processing applications and free-space---fiber-coupled hybrid optical sensors. The analog-digital hybrid signal processing applications include a high-performance analog-digital hybrid MEMS variable optical attenuator that can simultaneously provide high dynamic range as well as high resolution attenuation controls; an analog-digital hybrid MEMS beam profiler that allows high-power watt-level laser beam profiling and also provides both submicron-level high resolution and wide area profiling coverage; and all optical transversal RF filters that operate on the principle of broadband optical spectral control using MEMS and/or Acousto-Optic tunable Filters (AOTF) devices which can provide continuous, digital or hybrid signal time delay and weight selection. The hybrid optical sensors presented in the thesis are extreme environment pressure sensors and dual temperature-pressure sensors. The sensors employ hybrid free-space and fiber-coupled techniques for remotely monitoring a system under simultaneous extremely high temperatures and pressures.

Proper alignment of the microscope.

PubMed

Rottenfusser, Rudi

2013-01-01

The light microscope is merely the first element of an imaging system in a research facility. Such a system may include high-speed and/or high-resolution image acquisition capabilities, confocal technologies, and super-resolution methods of various types. Yet more than ever, the proverb "garbage in-garbage out" remains a fact. Image manipulations may be used to conceal a suboptimal microscope setup, but an artifact-free image can only be obtained when the microscope is optimally aligned, both mechanically and optically. Something else is often overlooked in the quest to get the best image out of the microscope: Proper sample preparation! The microscope optics can only do its job when its design criteria are matched to the specimen or vice versa. The specimen itself, the mounting medium, the cover slip, and the type of immersion medium (if applicable) are all part of the total optical makeup. To get the best results out of a microscope, understanding the functions of all of its variable components is important. Only then one knows how to optimize these components for the intended application. Different approaches might be chosen to discuss all of the microscope's components. We decided to follow the light path which starts with the light source and ends at the camera or the eyepieces. To add more transparency to this sequence, the section up to the microscope stage was called the "Illuminating Section", to be followed by the "Imaging Section" which starts with the microscope objective. After understanding the various components, we can start "working with the microscope." To get the best resolution and contrast from the microscope, the practice of "Koehler Illumination" should be understood and followed by every serious microscopist. Step-by-step instructions as well as illustrations of the beam path in an upright and inverted microscope are included in this chapter. A few practical considerations are listed in Section 3. Copyright © 2013 Elsevier Inc. All rights reserved.

The virtual case: a new method to completely digitize cytological and histological slides.

PubMed

Demichelis, F; Barbareschi, M; Dalla Palma, P; Forti, S

2002-08-01

The purpose of this study was to present a new method for handling histological/cytological cases. Thanks to the introduction of information technology in pathology, including the amenities afforded by robotic microscopes and digital imaging, tissue slides can be represented and evaluated using digital techniques in order to construct virtual cases through completely automated procedures. A virtual case (VC) is composed of a collection of digital images representing a histological/cytological slide at all magnification levels together with all relevant clinical data. In the present study, we describe an automated system to manage robotic microscope and image acquisition for the proper construction of VCs. These can then be viewed on a computer by means of an interface ("user-friendly") that allows one to select the more appropriate fields and to examine them at different magnifications, rapidly going from panoramic views to high resolution and vice versa. In comparison with glass slides, VCs have several advantages arising from their digital nature and can be considered a common platform for a wide range of applications such as teleconsultation, education, research, and quality control and proficiency tests.

Three-dimensional automated nanoparticle tracking using Mie scattering in an optical microscope.

PubMed

Gineste, J-M; Macko, P; Patterson, E A; Whelan, M P

2011-08-01

The forward scattering of light in a conventional inverted optical microscope by nanoparticles ranging in diameter from 10 to 50nm has been used to automatically and quantitatively identify and track their location in three-dimensions with a temporal resolution of 200ms. The standard deviation of the location of nominally stationary 50-nm-diameter nanoparticles was found to be about 50nm along the light path and about 5nm in the plane perpendicular to the light path. The method is based on oscillating the microscope objective along the light path using a piezo actuator and acquiring images with the condenser aperture closed to a minimum to enhance the effects of diffraction. Data processing in the time and spatial domains allowed the location of particles to be obtained automatically so that the technique has potential applications both in the processing of nanoparticles and in their use in a variety of fields including nanobiotechnology, pharmaceuticals and food processing where a simple optical microscope maybe preferred for a variety of reasons. © 2011 The Authors Journal of Microscopy © 2011 Royal Microscopical Society.

Programmable Colored Illumination Microscopy (PCIM): A practical and flexible optical staining approach for microscopic contrast enhancement

NASA Astrophysics Data System (ADS)

Zuo, Chao; Sun, Jiasong; Feng, Shijie; Hu, Yan; Chen, Qian

2016-03-01

Programmable colored illumination microscopy (PCIM) has been proposed as a flexible optical staining technique for microscopic contrast enhancement. In this method, we replace the condenser diaphragm of a conventional microscope with a programmable thin film transistor-liquid crystal display (TFT-LCD). By displaying different patterns on the LCD, numerous established imaging modalities can be realized, such as bright field, dark field, phase contrast, oblique illumination, and Rheinberg illuminations, which conventionally rely on intricate alterations in the respective microscope setups. Furthermore, the ease of modulating both the color and the intensity distribution at the aperture of the condenser opens the possibility to combine multiple microscopic techniques, or even realize completely new methods for optical color contrast staining, such as iridescent dark-field and iridescent phase-contrast imaging. The versatility and effectiveness of PCIM is demonstrated by imaging of several transparent colorless specimens, such as unstained lung cancer cells, diatom, textile fibers, and a cryosection of mouse kidney. Finally, the potentialities of PCIM for RGB-splitting imaging with stained samples are also explored by imaging stained red blood cells and a histological section.

A dark-field microscope for background-free detection of resonance fluorescence from single semiconductor quantum dots operating in a set-and-forget mode

NASA Astrophysics Data System (ADS)

Kuhlmann, Andreas V.; Houel, Julien; Brunner, Daniel; Ludwig, Arne; Reuter, Dirk; Wieck, Andreas D.; Warburton, Richard J.

2013-07-01

Optically active quantum dots, for instance self-assembled InGaAs quantum dots, are potentially excellent single photon sources. The fidelity of the single photons is much improved using resonant rather than non-resonant excitation. With resonant excitation, the challenge is to distinguish between resonance fluorescence and scattered laser light. We have met this challenge by creating a polarization-based dark-field microscope to measure the resonance fluorescence from a single quantum dot at low temperature. We achieve a suppression of the scattered laser exceeding a factor of 107 and background-free detection of resonance fluorescence. The same optical setup operates over the entire quantum dot emission range (920-980 nm) and also in high magnetic fields. The major development is the outstanding long-term stability: once the dark-field point has been established, the microscope operates for days without alignment. The mechanical and optical designs of the microscope are presented, as well as exemplary resonance fluorescence spectroscopy results on individual quantum dots to underline the microscope's excellent performance.

Accuracy and eligibility of CBCT to digitize dental plaster casts.

PubMed

Becker, Kathrin; Schmücker, Ulf; Schwarz, Frank; Drescher, Dieter

2018-05-01

Software-based dental planning requires digital casts and oftentimes cone-beam computed tomography (CBCT) radiography. However, buying a dedicated model digitizing device can be expensive and might not be required. The present study aimed to assess whether digital models derived from CBCT and models digitized using a dedicated optical device are of comparable accuracy. A total of 20 plaster casts were digitized with eight CBCT and five optical model digitizers. Corresponding models were superimposed using six control points and subsequent iterative closest point matching. Median distances were calculated among all registered models. Data were pooled per scanner and model. Boxplots were generated, and the paired t test, a Friedman test, and a post-hoc Nemenyi test were employed for statistical comparison. Results were found significant at p < 0.05. All CBCT devices allowed the digitization of plaster casts, but failed to reach the accuracy of the dedicated model digitizers (p < 0.001). Median distances between CBCT and optically digitized casts were 0.064 + - 0.005 mm. Qualitative differences among the CBCT systems were detected (χ 2  = 78.07, p < 0.001), and one CBCT providing a special plaster cast digitization mode was found superior to the competitors (p < 0.05). CBCT systems failed to reach the accuracy from optical digitizers, but within the limits of the study, accuracy appeared to be sufficient for digital planning and forensic purposes. Most CBCT systems enabled digitization of plaster casts, and accuracy was found sufficient for digital planning and storage purposes.

Digital learning programs - competition for the classical microscope?

PubMed Central

Schmidt, Peter

2013-01-01

The development of digital media has been impressive in recent years which is also among the reason for their increasing use in academic teaching. This is especially true for teaching Anatomy and Histology in the first two years in medical and dental curricula. Modern digital technologies allow for efficient, affordable and easily accessible distribution of histological images in high quality. Microscopy depends almost exclusively on such images. Since 20 years numerous digital teaching systems have been developed for this purpose. Respective developments have changed the ways students acquire knowledge and prepare for exams. Teaching staff should adapt lectures, seminars and labs accordingly. As a first step, a collection of high resolution digital microscopic slides was made available for students at the Friedrich-Schiller-University in Jena. The aim of the present study was to evaluate the importance of conventional light microscopy and related technologies in current and future medical and dental education aswell. A survey was done among 172 medical and dental students at the Friedrich-Schiller-University Jena. 51% of students use now frequently new digital media for learning histology in contrast to 5% in the year 2000 [1]. Digital media including Internet, CD- based learning combined with social networks successfully compete with classical light microscopy. PMID:23467698

Bi-directional transmission of molecular information by photon or electron beams passing in the close vicinity of specific molecules, and its clinical and basic research applications: 1) Diagnosis of humans or animal patients without any direct contact; 2) Light microscopic and electron microscopic localization of neuro-transmitters, heavy metals, Oncogen C-fos (AB2), etc. of intracellular fine structures of normal and abnormal single cells using light or electro-microscopic indirect Bi-Digital O-Ring Test.

PubMed

Omura, Y; Losco, M; Omura, A K; Takeshige, C; Hisamitsu, T; Nakajima, H; Soejima, K; Yamamoto, S; Ishikawa, H; Kagoshima, T

1992-01-01

In 1985, Omura, Y. discovered that, when specific molecules were placed anywhere in the close vicinity of the path of a light beam (laser), their molecular information, as well as information on electrical & magnetic fields, is transmitted bi-directionally along the path of this light beam. Namely, this information is transmitted in the direction the light beam is projected and towards the direction from which the light beam is coming. This finding was applied to the following clinical and basic research: 1) In the past, using indirect Bi-Digital O-Ring Test, human or animal patients were diagnosed through an intermediate third person holding a good electrical conducting probe, the tip of which was touching the part of the patient to be examined. However, in order to diagnose the patient in isolation from a distance, or a dangerous or unmanagable unanesthesized animal, such as a lion or tiger, the author succeeded in making a diagnosis by replacing the metal conducting probe with a soft laser beam which is held by the one hand of the third person whose index finger is placed in close vicinity of the laser beam generated by a battery-powered penlight-type solid state laser generator. Thus, diagnosis within visible distance, without direct patient contact, became a reality. 2) Using a projection light microscope, by giving indirect Bi-Digital O-Ring Test while contacting with a fine electro-conductive probe on the magnified fine structure of normal and abnormal cells, various normal and abnormal intracellular substances were localized through a third person holding a pure reference control substance with the same hand that is holding the probe as an intermediary for the indirect Bi-Digital O-Ring Test. Instead of the photon beam in a light microscope, the author found that, using an electron beam passing through the close vicinity of specific molecules of specimens in an electron microscope, the molecular information is transmitted to the magnified fluorescent screen, and an indirect Bi-Digital O-Ring Test could be performed through a projected penlight-type solid state soft laser beam on the magnified intracellular structure through an observation glass window. Using the magnified fine structure of the cells, by either a light projection microscopic field or electron microscope, in various cancer cells of both humans and animals, Oncogen C-fos (AB2) and mercury were found inside of the nucleus. Integrin alpha 5 beta 1 was found on cell membranes and nuclear cell membranes of cancer cells. Acetylcholine was not found anywhere within cancer cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Variable Magnification With Kirkpatrick-Baez Optics for Synchrotron X-Ray Microscopy

PubMed Central

Jach, Terrence; Bakulin, Alex S.; Durbin, Stephen M.; Pedulla, Joseph; Macrander, Albert

2006-01-01

We describe the distinction between the operation of a short focal length x-ray microscope forming a real image with a laboratory source (convergent illumination) and with a highly collimated intense beam from a synchrotron light source (Köhler illumination). We demonstrate the distinction with a Kirkpatrick-Baez microscope consisting of short focal length multilayer mirrors operating at an energy of 8 keV. In addition to realizing improvements in the resolution of the optics, the synchrotron radiation microscope is not limited to the usual single magnification at a fixed image plane. Higher magnification images are produced by projection in the limit of geometrical optics with a collimated beam. However, in distinction to the common method of placing the sample behind the optical source of a diverging beam, we describe the situation in which the sample is located in the collimated beam before the optical element. The ultimate limits of this magnification result from diffraction by the specimen and are determined by the sample position relative to the focal point of the optic. We present criteria by which the diffraction is minimized. PMID:27274930

Enhancing performance of LCoS-SLM as adaptive optics by using computer-generated holograms modulation software

NASA Astrophysics Data System (ADS)

Tsai, Chun-Wei; Lyu, Bo-Han; Wang, Chen; Hung, Cheng-Chieh

2017-05-01

We have already developed multi-function and easy-to-use modulation software that was based on LabVIEW system. There are mainly four functions in this modulation software, such as computer generated holograms (CGH) generation, CGH reconstruction, image trimming, and special phase distribution. Based on the above development of CGH modulation software, we could enhance the performance of liquid crystal on silicon - spatial light modulator (LCoSSLM) as similar as the diffractive optical element (DOE) and use it on various adaptive optics (AO) applications. Through the development of special phase distribution, we are going to use the LCoS-SLM with CGH modulation software into AO technology, such as optical microscope system. When the LCOS-SLM panel is integrated in an optical microscope system, it could be placed on the illumination path or on the image forming path. However, LCOS-SLM provides a program-controllable liquid crystal array for optical microscope. It dynamically changes the amplitude or phase of light and gives the obvious advantage, "Flexibility", to the system

Radiation pressure excitation of Low Temperature Atomic Force & Magnetic Force Microscope (LT-AFM/MFM) for Imaging

NASA Astrophysics Data System (ADS)

Karci, Ozgur; Celik, Umit; Oral, Ahmet; NanoMagnetics Instruments Ltd. Team; Middle East Tech Univ Team

2015-03-01

We describe a novel method for excitation of Atomic Force Microscope (AFM) cantilevers by means of radiation pressure for imaging in an AFM for the first time. Piezo excitation is the most common method for cantilever excitation, but it may cause spurious resonance peaks. A fiber optic interferometer with 1310 nm laser was used both to measure the deflection of cantilever and apply a force to the cantilever in a LT-AFM/MFM from NanoMagnetics Instruments. The laser power was modulated at the cantilever`s resonance frequency by a digital Phase Lock Loop (PLL). The force exerted by the radiation pressure on a perfectly reflecting surface by a laser beam of power P is F = 2P/c. We typically modulate the laser beam by ~ 800 μW and obtain 10nm oscillation amplitude with Q ~ 8,000 at 2.5x10-4 mbar. The cantilever's stiffness can be accurately calibrated by using the radiation pressure. We have demonstrated performance of the radiation pressure excitation in AFM/MFM by imaging a hard disk sample between 4-300K and Abrikosov vortex lattice in BSCCO single crystal at 4K to for the first time.

