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Sample records for applications reflectance confocal

  1. Comparison of mouse mammary gland imaging techniques and applications: Reflectance confocal microscopy, GFP Imaging, and ultrasound

    PubMed Central

    Tilli, Maddalena T; Parrish, Angela R; Cotarla, Ion; Jones, Laundette P; Johnson, Michael D; Furth, Priscilla A

    2008-01-01

    Background Genetically engineered mouse models of mammary gland cancer enable the in vivo study of molecular mechanisms and signaling during development and cancer pathophysiology. However, traditional whole mount and histological imaging modalities are only applicable to non-viable tissue. Methods We evaluated three techniques that can be quickly applied to living tissue for imaging normal and cancerous mammary gland: reflectance confocal microscopy, green fluorescent protein imaging, and ultrasound imaging. Results In the current study, reflectance confocal imaging offered the highest resolution and was used to optically section mammary ductal structures in the whole mammary gland. Glands remained viable in mammary gland whole organ culture when 1% acetic acid was used as a contrast agent. Our application of using green fluorescent protein expressing transgenic mice in our study allowed for whole mammary gland ductal structures imaging and enabled straightforward serial imaging of mammary gland ducts in whole organ culture to visualize the growth and differentiation process. Ultrasound imaging showed the lowest resolution. However, ultrasound was able to detect mammary preneoplastic lesions 0.2 mm in size and was used to follow cancer growth with serial imaging in living mice. Conclusion In conclusion, each technique enabled serial imaging of living mammary tissue and visualization of growth and development, quickly and with minimal tissue preparation. The use of the higher resolution reflectance confocal and green fluorescent protein imaging techniques and lower resolution ultrasound were complementary. PMID:18215290

  2. A handheld laser scanning confocal reflectance imagingconfocal Raman microspectroscopy system

    PubMed Central

    Patil, Chetan A.; Arrasmith, Christopher L.; Mackanos, Mark A.; Dickensheets, David L.; Mahadevan-Jansen, Anita

    2012-01-01

    Confocal reflectance microscopy and confocal Raman spectroscopy have shown potential for non-destructive analysis of samples at micron-scale resolutions. Current studies utilizing these techniques often employ large bench-top microscopes, and are not suited for use outside of laboratory settings. We have developed a microscope which combines laser scanning confocal reflectance imaging and confocal Raman spectroscopy into a compact handheld probe that is capable of high-resolution imaging and spectroscopy in a variety of settings. The compact size of the probe is largely due to the use of a MEMS mirror for beam scanning. The probe is capable of axial resolutions of up to 4 ?m for the confocal imaging channel and 10 ?m for the confocal Raman spectroscopy channel. Here, we report instrument design, characterize optical performance, and provide images and spectra from normal skin to demonstrate the instruments capabilities for clinical diagnostics. PMID:22435097

  3. Pupil engineering for a confocal reflectance line-scanning microscope

    NASA Astrophysics Data System (ADS)

    Patel, Yogesh G.; Rajadhyaksha, Milind; DiMarzio, Charles A.

    2011-03-01

    Confocal reflectance microscopy may enable screening and diagnosis of skin cancers noninvasively and in real-time, as an adjunct to biopsy and pathology. Current confocal point-scanning systems are large, complex, and expensive. A confocal line-scanning microscope, utilizing a of linear array detector can be simpler, smaller, less expensive, and may accelerate the translation of confocal microscopy in clinical and surgical dermatology. A line scanner may be implemented with a divided-pupil, half used for transmission and half for detection, or with a full-pupil using a beamsplitter. The premise is that a confocal line-scanner with either a divided-pupil or a full-pupil will provide high resolution and optical sectioning that would be competitive to that of the standard confocal point-scanner. We have developed a confocal line-scanner that combines both divided-pupil and full-pupil configurations. This combined-pupil prototype is being evaluated to determine the advantages and limitations of each configuration for imaging skin, and comparison of performance to that of commercially available standard confocal point-scanning microscopes. With the combined configuration, experimental evaluation of line spread functions (LSFs), contrast, signal-to-noise ratio, and imaging performance is in progress under identical optical and skin conditions. Experimental comparisons between divided-pupil and full-pupil LSFs will be used to determine imaging performance. Both results will be compared to theoretical calculations using our previously reported Fourier analysis model and to the confocal point spread function (PSF). These results may lead to a simpler class of confocal reflectance scanning microscopes for clinical and surgical dermatology.

  4. Laser reflection differential confocal large-radius measurement.

    PubMed

    Zhao, Weiqian; Zhang, Xin; Wang, Yun; Qiu, Lirong

    2015-11-01

    A laser reflection differential confocal large-radius measurement (RDCLRM) method is proposed to meet the requirements of high-precision measurement for a large radius of curvature (ROC). The RDCLRM identifies the converging point of the multiply reflected test beam by using the differential confocal focusing technology. It then measures the distance between the positions of the test lens corresponding to these converging points for different reflection times. Therefore, a precise and high-efficiency measurement of a large ROC is achieved with a shorter measurement lightpath. The theoretical analyses and preliminary experimental results indicate that RDCLRM has a relative expanded uncertainty of better than 0.005% (k=2). PMID:26560586

  5. Reflectance confocal microscopy: an effective diagnostic tool for dermatophytic infections.

    PubMed

    Friedman, Daniel; Friedman, Peter C; Gill, Melissa

    2015-02-01

    Current methods for diagnosing dermatophytic infections have various drawbacks. Analysis via skin scrapings and biopsies can be invasive and/or take too long to yield results. Reflectance confocal microscopy (RCM) is an emerging in vivo imaging technology that can potentially be used to diagnose cutaneous dermatophytic infections. This modality provides high-resolution images of the skin extending to the level of the superficial reticular dermis that could reveal the presence of fungal hyphae. In this retrospective chart review, we investigated the application of RCM as a diagnostic tool in the setting of a private practice. Images were used to diagnose dermatophyte infections and the results were compared to those of other established diagnostic methods. We found RCM to be a potentially effective and highly sensitive tool in the diagnosis of cutaneous dermatophytic infections. PMID:25750962

  6. Spectral confocal reflection microscopy using a white light source

    NASA Astrophysics Data System (ADS)

    Booth, M.; Juškaitis, R.; Wilson, T.

    2008-08-01

    We present a reflection confocal microscope incorporating a white light supercontinuum source and spectral detection. The microscope provides images resolved spatially in three-dimensions, in addition to spectral resolution covering the wavelength range 450-650nm. Images and reflection spectra of artificial and natural specimens are presented, showing features that are not normally revealed in conventional microscopes or confocal microscopes using discrete line lasers. The specimens include thin film structures on semiconductor chips, iridescent structures in Papilio blumei butterfly scales, nacre from abalone shells and opal gemstones. Quantitative size and refractive index measurements of transparent beads are derived from spectral interference bands.

  7. Reflectance confocal microscopy of red blood cells: simulation and experiment

    PubMed Central

    Zeidan, Adel; Yelin, Dvir

    2015-01-01

    Measuring the morphology of red blood cells is important for clinical diagnosis, providing valuable indications on a patient’s health. In this work, we have simulated the appearance of normal red blood cells under a reflectance confocal microscope and discovered unique relations between the morphological parameters and the resulting characteristic interference patterns of the cell. The simulation results showed good agreement with in vitro reflectance confocal images of red blood cells, acquired using spectrally encoded flow cytometry that imaged the cells in a linear flow without artificial staining. By matching the simulated patterns to confocal images of the cells, this method could be used for measuring cell morphology in three dimensions and for studying their physiology. PMID:26600999

  8. FOOD SURFACE TEXTURE MEASUREMENT USING REFLECTIVE CONFOCAL LASER SCANNING MICROSCOPY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Confocal laser scanning microscopy (CLSM) was used in the reflection mode to characterize the surface texture (roughness) of sliced food surfaces. Sandpapers of grit size between 150 and 600 were used as the height reference to standardize the CLSM hardware settings. Sandpaper particle sizes were v...

  9. A laser reflection confocal large-radius measurement

    NASA Astrophysics Data System (ADS)

    Zhang, Xin; Qiu, Lirong; Li, Zhigang; Zhao, Weiqian

    2015-12-01

    We propose a new laser reflection confocal large-radius measurement (RCLRM) method. By utilizing the precise correspondence relationship between the peak point of the confocal curve and the convergence point of the multi-reflected measuring beam, we identify the position of the test lens. With a distance interferometer, we obtain the position variation of the test lens with different reflection times. Therefore, a fast and precise large-radius measurement is achieved with a shorter measuring system. Additionally, the RCLRM significantly enhances the measurement accuracy by using conic fitting. The theoretical analyses and experiments indicate that the relative expanded uncertainty is better than 0.008% (k  =  2).

  10. Confocal Endomicroscopy: Instrumentation and Medical Applications

    PubMed Central

    Jabbour, Joey M.; Saldua, Meagan A.; Bixler, Joel N.; Maitland, Kristen C.

    2013-01-01

    Advances in fiber optic technology and miniaturized optics and mechanics have propelled confocal endomicroscopy into the clinical realm. This high resolution, non-invasive imaging technology provides the ability to microscopically evaluate cellular and sub-cellular features in tissue in vivo by optical sectioning. Because many cancers originate in epithelial tissues accessible by endoscopes, confocal endomicroscopy has been explored to detect regions of possible neoplasia at an earlier stage by imaging morphological features in vivo that are significant in histopathologic evaluation. This technique allows real-time assessment of tissue which may improve diagnostic yield by guiding biopsy. Research and development continues to reduce the overall size of the imaging probe, increase the image acquisition speed, and improve resolution and field of view of confocal endomicroscopes. Technical advances will continue to enable application to less accessible organs and more complex systems in the body. Lateral and axial resolutions down to 0.5 μm and 3 μm, respectively, field of view as large as 800×450 μm, and objective lens and total probe outer diameters down to 350 μm and 1.25 mm, respectively, have been achieved. We provide a review of the historical developments of confocal imaging in vivo, the evolution of endomicroscope instrumentation, and the medical applications of confocal endomicroscopy. PMID:21994069

  11. A Pulse Coupled Neural Network Segmentation Algorithm for Reflectance Confocal Images of Epithelial Tissue

    PubMed Central

    Malik, Bilal H.; Jabbour, Joey M.; Maitland, Kristen C.