Leaf epidermis images for robust identification of plants

PubMed Central

da Silva, Núbia Rosa; Oliveira, Marcos William da Silva; Filho, Humberto Antunes de Almeida; Pinheiro, Luiz Felipe Souza; Rossatto, Davi Rodrigo; Kolb, Rosana Marta; Bruno, Odemir Martinez

2016-01-01

This paper proposes a methodology for plant analysis and identification based on extracting texture features from microscopic images of leaf epidermis. All the experiments were carried out using 32 plant species with 309 epidermal samples captured by an optical microscope coupled to a digital camera. The results of the computational methods using texture features were compared to the conventional approach, where quantitative measurements of stomatal traits (density, length and width) were manually obtained. Epidermis image classification using texture has achieved a success rate of over 96%, while success rate was around 60% for quantitative measurements taken manually. Furthermore, we verified the robustness of our method accounting for natural phenotypic plasticity of stomata, analysing samples from the same species grown in different environments. Texture methods were robust even when considering phenotypic plasticity of stomatal traits with a decrease of 20% in the success rate, as quantitative measurements proved to be fully sensitive with a decrease of 77%. Results from the comparison between the computational approach and the conventional quantitative measurements lead us to discover how computational systems are advantageous and promising in terms of solving problems related to Botany, such as species identification. PMID:27217018

Nonlinear optical microscopy for immunoimaging: a custom optimized system of high-speed, large-area, multicolor imaging

PubMed Central

Li, Hui; Cui, Quan; Zhang, Zhihong; Luo, Qingming

2015-01-01

Background The nonlinear optical microscopy has become the current state-of-the-art for intravital imaging. Due to its advantages of high resolution, superior tissue penetration, lower photodamage and photobleaching, as well as intrinsic z-sectioning ability, this technology has been widely applied in immunoimaging for a decade. However, in terms of monitoring immune events in native physiological environment, the conventional nonlinear optical microscope system has to be optimized for live animal imaging. Generally speaking, three crucial capabilities are desired, including high-speed, large-area and multicolor imaging. Among numerous high-speed scanning mechanisms used in nonlinear optical imaging, polygon scanning is not only linearly but also dispersion-freely with high stability and tunable rotation speed, which can overcome disadvantages of multifocal scanning, resonant scanner and acousto-optical deflector (AOD). However, low frame rate, lacking large-area or multicolor imaging ability make current polygonbased nonlinear optical microscopes unable to meet the requirements of immune event monitoring. Methods We built up a polygon-based nonlinear optical microscope system which was custom optimized for immunoimaging with high-speed, large-are and multicolor imaging abilities. Results Firstly, we validated the imaging performance of the system by standard methods. Then, to demonstrate the ability to monitor immune events, migration of immunocytes observed by the system based on typical immunological models such as lymph node, footpad and dorsal skinfold chamber are shown. Finally, we take an outlook for the possible advance of related technologies such as sample stabilization and optical clearing for more stable and deeper intravital immunoimaging. Conclusions This study will be helpful for optimizing nonlinear optical microscope to obtain more comprehensive and accurate information of immune events. PMID:25694951

Analysis of slide exploration strategy of cytologists when reading digital slides

NASA Astrophysics Data System (ADS)

Pantanowitz, Liron; Parwani, Anil; Tseytlin, Eugene; Mello-Thoms, Claudia

2012-02-01

Cytology is the sub-domain of Pathology that deals mainly with the diagnosis of cellular changes caused by disease. Current clinical practice involves a cytotechnologist that manually screens glass slides containing fixed cytology material using a light microscope. Screened slides are then forwarded to a specialized pathologist, a cytopathologist, for microscopic review and final diagnostic interpretation. If no abnormalities are detected, the specimen is interpreted as "normal", otherwise the abnormalities are marked with a pen on the glass slide by the cytotechnologist and then are used to render a diagnosis. As Pathology is migrating towards a digital environment it is important to determine whether these crucial screening and diagnostic tasks can be performed as well using digital slides as the current practice with glass slides. The purpose of this work is to make this assessment, by using a set of digital slides depicting cytological materials of different disease processes in several organs, and then to analyze how different cytologists including cytotechnologists, cytopathologists and cytotechnology-trainees explored the digital slides. We will (1) collect visual search data from the cytologists as they navigate the digital slides, as well as record any electronic marks (annotations) made by the cytologists; (2) convert the dynamic visual search data into a static representation of the observers' exploration strategy using 'search maps'; and (3) determine slide coverage, per viewing magnification range, for each group. We have developed a virtual microscope to collect this data, and this interface allows for interactive navigation of the virtual slide (including panning and zooming), as well as annotation of reportable findings. Furthermore, all interactions with the interface are time stamped, which allows us to recreate the cytologists' search strategy.

Comparison of a digital and an optical analogue hand-held refractometer for the measurement of canine urine specific gravity.

PubMed

Paris, J K; Bennett, A D; Dodkin, S J; Gunn-Moore, D A

2012-05-05

Urine specific gravity (USG) is used clinically as a measure of urine concentration, and is routinely assessed by refractometry. A comparison between optical analogue and digital refractometers for evaluation of canine urine has not been reported. The aim of this study was to compare a digital and an optical analogue hand-held refractometer for the measurement of canine USG, and to assess correlation with urine osmolality. Prospective study. Free-catch urine samples were collected from 285 hospitalised adult dogs, and paired USG readings were obtained with a digital and an optical analogue refractometer. In 50 dogs, urine osmolality was also measured using a freezing point depression osmometer. There was a small but statistically significant difference between the two refractometers (P<0.001), with the optical analogue refractometer reading higher than the digital refractometer (mean difference 0.0006, sd 0.0012). Paired refractometer measurements varied by <0.002 in 91.5 per cent of cases. The optical analogue and digital refractometer readings showed excellent correlation with osmolality (r=0.980 and r=0.977, respectively, P<0.001 in both cases). Despite statistical significance, the difference between the two refractometers is unlikely to be clinically significant. Both instruments provide an accurate assessment of USG in dogs.

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.

Magnetically tunable oil droplet lens of deep-sea shrimp

NASA Astrophysics Data System (ADS)

Iwasaka, M.; Hirota, N.; Oba, Y.

2018-05-01

In this study, the tunable properties of a bio-lens from a deep-sea shrimp were investigated for the first time using magnetic fields. The skin of the shrimp exhibited a brilliantly colored reflection of incident white light. The light reflecting parts and the oil droplets in the shrimp's skin were observed in a glass slide sample cell using a digital microscope that operated in the bore of two superconducting magnets (maximum strengths of 5 and 13 T). In the ventral skin of the shrimp, which contained many oil droplets, some comparatively large oil droplets (50 to 150 μm in diameter) were present. A distinct response to magnetic fields was found in these large oil droplets. Further, the application of the magnetic fields to the sample cell caused a change in the size of the oil droplets. The phenomena observed in this work indicate that the oil droplets of deep sea shrimp can act as lenses in which the optical focusing can be modified via the application of external magnetic fields. The results of this study will make it possible to fabricate bio-inspired soft optical devices in future.

Qualitative and quantitative interpretation of SEM image using digital image processing.

PubMed

Saladra, Dawid; Kopernik, Magdalena

2016-10-01

The aim of the this study is improvement of qualitative and quantitative analysis of scanning electron microscope micrographs by development of computer program, which enables automatic crack analysis of scanning electron microscopy (SEM) micrographs. Micromechanical tests of pneumatic ventricular assist devices result in a large number of micrographs. Therefore, the analysis must be automatic. Tests for athrombogenic titanium nitride/gold coatings deposited on polymeric substrates (Bionate II) are performed. These tests include microshear, microtension and fatigue analysis. Anisotropic surface defects observed in the SEM micrographs require support for qualitative and quantitative interpretation. Improvement of qualitative analysis of scanning electron microscope images was achieved by a set of computational tools that includes binarization, simplified expanding, expanding, simple image statistic thresholding, the filters Laplacian 1, and Laplacian 2, Otsu and reverse binarization. Several modifications of the known image processing techniques and combinations of the selected image processing techniques were applied. The introduced quantitative analysis of digital scanning electron microscope images enables computation of stereological parameters such as area, crack angle, crack length, and total crack length per unit area. This study also compares the functionality of the developed computer program of digital image processing with existing applications. The described pre- and postprocessing may be helpful in scanning electron microscopy and transmission electron microscopy surface investigations. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

Modified signed-digit trinary arithmetic by using optical symbolic substitution.

PubMed

Awwal, A A; Islam, M N; Karim, M A

1992-04-10

Carry-free addition and borrow-free subtraction of modified signed-digit trinary numbers with optical symbolic substitution are presented. The proposed two-step and three-step algorithms can be easily implemented by using phase-only holograms, optical content-addressable memories, a multichannel correlator, or a polarization-encoded optical shadow-casting system.

Modified signed-digit trinary arithmetic by using optical symbolic substitution

NASA Astrophysics Data System (ADS)

Awwal, A. A. S.; Islam, M. N.; Karim, M. A.

1992-04-01

Carry-free addition and borrow-free subtraction of modified signed-digit trinary numbers with optical symbolic substitution are presented. The proposed two-step and three-step algorithms can be easily implemented by using phase-only holograms, optical content-addressable memories, a multichannel correlator, or a polarization-encoded optical shadow-casting system.

Systems and methods for free space optical communication

DOEpatents

Harper, Warren W [Benton City, WA; Aker, Pamela M [Richland, WA; Pratt, Richard M [Richland, WA

2011-05-10

Free space optical communication methods and systems, according to various aspects are described. The methods and systems are characterized by transmission of data through free space with a digitized optical signal acquired using wavelength modulation, and by discrimination between bit states in the digitized optical signal using a spectroscopic absorption feature of a chemical substance.

Digital adaptive optics confocal microscopy based on iterative retrieval of optical aberration from a guidestar hologram

PubMed Central

Liu, Changgeng; Thapa, Damber; Yao, Xincheng

2017-01-01

Guidestar hologram based digital adaptive optics (DAO) is one recently emerging active imaging modality. It records each complex distorted line field reflected or scattered from the sample by an off-axis digital hologram, measures the optical aberration from a separate off-axis digital guidestar hologram, and removes the optical aberration from the distorted line fields by numerical processing. In previously demonstrated DAO systems, the optical aberration was directly retrieved from the guidestar hologram by taking its Fourier transform and extracting the phase term. For the direct retrieval method (DRM), when the sample is not coincident with the guidestar focal plane, the accuracy of the optical aberration retrieved by DRM undergoes a fast decay, leading to quality deterioration of corrected images. To tackle this problem, we explore here an image metrics-based iterative method (MIM) to retrieve the optical aberration from the guidestar hologram. Using an aberrated objective lens and scattering samples, we demonstrate that MIM can improve the accuracy of the retrieved aberrations from both focused and defocused guidestar holograms, compared to DRM, to improve the robustness of the DAO. PMID:28380937

Microscopic Studies of Quantum Phase Transitions in Optical Lattices

NASA Astrophysics Data System (ADS)

Bakr, Waseem S.