    2015-01-01

    Automatic segmentation of nuclei in reflectance confocal microscopy images is critical for visualization and rapid quantification of nuclear-to-cytoplasmic ratio, a useful indicator of epithelial precancer. Reflectance confocal microscopy can provide three-dimensional imaging of epithelial tissue in vivo with sub-cellular resolution. Changes in nuclear density or nuclear-to-cytoplasmic ratio as a function of depth obtained from confocal images can be used to determine the presence or stage of epithelial cancers. However, low nuclear to background contrast, low resolution at greater imaging depths, and significant variation in reflectance signal of nuclei complicate segmentation required for quantification of nuclear-to-cytoplasmic ratio. Here, we present an automated segmentation method to segment nuclei in reflectance confocal images using a pulse coupled neural network algorithm, specifically a spiking cortical model, and an artificial neural network classifier. The segmentation algorithm was applied to an image model of nuclei with varying nuclear to background contrast. Greater than 90% of simulated nuclei were detected for contrast of 2.0 or greater. Confocal images of porcine and human oral mucosa were used to evaluate application to epithelial tissue. Segmentation accuracy was assessed using manual segmentation of nuclei as the gold standard. PMID:25816131

  12. Emulation and design of terahertz reflection-mode confocal scanning microscopy based on virtual pinhole

    NASA Astrophysics Data System (ADS)

    Yang, Yong-fa; Li, Qi

    2014-12-01

    In the practical application of terahertz reflection-mode confocal scanning microscopy, the size of detector pinhole is an important factor that determines the performance of spatial resolution characteristic of the microscopic system. However, the use of physical pinhole brings some inconvenience to the experiment and the adjustment error has a great influence on the experiment result. Through reasonably selecting the parameter of matrix detector virtual pinhole (VPH), it can efficiently approximate the physical pinhole. By using this approach, the difficulty of experimental calibration is reduced significantly. In this article, an imaging scheme of terahertz reflection-mode confocal scanning microscopy that is based on the matrix detector VPH is put forward. The influence of detector pinhole size on the axial resolution of confocal scanning microscopy is emulated and analyzed. Then, the parameter of VPH is emulated when the best axial imaging performance is reached.

  13. Distinct melanoma types based on reflectance confocal microscopy.

    PubMed

    Pellacani, Giovanni; De Pace, Barbara; Reggiani, Camilla; Cesinaro, Anna Maria; Argenziano, Giuseppe; Zalaudek, Iris; Soyer, H Peter; Longo, Caterina

    2014-06-01

    Distinct melanoma types exist in relation to patient characteristics, tumor morphology, histopathologic aspects and genetic background. A new diagnostic imaging tool, reflectance confocal microscopy (RCM), allows in vivo analysis of a given lesion with nearly histologic resolution while offering a dynamic view of the tissue in its 'natural' environment. The aim of this study was to analyse cell morphology of consecutive melanomas as they appear on RCM and to correlate morphology with tumor and patient characteristics. One hundred melanomas were visualized by RCM before excision. Clinical data, confocal features and histologic criteria were analysed. Four types of melanomas were identified as follows: (i) Melanomas with a predominantly dendritic cell population ('dendritic-cell melanomas') typically were thin by Breslow index; (ii) Melanomas typified by roundish melanocytes were smaller in size than dendritic cell MMs, but thicker by Breslow index, and predominantly occurred in patients with a high nevus count; (iii) Melanomas characterized by dermal nesting proliferation usually were thick by Breslow index at the time of diagnosis, although frequently smaller in size compared with the other types; and (iv) combined type melanomas may represent an evolution of dendritic cell and/or round cell types. Integration of confocal microscopy with clinical and histologic aspects may help in identifying and managing distinct tumors. PMID:24750486

  14. Lens central thickness measurement by laser reflection-confocal technology

    NASA Astrophysics Data System (ADS)

    Qiu, Lirong; Guo, Yongkui; Zhao, Weiqian; Xiao, Yang

    2015-08-01

    A new laser reflection-confocal thickness measurement (LRCTM) method is proposed for the reference lens central thickness calibration of the combined focal-length. LRCTM uses the reflector to reflect the convergent beam that come from the test lens to precisely identify the vertexes of test lens first and last surface, then uses ray tracing facet iterative algorithm to obtain lens central thickness. The test lens is put in the parallel light which makes its coaxality easier to adjust, and the optical path can be shortened with the reflector reflecting the convergent beam. LRCTM has high precision and concise structure, and it is suitable to be applied in the engineering. Preliminary experiments and analysis indicate that the relative measurement accuracy can be better than 0.03%.

  15. Confocal scanning beam laser microscope/macroscope: applications in fluorescence

    NASA Astrophysics Data System (ADS)

    Dixon, Arthur E.; Damaskinos, Savvas; Ribes, Alfonso

    1996-03-01

    A new confocal scanning beam laser microscope/macroscope is described that combines the rapid scan of a scanning beam laser microscope with the large specimen capability of a scanning stage microscope. This instrument combines an infinity-corrected confocal scanning laser microscope with a scanning laser macroscope that uses a telecentric f*(Theta) laser scan lens to produce a confocal imaging system with a resolution of 0.25 microns at a field of view of 25 microns and 5 microns at a field of view of 75,000 microns. The frame rate is 5 seconds per frame for a 512 by 512 pixel image, and 25 seconds for a 2048 by 2048 pixel image. Applications in fluorescence are discussed that focus on two important advantages of the instrument over a confocal scanning laser microscope: an extremely wide range of magnification, and the ability to image very large specimens. Examples are presented of fluorescence and reflected-light images of high quality printing, fluorescence images of latent fingerprints, packaging foam, and confocal autofluorescence images of a cricket.

  16. Segmentation of skin strata in reflectance confocal microscopy depth stacks

    NASA Astrophysics Data System (ADS)

    Hames, Samuel C.; Ardigò, Marco; Soyer, H. Peter; Bradley, Andrew P.; Prow, Tarl W.

    2015-03-01

    Reflectance confocal microscopy is an emerging tool for imaging human skin, but currently requires expert human assessment. To overcome the need for human experts it is necessary to develop automated tools for automatically assessing reflectance confocal microscopy imagery. This work presents a novel approach to this task, using a bag of visual words approach to represent and classify en-face optical sections from four distinct strata of the skin. A dictionary of representative features is learned from whitened and normalised patches using hierarchical spherical k-means. Each image is then represented by extracting a dense array of patches and encoding each with the most similar element in the dictionary. Linear discriminant analysis is used as a simple linear classifier. The proposed framework was tested on 308 depth stacks from 54 volunteers. Parameters are tuned using 10 fold cross validation on a training sub-set of the data, and final evaluation was performed on a held out test set. The proposed method generated physically plausible profiles of the distinct strata of human skin, and correctly classified 81.4% of sections in the test set.

  17. Confocal microscopy patterns in nonmelanoma skin cancer and clinical applications.

    PubMed

    Gonzlez, S; Snchez, V; Gonzlez-Rodrguez, A; Parrado, C; Ullrich, M

    2014-06-01

    Reflectance confocal microscopy is currently the most promising noninvasive diagnostic tool for studying cutaneous structures between the stratum corneum and the superficial reticular dermis. This tool gives real-time images parallel to the skin surface; the microscopic resolution is similar to that of conventional histology. Numerous studies have identified the main confocal features of various inflammatory skin diseases and tumors, demonstrating the good correlation of these features with certain dermatoscopic patterns and histologic findings. Confocal patterns and diagnostic algorithms have been shown to have high sensitivity and specificity in melanoma and nonmelanoma skin cancer. Possible present and future applications of this noninvasive technology are wide ranging and reach beyond its use in noninvasive diagnosis. This tool can also be used, for example, to evaluate dynamic skin processes that occur after UV exposure or to assess tumor response to noninvasive treatments such as photodynamic therapy. We explain the characteristic confocal features found in the main nonmelanoma skin tumors and discuss possible applications for this novel diagnostic technique in routine dermatology practice. PMID:24002008

  18. Combined FLIM and reflectance confocal microscopy for epithelial imaging

    NASA Astrophysics Data System (ADS)

    Jabbour, Joey M.; Cheng, Shuna; Shrestha, Sebina; Malik, Bilal; Jo, Javier A.; Applegate, Brian; Maitland, Kristen C.

    2012-03-01

    Current methods for detection of oral cancer lack the ability to delineate between normal and precancerous tissue with adequate sensitivity and specificity. The usual diagnostic mechanism involves visual inspection and palpation followed by tissue biopsy and histopathology, a process both invasive and time-intensive. A more sensitive and objective screening method can greatly facilitate the overall process of detection of early cancer. To this end, we present a multimodal imaging system with fluorescence lifetime imaging (FLIM) for wide field of view guidance and reflectance confocal microscopy for sub-cellular resolution imaging of epithelial tissue. Moving from a 12 x 12 mm2 field of view with 157 m lateral resolution using FLIM to 275 x 200 ?m2 with lateral resolution of 2.2 ?m using confocal microscopy, hamster cheek pouch model is imaged both in vivo and ex vivo. The results indicate that our dual modality imaging system can identify and distinguish between different tissue features, and, therefore, can potentially serve as a guide in early oral cancer detection..

  19. Dual-axes confocal reflectance microscope for distinguishing colonic neoplasia

    PubMed Central

    Liu, Jonathan T. C.; Mandella, Michael J.; Friedland, Shai; Soetikno, Roy; Crawford, James M.; Contag, Christopher H.; Kino, Gordon S.; Wang, Thomas D.

    2007-01-01

    A dual-axes confocal reflectance microscope has been developed that utilizes a narrowband laser at 1310 nm to achieve high axial resolution, image contrast, field of view, and tissue penetration for distinguishing among normal, hyperplastic, and dysplastic colonic mucosa ex vivo. Light is collected off-axis using a low numerical aperture objective to obtain vertical image sections, with 4- to 5-?m resolution, at tissue depths up to 610 ?m. Post-objective scanning enables a large field of view (610 640 ?m), and balanced-heterodyne detection provides sensitivity to collect vertical sections at one frame per second. System optics are optimized to effectively reject out-of-focus scattered light without use of a low-coherence gate. This design is scalable to millimeter dimensions, and the results demonstrate the potential for a miniature instrument to detect precancerous tissues, and hence to perform in vivo histopathology. PMID:17092168

  20. The use of reflectance confocal microscopy for examination of benign and malignant skin tumors

    PubMed Central

    Wielowieyska-Szybińska, Dorota; Białek-Galas, Kamila; Podolec, Katarzyna

    2014-01-01

    Reflectance confocal microscopy (RCM) is a modern, non-invasive diagnostic method that enables real-time imaging of epidermis and upper layers of the dermis with a nearly histological precision and high contrast. The application of this technology in skin imaging in the last few years has resulted in the progress of dermatological diagnosis, providing virtual access to the living skin erasing the need for conventional histopathology. The RCM has a potential of wide application in the dermatological diagnostic process with a particular reference to benign and malignant skin tumors. This article provides a summary of the latest reports and previous achievements in the field of RCM application in the diagnostic process of skin neoplasms. A range of dermatological indications and general characteristics of confocal images in various types of tumors are presented. PMID:25610353

  1. Biological applications of confocal fluorescence polarization microscopy

    NASA Astrophysics Data System (ADS)

    Bigelow, Chad E.

    Fluorescence polarization microscopy is a powerful modality capable of sensing changes in the physical properties and local environment of fluorophores. In this thesis we present new applications for the technique in cancer diagnosis and treatment and explore the limits of the modality in scattering media. We describe modifications to our custom-built confocal fluorescence microscope that enable dual-color imaging, optical fiber-based confocal spectroscopy and fluorescence polarization imaging. Experiments are presented that indicate the performance of the instrument for all three modalities. The limits of confocal fluorescence polarization imaging in scattering media are explored and the microscope parameters necessary for accurate polarization images in this regime are determined. A Monte Carlo routine is developed to model the effect of scattering on images. Included in it are routines to track the polarization state of light using the Mueller-Stokes formalism and a model for fluorescence generation that includes sampling the excitation light polarization ellipse, Brownian motion of excited-state fluorophores in solution, and dipole fluorophore emission. Results from this model are compared to experiments performed on a fluorophore-embedded polymer rod in a turbid medium consisting of polystyrene microspheres in aqueous suspension. We demonstrate the utility of the fluorescence polarization imaging technique for removal of contaminating autofluorescence and for imaging photodynamic therapy drugs in cell monolayers. Images of cells expressing green fluorescent protein are extracted from contaminating fluorescein emission. The distribution of meta-tetrahydroxypheny1chlorin in an EMT6 cell monolayer is also presented. A new technique for imaging enzyme activity is presented that is based on observing changes in the anisotropy of fluorescently-labeled substrates. Proof-of-principle studies are performed in a model system consisting of fluorescently labeled bovine serum albumin attached to sepharose beads. The action of trypsin and proteinase K on the albumin is monitored to demonstrate validity of the technique. Images of the processing of the albumin in J774 murine macrophages are also presented indicating large intercellular differences in enzyme activity. Future directions for the technique are also presented, including the design of enzyme probes specific for prostate specific antigen based on fluorescently-labeled dendrimers. A technique for enzyme imaging based on extracellular autofluorescence is also proposed.

  2. Genital warts: comparing clinical findings to dermatoscopic aspects, in vivo reflectance confocal features and histopathologic exam*

    PubMed Central

    Veasey, John Verrinder; Framil, Valria Maria de Souza; Nadal, Sidney Roberto; Marta, Alessandra Cristine; Lellis, Rute Facchini

    2014-01-01

    Genital warts can be diagnosed through physical examination and confirmed by histopathology. Noninvasive methods are useful for ruling out other diagnoses with no harm to the patient. In this study the clinical findings were compared to dermoscopy, reflectance confocal microscopy (RCM), and to histopathology findings, in order to determine possible patterns that can aid diagnosis of the lesion. It was possible to identify structural changes on reflectance confocal microscopy that are already known by dermoscopy, in addition to cellular changes previously seen only by histopathological examination. This study shows the use of reflectance confocal microscopy in cases of genital warts, providing important information that can be used in further studies. PMID:24626658

  3. Character research on 2.52 terahertz coaxial reflection-mode confocal scanning microscopic imaging

    NASA Astrophysics Data System (ADS)

    Yang, Yong-fa; Li, Qi; Hu, Jia-qi

    2014-12-01

    The technology of terahertz (THz) is a major research area in the 21st century. THz imaging is an important research direction. The single-frequency continuous-wave THz technology is combined with coaxial reflection-mode confocal scanning microscopic imaging in this article. Under the given system parameters, the transverse response character of 2.52THz (118.83?m) coaxial reflection-mode confocal scanning microscopic imaging is emulated and analyzed. The results of emulation show that coaxial reflection-mode confocal scanning microscopic imaging is feasible in THz region.

  4. Visualization of the microtubules of glutaraldehyde-fixed cells by reflection-enhanced backscatter confocal microscopy.

    PubMed

    Keith, Charles H; Farmer, Mark A

    2006-04-01

    Performing reflection-mode (backscatter-mode) confocal microscopy on cells growing on reflective substrates gives images that have improved contrast and are more easily interpreted than standard reflection-mode confocal micrographs (Keith et al., 1998). However, a number of factors degrade the quality of images taken with the highest-resolution microscope objectives in this technique. We here describe modifications to reflection-enhanced backscatter confocal microscopy that (partially) overcome these factors. With these modifications of the technique, it is possible to visualize structures the size-and refractility-of individual microtubules in intact cells. Additionally, we demonstrate that this technique, in common with fluorescence techniques such as standing wave widefield fluorescence microscopy and 4-Pi confocal microscopy, offers improved resolution in the Z-direction. PMID:17481347

  5. Two-photon fluorescence and confocal reflected light imaging of thick tissue structures

    NASA Astrophysics Data System (ADS)

    Kim, Ki H.; So, Peter T. C.; Kochevar, Irene E.; Masters, Barry R.; Gratton, Enrico

    1998-04-01

    The technology of two-photon excitation has opened a window of opportunity for developing non-invasive medical diagnostic tools capable of monitoring thick tissue biochemical states. Using cellular endogenous chromophores, (beta) -nicotinamide- adenine dinucleotide phosphate [NAD(P)H], the cellular metabolic rates in living human skin were determined. Although important functional information can be obtained from the fluorescence spectroscopy of endogenous chromophores, these chromophores are rather poor contrast enhancing agent for mapping cellular morphology. First, most endogenous chromophores are confined to the cellular cytoplasm which prevents the visualization of other cellular organelles. Second, there is significant variability in the distribution and the quantum yield of endogenous chromophores which depends on tissue biochemistry but prevents consistent comparison of cellular morphology. On the other hand, the deep tissue cellular morphology has been imaged with excellent resolution using reflected light confocal microscopy. In reflected light microscopy, the image contrast originates from the index of refraction differences of the cellular structures. The organelle boundaries with significant index differences such as the plasma membrane and the nucleus envelope can be consistently visualized. A combination of morphological and functional information is required for a thorough tissue study. This presentation describes the development of a new microscope which is capable of simultaneously collecting both two-photon fluorescence and confocal reflected light signals. Promising biomedical applications include the non-invasive diagnosis of skin cancer and the study of wound healing.

  6. Reflectance confocal endomicroscope with optical axial scanning for in vivo imaging of the oral mucosa

    PubMed Central

    Jabbour, Joey M.; Bentley, Julie L.; Malik, Bilal H.; Nemechek, John; Warda, John; Cuenca, Rodrigo; Cheng, Shuna; Jo, Javier A.; Maitland, Kristen C.

    2014-01-01

    This paper presents the design and evaluation of a reflectance confocal laser endomicroscope using a miniature objective lens within a rigid probe in conjunction with an electrically tunable lens for axial scanning. The miniature lens was characterized alone as well as in the endoscope across a 200 µm axial scan range using the tunable lens. The ability of the confocal endoscope to probe the human oral cavity is demonstrated by imaging of the oral mucosa in vivo. The results indicate that reflectance confocal endomicroscopy has the potential to be used in a clinical setting and guide diagnostic evaluation of biological tissue. PMID:25426310

  7. [Current application of confocal laser scanning microscope (CLSM) in stomatology].