2011-12-01

In this thesis, I report on experiments that microscopically probe quantum phase transitions of ultracold atoms in optical lattices. We have developed a "quantum gas microscope" that allowed, for the first time, optical imaging and manipulation of single atoms in a quantum-degenerate gas on individual sites of an optical lattice. This system acts as a quantum simulator of strongly correlated materials, which are currently the subject of intense research because of the technological potential of high--T c superconductors and spintronic materials. We have used our microscope to study the superfluid to Mott insulator transition in bosons and a magnetic quantum phase transition in a spin system. In our microscopic study of the superfluid-insulator transition, we have characterized the on-site number statistics in a space- and time-resolved manner. We observed Mott insulators with fidelities as high as 99%, corresponding to entropies of 0.06kB per particle. We also measured local quantum dynamics and directly imaged the shell structure of the Mott insulator. I report on the first quantum magnetism experiments in optical lattices. We have realized a quantum Ising chain in a magnetic field, and observed a quantum phase transition between a paramagnet and antiferromagnet. We achieved strong spin interactions by encoding spins in excitations of a Mott insulator in a tilted lattice. We detected the transition by measuring the total magnetization of the system across the transition using in-situ measurements as well as the Neel ordering in the antiferromagnetic state using noise-correlation techniques. We characterized the dynamics of domain formation in the system. The spin mapping introduced opens up a new path to realizing more exotic states in optical lattices including spin liquids and quantum valence bond solids. As our system sizes become larger, simulating their physics on classical computers will require exponentially larger resources because of entanglement build-up near a quantum phase transition. We have demonstrated a quantum simulator in which all degrees of freedom can be read out microscopically, allowing the simulation of quantum many-body systems with manageable resources. More generally, the ability to image and manipulate individual atoms in optical lattices opens an avenue towards scalable quantum computation.

Tele-manufactured affordable smartphone anterior segment microscope.

PubMed

Chiong, Hong Sheng; Fang, Joyce Lim Luann; Wilson, Graham

2016-11-01

The recent advances in mobile technology have made the smartphone a powerful and accessible tool. This article describe the development of a novel smartphone-based anterior segment microscope that is compatible with tele-manufacturing. The anterior segment microscope is equipped with both cobalt-blue and red-free filters that can be used for clinical photo-documentation. The digital files of the microscope are transferrable and compatible with additive-manufacturing. Therefore, the entire device can be locally manufactured with rapid prototyping techniques such as 3D printing. © 2016 Optometry Australia.

Development and Optical Testing of the Camera, Hand Lens, and Microscope Probe with Scannable Laser Spectroscopy (CHAMP-SLS)

NASA Technical Reports Server (NTRS)

Mungas, Greg S.; Gursel, Yekta; Sepulveda, Cesar A.; Anderson, Mark; La Baw, Clayton; Johnson, Kenneth R.; Deans, Matthew; Beegle, Luther; Boynton, John

2008-01-01

Conducting high resolution field microscopy with coupled laser spectroscopy that can be used to selectively analyze the surface chemistry of individual pixels in a scene is an enabling capability for next generation robotic and manned spaceflight missions, civil, and military applications. In the laboratory, we use a range of imaging and surface preparation tools that provide us with in-focus images, context imaging for identifying features that we want to investigate at high magnification, and surface-optical coupling that allows us to apply optical spectroscopic analysis techniques for analyzing surface chemistry particularly at high magnifications. The camera, hand lens, and microscope probe with scannable laser spectroscopy (CHAMP-SLS) is an imaging/spectroscopy instrument capable of imaging continuously from infinity down to high resolution microscopy (resolution of approx. 1 micron/pixel in a final camera format), the closer CHAMP-SLS is placed to a feature, the higher the resultant magnification. At hand lens to microscopic magnifications, the imaged scene can be selectively interrogated with point spectroscopic techniques such as Raman spectroscopy, microscopic Laser Induced Breakdown Spectroscopy (micro-LIBS), laser ablation mass-spectrometry, Fluorescence spectroscopy, and/or Reflectance spectroscopy. This paper summarizes the optical design, development, and testing of the CHAMP-SLS optics.

Five years of experience teaching pathology to dental students using the WebMicroscope

PubMed Central

2011-01-01

Background We describe development and evaluation of the user-friendly web based virtual microscopy - WebMicroscope for teaching and learning dental students basic and oral pathology. Traditional students microscopes were replaced by computer workstations. Methods The transition of the basic and oral pathology courses from light to virtual microscopy has been completed gradually over a five-year period. A pilot study was conducted in academic year 2005/2006 to estimate the feasibility of integrating virtual microscopy into a traditional light microscopy-based pathology course. The entire training set of glass slides was subsequently converted to virtual slides and placed on the WebMicroscope server. Giving access to fully digitized slides on the web with a browser and a viewer plug-in, the computer has become a perfect companion of the student. Results The study material consists now of over 400 fully digitized slides which covering 15 entities in basic and systemic pathology and 15 entities in oral pathology. Digitized slides are linked with still macro- and microscopic images, organized with clinical information into virtual cases and supplemented with text files, syllabus, PowerPoint presentations and animations on the web, serving additionally as material for individual studies. After their examinations, the students rated the use of the software, quality of the images, the ease of handling the images, and the effective use of virtual slides during the laboratory practicals. Responses were evaluated on a standardized scale. Because of the positive opinions and support from the students, the satisfaction surveys had shown a progressive improvement over the past 5 years. The WebMicroscope as a didactic tool for laboratory practicals was rated over 8 on a 1-10 scale for basic and systemic pathology and 9/10 for oral pathology especially as various students’ suggestions were implemented. Overall, the quality of the images was rated as very good. Conclusions An overwhelming majority of our students regarded a possibility of using virtual slides at their convenience as highly desirable. Our students and faculty consider the use of the virtual microscope for the study of basic as well as oral pathology as a significant improvement over the light microscope. PMID:21489183

Optical Coherence Tomography–Enhanced Microlaryngoscopy: Preliminary Report of a Noncontact Optical Coherence Tomography System Integrated With a Surgical Microscope

PubMed Central

Vokes, David E.; Jackson, Ryan; Guo, Shuguang; Perez, Jorge A.; Su, Jianping; Ridgway, James M.; Armstrong, William B.; Chen, Zhongping; Wong, Brian J. F.

2014-01-01

Objectives Optical coherence tomography (OCT) is a new imaging modality that uses near-infrared light to produce cross-sectional images of tissue with a resolution approaching that of light microscopy. We have previously reported use of OCT imaging of the vocal folds (VFs) during direct laryngoscopy with a probe held in contact or near-contact with the VFs. This aim of this study was to develop and evaluate a novel OCT system integrated with a surgical microscope to allow hands-free OCT imaging of the VFs, which could be performed simultaneously with microscopic visualization. Methods We performed a prospective evaluation of a new method of acquiring OCT images of the VFs. Results An OCT system was successfully integrated with a surgical microscope to permit noncontact OCT imaging of the VFs of 10 patients. With this novel device we were able to identify VF epithelium and lamina propria; however, the resolution was reduced compared to that achieved with the standard contact or near-contact OCT. Conclusions Optical coherence tomography is able to produce high-resolution images of vocal fold mucosa to a maximum depth of 1.6 mm. It may be used in the diagnosis of VF lesions, particularly early squamous cell carcinoma, in which OCT can show disruption of the basement membrane. Mounting the OCT device directly onto the operating microscope allows hands-free noncontact OCT imaging and simultaneous conventional microscopic visualization of the VFs. However, the lateral resolution of the OCT microscope system is 50 µm, in contrast to the conventional handheld probe system (10 µm). Although such images at this resolution are still useful clinically, improved resolution would enhance the system’s performance, potentially enabling real-time OCT-guided microsurgery of the larynx. PMID:18700431

Wavelet processing and digital interferometric contrast to improve reconstructions from X-ray Gabor holograms.

PubMed

Aguilar, Juan C; Misawa, Masaki; Matsuda, Kiyofumi; Suzuki, Yoshio; Takeuchi, Akihisa; Yasumoto, Masato

2018-05-01

In this work, the application of an undecimated wavelet transformation together with digital interferometric contrast to improve the resulting reconstructions in a digital hard X-ray Gabor holographic microscope is shown. Specifically, the starlet transform is used together with digital Zernike contrast. With this contrast, the results show that only a small set of scales from the hologram are, in effect, useful, and it is possible to enhance the details of the reconstruction.

CMOS-compatible InP/InGaAs digital photoreceiver

DOEpatents

Lovejoy, Michael L.; Rose, Benny H.; Craft, David C.; Enquist, Paul M.; Slater, Jr., David B.

1997-01-01

A digital photoreceiver is formed monolithically on an InP semiconductor substrate and comprises a p-i-n photodetector formed from a plurality of InP/InGaAs layers deposited by an epitaxial growth process and an adjacent heterojunction bipolar transistor (HBT) amplifier formed from the same InP/InGaAs layers. The photoreceiver amplifier operates in a large-signal mode to convert a detected photocurrent signal into an amplified output capable of directly driving integrated circuits such as CMOS. In combination with an optical transmitter, the photoreceiver may be used to establish a short-range channel of digital optical communications between integrated circuits with applications to multi-chip modules (MCMs). The photoreceiver may also be used with fiber optic coupling for establishing longer-range digital communications (i.e. optical interconnects) between distributed computers or the like. Arrays of digital photoreceivers may be formed on a common substrate for establishing a plurality of channels of digital optical communication, with each photoreceiver being spaced by less than about 1 mm and consuming less than about 20 mW of power, and preferably less than about 10 mW. Such photoreceiver arrays are useful for transferring huge amounts of digital data between integrated circuits at bit rates of up to about 1000 Mb/s or more.

A home-built digital optical MRI console using high-speed serial links.

PubMed

Tang, Weinan; Wang, Weimin; Liu, Wentao; Ma, Yajun; Tang, Xin; Xiao, Liang; Gao, Jia-Hong

2015-08-01

To develop a high performance, cost-effective digital optical console for scalable multichannel MRI. The console system was implemented with flexibility and efficiency based on a modular architecture with distributed pulse sequencers. High-speed serial links were optimally utilized to interconnect the system, providing fast digital communication with a multi-gigabit data rate. The conventional analog radio frequency (RF) chain was replaced with a digital RF manipulation. The acquisition electronics were designed in close proximity to RF coils and preamplifiers, using a digital optical link to transmit the MR signal. A prototype of the console was constructed with a broad frequency range from direct current to 100 MHz. A temporal resolution of 1 μs was achieved for both the RF and gradient operations. The MR signal was digitized in the scanner room with an overall dynamic range between 16 and 24 bits and was transmitted to a master controller over a duplex optic fiber with a high data rate of 3.125 gigabits per second. High-quality phantom and human images were obtained using the prototype on both 0.36T and 1.5T clinical MRI scanners. A homemade digital optical MRI console with high-speed serial interconnection has been developed to better serve imaging research and clinical applications. © 2014 Wiley Periodicals, Inc.

CMOS-compatible InP/InGaAs digital photoreceiver

DOEpatents

Lovejoy, M.L.; Rose, B.H.; Craft, D.C.; Enquist, P.M.; Slater, D.B. Jr.

1997-11-04

A digital photoreceiver is formed monolithically on an InP semiconductor substrate and comprises a p-i-n photodetector formed from a plurality of InP/InGaAs layers deposited by an epitaxial growth process and an adjacent heterojunction bipolar transistor (HBT) amplifier formed from the same InP/InGaAs layers. The photoreceiver amplifier operates in a large-signal mode to convert a detected photocurrent signal into an amplified output capable of directly driving integrated circuits such as CMOS. In combination with an optical transmitter, the photoreceiver may be used to establish a short-range channel of digital optical communications between integrated circuits with applications to multi-chip modules (MCMs). The photoreceiver may also be used with fiber optic coupling for establishing longer-range digital communications (i.e. optical interconnects) between distributed computers or the like. Arrays of digital photoreceivers may be formed on a common substrate for establishing a plurality of channels of digital optical communication, with each photoreceiver being spaced by less than about 1 mm and consuming less than about 20 mW of power, and preferably less than about 10 mW. Such photoreceiver arrays are useful for transferring huge amounts of digital data between integrated circuits at bit rates of up to about 1,000 Mb/s or more. 4 figs.

Optics of high-performance electron microscopes*

PubMed Central

Rose, H H

2008-01-01

During recent years, the theory of charged particle optics together with advances in fabrication tolerances and experimental techniques has lead to very significant advances in high-performance electron microscopes. Here, we will describe which theoretical tools, inventions and designs have driven this development. We cover the basic theory of higher-order electron optics and of image formation in electron microscopes. This leads to a description of different methods to correct aberrations by multipole fields and to a discussion of the most advanced design that take advantage of these techniques. The theory of electron mirrors is developed and it is shown how this can be used to correct aberrations and to design energy filters. Finally, different types of energy filters are described. PMID:27877933

Using Digital Microscopy

ERIC Educational Resources Information Center

Travaille, Madelaine; Adams, Sandra D.

2006-01-01

Studying "Caenorhabditis elegans" ("C. elegans") live cultures provides excellent opportunities for authentic inquiry in a high school anatomy and physiology or other biology lab course. Using a digital dissection microscope, a student can photograph the organism during various stages of development and study and analyze the images. In this…

Code conversion from signed-digit to complement representation based on look-ahead optical logic operations

NASA Astrophysics Data System (ADS)

Li, Guoqiang; Qian, Feng

2001-11-01

We present, for the first time to our knowledge, a generalized lookahead logic algorithm for number conversion from signed-digit to complement representation. By properly encoding the signed-digits, all the operations are performed by binary logic, and unified logical expressions can be obtained for conversion from modified-signed- digit (MSD) to 2's complement, trinary signed-digit (TSD) to 3's complement, and quarternary signed-digit (QSD) to 4's complement. For optical implementation, a parallel logical array module using an electron-trapping device is employed and experimental results are shown. This optical module is suitable for implementing complex logic functions in the form of the sum of the product. The algorithm and architecture are compatible with a general-purpose optoelectronic computing system.

Implementing digital technology to enhance student learning of pathology.