    PubMed

    Zhang, Yu-sen; Li, Ning-yi

    2007-04-01

    Confocal laser scanning microscopy is one kind of modern Hi-tech on the basis of confocal imaging which is characterized by depth discrimination capability. It has been widely used in the field of stomatology due to its great advantages of non-destructive and non-invasive optical sectioning and three-dimensional reconstruction of the vital objects, in situ and dynamic real-time observation of the tissues and cells can be performed at high resolution. This paper reviews the fundamentals of confocal imaging and the application of CLSM in the fields of dental material, caries, dentin bonding interface and other basic researches in stomatology in recent years. PMID:17546397

  8. Confocal and Two-Photon Microscopy: Foundations, Applications and Advances

    NASA Astrophysics Data System (ADS)

    Diaspro, Alberto

    2001-11-01

    Confocal and Two-Photon Microscopy Foundations, Applications, and Advances Edited by Alberto Diaspro Confocal and two-photon fluorescence microscopy has provided researchers with unique possibilities of three-dimensional imaging of biological cells and tissues and of other structures such as semiconductor integrated circuits. Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances provides clear, comprehensive coverage of basic foundations, modern applications, and groundbreaking new research developments made in this important area of microscopy. Opening with a foreword by G. J. Brakenhoff, this reference gathers the work of an international group of renowned experts in chapters that are logically divided into balanced sections covering theory, techniques, applications, and advances, featuring: In-depth discussion of applications for biology, medicine, physics, engineering, and chemistry, including industrial applications Guidance on new and emerging imaging technology, developmental trends, and fluorescent molecules Uniform organization and review-style presentation of chapters, with an introduction, historical overview, methodology, practical tips, applications, future directions, chapter summary, and bibliographical references Companion FTP site with full-color photographs The significant experience of pioneers, leaders, and emerging scientists in the field of confocal and two-photon excitation microscopy Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances is invaluable to researchers in the biological sciences, tissue and cellular engineering, biophysics, bioengineering, physics of matter, and medicine, who use these techniques or are involved in developing new commercial instruments.

  9. Fluorescence lifetime imaging and reflectance confocal microscopy for multiscale imaging of oral precancer

    NASA Astrophysics Data System (ADS)

    Jabbour, Joey M.; Cheng, Shuna; Malik, Bilal H.; Cuenca, Rodrigo; Jo, Javier A.; Wright, John; Cheng, Yi-Shing Lisa; Maitland, Kristen C.

    2013-04-01

    Optical imaging techniques using a variety of contrast mechanisms are under evaluation for early detection of epithelial precancer; however, tradeoffs in field of view (FOV) and resolution may limit their application. Therefore, we present a multiscale multimodal optical imaging system combining macroscopic biochemical imaging of fluorescence lifetime imaging (FLIM) with subcellular morphologic imaging of reflectance confocal microscopy (RCM). The FLIM module images a 1616 mm2 tissue area with 62.5 ?m lateral and 320 ps temporal resolution to guide cellular imaging of suspicious regions. Subsequently, coregistered RCM images are acquired at 7 Hz with 400 ?m diameter FOV, <1 ?m lateral and 3.5 ?m axial resolution. FLIM-RCM imaging was performed on a tissue phantom, normal porcine buccal mucosa, and a hamster cheek pouch model of oral carcinogenesis. While FLIM is sensitive to biochemical and macroscopic architectural changes in tissue, RCM provides images of cell nuclear morphology, all key indicators of precancer progression.

  10. Consistency and distribution of reflectance confocal microscopy features for diagnosis of cutaneous T cell lymphoma.

    PubMed

    Lange-Asschenfeldt, Susanne; Babilli, Jasmin; Beyer, Marc; Rus-Diaz, Francisca; Gonzlez, Salvador; Stockfleth, Eggert; Ulrich, Martina

    2012-01-01

    Reflectance confocal microscopy (RCM) represents a noninvasive imaging technique that has previously been used for characterization of mycosis fungoides (MF) in a pilot study. We aimed to test the applicability of RCM for diagnosis and differential diagnosis of MF in a clinical study. A total of 39 test sites of 15 patients with a biopsy-proven diagnosis of either MF, parapsoriasis, Se?zary syndrome, or lymphomatoid papulosis were analyzed for presence and absence of RCM features of MF. Cochran and Chi(2) analysis were applied to test the concordance between investigators and the distribution of RCM features, respectively. For selected parameters, the Cochran analysis showed good concordance between investigators. Inter-observer reproducibility was highest for junctional atypical lymphocytes, architectural disarray, and spongiosis. Similarly, Chi(2) analysis demonstrated that selected features were present at particularly high frequency in individual skin diseases, with values ranging from 73% to 100% of all examined cases. PMID:22352651

  11. Fluorescence lifetime imaging and reflectance confocal microscopy for multiscale imaging of oral precancer

    PubMed Central

    Jabbour, Joey M.; Cheng, Shuna; Malik, Bilal H.; Cuenca, Rodrigo; Jo, Javier A.; Wright, John; Cheng, Yi-Shing Lisa

    2013-01-01

    Abstract. Optical imaging techniques using a variety of contrast mechanisms are under evaluation for early detection of epithelial precancer; however, tradeoffs in field of view (FOV) and resolution may limit their application. Therefore, we present a multiscale multimodal optical imaging system combining macroscopic biochemical imaging of fluorescence lifetime imaging (FLIM) with subcellular morphologic imaging of reflectance confocal microscopy (RCM). The FLIM module images a 1616??mm2 tissue area with 62.5?m lateral and 320ps temporal resolution to guide cellular imaging of suspicious regions. Subsequently, coregistered RCM images are acquired at 7Hz with 400?m diameter FOV, <1???m lateral and 3.5?m axial resolution. FLIM-RCM imaging was performed on a tissue phantom, normal porcine buccal mucosa, and a hamster cheek pouch model of oral carcinogenesis. While FLIM is sensitive to biochemical and macroscopic architectural changes in tissue, RCM provides images of cell nuclear morphology, all key indicators of precancer progression. PMID:23595826

  12. Consistency and distribution of reflectance confocal microscopy features for diagnosis of cutaneous T cell lymphoma

    NASA Astrophysics Data System (ADS)

    Lange-Asschenfeldt, Susanne; Babilli, Jasmin; Beyer, Marc; Ríus-Diaz, Francisca; González, Salvador; Stockfleth, Eggert; Ulrich, Martina

    2012-01-01

    Reflectance confocal microscopy (RCM) represents a noninvasive imaging technique that has previously been used for characterization of mycosis fungoides (MF) in a pilot study. We aimed to test the applicability of RCM for diagnosis and differential diagnosis of MF in a clinical study. A total of 39 test sites of 15 patients with a biopsy-proven diagnosis of either MF, parapsoriasis, Sézary syndrome, or lymphomatoid papulosis were analyzed for presence and absence of RCM features of MF. Cochran and Chi2 analysis were applied to test the concordance between investigators and the distribution of RCM features, respectively. For selected parameters, the Cochran analysis showed good concordance between investigators. Inter-observer reproducibility was highest for junctional atypical lymphocytes, architectural disarray, and spongiosis. Similarly, Chi2 analysis demonstrated that selected features were present at particularly high frequency in individual skin diseases, with values ranging from 73% to 100% of all examined cases.

  13. Anti-translational research: from the bedside back to the bench for reflectance confocal microscopy

    NASA Astrophysics Data System (ADS)

    Gareau, Daniel

    2014-03-01

    The reflectance confocal microscope has made translational progress in dermatology. 0.5 micrometer lateral resolution, 0.75mm field-of-view and excellent temporal resolution at ~15 frames/second serve the VivaScope well in the clinic, but it may be overlooked in basic research. This work reviews high spatiotemporal confocal microscopy and presents images acquired of various samples: zebra fish embryo where melanocytes with excellent contrast overly the spinal column, chicken embryo, where myocardium is seen moving at 15 frames/ second, calcium spikes in dendrites (fluorescence mode) just beyond the temporal resolution, and human skin where blood cells race through the artereovenous microvasculature. For an introduction to confocal microscopy, see: http://dangareau.net.s69818.gridserver.com/science/confocal-microscopy

  14. Case Report: melanoma and melanocytic nevus differentiation with reflectance confocal microscopy.

    PubMed Central

    ?udzik, Joanna; Witkowski, Alexander M; Pellacani, Giovanni

    2015-01-01

    Historically, melanoma has been typically diagnosed by naked-eye examination and confirmed with invasive biopsy. However, recently the use of reflectance confocal microscopy enables non-invasive bedside diagnosis of clinically equivocal lesions. We present a case in which reflectance confocal microscopy was used to evaluate two skin lesions in the same patient confirming the diagnosis of a melanoma and potentially avoiding invasive biopsy in the second benign melanocytic lesion. Clinicians should be aware of the availability of new non-invasive technologies that can aid in early diagnosis of malignant skin tumors and potentially reduce the number of benign lesion excisions. PMID:26236471

  15. High speed 3D surface profile without axial scanning: dual-detection confocal reflectance microscopy

    NASA Astrophysics Data System (ADS)

    Lee, Dong-Ryoung; Kim, Young-Duk; Gweon, Dae-Gab; Yoo, Hongki

    2014-12-01

    We propose dual-detection confocal reflectance microscopy (DDCRM) for high-speed 3D surface profiling. In comparison with conventional confocal microscopy, DDCRM can realize surface profiling without axial scanning. DDCRM is composed of two point detectors, each with a pinhole of different size. The ratio of the axial response curves measured by the two detectors provides the relationship between the axial position of the sample and the ratio of the intensity signals. Furthermore, DDCRM has a normalizing effect which allows this method to accurately measure the height of samples with various reflectance characteristics.

  16. Microscopic heterogeneity vs. macroscopic homogeneity in tissue phantoms using reflectance-mode confocal scanning laser microscopy

    NASA Astrophysics Data System (ADS)

    Samatham, Ravikant; Levitz, David; Fletcher, Reid; Fu, Yongji; Jacques, Steven L.

    2008-02-01

    Preparation of phantoms with reproducible and homogenous optical properties is tricky. The microscopic heterogeneity and macroscopic homogeneity of tissue phantoms were compared using reflectance-mode confocal laser scanning microscopy. Tissue phantoms were prepared using polystyrene microspheres as scattering medium in aqueous and gel matrix. Uniform distribution of microparticles in phantoms was evaluated by confocal imaging. Comparison of the heterogeneity of the phantoms was accomplished based on microscopic optical scattering properties. Distribution of optical properties at the microscopic levels was determined by a simple theory developed based on the depth-dependent decay of the reflectance-mode confocal signal. The variability of these optical properties is correlated to heterogeneity of the phantom. These microscopic properties were compared with macroscopic properties determined by ballistic transmission experiment. This enabled to optimize the phantom preparation procedure.

  17. Characterization of hydrogel microstructure using laser tweezers particle tracking and confocal reflection imaging

    PubMed Central

    Kotlarchyk, M A; Botvinick, E L; Putnam, A J

    2010-01-01

    Hydrogels are commonly used as extracellular matrix mimetics for applications in tissue engineering and increasingly as cell culture platforms with which to study the influence of biophysical and biochemical cues on cell function in 3D. In recent years, a significant number of studies have focused on linking substrate mechanical properties to cell function using standard methodologies to characterize the bulk mechanical properties of the hydrogel substrates. However, current understanding of the correlations between the microstructural mechanical properties of hydrogels and cell function in 3D is poor, in part because of a lack of appropriate techniques. Here we have utilized a laser tracking system, based on passive optical microrheology instrumentation, to characterize the microstructure of viscoelastic fibrin clots. Trajectories and mean square displacements were observed as bioinert PEGylated (PEG: polyethylene glycol) microspheres (1, 2 or 4.7 ?m in diameter) diffused within confined pores created by the protein phase of fibrin hydrogels. Complementary confocal reflection imaging revealed microstructures comprised of a highly heterogeneous fibrin network with a wide range of pore sizes. As the protein concentration of fibrin gels was increased, our quantitative laser tracking measurements showed a corresponding decrease in particle mean square displacements with greater resolution and sensitivity than conventional imaging techniques. This platform-independent method will enable a more complete understanding of how changes in substrate mechanical properties simultaneously influence other microenvironmental parameters in 3D cultures. PMID:20877437

  18. Dye-enhanced reflectance and fluorescence confocal microscopy as an optical pathology tool

    NASA Astrophysics Data System (ADS)

    Yaroslavsky, Anna N.; Salomatina, Elena; Novak, John; Amat-Roldan, Ivan; Castano, Ana; Hamblin, Michael

    2006-02-01

    Early detection and precise excision of neoplasms are imperative requirements for successful cancer treatment. In this study we evaluated the use of dye-enhanced confocal microscopy as an optical pathology tool in the ex vivo trial with fresh thick non-melanoma skin cancer excisions and in vivo trial with B16F10 melanoma cancer in mice. For the experiments the tumors were rapidly stained using aqueous solutions of either toluidine blue or methylene blue and imaged using multimodal confocal microscope. Reflectance images were acquired at the wavelengths of 630nm and 650 nm. Fluorescence was excited at 630 nm and 650 nm. Fluorescence emission was registered in the range between 680 nm and 710 nm. The images were compared to the corresponding en face frozen H&E sections. The results of the study indicate confocal images of stained cancerous tissue closely resemble corresponding H&E sections both in vivo and in vitro. This remarkable similarity enables interpretation of confocal images in a manner similar to that of histopathology. The developed technique may provide an efficient real-time optical tool for detecting skin pathology.

  19. Learning Reflectance Confocal Microscopy of Melanocytic Skin Lesions through Histopathologic Transversal Sections

    PubMed Central

    Braga, Juliana Casagrande Tavoloni; Macedo, Mariana Petaccia; Pinto, Clovis; Duprat, Joo; Begnami, Maria Dirlei; Pellacani, Giovanni; Rezze, Gisele Gargantini

    2013-01-01

    Histopathologic interpretation of dermoscopic and reflectance confocal microscopy (RCM) features of cutaneous melanoma was timidly carried out using perpendicular histologic sections, which does not mimic the same plane of the image achieved at both techniques (horizontal plane). The aim of this study was to describe the transverse histologic sections research technique and correlate main dermoscopic features characteristic of cutaneous melanoma (atypical network, irregular globules and pseudopods) with RCM and histopathology in perpendicular and transverse sections in order to offer a more precise interpretation of in vivo detectable features. Four melanomas and 2 nevi with different dermoscopic clues have been studied. Lesion areas that showed characteristic dermoscopic features were imaged by dermoscopy and confocal microscopy and directly correlated with histopathology in perpendicular and transverse sections. We presented the possibility to perform transverse sections as a new approach to understand RCM features. Atypical network showed different aspects in the 2 melanomas: in one case it was characterized by pleomorphic malignant melanocytes with tendency to form aggregates, whereas in the other elongated dendritic cells crowded around dermal papillae, some of them forming bridges that resembled the mitochondrial aspect at confocal and histopathology transversal sections. Pigment globules in melanomas and nevi differed for the presence of large atypical cells in the former, and pseudopods showed up as elongated nests protruded toward the periphery of the lesion. Transverse histologic research sections have a consistent dermoscopic and confocal correlate, and it may represent an help in confocal feature interpretation and an advance in improving melanoma diagnosis and knowledge of the biology of melanocytic lesions. PMID:24339910

  20. Near-IR fluorescence and reflectance confocal microscopy for imaging of quantum dots in mammalian skin

    PubMed Central

    Mortensen, Luke J.; Glazowski, Christopher E.; Zavislan, James M.; DeLouise, Lisa A.

    2011-01-01

    Understanding the skin penetration of nanoparticles (NPs) is an important concern due to the increasing presence of NPs in consumer products, including cosmetics. Technical challenges have slowed progress in evaluating skin barrier and NP factors that contribute to skin penetration risk. To limit sampling error and other problems associated with histological processing, many researchers are implementing whole tissue confocal or multiphoton microscopies. This work introduces a fluorescence and reflectance confocal microscopy system that utilizes near-IR excitation and emission to detect near-IR lead sulfide quantum dots (QDs) through ex vivo human epidermis. We provide a detailed prediction and experimental analysis of QD detection sensitivity and demonstrate detection of QD skin penetration in a barrier disrupted model. The unique properties of near-IR lead-based QDs will enable future studies that examine the impact of further barrier-disrupting agents on skin penetration of QDs and elucidate mechanistic insight into QD tissue interactions at the cellular level. PMID:21698023

  1. Measuring the lens focal length by laser reflection-confocal technology.

    PubMed

    Yang, Jiamiao; Qiu, Lirong; Zhao, Weiqian; Shao, Rongjun; Li, Zhigang

    2013-06-01

    A laser reflection-confocal focal-length measurement (LRCFM) is proposed for the high-accuracy measurement of lens focal length. LRCFM uses the peak points of confocal response curves to precisely identify the lens focus and vertex of the lens last surface. LRCFM then accurately measures the distance between the two positions to determine the lens focal length. LRCFM uses conic fitting, which significantly enhances measurement accuracy by inhibiting the influence of environmental disturbance and system noise on the measurement results. The experimental results indicate that LRCFM has a relative expanded uncertainty of less than 0.0015%. Compared with existing measurement methods, LRCFM has high accuracy and a concise structure. Thus, LRCFM is a feasible method for high-accuracy focal-length measurements. PMID:23736337

  2. Topographic variations in normal skin, as viewed by in vivo reflectance confocal microscopy.

    PubMed

    Huzaira, M; Rius, F; Rajadhyaksha, M; Anderson, R R; Gonzlez, S

    2001-06-01

    Near-infrared confocal microscopy is a new tool that provides skin images in vivo, with high resolution and contrast at a specific depth. Regional variations in live human skin viewed by confocal microscope have not been studied so far. In vivo reflectance confocal microscopy was performed in 10 adults (eight males, two females) of various skin phototypes. Six topographic sites were studied in each subject: forehead, cheek, inner and outer forearm surfaces, lower back and leg. Epidermal thickness at suprapapillary epidermal plates and rete pegs was measured during real-time imaging and the number and diameter of epidermal keratinocytes in each epidermal cell layer as well as the characteristics of dermal papillae were defined from the grabbed images. Stratum corneum appeared brighter in sun-exposed than in sun-protected areas and particularly pronounced in heavily pigmented individuals. The epidermal thickness at rete pegs, but not the suprapapillary epidermal plate, was greater in sun-exposed areas than in sun-protected sites except forearm flexor surface. The en face numerical density of granular keratinocytes is greater on the face as compared with all other sites, whereas the surface density of spinous keratinocytes is greater on sun-protected sites. Additionally, the number of basal keratinocytes per millimeter length of dermoepidermal junction is greater in sun exposed areas. Interestingly, the dermal papillae shape varies and their sizes increase in circumference from sun-exposed to sun-protected sites, as observed at a specific depth below the stratum corneum. In summary, our results demonstrate that near infra-red reflectance confocal microscopy is a feasible tool for microscopic analysis of skin morphometry in vivo. PMID:11407970

  3. Evaluation of dermal extracellular matrix and epidermal-dermal junction modifications using matrix-assisted laser desorption/ionization mass spectrometric imaging, in vivo reflectance confocal microscopy, echography, and histology: effect of age and peptide applications.