PubMed

Farah, C S; Maybury, T

2009-08-01

The introduction of digital technologies into the dental curriculum is an ongoing feature of broader changes going on in tertiary education. This report examines the introduction of digital virtual microscopy technology into the curriculum of the School of Dentistry at the University of Queensland (UQ) in Brisbane, Australia. Sixty students studying a course in pathology in 2005 were introduced to virtual microscopy technology alongside the more traditional light microscope and then asked to evaluate their own learning outcomes from this technology via a structured 5-point LIKART survey. A wide variety of questions dealing the pedagogic implications of the introduction of virtual microscopy into pathology were asked of students with the overall result being that it positively enhanced their learning of pathology via digital microscopic means. The success of virtual microscopy in dentistry at UQ is then discussed in the larger context of changes going on in tertiary education. In particular, the change from the print-literate tradition to the electronic one, that is from 'literacy to electracy'. Virtual microscopy is designated as a component of this transformation to electracy. Whilst traditional microscopic skills may still be valued in dental curricula, the move to virtual microscopy and computer-assisted, student-centred learning of pathology appears to enhance the learning experience in relation to its effectiveness in helping students engage and interact with the course material.

Using Light Scattering to Track, Characterize and Manipulate Colloids

NASA Astrophysics Data System (ADS)

van Oostrum, P. D. J.

2011-03-01

A new technique is developed to analyze in-line Digital Holographic Microscopy images, making it possible to characterize, and track colloidal particles in three dimensions at unprecedented accuracy. We took digital snapshots of the interference pattern between the light scattered by micrometer particles and the unaltered portion of a laser beam that was used to illuminate dilute colloidal dispersions on a light microscope in transmission mode. We numerically fit Mie-theory for the light-scattering by micrometer sized particles to these experimental in-line holograms. The fit values give the position in three dimensions with an accuracy of a few nanometers in the lateral directions and several tens of nanometers in the axial direction. The individual particles radii and refractive indices could be determined to within tens of nanometers and a few hundredths respectively. By using a fast CCD camera, we can track particles with millisecond resolution in time which allows us to study dynamical properties such as the hydrodynamic radius and the sedimentation coefficient. The scattering behavior of the particles that we use to track and characterize colloidal particles makes it possible to exert pico-Newton forces on them close to a diffraction limited focus. When these effects are used to confine colloids in space, this technique is called Optical Tweezers. Both by numerical calculations and by experiments, we explore the possibilities of optical tweezers in soft condensed matter research. Using optical tweezers we placed multiple particles in interesting configurations to measure the interaction forces between them. The interaction forces were Yukawa-like screened charge repulsions. Careful timing of the blinking of time-shared optical tweezers and of the recording of holographic snapshots, we were able to measure interaction forces with femto-Newton accuracy from an analysis of (driven) Brownian motion. Forces exerted by external fields such as electric fields and gravity were measured as well. We induced electric dipoles in colloidal particles by applying radio frequency electric fields. Dipole induced strings of particles were formed and made permanent by van der Waals attractions or thermal annealing. Such colloidal strings form colloidal analogues of charged and un-charged (bio-) polymers. The diffusion and bending behavior of such strings was probed using DHM and optical tweezers.

Matrix-vector multiplication using digital partitioning for more accurate optical computing

NASA Technical Reports Server (NTRS)

Gary, C. K.

1992-01-01

Digital partitioning offers a flexible means of increasing the accuracy of an optical matrix-vector processor. This algorithm can be implemented with the same architecture required for a purely analog processor, which gives optical matrix-vector processors the ability to perform high-accuracy calculations at speeds comparable with or greater than electronic computers as well as the ability to perform analog operations at a much greater speed. Digital partitioning is compared with digital multiplication by analog convolution, residue number systems, and redundant number representation in terms of the size and the speed required for an equivalent throughput as well as in terms of the hardware requirements. Digital partitioning and digital multiplication by analog convolution are found to be the most efficient alogrithms if coding time and hardware are considered, and the architecture for digital partitioning permits the use of analog computations to provide the greatest throughput for a single processor.

Digitized locksmith forensics: automated detection and segmentation of toolmarks on highly structured surfaces

NASA Astrophysics Data System (ADS)

Clausing, Eric; Vielhauer, Claus

2014-02-01

Locksmith forensics is an important area in crime scene forensics. Due to new optical, contactless, nanometer range sensing technology, such traces can be captured, digitized and analyzed more easily allowing a complete digital forensic investigation. In this paper we present a significantly improved approach for the detection and segmentation of toolmarks on surfaces of locking cylinder components (using the example of the locking cylinder component 'key pin') acquired by a 3D Confocal Laser Scanning Microscope. This improved approach is based on our prior work1 using a block-based classification approach with textural features. In this prior work1 we achieve a solid detection rate of 75-85% for the detection of toolmarks originating from illegal opening methods. Here, in this paper we improve, expand and fuse this prior approach with additional features from acquired surface topography data, color data and an image processing approach using adapted Gabor filters. In particular we are able of raising the detection and segmentation rates above 90% with our test set of 20 key pins with approximately 700 single toolmark traces of four different opening methods. We can provide a precise pixel- based segmentation as opposed to the rather imprecise segmentation of our prior block-based approach and as the use of the two additional data types (color and especially topography) require a specific pre-processing, we furthermore propose an adequate approach for this purpose.

Design, Fabrication and Testing of Multilayer Coated X-Ray Optics for the Water Window Imaging X-Ray Microscope

NASA Technical Reports Server (NTRS)

Spencer, Dwight C.

1996-01-01

Hoover et. al. built and tested two imaging Schwarzschild multilayer microscopes. These instruments were constructed as prototypes for the "Water Window Imaging X-Ray Microscope," which is a doubly reflecting, multilayer x-ray microscope configured to operate within the "water window." The "water window" is the narrow region of the x-ray spectrum between the K absorption edges of oxygen (lamda = 23.3 Angstroms) and of carbon (lamda = 43.62 Angstroms), where water is relatively highly transmissive and carbon is highly absorptive. This property of these materials, thus permits the use of high resolution multilayer x-ray microscopes for producing high contrast images of carbon-based structures within the aqueous physiological environments of living cells. We report the design, fabrication and testing of multilayer optics that operate in this regime.

Hyperspectral microscope for in vivo imaging of microstructures and cells in tissues

DOEpatents

Demos,; Stavros, G [Livermore, CA

2011-05-17

An optical hyperspectral/multimodal imaging method and apparatus is utilized to provide high signal sensitivity for implementation of various optical imaging approaches. Such a system utilizes long working distance microscope objectives so as to enable off-axis illumination of predetermined tissue thereby allowing for excitation at any optical wavelength, simplifies design, reduces required optical elements, significantly reduces spectral noise from the optical elements and allows for fast image acquisition enabling high quality imaging in-vivo. Such a technology provides a means of detecting disease at the single cell level such as cancer, precancer, ischemic, traumatic or other type of injury, infection, or other diseases or conditions causing alterations in cells and tissue micro structures.

Optically-synchronized encoder and multiplexer scheme for interleaved photonics analog-to-digital conversion

NASA Astrophysics Data System (ADS)

Villa, Carlos; Kumavor, Patrick; Donkor, Eric

2008-04-01

Photonics Analog-to-Digital Converters (ADCs) utilize a train of optical pulses to sample an electrical input waveform applied to an electrooptic modulator or a reverse biased photodiode. In the former, the resulting train of amplitude-modulated optical pulses is detected (converter to electrical) and quantized using a conversional electronics ADC- as at present there are no practical, cost-effective optical quantizers available with performance that rival electronic quantizers. In the latter, the electrical samples are directly quantized by the electronics ADC. In both cases however, the sampling rate is limited by the speed with which the electronics ADC can quantize the electrical samples. One way to increase the sampling rate by a factor N is by using the time-interleaved technique which consists of a parallel array of N electrical ADC converters, which have the same sampling rate but different sampling phase. Each operating at a quantization rate of fs/N where fs is the aggregated sampling rate. In a system with no real-time operation, the N channels digital outputs are stored in memory, and then aggregated (multiplexed) to obtain the digital representation of the analog input waveform. Alternatively, for real-time operation systems the reduction of storing time in the multiplexing process is desired to improve the time response of the ADC. The complete elimination of memories come expenses of concurrent timing and synchronization in the aggregation of the digital signal that became critical for a good digital representation of the analog signal waveform. In this paper we propose and demonstrate a novel optically synchronized encoder and multiplexer scheme for interleaved photonics ADCs that utilize the N optical signals used to sample different phases of an analog input signal to synchronize the multiplexing of the resulting N digital output channels in a single digital output port. As a proof of concept, four 320 Megasamples/sec 12-bit of resolution digital signals were multiplexed to form an aggregated 1.28 Gigasamples/sec single digital output signal.

Silicon photonics devices for metro applications

NASA Astrophysics Data System (ADS)

Fukuda, H.; Kikuchi, K.; Jizodo, M.; Kawamura, Y.; Takeda, K.; Honda, K.

2017-01-01

Digital coherent technology is considered an attractive way of realizing both high-speed metro links and long distance transmissions. In metro areas, there is a strong demand for a smaller, faster transceiver module. This demand is mainly driven by the rapidly increasing data center interconnection traffic, where transmission capacity per faceplane is a key feature. Therefore, optical integration technology is desired. Since compensation in digital coherent technology is performed in the electrical or digital domain, users can deal with those optics performances that are not compensated for digitally. This means using a new material that cannot provide perfect characteristics but that is suitable for miniaturization and integration is possible. Silicon photonics (SiPh) is considered an attractive technology that would enable the significant miniaturization of optical circuits and be capable of optical integration with high manufacturability. While SiPh-based devices have begun to be deployed for very short or short reach links on the basis of direct detection technology, their digital coherent applications have recently been investigated in view of their integration capability. This paper describes recent progress on SiPh-based integrated optical devices for high-speed digital coherent transceivers targeting metro links. An optical modulator and receiver with related circuits have been integrated into a single SiPh chip. TEC-free operation under non-hermetic conditions and the direct attachment of optical fibers have both been realized. Very thin and small packaging with sufficient performance has been demonstrated by using the SiPh chip co-packaged with high-speed ICs.

Navigation and Image Injection for Control of Bone Removal and Osteotomy Planes in Spine Surgery.

PubMed

Kosterhon, Michael; Gutenberg, Angelika; Kantelhardt, Sven Rainer; Archavlis, Elefterios; Giese, Alf

2017-04-01

In contrast to cranial interventions, neuronavigation in spinal surgery is used in few applications, not tapping into its full technological potential. We have developed a method to preoperatively create virtual resection planes and volumes for spinal osteotomies and export 3-D operation plans to a navigation system controlling intraoperative visualization using a surgical microscope's head-up display. The method was developed using a Sawbone ® model of the lumbar spine, demonstrating feasibility with high precision. Computer tomographic and magnetic resonance image data were imported into Amira ® , a 3-D visualization software. Resection planes were positioned, and resection volumes representing intraoperative bone removal were defined. Fused to the original Digital Imaging and Communications in Medicine data, the osteotomy planes were exported to the cranial version of a Brainlab ® navigation system. A navigated surgical microscope with video connection to the navigation system allowed intraoperative image injection to visualize the preplanned resection planes. The workflow was applied to a patient presenting with a congenital hemivertebra of the thoracolumbar spine. Dorsal instrumentation with pedicle screws and rods was followed by resection of the deformed vertebra guided by the in-view image injection of the preplanned resection planes into the optical path of a surgical microscope. Postoperatively, the patient showed no neurological deficits, and the spine was found to be restored in near physiological posture. The intraoperative visualization of resection planes in a microscope's head-up display was found to assist the surgeon during the resection of a complex-shaped bone wedge and may help to further increase accuracy and patient safety. Copyright © 2017 by the Congress of Neurological Surgeons

Resolution and throughput optimized intraoperative spectrally encoded coherence tomography and reflectometry (iSECTR) for multimodal imaging during ophthalmic microsurgery

NASA Astrophysics Data System (ADS)

Malone, Joseph D.; El-Haddad, Mohamed T.; Leeburg, Kelsey C.; Terrones, Benjamin D.; Tao, Yuankai K.

2018-02-01

Limited visualization of semi-transparent structures in the eye remains a critical barrier to improving clinical outcomes and developing novel surgical techniques. While increases in imaging speed has enabled intraoperative optical coherence tomography (iOCT) imaging of surgical dynamics, several critical barriers to clinical adoption remain. Specifically, these include (1) static field-of-views (FOVs) requiring manual instrument-tracking; (2) high frame-rates require sparse sampling, which limits FOV; and (3) small iOCT FOV also limits the ability to co-register data with surgical microscopy. We previously addressed these limitations in image-guided ophthalmic microsurgery by developing microscope-integrated multimodal intraoperative swept-source spectrally encoded scanning laser ophthalmoscopy and optical coherence tomography. Complementary en face images enabled orientation and coregistration with the widefield surgical microscope view while OCT imaging enabled depth-resolved visualization of surgical instrument positions relative to anatomic structures-of-interest. In addition, we demonstrated novel integrated segmentation overlays for augmented-reality surgical guidance. Unfortunately, our previous system lacked the resolution and optical throughput for in vivo retinal imaging and necessitated removal of cornea and lens. These limitations were predominately a result of optical aberrations from imaging through a shared surgical microscope objective lens, which was modeled as a paraxial surface. Here, we present an optimized intraoperative spectrally encoded coherence tomography and reflectometry (iSECTR) system. We use a novel lens characterization method to develop an accurate model of surgical microscope objective performance and balance out inherent aberrations using iSECTR relay optics. Using this system, we demonstrate in vivo multimodal ophthalmic imaging through a surgical microscope

Skin suturing and cortical surface viral infusion improves imaging of neuronal ensemble activity with head-mounted miniature microscopes.