    PubMed

    Mondon, Philippe; Hillion, Mlanie; Peschard, Olivier; Andre, Nada; Marchand, Thibault; Doridot, Emmanuel; Feuilloley, Marc Gj; Pionneau, Cdric; Chardonnet, Solenne

    2015-06-01

    This study was conducted to establish a new methodology for evaluating elements of dermal extracellular matrix (ECM), of epidermal-dermal junction (EDJ), and effects of molecules which can modulate their synthesis. This methodology is based on matrix-assisted laser desorption/ionization mass spectrometric imaging (MALDI-MSI). In vivo reflectance confocal microscopy (in vivo RCM) and echography were also used. Using immunohistochemistry methods on explants, age-related modification data were obtained for selected dermal ECM and EDJ proteins (collagen I, collagen IV, collagen VII, collagen XVII, nidogen I, decorin/decorunt) and used as reference for MALDI-MSI studies. A methodology was developed with MALDI-MSI to map epidermis and dermis proteins. Then MALDI-MSI was used to study age modifications. In vivo RCM and high-frequency ultrasounds were used to evaluate ECM and EDJ undulation modifications caused by aging. Anti-aging molecule evaluations were performed with a blend of palmitoyl oligopeptide and palmitoyl tetrapeptide-7. Immunohistochemistry studies demonstrated that the selected proteins were found to be less abundant in aged group explants vs. young group except for decorin. MALDI-MSI studies correlated the results obtained for decorin. In vivo RCM measurements indicated a decrease of EDJ undulation depth with age and ECM modifications in the upper part of dermis. Echography demonstrated that the peptide blend reduced subepidermal low-echogenic band thickness and improved its density. In vivo RCM studies indicated that the peptides improved the ECM structure vs. placebo. This preliminary MALDI-MSI study raised some technical difficulties that were overcome. Further studies will be conducted to identify more proteins and to demonstrate the interest of this method for cosmetic evaluations. PMID:25817264

  4. Conventional and confocal epi-reflection and fluorescence microscopy of the rat kidney in vivo.

    PubMed

    Boyde, A; Capasso, G; Unwin, R J

    1998-01-01

    To visualize superficial and accessible renal tubule cells functioning in situ and to relate what we can 'see' to what we know of their function from more invasive in vivo or less direct in vitro studies means applying and adapting recent advances in epifluorescence and confocal microscopy to improve image resolution and to combine this with the use of fluorescent labels to monitor the handling of specific molecules by the proximal and distal renal tubule cells in vivo. Doing this in living tissue is novel, especially in the kidney. Application of confocal microscopy to the imaging of living tissue, as opposed to isolated cells, has not been widely reported. The kidney surface has been imaged before using the confocal microscope and in preliminary studies we have extended this by using a different confocal system with and without fluorescence. While the studies published up to now have been morphological, comparing standard renal (structural) histology of surface glomeruli and renal tubules with the corresponding in vivo confocal images, more dynamic, real-time studies have been limited. Individual red blood cells can be seen flowing around the peritubule capillary network and nucleated white blood cells can also be distinguished. Tubule cells, endothelial cells, the proximal tubule cell brush border and cell mitochondria can be visualized. Filtration and secretion can be observed, and the early and late parts of the proximal tubule distinguished, and the distal tubule recognized. Localization of fluorescently labeled insulin to the luminal brush border and progressive uptake of label and distribution within proximal tubule cells toward the basolateral (blood side) membrane can be demonstrated. The possibility of monitoring hemodynamic changes and tracking the filtration, uptake, secretion and absorption of fluorescently tagged molecules, as well as intracellular fluorescence, e.g. calcium or pH, is an exciting prospect and is ripe for detailed exploration. PMID:9730655

  5. High numerical aperture injection-molded miniature objective for fiber-optic confocal reflectance microscopy

    NASA Astrophysics Data System (ADS)

    Chidley, Matthew Douglas

    This dissertation presents the design of a miniature injection-molded objective lens for a fiber-optic confocal reflectance microscope. This is part of an effort to demonstrate the ability to fabricate 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 enable large-scale clinical screening and detection of early cancers and pre-cancerous lesions. This five lens plastic objective has been tested as a stand-alone optical system and has been coupled to 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. An optical-bench testing system was constructed to allow interactive alignment during testing. The modulation transfer function (MTF) of the miniature objective lens is determined using the slanted-edge method. A custom MATLAB program, edgeMTF, was written to collect, analyze, and record test data. An estimated Strehl ratio of 0.64 and an MTF value of 0.70, at the fiber-optic bundle Nyquist frequency, have been obtained. The main performance limitations of the miniature objective are mechanical alignment and flow-induced birefringence. Annealing and experimental injection molding runs were conducted in effort to reduce birefringence.

  6. High-precision confocal reflection measurement for two dimensional refractive index mapping of optical fibers

    NASA Astrophysics Data System (ADS)

    Raisin, Philippe; Scheuner, Jonas; Romano, Valerio; Ryser, Manuel

    2015-05-01

    We introduce a new fiber-optical approach for reflection based refractive index mapping. Our approach leads to improved stability and reliability over existing free-space confocal instruments and significantly cuts alignment efforts and reduces the number of components needed. Other than properly cleaved fiber end-faces, this setup requires no additional sample preparation. The instrument is calibrated by means of a set of samples with known refractive indices. The index steps of commercially available fibers are measured accurately down to < 10-3. The precision limit of the instrument is currently of the order of 10-4.

  7. Living Matter Observations with a Novel Hyperspectral Supercontinuum Confocal Microscope for VIS to Near-IR Reflectance Spectroscopy

    PubMed Central

    Bertani, Francesca R.; Ferrari, Luisa; Mussi, Valentina; Botti, Elisabetta; Costanzo, Antonio; Selci, Stefano

    2013-01-01

    A broad range hyper-spectroscopic microscope fed by a supercontinuum laser source and equipped with an almost achromatic optical layout is illustrated with detailed explanations of the design, implementation and data. The real novelty of this instrument, a confocal spectroscopic microscope capable of recording high resolution reflectance data in the VIS-IR spectral range from about 500 nm to 2.5 μm wavelengths, is the possibility of acquiring spectral data at every physical point as defined by lateral coordinates, X and Y, as well as at a depth coordinate, Z, as obtained by the confocal optical sectioning advantage. With this apparatus we collect each single scanning point as a whole spectrum by combining two linear spectral detector arrays, one CCD for the visible range, and one InGaAs infrared array, simultaneously available at the sensor output channel of the home made instrument. This microscope has been developed for biomedical analysis of human skin and other similar applications. Results are shown illustrating the technical performances of the instrument and the capability in extracting information about the composition and the structure of different parts or compartments in biological samples as well as in solid statematter. A complete spectroscopic fingerprinting of samples at microscopic level is shown possible by using statistical analysis on raw data or analytical reflectance models based on Abelés matrix transfer methods. PMID:24233077

  8. The application of dermal papillary rings in dermatology by in vivo confocal laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Xiang, W. Z.; Xu, A. E.; Xu, J.; Bi, Z. G.; Shang, Y. B.; Ren, Q. S.

    2010-08-01

    Confocal laser scanning microscopy (CLSM) allows noninvasive visualization of human skin in vivo, without needing to fix or section the tissue. Melanocytes and pigmented keratinocytes at the level of the basal layer form bright dermal papillary rings which are readily amenable to identify in confocal images. Our purpose was to explore the role of dermal papillary rings in assessment of lesion location, the diagnosis, differential diagnosis of lesions and assessment of therapeutic efficacy by in vivo CLSM. Seventy-one patients were imaged with the VivaScope 1500 reflectance confocal microscope provided by Lucid, Inc. The results indicate that dermal papillary rings can assess the location of lesion; the application of dermal papillary rings can provide diagnostic support and differential diagnosis for vitiligo, nevus depigmentosus, tinea versicolor, halo nevus, common nevi, and assess the therapeutic efficacy of NBUVB phototherapy plus topical 0.1 percent tacrolimus ointment for vitiligo. In conclusion, our findings indicate that the dermal papillary rings play an important role in the assessment the location of lesion, diagnosis, differential diagnosis of lesions and assessment of therapeutic efficacy by in vivo CLSM. CLSM may be a promising tool for noninvasive examination in dermatology. However, larger studies are needed to expand the application of dermal papillary rings in dermatology.

  9. Performance of full-pupil line-scanning reflectance confocal microscopy in human skin and oral mucosa in vivo

    PubMed Central

    Larson, Bjorg; Abeytunge, Sanjeewa; Rajadhyaksha, Milind

    2011-01-01

    Point-scanning reflectance confocal microscopes continue to be successfully translated for detection of skin cancer. Line-scanning, with the use of a single scanner and a linear-array detector, offers a potentially smaller, simpler and lower cost alternative approach, to accelerate widespread dissemination into the clinic. However, translation will require an understanding of imaging performance deep within scattering and aberrating human tissues. We report the results of an investigation of the performance of a full-pupil line-scanning reflectance confocal microscope in human skin and oral mucosa, in terms of resolution, optical sectioning, contrast, signal-to-noise ratio, imaging and the effect of speckle noise. PMID:21750780

  10. Virtual pinhole confocal microscope

    SciTech Connect

    George, J.S.; Rector, D.M.; Ranken, D.M.; Peterson, B.; Kesteron, J.

    1999-06-01

    Scanned confocal microscopes enhance imaging capabilities, providing improved contrast and image resolution in 3-D, but existing systems have significant technical shortcomings and are expensive. Researchers at Los Alamos National Laboratory have developed a novel approach--virtual pinhole confocal microscopy--that uses state of the art illumination, detection, and data processing technologies to produce an imager with a number of advantages: reduced cost, faster imaging, improved efficiency and sensitivity, improved reliability and much greater flexibility. Work at Los Alamos demonstrated proof of principle; prototype hardware and software have been used to demonstrate technical feasibility of several implementation strategies. The system uses high performance illumination, patterned in time and space. The authors have built functional confocal imagers using video display technologies (LCD or DLP) and novel scanner based on a micro-lens array. They have developed a prototype system for high performance data acquisition and processing, designed to support realtime confocal imaging. They have developed algorithms to reconstruct confocal images from a time series of spatially sub-sampled images; software development remains an area of active development. These advances allow the collection of high quality confocal images (in fluorescence, reflectance and transmission modes) with equipment that can inexpensively retrofit to existing microscopes. Planned future extensions to these technologies will significantly enhance capabilities for microscopic imaging in a variety of applications, including confocal endoscopy, and confocal spectral imaging.

  11. A comparison study of detecting gold nanorods in living cells with confocal reflectance microscopy and two-photon fluorescence microscopy.

    PubMed

    Zhou, Y; Wu, X; Wang, T; Ming, T; Wang, P N; Zhou, L W; Chen, J Y

    2010-02-01

    Two-photon fluorescence microscopy and confocal reflectance microscopy were compared to detect intracellular gold nanorods in rat basophilic leukaemia cells. The two-photon photoluminescence images of gold nanorods were acquired by an 800 nm fs laser with the power of milliwatts. The advantages of the obtained two-photon photoluminescence images are high spatial resolution and reduced background. However, a remarkable photothermal effect on cells was seen after 30 times continuous scanning of the femto-second laser, potentially affecting the subcellular localization pattern of the nanorods. In the case of confocal reflectance microscopy the images of gold nanorods can be obtained with the power of light source as low as microwatts, thus avoiding the photothermal effect, but the resolution of such images is reduced. We have noted that confocal reflectance images of cellular gold nanorods achieved with 50 microW 800 nm fs have a relatively poor resolution, whereas the 50 microW 488 nm CW laser can acquire reasonably satisfactory 3D reflectance images with improved resolution because of its shorter wavelength. Therefore, confocal reflectance microscopy may also be a suitable means to image intracellular gold nanorods with the advantage of reduced photothermal effect. PMID:20096050

  12. Investigation by in vivo reflectance confocal microscopy: melanocytes at the edges of solar lentigines.

    PubMed

    Nakajima, Akiko; Funasaka, Yoko; Kawana, Seiji

    2012-07-01

    In vivo reflectance confocal microscopy (RCM) provides high-resolution, real-time optical sections of the skin in a non-invasive manner, allowing visualization of the skin in its native state. Highly reflective skin components including melanin, collagen and keratin appear bright (white) in RCM images. RCM examination of solar lentigines is known to show features that correlate well with histologic findings such as supranuclear melanin caps, but there are a limited number of reports on melanocyte dendrites. In this study, we utilized RCM to investigate the melanocyte dendricity and distribution within solar lentigines. Seventeen healthy Japanese females who had fairly large solar lentigines on their faces were recruited to join our clinical study, and we examined them by using RCM on their non-lesional areas, and the inside and the outer rim of the lesional areas. As a result, we discovered that dendritic melanocytes were rarely seen in the center of a solar lentigo (SL), but were seen at a very high frequency in the outer rim of a SL. The results suggest that the melanocytes are more active at the edge of a SL, produce more melanin, and often spread their dendrites widely in a horizontal direction. The findings in this report might shed light on the dynamic pathomechanisms of solar lentigines in vivo. PMID:22626466

  13. Time-lapse confocal reflection microscopy of collagen fibrillogenesis and extracellular matrix assembly in vitro.

    PubMed

    Brightman, A O; Rajwa, B P; Sturgis, J E; McCallister, M E; Robinson, J P; Voytik-Harbin, S L

    2000-09-01

    The development of the next generation of biomaterials for restoration of tissues and organs (i.e., tissue engineering) requires a better understanding of the extracellular matrix (ECM) and its interaction with cells. Extracellular matrix is a macromolecular assembly of natural biopolymers including collagens, glycosaminoglycans (GAGs), proteoglycans (PGs), and glycoproteins. Interestingly, several ECM components have the ability to form three-dimensional (3D), supramolecular matrices (scaffolds) in vitro by a process of self-directed polymerization, "self-assembly". It has been shown previously that 3D matrices with distinct architectural and biological properties can be formed from either purified type I collagen or a complex mixture of interstitial ECM components derived from intestinal submucosa. Unfortunately, many of the imaging and analysis techniques available to study these matrices either are unable to provide insight into 3D preparations or demand efforts that are often prohibitory to observations of living, dynamic systems. This is the first report on the use of reflection imaging at rapid time intervals combined with laser-scanning confocal microscopy for analysis of structural properties and kinetics of collagen and ECM assembly in 3D. We compared time-lapse confocal reflection microscopy (TL-CRM) with a well-established spectrophotometric method for determining the self-assembly properties of both purified type I collagen and soluble interstitial ECM. While both TL-CRM and spectrophotometric techniques provided insight into the kinetics of the polymerization process, only TL-CRM allowed qualitative and quantitative evaluation of the structural parameters (e.g., fibril diameter) and 3D organization (e.g., fibril density) of component fibrils over time. Matrices formed from the complex mixture of soluble interstitial ECM components showed an increased rate of assembly, decreased opacity, decreased fibril diameter, and increased fibril density compared to that of purified type I collagen. These results suggested that the PG/GAG components of soluble interstitial ECM were affecting the polymerization of the component collagens. Therefore, the effects of purified and complex mixtures of PG/GAG components on the assembly properties of type I collagen and interstitial ECM were evaluated. The data confirmed that the presence of PG/GAG components altered the kinetics and the 3D fibril morphology of assembled matrices. In summary, TL-CRM was demonstrated to be a new and useful technique for analysis of the 3D assembly properties of collagen and other natural biopolymers which requires no specimen fixation and/or staining. PMID:10861383

  14. Image segmentation for integrated multiphoton microscopy and reflectance confocal microscopy imaging of human skin in vivo

    PubMed Central

    Chen, Guannan; Lui, Harvey

    2015-01-01

    Background Non-invasive cellular imaging of the skin in vivo can be achieved in reflectance confocal microscopy (RCM) and multiphoton microscopy (MPM) modalities to yield complementary images of the skin based on different optical properties. One of the challenges of in vivo microscopy is the delineation (i.e., segmentation) of cellular and subcellular architectural features. Methods In this work we present a method for combining watershed and level-set models for segmentation of multimodality images obtained by an integrated MPM and RCM imaging system from human skin in vivo. Results Firstly, a segmentation model based on watershed is introduced for obtaining the accurate structure of cell borders from the RCM image. Secondly,, a global region based energy level-set model is constructed for extracting the nucleus of each cell from the MPM image. Thirdly, a local region-based Lagrange Continuous level-set approach is used for segmenting cytoplasm from the MPM image. Conclusions Experimental results demonstrated that cell borders from RCM image and boundaries of cytoplasm and nucleus from MPM image can be obtained by our segmentation method with better accuracy and effectiveness. We are planning to use this method to perform quantitative analysis of MPM and RCM images of in vivo human skin to study the variations of cellular parameters such as cell size, nucleus size and other mophormetric features with skin pathologies. PMID:25694949

  15. In Vivo Reflectance Confocal Microscopy of Basal Cell Carcinoma: Feasibility of Preoperative Mapping of Cancer Margins

    PubMed Central

    Pan, Zhan-Yan; Lin, Jing-Ran; Cheng, Ting-Ting; Wu, Jia-Qiang; Wu, Wen-Yu

    2012-01-01

    Reflectance confocal microscopy (RCM) images skin at cellular resolution and has shown utility for the diagnosis of nonmelanoma skin cancer in vivo. It has the potential to define lesion margins before surgical therapy. Objectives To investigate the feasibility of RCM in defining the margins of basal cell carcinoma before surgery. Methods The margins of 10 lesions were evaluated using RCM. Biopsies of the margins were used to confirm the results. A protocol was constructed to define margins. RCM was used to delineate preoperative surgical margins in 13 patients. Intraoperative frozen biopsy was used to confirm the margins. Results In seven of 10 (70.0%) cases, the margins of the cancer were identified suing RCM. The tumor island was the critical feature in identifying the margins. In 12 of 13 (92.3%) cases, frozen biopsy corroborated that the surgical margins delineated by RCM were clear. Conclusion RCM imaging of the margins is feasible and demonstrates the possibility of preoperative mapping of cancer margins. PMID:23039159

  16. Automated Delineation of Dermal-Epidermal Junction In Reflectance Confocal Microscopy Image Stacks Of Human Skin

    PubMed Central

    Park, Brian

    2014-01-01

    Reflectance confocal microscopy (RCM) images skin non-invasively, with optical sectioning and nuclear-level resolution comparable to that of pathology. Based on assessment of the dermal-epidermal junction (DEJ) and morphologic features in its vicinity, skin cancer can be diagnosed in vivo with high sensitivity and specificity. However, the current visual, qualitative approach for reading images leads to subjective variability in diagnosis. We hypothesize that machine learning-based algorithms may enable a more quantitative, objective approach. Testing and validation was performed with two algorithms that can automatically delineate the DEJ in RCM stacks of normal human skin. The test set was composed of 15 fair and 15 dark skin stacks (30 subjects) with expert labellings. In dark skin, in which the contrast is high due to melanin, the algorithm produced an average error of 7.9±6.4μm. In fair skin, the algorithm delineated the DEJ as a transition zone, with average error of 8.3±5.8μm for the epidermis-to-transition zone boundary and 7.6±5.6μm for the transition zone-to-dermis. Our results suggest that automated algorithms may quantitatively guide the delineation of the DEJ, to assist in objective reading of RCM images. Further development of such algorithms may guide assessment of abnormal morphological features at the DEJ. PMID:25184959

  17. Combined reflectance confocal microscopy/optical coherence tomography imaging for skin burn assessment

    PubMed Central

    Iftimia, Nicusor; Ferguson, R. Daniel; Mujat, Mircea; Patel, Ankit H.; Zhang, Ellen Ziyi; Fox, William; Rajadhyaksha, Milind

    2013-01-01

    A combined high-resolution reflectance confocal microscopy (RCM)/optical coherence tomography (OCT) instrument for assessing skin burn gravity has been built and tested. This instruments allows for visualizing skin intracellular details with submicron resolution in the RCM mode and morphological and birefringence modifications to depths on the order of 1.2 mm in the OCT mode. Preliminary testing of the dual modality imaging approach has been performed on the skin of volunteers with some burn scars and on normal and thermally-injured Epiderm FTTM skin constructs. The initial results show that these two optical technologies have complementary capabilities that can offer the clinician a set of clinically comprehensive parameters: OCT helps to visualize deeper burn injuries and possibly quantify collagen destruction by measuring skin birefringence, while RCM provides submicron details of the integrity of the epidermal layer and identifies the presence of the superficial blood flow in the upper dermis. Therefore, the combination of these two technologies within the same instrument may provide a more comprehensive set of parameters that may help clinicians to more objectively and nonivasively assess burn injury gravity by determining tissue structural integrity and viability. PMID:23667785