PubMed

Li, Xinjian; Cao, Vania Y; Zhang, Wenyu; Mastwal, Surjeet S; Liu, Qing; Otte, Stephani; Wang, Kuan Hong

2017-11-01

In vivo optical imaging of neural activity provides important insights into brain functions at the single-cell level. Cranial windows and virally delivered calcium indicators are commonly used for imaging cortical activity through two-photon microscopes in head-fixed animals. Recently, head-mounted one-photon microscopes have been developed for freely behaving animals. However, minimizing tissue damage from the virus injection procedure and maintaining window clarity for imaging can be technically challenging. We used a wide-diameter glass pipette at the cortical surface for infusing the viral calcium reporter AAV-GCaMP6 into the cortex. After infusion, the scalp skin over the implanted optical window was sutured to facilitate postoperative recovery. The sutured scalp was removed approximately two weeks later and a miniature microscope was attached above the window to image neuronal activity in freely moving mice. We found that cortical surface virus infusion efficiently labeled neurons in superficial layers, and scalp skin suturing helped to maintain the long-term clarity of optical windows. As a result, several hundred neurons could be recorded in freely moving animals. Compared to intracortical virus injection and open-scalp postoperative recovery, our methods minimized tissue damage and dura overgrowth underneath the optical window, and significantly increased the experimental success rate and the yield of identified neurons. Our improved cranial surgery technique allows for high-yield calcium imaging of cortical neurons with head-mounted microscopes in freely behaving animals. This technique may be beneficial for other optical applications such as two-photon microscopy, multi-site imaging, and optogenetic modulation. Published by Elsevier B.V.

Simple and robust image-based autofocusing for digital microscopy.

PubMed

Yazdanfar, Siavash; Kenny, Kevin B; Tasimi, Krenar; Corwin, Alex D; Dixon, Elizabeth L; Filkins, Robert J

2008-06-09

A simple image-based autofocusing scheme for digital microscopy is demonstrated that uses as few as two intermediate images to bring the sample into focus. The algorithm is adapted to a commercial inverted microscope and used to automate brightfield and fluorescence imaging of histopathology tissue sections.

Digital Timing Recovery for High Speed Optical Drives

NASA Astrophysics Data System (ADS)

Ko, Seok Jun; Kim, Pan Soo; Choi, Hyung Jin; Lee, Jae-Wook

2002-03-01

A new digital timing recovery scheme for the optical drive system is presented. By comparative simulations using digital versatile disc (DVD) patterns with marginal input conditions, the proposed algorithm shows enhanced performances in jitter variance and signal-to-noise ratio (SNR) margin by four times and 3 [dB], respectively.

Densely packed beta-structure at the protein-lipid interface of porin is revealed by high-resolution cryo-electron microscopy.

PubMed

Sass, H J; Büldt, G; Beckmann, E; Zemlin, F; van Heel, M; Zeitler, E; Rosenbusch, J P; Dorset, D L; Massalski, A

1989-09-05

Porin is an integral membrane protein that forms channels across the outer membrane of Escherichia coli. Electron microscopic studies of negatively stained two-dimensional porin crystals have shown three stain accumulations per porin trimer, revealing the locations of pores spanning the membrane. In this study, reconstituted porin lattices embedded in glucose were investigated using the low-dose technique on a cryo-electron microscope equipped with a helium-cooled superconducting objective lens. The specimen temperature was maintained at 5 K to yield an improved microscopic and specimen stability. Under these conditions, we obtained for the first time electron diffraction patterns from porin lattices to a resolution of 3.2 A and images showing optical diffraction up to a resolution of 4.9 A. Applying correlation averaging techniques to the digitized micrographs, we were able to reconstruct projected images of the porin trimer to a resolution of up to 3.5 A. In the final projection maps, amplitudes from electron diffraction and phases from these images were combined. The predominant feature is a high-density narrow band (about 6 A in thickness) that delineates the outer perimeter of the trimer. Since the molecule consists of almost exclusively beta-sheet structure, as revealed by spectroscopic data, we conclude that this band is a cylindrical beta-pleated sheet crossing the membrane nearly perpendicularly to its plane. Another intriguing finding is a low-density area (about 70 A2) situated in the centre of the trimer.

In vivo imaging of oral neoplasia using a miniaturized fiber optic confocal reflectance microscope.

PubMed

Maitland, Kristen C; Gillenwater, Ann M; Williams, Michelle D; El-Naggar, Adel K; Descour, Michael R; Richards-Kortum, Rebecca R

2008-11-01

The purpose of this study was to determine whether in vivo images of oral mucosa obtained with a fiber optic confocal reflectance microscope could be used to differentiate normal and neoplastic tissues. We imaged 20 oral sites in eight patients undergoing surgery for squamous cell carcinoma. Normal and abnormal areas within the oral cavity were identified clinically, and real-time videos of each site were obtained in vivo using a fiber optic confocal reflectance microscope. Following imaging, each site was biopsied and submitted for histopathologic examination. We identified distinct features, such as nuclear irregularity and spacing, which can be used to qualitatively differentiate between normal and abnormal tissue. Representative confocal images of normal, pre-neoplastic, and neoplastic oral tissue are presented. Previous work using much larger microscopes has demonstrated the ability of confocal reflectance microscopy to image cellular and tissue architecture in situ. New advances in technology have enabled miniaturization of imaging systems for in vivo use.

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

Kuhlmann, Andreas V.; Houel, Julien; Warburton, Richard J.

Optically active quantum dots, for instance self-assembled InGaAs quantum dots, are potentially excellent single photon sources. The fidelity of the single photons is much improved using resonant rather than non-resonant excitation. With resonant excitation, the challenge is to distinguish between resonance fluorescence and scattered laser light. We have met this challenge by creating a polarization-based dark-field microscope to measure the resonance fluorescence from a single quantum dot at low temperature. We achieve a suppression of the scattered laser exceeding a factor of 10{sup 7} and background-free detection of resonance fluorescence. The same optical setup operates over the entire quantum dotmore » emission range (920–980 nm) and also in high magnetic fields. The major development is the outstanding long-term stability: once the dark-field point has been established, the microscope operates for days without alignment. The mechanical and optical designs of the microscope are presented, as well as exemplary resonance fluorescence spectroscopy results on individual quantum dots to underline the microscope's excellent performance.« less

Sensing of Streptococcus mutans by microscopic imaging ellipsometry

NASA Astrophysics Data System (ADS)

Khaleel, Mai Ibrahim; Chen, Yu-Da; Chien, Ching-Hang; Chang, Yia-Chung

2017-05-01

Microscopic imaging ellipsometry is an optical technique that uses an objective and sensing procedure to measure the ellipsometric parameters Ψ and Δ in the form of microscopic maps. This technique is well known for being noninvasive and label-free. Therefore, it can be used to detect and characterize biological species without any impact. Microscopic imaging ellipsometry was used to measure the optical response of dried Streptococcus mutans cells on a glass substrate. The ellipsometric Ψ and Δ maps were obtained with the Optrel Multiskop system for specular reflection in the visible range (λ=450 to 750 nm). The Ψ and Δ images at 500, 600, and 700 nm were analyzed using three different theoretical models with single-bounce, two-bounce, and multibounce light paths to obtain the optical constants and height distribution. The obtained images of the optical constants show different aspects when comparing the single-bounce analysis with the two-bounce or multibounce analysis in detecting S. mutans samples. Furthermore, the height distributions estimated by two-bounce and multibounce analyses of S. mutans samples were in agreement with the thickness values measured by AFM, which implies that the two-bounce and multibounce analyses can provide information complementary to that obtained by a single-bounce light path.

OPTOELECTRONICS, FIBER OPTICS, AND OTHER ASPECTS OF QUANTUM ELECTRONICS: Nonlinear optical devices: basic elements of a future optical digital computer?

NASA Astrophysics Data System (ADS)

Fischer, R.; Müller, R.

1989-08-01

It is shown that nonlinear optical devices are the most promising elements for an optical digital supercomputer. The basic characteristics of various developed nonlinear elements are presented, including bistable Fabry-Perot etalons, interference filters, self-electrooptic effect devices, quantum-well devices utilizing transitions between the lowest electron states in the conduction band of GaAs, etc.

Optical XOR gate

DOEpatents

Vawter, G. Allen

2013-11-12

An optical XOR gate is formed as a photonic integrated circuit (PIC) from two sets of optical waveguide devices on a substrate, with each set of the optical waveguide devices including an electroabsorption modulator electrically connected in series with a waveguide photodetector. The optical XOR gate utilizes two digital optical inputs to generate an XOR function digital optical output. The optical XOR gate can be formed from III-V compound semiconductor layers which are epitaxially deposited on a III-V compound semiconductor substrate, and operates at a wavelength in the range of 0.8-2.0 .mu.m.

Optical NOR gate

DOEpatents

Skogen, Erik J [Albuquerque, NM; Tauke-Pedretti, Anna [Albuquerque, NM

2011-09-06

An optical NOR gate is formed from two pair of optical waveguide devices on a substrate, with each pair of the optical waveguide devices consisting of an electroabsorption modulator electrically connected in series with a waveguide photodetector. The optical NOR gate utilizes two digital optical inputs and a continuous light input to provide a NOR function digital optical output. The optical NOR gate can be formed from III-V compound semiconductor layers which are epitaxially deposited on a III-V compound semiconductor substrate, and operates at a wavelength in the range of 0.8-2.0 .mu.m.

A Review of Adaptive Optics Optical Coherence Tomography: Technical Advances, Scientific Applications, and the Future.

PubMed

Jonnal, Ravi S; Kocaoglu, Omer P; Zawadzki, Robert J; Liu, Zhuolin; Miller, Donald T; Werner, John S

2016-07-01

Optical coherence tomography (OCT) has enabled "virtual biopsy" of the living human retina, revolutionizing both basic retina research and clinical practice over the past 25 years. For most of those years, in parallel, adaptive optics (AO) has been used to improve the transverse resolution of ophthalmoscopes to foster in vivo study of the retina at the microscopic level. Here, we review work done over the last 15 years to combine the microscopic transverse resolution of AO with the microscopic axial resolution of OCT, building AO-OCT systems with the highest three-dimensional resolution of any existing retinal imaging modality. We surveyed the literature to identify the most influential antecedent work, important milestones in the development of AO-OCT technology, its applications that have yielded new knowledge, research areas into which it may productively expand, and nascent applications that have the potential to grow. Initial efforts focused on demonstrating three-dimensional resolution. Since then, many improvements have been made in resolution and speed, as well as other enhancements of acquisition and postprocessing techniques. Progress on these fronts has produced numerous discoveries about the anatomy, function, and optical properties of the retina. Adaptive optics OCT continues to evolve technically and to contribute to our basic and clinical knowledge of the retina. Due to its capacity to reveal cellular and microscopic detail invisible to clinical OCT systems, it is an ideal companion to those instruments and has the demonstrable potential to produce images that can guide the interpretation of clinical findings.

Optical Signal Processing: Poisson Image Restoration and Shearing Interferometry

NASA Technical Reports Server (NTRS)

Hong, Yie-Ming

1973-01-01

Optical signal processing can be performed in either digital or analog systems. Digital computers and coherent optical systems are discussed as they are used in optical signal processing. Topics include: image restoration; phase-object visualization; image contrast reversal; optical computation; image multiplexing; and fabrication of spatial filters. Digital optical data processing deals with restoration of images degraded by signal-dependent noise. When the input data of an image restoration system are the numbers of photoelectrons received from various areas of a photosensitive surface, the data are Poisson distributed with mean values proportional to the illuminance of the incoherently radiating object and background light. Optical signal processing using coherent optical systems is also discussed. Following a brief review of the pertinent details of Ronchi's diffraction grating interferometer, moire effect, carrier-frequency photography, and achromatic holography, two new shearing interferometers based on them are presented. Both interferometers can produce variable shear.

Optical analysis of a compound quasi-microscope for planetary landers

NASA Technical Reports Server (NTRS)

Wall, S. D.; Burcher, E. E.; Huck, F. O.

1974-01-01

A quasi-microscope concept, consisting of facsimile camera augmented with an auxiliary lens as a magnifier, was introduced and analyzed. The performance achievable with this concept was primarily limited by a trade-off between resolution and object field; this approach leads to a limiting resolution of 20 microns when used with the Viking lander camera (which has an angular resolution of 0.04 deg). An optical system is analyzed which includes a field lens between camera and auxiliary lens to overcome this limitation. It is found that this system, referred to as a compound quasi-microscope, can provide improved resolution (to about 2 microns ) and a larger object field. However, this improvement is at the expense of increased complexity, special camera design requirements, and tighter tolerances on the distances between optical components.

Surface Inspection Tool for Optical Detection of Surface Defects

NASA Technical Reports Server (NTRS)

Nurge, Mark; Youngquist, Robert; Dyer, Dustin

2013-01-01

The Space Shuttle Orbiter windows were damaged both by micrometeor impacts and by handling, and required careful inspection before they could be reused. The launch commit criteria required that no defect be deeper than a critical depth. The shuttle program used a refocus microscope to perform a quick pass/fail determination, and then followed up with mold impressions to better quantify any defect. However, the refocus microscope is slow and tedious to use due to its limited field of view, only focusing on one small area of glass at a time. Additionally, the unit is bulky and unable to be used in areas with tight access, such as defects near the window frame or on the glass inside the Orbiter due to interference with the dashboard. The surface inspection tool is a low-profile handheld instrument that provides two digital video images on a computer for monitoring surface defects. The first image is a wide-angle view to assist the user in locating defects. The second provides an enlarged view of a defect centered in the window of the first image. The focus is adjustable for each of the images. However, the enlarged view was designed to have a focal plane with a short depth. This allows the user to get a feel for the depth of different parts of the defect under inspection as the focus control is varied. A light source is also provided to illuminate the defect, precluding the need for separate lighting tools. The software provides many controls to adjust image quality, along with the ability to zoom digitally the images and to capture and store them for later processing.

EDITORIAL: Optical tomography and digital holography

NASA Astrophysics Data System (ADS)

Coupland, Jeremy; Lobera, Julia

2008-07-01

The articles in this special feature in Measurement Science and Technology concern exciting new developments in the field of digital holography—the process of electronically recording and numerically reconstructing an optical field [1]. Making use of the enormous advances in digital imaging and computer technology, digital holography is presented in a range of applications from fluid flow measurement and structural analysis to medical imaging. The science of digital holography rests on the foundations of optical holography, on the work of Gabor in the late 1940s, and on the development of laser sources in the 1960s, which made his vision a practical reality [2]. Optical holography, however, uses a photosensitive material, both to record a latent image and subsequently to behave as a diffractive optical element with which to reconstruct the incident field. In this way display holograms, using silver halide materials for example, can produce life-size images that are virtually indistinguishable from the object itself [3]. Digital holography, in contrast, separates the steps of recording and reconstruction, and the final image is most often in the form of a 3D computer model. Of course, television cameras have been used from the beginnings of holography to record interferometric images. However, the huge disparity between the resolution of holographic recording materials (more than 3000 cycles/mm) and television cameras (around 50 cycles/mm) was raised as a major concern by early researchers. TV holography, as it was sometimes called, generally recorded low numerical aperture (NA) holograms producing images with characteristically large speckle and was therefore more often referred to as electronic speckle pattern interferomery (ESPI) [4]. It is possible, however, to record large NA holograms on a sensor with restricted resolution by using an objective lens or a diverging reference wave [5]. This is generally referred to as digital holographic microscopy (DHM) since the resolution now places a limit on the size of the object that can be recorded. Some 60 years after the pioneering work of Gabor, digital imaging and associated computer technology offers a step change in capability with which to further exploit holography. Modern image sensors are now available with almost 30 million photosensitive elements, which corresponds to a staggering 100-fold increase compared to standard television images. At the same time personal computers have been optimized for imaging and graphics applications and this allows more sophisticated algorithms to be used in the reconstruction process. Although resolution still falls short of the materials used for optical holography, the ability to process data numerically generally outweighs this drawback and presents us with a host of new opportunities. Faced with the ability to record and process holograms numerically, it is natural to ask the question 'what information is present within recordings of scattered light?'. In fact this question could be posed by anyone using light, or indeed any other wave disturbance, for measurement purposes. For the case of optical holography, Wolf published his answer in 1969 [6], showing that for the case of weak scattering (small perturbations) and plane wave illumination, the amplitude and phase of each plane wave within the scattered field are proportional to those of a periodic variation in the refractive index contrast (i.e. a Bragg grating). This Fourier decomposition of the object was published almost simultaneously by Dandliker and Weiss [7], who also provided a graphical illustration of the technique. These works are the basis of optical tomography and provide us with the link between holographic data and 3D form. Digital holographic reconstruction and optical tomography was the theme of an international workshop [8] held in Loughborough in 2007, and many of the topics debated at the workshop have become the subject of the papers in this issue. In general terms the papers we present describe closely related holographic techniques that address application areas within the field of engineering. The application of digital holography to 3D fluid flow measurement is addressed by several authors. Salah et al demonstrate the simplicity of digital holography with an in-line multiple exposure holographic system using a low-cost laser diode. Soria and Atkinson discuss limitations of low NA holography in fluid velocimetry and demonstrate the potential of a multiple camera, in-line technique which they call Tomographic Digital Holographic Particle Image Velocimetry (Tomo-HPIV). Problems caused by the twin images (real and virtual) of in-line HPIV are described by Ooms et al. It is shown how sign ambiguity can be eliminated and bias errors suppressed by the application of a suitable threshold in piecewise correlation of the reconstructed field. Denis et al explain the problem of twin image removal as a deconvolution process and compare suppression algorithms based on wavelet decomposition. This process can be considered as an inverse problem and the benefits of this approach are discussed with reference to particulate holograms by Gire et al. Of course, the twin image problem can be solved by off-axis holographic geometries which, in effect, add a carrier modulation. Arroyo presents a comparison of carrier modulation strategies that have been presented in the literature and shows circumstances in which the information in each of the real and virtual images can be separated when the sensor resolution is less than that required by the NA of the objective. State-of-the-art digital holographic microscopy (DHM) is presented by Kühn et al. This paper uses an off-axis geometry that simultaneously records images at two wavelengths. The microscope allows the surface profile to be measured from a single recording and sub-nanometre axial resolution is demonstrated. Another interesting application of DHM is addressed by Grilli et al. They report a transmission set-up to investigate poling in a lithium niobate crystal. Developments in the field of optical tomography are covered by the majority of the papers in this issue. The paper by Debailleul et al shows the differences between images reconstructed from a single holographic recording and those synthesized from a series of holograms made with different plane wave illumination. This is optical diffraction tomography (ODT), the original method discussed by Wolf that is characterized by large NA and monochromatic illumination. An alternative strategy is to synthesize the image from holograms made at several wavelengths with low NA optics. This can be done either by sweeping the source or detector response or the reference path in a white light interferometer. These methods are called spectral domain and temporal domain optical coherence tomography (SD-ODT and TD-OCT) respectively. SD-OCT is illustrated in the paper by Potcoava and Kim for biomedical applications. SD- and TD-OCT are compared with confocal microscopy in the paper by Stifter et al. The huge potential of OCT as a diagnostic in polymer and composite materials is apparent from this work. There are clearly many different ways to implement optical tomography, and several established techniques, such as scanning white light interferometry (SWLI) and confocal microscopy, can be considered to be tomographic processes. We present two papers in this issue. The first attempts to bring together the topics of holography, microscopy and tomography within the framework of linear systems theory. It is shown that the images (or interferograms) produced by these instruments can be considered as estimates of refractive index contrast that are obtained using a linear inversion of the scattered field data. It is noted, however, that this is only strictly correct for the case of weak scattering and this is only a crude approximation for many cases of practical interest. The second paper that we present illustrates this for the case of mono-disperse particles in air. Here the number density of the particles is such that multiple scattering is prevalent; however, a priori knowledge of particle size and refractive index allows individual particles to be located accurately. In general, reconstruction can be thought of as a nonlinear optimization process that is used to discover the object which best explains the measured field and is consistent with a priori information. As Gire et al point out in their article, a priori knowledge can also be used to overcome the Nyquist sampling criteria. Although some caution should be exercised (for example, it is not usually possible to decide whether a given solution is unique), it is interesting to note that despite the disparity in resolution, digital holography and computer technology might yet create 3D images of greater clarity than the best optical holograms. References [1] Schnars U and Jueptner W 2005 Digital Holography (Berlin: Springer) ISBN: 978 3 540 21934 7 [2] Gabor D 1948 A new microscopic principle Nature 161 777-8 [3] Bjelkhagen H I 1993 Silver-Halide Recording Materials (Berlin: Springer) ISBN 3 540 58619 9 [4] Leendertz J A 1970 Interferometric displacement measurement on scattering surfaces utilizing speckle effect J. Phys. E: Sci. Instrum. 3 214-8 [5] Marquet P, Rappaz B, Magistretti P J, Cuche E, Emery Y, Colomb T and Depeursinge C 2005 Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy Opt. Lett. 30 468-70 [6] Wolf E 1969 Three-dimensional structure determination of semi-transparent objects from holographic data Opt. Commun. 1 153-6 [7] Dandliker R and Weiss K 1970 Reconstruction of the three-dimensional refractive index from scattered waves Opt. Commun. 1 323-8 [8] Coupland J and Lobera J 2007 International Workshop on Digital Holographic Reconstruction and Optical Tomography for Engineering Applications ISBN 978 0 947974 56 5

Integrated Real-Time Control and Imaging System for Microbiorobotics and Nanobiostructures

DTIC Science & Technology

2016-01-11

kit with a control board and ALP 4.1 basic controller suite. The digital micromirror device is the highest resolution 16:9 aspect ratio system. This...in Figure 1, consisted of the following: (1) digital micromirror device (DMD) and controller, (2) an inverted epifluorescence microscope with a flat...accompanying control board and ALP 4.1 basic controller suite. The digital micromirror device is currently the highest commercially available

Research of optical coherence tomography microscope based on CCD detector

NASA Astrophysics Data System (ADS)

Zhang, Hua; Xu, Zhongbao; Zhang, Shuomo

2008-12-01

The reference wave phase was modulated with a sinusoidal vibrating mirror attached to a Piezoelectric Transducer (PZT), the integration was performed by a CCD, and the charge storage period of the CCD image sensor was one-quarter period of the sinusoidal phase modulation. With the frequency- synchronous detection technique, four images (four frames of interference pattern) were recorded during one period of the phase modulation. In order to obtain the optimum modulation parameter, the values of amplitude and phase of the sinusoidal phase modulation were determined by considering the measurement error caused by the additive noise contained in the detected values. The PZT oscillation was controlled by a closed loop control system based on PID controller. An ideal discrete digital sine function at 50Hz with adjustable amplitude was used to adjust the vibrating of PZT, and a digital phase shift techniques was used to adjust vibrating phase of PZT so that the phase of the modulation could reach their optimum values. The CCD detector was triggered with software at 200Hz. Based on work above a small coherent signal masked by the preponderant incoherent background with a CCD detector was obtained.

A new computerized moving stage for optical microscopes

NASA Astrophysics Data System (ADS)

Hatiboglu, Can Ulas; Akin, Serhat

2004-06-01

Measurements of microscope stage movements in the x and y directions are of importance for some stereological methods. Traditionally, the length of stage movements is measured with differing precision and accuracy using a suitable motorized stage, a microscope and software. Such equipment is generally expensive and not readily available in many laboratories. One other challenging problem is the adaptability to available microscope systems which weakens the possibility of the equipment to be used with any kind of light microscope. This paper describes a simple and cheap programmable moving stage that can be used with the available microscopes in the market. The movements of the stage are controlled by two servo-motors and a controller chip via a Java-based image processing software. With the developed motorized stage and a microscope equipped with a CCD camera, the software allows complete coverage of the specimens with minimum overlap, eliminating the optical strain associated with counting hundreds of images through an eyepiece, in a quick and precise fashion. The uses and the accuracy of the developed stage are demonstrated using thin sections obtained from a limestone core plug.

Enhancing the performance of the light field microscope using wavefront coding.

PubMed

Cohen, Noy; Yang, Samuel; Andalman, Aaron; Broxton, Michael; Grosenick, Logan; Deisseroth, Karl; Horowitz, Mark; Levoy, Marc

2014-10-06

Light field microscopy has been proposed as a new high-speed volumetric computational imaging method that enables reconstruction of 3-D volumes from captured projections of the 4-D light field. Recently, a detailed physical optics model of the light field microscope has been derived, which led to the development of a deconvolution algorithm that reconstructs 3-D volumes with high spatial resolution. However, the spatial resolution of the reconstructions has been shown to be non-uniform across depth, with some z planes showing high resolution and others, particularly at the center of the imaged volume, showing very low resolution. In this paper, we enhance the performance of the light field microscope using wavefront coding techniques. By including phase masks in the optical path of the microscope we are able to address this non-uniform resolution limitation. We have also found that superior control over the performance of the light field microscope can be achieved by using two phase masks rather than one, placed at the objective's back focal plane and at the microscope's native image plane. We present an extended optical model for our wavefront coded light field microscope and develop a performance metric based on Fisher information, which we use to choose adequate phase masks parameters. We validate our approach using both simulated data and experimental resolution measurements of a USAF 1951 resolution target; and demonstrate the utility for biological applications with in vivo volumetric calcium imaging of larval zebrafish brain.

Scanning optical microscope with long working distance objective

DOEpatents

Cloutier, Sylvain G.

2010-10-19

A scanning optical microscope, including: a light source to generate a beam of probe light; collimation optics to substantially collimate the probe beam; a probe-result beamsplitter; a long working-distance, infinity-corrected objective; scanning means to scan a beam spot of the focused probe beam on or within a sample; relay optics; and a detector. The collimation optics are disposed in the probe beam. The probe-result beamsplitter is arranged in the optical paths of the probe beam and the resultant light from the sample. The beamsplitter reflects the probe beam into the objective and transmits resultant light. The long working-distance, infinity-corrected objective is also arranged in the optical paths of the probe beam and the resultant light. It focuses the reflected probe beam onto the sample, and collects and substantially collimates the resultant light. The relay optics are arranged to relay the transmitted resultant light from the beamsplitter to the detector.

Experimental stress–strain analysis of tapered silica optical fibers with nanofiber waist

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

Holleis, S.; Hoinkes, T.; Wuttke, C.

2014-04-21

We experimentally determine tensile force–elongation diagrams of tapered optical fibers with a nanofiber waist. The tapered optical fibers are produced from standard silica optical fibers using a heat and pull process. Both, the force–elongation data and scanning electron microscope images of the rupture points indicate a brittle material. Despite the small waist radii of only a few hundred nanometers, our experimental data can be fully explained by a nonlinear stress–strain model that relies on material properties of macroscopic silica optical fibers. This is an important asset when it comes to designing miniaturized optical elements as one can rely on themore » well-founded material characteristics of standard optical fibers. Based on this understanding, we demonstrate a simple and non-destructive technique that allows us to determine the waist radius of the tapered optical fiber. We find excellent agreement with independent scanning electron microscope measurements of the waist radius.« less

Aqueous carrier waveguide in a flow cytometer

DOEpatents

Mariella, Jr., Raymond P.; van den Engh, Gerrit; Northrup, M. Allen

1995-01-01

The liquid of a flow cytometer itself acts as an optical waveguide, thus transmitting the light to an optical filter/detector combination. This alternative apparatus and method for detecting scattered light in a flow cytometer is provided by a device which views and detects the light trapped within the optical waveguide formed by the flow stream. A fiber optic or other light collecting device is positioned within the flow stream. This provides enormous advantages over the standard light collection technique which uses a microscope objective. The signal-to-noise ratio is greatly increased over that for right-angle-scattered light collected by a microscope objective, and the alignment requirements are simplified.

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

Attota, Ravikiran, E-mail: Ravikiran.attota@nist.gov; Dixson, Ronald G.

We experimentally demonstrate that the three-dimensional (3-D) shape variations of nanometer-scale objects can be resolved and measured with sub-nanometer scale sensitivity using conventional optical microscopes by analyzing 4-D optical data using the through-focus scanning optical microscopy (TSOM) method. These initial results show that TSOM-determined cross-sectional (3-D) shape differences of 30 nm–40 nm wide lines agree well with critical-dimension atomic force microscope measurements. The TSOM method showed a linewidth uncertainty of 1.22 nm (k = 2). Complex optical simulations are not needed for analysis using the TSOM method, making the process simple, economical, fast, and ideally suited for high volume nanomanufacturing process monitoring.

Optical microscope and tapered fiber coupling apparatus for a dilution refrigerator.

PubMed

MacDonald, A J R; Popowich, G G; Hauer, B D; Kim, P H; Fredrick, A; Rojas, X; Doolin, P; Davis, J P

2015-01-01

We have developed a system for tapered fiber measurements of optomechanical resonators inside a dilution refrigerator, which is compatible with both on- and off-chip devices. Our apparatus features full three-dimensional control of the taper-resonator coupling conditions enabling critical coupling, with an overall fiber transmission efficiency of up to 70%. Notably, our design incorporates an optical microscope system consisting of a coherent bundle of 37,000 optical fibers for real-time imaging of the experiment at a resolution of ∼1 μm. We present cryogenic optical and optomechanical measurements of resonators coupled to tapered fibers at temperatures as low as 9 mK.

Differential polarization nonlinear optical microscopy with adaptive optics controlled multiplexed beams.

PubMed

Samim, Masood; Sandkuijl, Daaf; Tretyakov, Ian; Cisek, Richard; Barzda, Virginijus

2013-09-09

Differential polarization nonlinear optical microscopy has the potential to become an indispensable tool for structural investigations of ordered biological assemblies and microcrystalline aggregates. Their microscopic organization can be probed through fast and sensitive measurements of nonlinear optical signal anisotropy, which can be achieved with microscopic spatial resolution by using time-multiplexed pulsed laser beams with perpendicular polarization orientations and photon-counting detection electronics for signal demultiplexing. In addition, deformable membrane mirrors can be used to correct for optical aberrations in the microscope and simultaneously optimize beam overlap using a genetic algorithm. The beam overlap can be achieved with better accuracy than diffraction limited point-spread function, which allows to perform polarization-resolved measurements on the pixel-by-pixel basis. We describe a newly developed differential polarization microscope and present applications of the differential microscopy technique for structural studies of collagen and cellulose. Both, second harmonic generation, and fluorescence-detected nonlinear absorption anisotropy are used in these investigations. It is shown that the orientation and structural properties of the fibers in biological tissue can be deduced and that the orientation of fluorescent molecules (Congo Red), which label the fibers, can be determined. Differential polarization microscopy sidesteps common issues such as photobleaching and sample movement. Due to tens of megahertz alternating polarization of excitation pulses fast data acquisition can be conveniently applied to measure changes in the nonlinear signal anisotropy in dynamically changing in vivo structures.

Optical triple-in digital logic using nonlinear optical four-wave mixing

NASA Astrophysics Data System (ADS)

Widjaja, Joewono; Tomita, Yasuo

1995-08-01

A new programmable optical processor is proposed for implementing triple-in combinatorial digital logic that uses four-wave mixing. Binary-coded decimal-to-octal decoding is experimentally demonstrated by use of a photorefractive BaTiO 3 crystal. The result confirms the feasibility of the proposed system.

Design and analysis of a fast, two-mirror soft-x-ray microscope

NASA Technical Reports Server (NTRS)

Shealy, D. L.; Wang, C.; Jiang, W.; Jin, L.; Hoover, R. B.

1992-01-01

During the past several years, a number of investigators have addressed the design, analysis, fabrication, and testing of spherical Schwarzschild microscopes for soft-x-ray applications using multilayer coatings. Some of these systems have demonstrated diffraction limited resolution for small numerical apertures. Rigorously aplanatic, two-aspherical mirror Head microscopes can provide near diffraction limited resolution for very large numerical apertures. The relationships between the numerical aperture, mirror radii and diameters, magnifications, and total system length for Schwarzschild microscope configurations are summarized. Also, an analysis of the characteristics of the Head-Schwarzschild surfaces will be reported. The numerical surface data predicted by the Head equations were fit by a variety of functions and analyzed by conventional optical design codes. Efforts have been made to determine whether current optical substrate and multilayer coating technologies will permit construction of a very fast Head microscope which can provide resolution approaching that of the wavelength of the incident radiation.

Bio-inspired multi-mode optic flow sensors for micro air vehicles

NASA Astrophysics Data System (ADS)

Park, Seokjun; Choi, Jaehyuk; Cho, Jihyun; Yoon, Euisik

2013-06-01

Monitoring wide-field surrounding information is essential for vision-based autonomous navigation in micro-air-vehicles (MAV). Our image-cube (iCube) module, which consists of multiple sensors that are facing different angles in 3-D space, can be applied to the wide-field of view optic flows estimation (μ-Compound eyes) and to attitude control (μ- Ocelli) in the Micro Autonomous Systems and Technology (MAST) platforms. In this paper, we report an analog/digital (A/D) mixed-mode optic-flow sensor, which generates both optic flows and normal images in different modes for μ- Compound eyes and μ-Ocelli applications. The sensor employs a time-stamp based optic flow algorithm which is modified from the conventional EMD (Elementary Motion Detector) algorithm to give an optimum partitioning of hardware blocks in analog and digital domains as well as adequate allocation of pixel-level, column-parallel, and chip-level signal processing. Temporal filtering, which may require huge hardware resources if implemented in digital domain, is remained in a pixel-level analog processing unit. The rest of the blocks, including feature detection and timestamp latching, are implemented using digital circuits in a column-parallel processing unit. Finally, time-stamp information is decoded into velocity from look-up tables, multiplications, and simple subtraction circuits in a chip-level processing unit, thus significantly reducing core digital processing power consumption. In the normal image mode, the sensor generates 8-b digital images using single slope ADCs in the column unit. In the optic flow mode, the sensor estimates 8-b 1-D optic flows from the integrated mixed-mode algorithm core and 2-D optic flows with an external timestamp processing, respectively.

Optical sectioning microscopes with no moving parts using a micro-stripe array light emitting diode.

PubMed

Poher, V; Zhang, H X; Kennedy, G T; Griffin, C; Oddos, S; Gu, E; Elson, D S; Girkin, M; French, P M W; Dawson, M D; Neil, M A

2007-09-03

We describe an optical sectioning microscopy system with no moving parts based on a micro-structured stripe-array light emitting diode (LED). By projecting arbitrary line or grid patterns onto the object, we are able to implement a variety of optical sectioning microscopy techniques such as grid-projection structured illumination and line scanning confocal microscopy, switching from one imaging technique to another without modifying the microscope setup. The micro-structured LED and driver are detailed and depth discrimination capabilities are measured and calculated.

4Pi Microscopy.

PubMed

Schmidt, Roman; Engelhardt, Johann; Lang, Marion

2013-01-01

Optical microscopy has become a key technology in the life sciences today. Its noninvasive nature provides access to the interior of intact and even living cells, where specific molecules can be precisely localized by fluorescent tagging. However, the attainable 3D resolution of an optical microscope has long been hampered by a comparatively poor resolution along the optic axis. By coherent focusing through two objective lenses, 4Pi microscopy improves the axial resolution by three- to fivefold. This primer is intended as a starting point for the design and operation of a 4Pi microscope of type A.

Operation of a wet near-field scanning optical microscope in stable zones by minimizing the resonance change of tuning forks.

PubMed

Park, Kyoung-Duck; Park, Doo Jae; Lee, Seung Gol; Choi, Geunchang; Kim, Dai-Sik; Byeon, Clare Chisu; Choi, Soo Bong; Jeong, Mun Seok

2014-02-21

A resonant shift and a decrease of resonance quality of a tuning fork attached to a conventional fiber optic probe in the vicinity of liquid is monitored systematically while varying the protrusion length and immersion depth of the probe. Stable zones where the resonance modification as a function of immersion depth is minimized are observed. A wet near-field scanning optical microscope (wet-NSOM) is operated for a sample within water by using such a stable zone.

Solid state optical microscope

DOEpatents

Young, I.T.

1983-08-09

A solid state optical microscope wherein wide-field and high-resolution images of an object are produced at a rapid rate by utilizing conventional optics with a charge-coupled photodiode array. A galvanometer scanning mirror, for scanning in one of two orthogonal directions is provided, while the charge-coupled photodiode array scans in the other orthogonal direction. Illumination light from the object is incident upon the photodiodes, creating packets of electrons (signals) which are representative of the illuminated object. The signals are then processed, stored in a memory, and finally displayed as a video signal. 2 figs.

Solid-state optical microscope

DOEpatents

Young, I.T.

1981-01-07

A solid state optical microscope is described wherein wide-field and high-resolution images of an object are produced at a rapid rate by utilizing conventional optics with a charge-coupled photodiode array. Means for scanning in one of two orthogonal directions are provided, while the charge-coupled photodiode array scans in the other orthogonal direction. Illumination light from the object is incident upon the photodiodes, creating packets of electrons (signals) which are representative of the illuminated object. The signals are then processed, stored in a memory, and finally displayed as a video signal.

Solid state optical microscope

DOEpatents

Young, Ian T.

1983-01-01

A solid state optical microscope wherein wide-field and high-resolution images of an object are produced at a rapid rate by utilizing conventional optics with a charge-coupled photodiode array. A galvanometer scanning mirror, for scanning in one of two orthogonal directions is provided, while the charge-coupled photodiode array scans in the other orthogonal direction. Illumination light from the object is incident upon the photodiodes, creating packets of electrons (signals) which are representative of the illuminated object. The signals are then processed, stored in a memory, and finally displayed as a video signal.

Implementation of large-scale routine diagnostics using whole slide imaging in Sweden: Digital pathology experiences 2006-2013

PubMed Central

Thorstenson, Sten; Molin, Jesper; Lundström, Claes

2014-01-01

Recent technological advances have improved the whole slide imaging (WSI) scanner quality and reduced the cost of storage, thereby enabling the deployment of digital pathology for routine diagnostics. In this paper we present the experiences from two Swedish sites having deployed routine large-scale WSI for primary review. At Kalmar County Hospital, the digitization process started in 2006 to reduce the time spent at the microscope in order to improve the ergonomics. Since 2008, more than 500,000 glass slides have been scanned in the routine operations of Kalmar and the neighboring Linköping University Hospital. All glass slides are digitally scanned yet they are also physically delivered to the consulting pathologist who can choose to review the slides on screen, in the microscope, or both. The digital operations include regular remote case reporting by a few hospital pathologists, as well as around 150 cases per week where primary review is outsourced to a private clinic. To investigate how the pathologists choose to use the digital slides, a web-based questionnaire was designed and sent out to the pathologists in Kalmar and Linköping. The responses showed that almost all pathologists think that ergonomics have improved and that image quality was sufficient for most histopathologic diagnostic work. 38 ± 28% of the cases were diagnosed digitally, but the survey also revealed that the pathologists commonly switch back and forth between digital and conventional microscopy within the same case. The fact that two full-scale digital systems have been implemented and that a large portion of the primary reporting is voluntarily performed digitally shows that large-scale digitization is possible today. PMID:24843825

Shape oscillations of microparticles on an optical microscope stage.

PubMed

Zhu, Z M; Apfel, R E

1985-11-01

A modulated acoustic radiation pressure technique to produce quadrupole shape oscillations of drops ranging in diameter from 50-220 micron has been used by us. These drops have been suspended by acoustic levitation in a small chamber mounted on a stage of an optical microscope, which allowed easy viewing. The fission of drops and the deformation of sea urchin eggs were also observed.

Joint Services Electronics Program Annual Progress Report.

DTIC Science & Technology

1987-10-15

polarizability of free carriers in the semiconductor perturb the index of refraction which can be detected in a Nomarski -type optical interferometer. For...interferomters. However, the charge probe relies on a different physical effect and operates by interferometrically detecting the phase change induced in an... Nomarski microscope systems. These techniques will be applied, eventually, in our real-time V.. scanning optical microscope described below. Recently

Hyperspectral microscope imaging methods to classify gram-positive and gram-negative foodborne pathogenic bacteria

USDA-ARS?s Scientific Manuscript database

An acousto-optic tunable filter-based hyperspectral microscope imaging method has potential for identification of foodborne pathogenic bacteria from microcolony rapidly with a single cell level. We have successfully developed the method to acquire quality hyperspectral microscopic images from variou...

Simultaneous acquisition of 3D shape and deformation by combination of interferometric and correlation-based laser speckle metrology.

PubMed

Dekiff, Markus; Berssenbrügge, Philipp; Kemper, Björn; Denz, Cornelia; Dirksen, Dieter

2015-12-01

A metrology system combining three laser speckle measurement techniques for simultaneous determination of 3D shape and micro- and macroscopic deformations is presented. While microscopic deformations are determined by a combination of Digital Holographic Interferometry (DHI) and Digital Speckle Photography (DSP), macroscopic 3D shape, position and deformation are retrieved by photogrammetry based on digital image correlation of a projected laser speckle pattern. The photogrammetrically obtained data extend the measurement range of the DHI-DSP system and also increase the accuracy of the calculation of the sensitivity vector. Furthermore, a precise assignment of microscopic displacements to the object's macroscopic shape for enhanced visualization is achieved. The approach allows for fast measurements with a simple setup. Key parameters of the system are optimized, and its precision and measurement range are demonstrated. As application examples, the deformation of a mandible model and the shrinkage of dental impression material are measured.

Digital holographic microscope for measuring three-dimensional particle distributions and motions.

PubMed

Sheng, Jian; Malkiel, Edwin; Katz, Joseph

2006-06-01

Better understanding of particle-particle and particle-fluid interactions requires accurate 3D measurements of particle distributions and motions. We introduce the application of in-line digital holographic microscopy as a viable tool for measuring distributions of dense micrometer (3.2 microm) and submicrometer (0.75 microm) particles in a liquid solution with large depths of 1-10 mm. By recording a magnified hologram, we obtain a depth of field of approximately 1000 times the object diameter and a reduced depth of focus of approximately 10 particle diameters, both representing substantial improvements compared to a conventional microscope and in-line holography. Quantitative information on depth of field, depth of focus, and axial resolution is provided. We demonstrate that digital holographic microscopy can resolve the locations of several thousand particles and can measure their motions and trajectories using cinematographic holography. A sample trajectory and detailed morphological information of a free-swimming copepod nauplius are presented.

Dynamic nano-imaging of label-free living cells using electron beam excitation-assisted optical microscope

PubMed Central

Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

2015-01-01

Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications. PMID:26525841

Dynamic nano-imaging of label-free living cells using electron beam excitation-assisted optical microscope.

PubMed

Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

2015-11-03

Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications.

Dynamic nano-imaging of label-free living cells using electron beam excitation-assisted optical microscope

NASA Astrophysics Data System (ADS)

Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

2015-11-01

Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications.

The Engineer Topographic Laboratories /ETL/ hybrid optical/digital image processor

NASA Astrophysics Data System (ADS)

Benton, J. R.; Corbett, F.; Tuft, R.

1980-01-01

An optical-digital processor for generalized image enhancement and filtering is described. The optical subsystem is a two-PROM Fourier filter processor. Input imagery is isolated, scaled, and imaged onto the first PROM; this input plane acts like a liquid gate and serves as an incoherent-to-coherent converter. The image is transformed onto a second PROM which also serves as a filter medium; filters are written onto the second PROM with a laser scanner in real time. A solid state CCTV camera records the filtered image, which is then digitized and stored in a digital image processor. The operator can then manipulate the filtered image using the gray scale and color remapping capabilities of the video processor as well as the digital processing capabilities of the minicomputer.

Apparatus for direct-to-digital spatially-heterodyned holography

DOEpatents

Thomas, Clarence E.; Hanson, Gregory R.

2006-12-12

An apparatus operable to record a spatially low-frequency heterodyne hologram including spatially heterodyne fringes for Fourier analysis includes: a laser; a beamsplitter optically coupled to the laser; an object optically coupled to the beamsplitter; a focusing lens optically coupled to both the beamsplitter and the object; a digital recorder optically coupled to the focusing lens; and a computer that performs a Fourier transform, applies a digital filter, and performs an inverse Fourier transform. A reference beam and an object beam are focused by the focusing lens at a focal plane of the digital recorder to form a spatially low-frequency heterodyne hologram including spatially heterodyne fringes for Fourier analysis which is recorded by the digital recorder, and the computer transforms the recorded spatially low-frequency heterodyne hologram including spatially heterodyne fringes and shifts axes in Fourier space to sit on top of a heterodyne carrier frequency defined by an angle between the reference beam and the object beam and cuts off signals around an original origin before performing the inverse Fourier transform.

Optical design of cipher block chaining (CBC) encryption mode by using digital holography

NASA Astrophysics Data System (ADS)

Gil, Sang Keun; Jeon, Seok Hee; Jung, Jong Rae; Kim, Nam

2016-03-01

We propose an optical design of cipher block chaining (CBC) encryption by using digital holographic technique, which has higher security than the conventional electronic method because of the analog-type randomized cipher text with 2-D array. In this paper, an optical design of CBC encryption mode is implemented by 2-step quadrature phase-shifting digital holographic encryption technique using orthogonal polarization. A block of plain text is encrypted with the encryption key by applying 2-step phase-shifting digital holography, and it is changed into cipher text blocks which are digital holograms. These ciphered digital holograms with the encrypted information are Fourier transform holograms and are recorded on CCDs with 256 gray levels quantized intensities. The decryption is computed by these encrypted digital holograms of cipher texts, the same encryption key and the previous cipher text. Results of computer simulations are presented to verify that the proposed method shows the feasibility in the high secure CBC encryption system.

[A digital micromirror device-based Hadamard transform near infrared spectrometer].

PubMed

Liu, Jia; Chen, Fen-Fei; Liao, Cheng-Sheng; Xu, Qian; Zeng, Li-Bo; Wu, Qiong-Shui

2011-10-01

A Hadamard transform near infrared spectrometer based on a digital micromirror device was constructed. The optical signal was collected by optical fiber, a grating was used for light diffraction, a digital micromirror device (DMD) was applied instead of traditional mechanical Hadamard masks for optical modulation, and an InGaAs near infrared detector was used as the optic sensor. The original spectrum was recovered by fast Hadamard transform algrithms. The advantages of the spectrometer, such as high resolution, signal-noise-ratio, stability, sensitivity and response speed were proved by experiments, which indicated that it is very suitable for oil and food-safety applications.

Measurement of the Resolution of the Optical Microscope.

ERIC Educational Resources Information Center

Bowlt, C.

1983-01-01

Outlines procedures demonstrating that the aperture of a microscope objective limits resolving power and then, by using ancillary measurements made with a calibrated graticule in the microscope eyepiece, that the experimentally determined value for the maximum resolving power of a given objective is close to the value predicted by theory. (JN)

A new 3D tracking method for cell mechanics investigation exploiting the capabilities of digital holography in microscopy

NASA Astrophysics Data System (ADS)

Miccio, L.; Memmolo, P.; Merola, F.; Fusco, S.; Netti, P. A.; Ferraro, P.

2014-03-01

A method for 3D tracking has been developed exploiting Digital Holography features in Microscopy (DHM). In the framework of self-consistent platform for manipulation and measurement of biological specimen we use DHM for quantitative and completely label free analysis of samples with low amplitude contrast. Tracking capability extend the potentiality of DHM allowing to monitor the motion of appropriate probes and correlate it with sample properties. Complete 3D tracking has been obtained for the probes avoiding the amplitude refocusing in traditional tracking processes. Moreover, in biology and biomedical research fields one of the main topic is the understanding of morphology and mechanics of cells and microorganisms. Biological samples present low amplitude contrast that limits the information that can be retrieved through optical bright-field microscope measurements. The main effect on light propagating in such objects is in phase. This is known as phase-retardation or phase-shift. DHM is an innovative and alternative approach in microscopy, it's a good candidate for no-invasive and complete specimen analysis because its main characteristic is the possibility to discern between intensity and phase information performing quantitative mapping of the Optical Path Length. In this paper, the flexibility of DH is employed to analyze cell mechanics of unstained cells subjected to appropriate stimuli. DHM is used to measure all the parameters useful to understand the deformations induced by external and controlled stresses on in-vitro cells. Our configuration allows 3D tracking of micro-particles and, simultaneously, furnish quantitative phase-contrast maps. Experimental results are presented and discussed for in vitro cells.

Separating higher-order nonlinearities in transient absorption microscopy

NASA Astrophysics Data System (ADS)

Wilson, Jesse W.; Anderson, Miguel; Park, Jong Kang; Fischer, Martin C.; Warren, Warren S.

2015-08-01

The transient absorption response of melanin is a promising optically-accessible biomarker for distinguishing malignant melanoma from benign pigmented lesions, as demonstrated by earlier experiments on thin sections from biopsied tissue. The technique has also been demonstrated in vivo, but the higher optical intensity required for detecting these signals from backscattered light introduces higher-order nonlinearities in the transient response of melanin. These components that are higher than linear with respect to the pump or the probe introduce intensity-dependent changes to the overall response that complicate data analysis. However, our data also suggest these nonlinearities might be advantageous to in vivo imaging, in that different types of melanins have different nonlinear responses. Therefore, methods to separate linear from nonlinear components in transient absorption measurements might provide additional information to aid in the diagnosis of melanoma. We will discuss numerical methods for analyzing the various nonlinear contributions to pump-probe signals, with the ultimate objective of real time analysis using digital signal processing techniques. To that end, we have replaced the lock-in amplifier in our pump-probe microscope with a high-speed data acquisition board, and reprogrammed the coprocessor field-programmable gate array (FPGA) to perform lock-in detection. The FPGA lock-in offers better performance than the commercial instrument, in terms of both signal to noise ratio and speed. In addition, the flexibility of the digital signal processing approach enables demodulation of more complicated waveforms, such as spread-spectrum sequences, which has the potential to accelerate microscopy methods that rely on slow relaxation phenomena, such as photo-thermal and phosphorescence lifetime imaging.

KEYNOTE ADDRESS: The role of standards in the emerging optical digital data disk storage systems market

NASA Astrophysics Data System (ADS)

Bainbridge, Ross C.

1984-09-01

The Institute for Computer Sciences and Technology at the National Bureau of Standards is pleased to cooperate with the International Society for Optical Engineering and to join with the other distinguished organizations in cosponsoring this conference on applications of optical digital data disk storage systems.

CytometryML and other data formats

NASA Astrophysics Data System (ADS)

Leif, Robert C.

2006-02-01

Cytology automation and research will be enhanced by the creation of a common data format. This data format would provide the pathology and research communities with a uniform way for annotating and exchanging images, flow cytometry, and associated data. This specification and/or standard will include descriptions of the acquisition device, staining, the binary representations of the image and list-mode data, the measurements derived from the image and/or the list-mode data, and descriptors for clinical/pathology and research. An international, vendor-supported, non-proprietary specification will allow pathologists, researchers, and companies to develop and use image capture/analysis software, as well as list-mode analysis software, without worrying about incompatibilities between proprietary vendor formats. Presently, efforts to create specifications and/or descriptions of these formats include the Laboratory Digital Imaging Project (LDIP) Data Exchange Specification; extensions to the Digital Imaging and Communications in Medicine (DICOM); Open Microscopy Environment (OME); Flowcyt, an extension to the present Flow Cytometry Standard (FCS); and CytometryML. The feasibility of creating a common data specification for digital microscopy and flow cytometry in a manner consistent with its use for medical devices and interoperability with both hospital information and picture archiving systems has been demonstrated by the creation of the CytometryML schemas. The feasibility of creating a software system for digital microscopy has been demonstrated by the OME. CytometryML consists of schemas that describe instruments and their measurements. These instruments include digital microscopes and flow cytometers. Optical components including the instruments' excitation and emission parts are described. The description of the measurements made by these instruments includes the tagged molecule, data acquisition subsystem, and the format of the list-mode and/or image data. Many of the CytometryML data-types are based on the Digital Imaging and Communications in Medicine (DICOM). Binary files for images and list-mode data have been created and read.

Applications and challenges of digital pathology and whole slide imaging.

PubMed

Higgins, C

2015-07-01

Virtual microscopy is a method for digitizing images of tissue on glass slides and using a computer to view, navigate, change magnification, focus and mark areas of interest. Virtual microscope systems (also called digital pathology or whole slide imaging systems) offer several advantages for biological scientists who use slides as part of their general, pharmaceutical, biotechnology or clinical research. The systems usually are based on one of two methodologies: area scanning or line scanning. Virtual microscope systems enable automatic sample detection, virtual-Z acquisition and creation of focal maps. Virtual slides are layered with multiple resolutions at each location, including the highest resolution needed to allow more detailed review of specific regions of interest. Scans may be acquired at 2, 10, 20, 40, 60 and 100 × or a combination of magnifications to highlight important detail. Digital microscopy starts when a slide collection is put into an automated or manual scanning system. The original slides are archived, then a server allows users to review multilayer digital images of the captured slides either by a closed network or by the internet. One challenge for adopting the technology is the lack of a universally accepted file format for virtual slides. Additional challenges include maintaining focus in an uneven sample, detecting specimens accurately, maximizing color fidelity with optimal brightness and contrast, optimizing resolution and keeping the images artifact-free. There are several manufacturers in the field and each has not only its own approach to these issues, but also its own image analysis software, which provides many options for users to enhance the speed, quality and accuracy of their process through virtual microscopy. Virtual microscope systems are widely used and are trusted to provide high quality solutions for teleconsultation, education, quality control, archiving, veterinary medicine, research and other fields.

Hartmann characterization of the PEEM-3 aberration-corrected X-ray photoemission electron microscope.

PubMed

Scholl, A; Marcus, M A; Doran, A; Nasiatka, J R; Young, A T; MacDowell, A A; Streubel, R; Kent, N; Feng, J; Wan, W; Padmore, H A

2018-05-01

Aberration correction by an electron mirror dramatically improves the spatial resolution and transmission of photoemission electron microscopes. We will review the performance of the recently installed aberration corrector of the X-ray Photoemission Electron Microscope PEEM-3 and show a large improvement in the efficiency of the electron optics. Hartmann testing is introduced as a quantitative method to measure the geometrical aberrations of a cathode lens electron microscope. We find that aberration correction leads to an order of magnitude reduction of the spherical aberrations, suggesting that a spatial resolution of below 100 nm is possible at 100% transmission of the optics when using x-rays. We demonstrate this improved performance by imaging test patterns employing element and magnetic contrast. Published by Elsevier B.V.