  18. Intraoperative imaging during Mohs surgery with reflectance confocal microscopy: initial clinical experience.

    PubMed

    Flores, Eileen S; Cordova, Miguel; Kose, Kivanc; Phillips, William; Rossi, Anthony; Nehal, Kishwer; Rajadhyaksha, Milind

    2015-06-01

    Mohs surgery for the removal of nonmelanoma skin cancers (NMSCs) is performed in stages, while being guided by the examination for residual tumor with frozen pathology. However, preparation of frozen pathology at each stage is time consuming and labor intensive. Real-time intraoperative reflectance confocal microscopy(RCM), combined with video mosaicking, may enable rapid detection of residual tumor directly in the surgical wounds on patients. We report our initial experience on 25 patients, using aluminum chloride for nuclear contrast. Imaging was performed in quadrants in the wound to simulate the Mohs surgeons examination of pathology. Images and videos of the epidermal and dermal margins were found to be of clinically acceptable quality. Bright nuclear morphology was identified at the epidermal margin and detectable in residual NMSC tumors. The presence of residual tumor and normal skin features could be detected in the peripheral and deep dermal margins. Intraoperative RCM imaging may enable detection of residual tumor directly on patients during Mohs surgery, and may serve as an adjunct for frozen pathology. Ultimately, for routine clinical utility, a stronger tumor-to-dermis contrast may be necessary, and also a smaller microscope with an automated approach for imaging in the entire wound in a rapid and controlled manner. PMID:25706821

  19. Intraoperative imaging during Mohs surgery with reflectance confocal microscopy: initial clinical experience

    NASA Astrophysics Data System (ADS)

    Flores, Eileen S.; Cordova, Miguel; Kose, Kivanc; Phillips, William; Rossi, Anthony; Nehal, Kishwer; Rajadhyaksha, Milind

    2015-06-01

    Mohs surgery for the removal of nonmelanoma skin cancers (NMSCs) is performed in stages, while being guided by the examination for residual tumor with frozen pathology. However, preparation of frozen pathology at each stage is time consuming and labor intensive. Real-time intraoperative reflectance confocal microscopy (RCM), combined with video mosaicking, may enable rapid detection of residual tumor directly in the surgical wounds on patients. We report our initial experience on 25 patients, using aluminum chloride for nuclear contrast. Imaging was performed in quadrants in the wound to simulate the Mohs surgeon's examination of pathology. Images and videos of the epidermal and dermal margins were found to be of clinically acceptable quality. Bright nuclear morphology was identified at the epidermal margin and detectable in residual NMSC tumors. The presence of residual tumor and normal skin features could be detected in the peripheral and deep dermal margins. Intraoperative RCM imaging may enable detection of residual tumor directly on patients during Mohs surgery, and may serve as an adjunct for frozen pathology. Ultimately, for routine clinical utility, a stronger tumor-to-dermis contrast may be necessary, and also a smaller microscope with an automated approach for imaging in the entire wound in a rapid and controlled manner.

  20. Feasibility of intraoperative imaging during Mohs surgery with reflectance confocal microscopy

    NASA Astrophysics Data System (ADS)

    Flores, Eileen S.; Cordova, Miguel; Kose, Kivanc; Phillips, William; Nehal, Kishwer; Rajadhyaksha, Milind

    2014-03-01

    Mohs surgery for the removal of non-melanoma skin cancers (NMSCs) is performed in stages, while being guided by the examination for residual tumor with frozen pathology. However, preparation of frozen pathology at each stage is timeconsuming and labor-intensive. Real-time intraoperative reflectance confocal microscopy (RCM) may enable rapid detection of residual tumor directly in surgical wounds on patients. We report initial feasibility on twenty-one patients, using 35% AlCl3 for nuclear contrast. Imaging was performed in quadrants in the wound, to simulate the Mohs surgeon's examination of pathology. Images and videos of the epidermal and dermal margins were found to be of clinically acceptable quality. Bright nuclear morphology was identified at the epidermal margin. The presence of residual BCC/SCC tumor and normal skin features could be detected in the peripheral and deep dermal margins. Nuclear morphology was detectable in residual BCC/SCC tumors. Intraoperative RCM imaging may enable detection of residual tumor, directly on Mohs patients, and may serve as an adjunct for frozen pathology. However, a stronger source of contrast will be necessary, and also a smaller device with an automated approach for imaging in the entire wound in a rapid and controlled manner for clinical utility.

  1. In vivo reflectance confocal microscopy detects pigmentary changes in melasma at a cellular level resolution.

    PubMed

    Kang, Hee Young; Bahadoran, Philippe; Suzuki, Itaru; Zugaj, Didier; Khemis, Abdallah; Passeron, Thierry; Andres, Philippe; Ortonne, Jean-Paul

    2010-08-01

    Melasma is a frequent pigmentary disorder caused by abnormal melanin deposits in the skin. In vivo reflectance confocal microscopy (RCM) is a repetitive imaging tool that provides real-time images of the skin at nearly histological resolution. As melanin is the strongest endogenous contrast in human skin, pigmentary disorders are the most suitable candidates for RCM examination but RCM features of melasma have never been reported. This study investigates the pilot use of RCM in melasma to provide a set of well-described morphological criteria with histological correlations. RCM images were acquired from melasma skin and compared to adjacent control skin in 26 patients. Skin biopsies were obtained from eight patients. In the epidermis, RCM showed in all patients a significant increase in hyperrefractile cobblestoning cells. These cells corresponded to hyperpigmented basal keratinocytes in histology. In six patients, dendritic cells corresponding to activated melanocytes were also found in the epidermis. In the dermis, RCM identified in nine patients plump bright cells corresponding to melanophages. Interestingly, for a given patient, the topographic distribution of melanophages in melasma lesions was very heterogeneous. RCM also showed a significant increase in solar elastosis and blood vessels in the dermis. RCM is a non-invasive technique that detects pigmentary changes in melasma at a cellular level resolution. Therefore, RCM provides an innovative way to classify melasma by pigment changes. PMID:20497220

  2. Establishing the dynamics of neutrophil accumulation in vivo by reflectance confocal microscopy.

    PubMed

    Wolberink, Eshter A W; Peppelman, Malou; van de Kerkhof, Peter C M; van Erp, Piet E J; Gerritsen, Marie-Jeanne P

    2014-03-01

    Reflectance confocal microscopy (RCM) is an imaging tool, which visualizes the epidermal skin layers in vivo with a cellular resolution. Neutrophil accumulation is a characteristic feature in psoriasis and is thought to play a role in the pathophysiology of psoriasis. Until now, imaging of neutrophil accumulation in vivo is not performed. We evaluated the dynamics of neutrophil migration in active psoriatic lesions by non-invasive RCM imaging. Additionally, we evaluated the time phasing and duration of neutrophil trafficking. We performed RCM imaging prior to the start of topical treatment and for seven consecutive days with a 24-h time interval at the Radboud University Medical Center, Nijmegen, the Netherlands. Twelve psoriatic lesions in three patients with a severe exacerbation of psoriasis were included. The four most active lesions were selected in each patient based on the highest degree of redness, induration and expansion in the previous 2 weeks. In all lesions, a cyclic pattern of neutrophil migration was observed, consisting of squirting papillae, transepidermal migration, accumulation in the stratum spinosum, accumulation in the stratum corneum and degeneration of the abscesses. The time interval of a neutrophil-trafficking cycle was 5-7 days and showed a synchronic time phasing. This study is the first to establish the dynamics and time phasing of neutrophil migration in vivo in psoriatic lesions. Previously reported theories were confirmed by these novel in vivo data. RCM might distinguish between active or chronic psoriatic areas, which might contribute to new insights into the pathogenesis of psoriasis. PMID:24521061

  3. Confocal fluorescence microscopy: some applications in bone cell biology.

    PubMed

    Jones, S J; Taylor, M L

    1990-05-01

    Three-dimensional information is necessary for the proper investigation of the interrelationships of bone cells, and of the complex interface between these cells and the bone matrix they form and destroy. The use of fluorescence confocal microscopy was explored in the determination of the distribution of immunolabelled actin and vinculin--cytoskeletal and attachment proteins--in isolated chick bone cells cultured on dentine, and in neonate rat and rabbit calvaria. Confocal imaging, compared with conventional fluorescence imaging, greatly enhanced the interpretation possible. PMID:2115085

  4. A new wide field-of-view confocal imaging system and its applications in drug discovery and pathology

    NASA Astrophysics Data System (ADS)

    Li, Gang; Damaskinos, Savvas; Dixon, Arthur E.; Lee, Lucy E. J.

    2005-11-01

    Conventional widefield light microscopy and confocal scanning microscopy have been indispensable for pathology and drug discovery research. Clinical specimens from diseased tissues are examined, new drug candidates are tested on drug targets, and the morphological and molecular biological changes of cells and tissues are observed. High throughput screening of drug candidates requires highly efficient screening instruments. A standard biomedical slide is 1 by 3 inches (25.4 by 76.2 mm) in size. A typical tissue specimen is 10 mm in diameter. To form a high resolution image of the entire specimen, a conventional widefield light microscope must acquire a large number of small images of the specimen, and then tile them together, which is tedious, inefficient and error-prone. A patented new wide field-of-view confocal scanning laser imaging system has been developed for tissue imaging, which is capable of imaging an entire microscope slide without tiling. It is capable of operating in brightfield, reflection and epi-fluorescence imaging modes. Three (red, green and blue (RGB)) lasers are used to produce brightfield and reflection images, and to excite various fluorophores. This new confocal system makes examination of large biomedical specimens more efficient, and makes fluorescence examination of large specimens possible for the first time without tiling. Description of the new confocal technology and applications of the imaging system in pathology and drug discovery research, for example, imaging large tissue specimens, tissue microarrays, and zebrafish sections, are reported in this paper.

  5. Development of a confocal optical system design for molecular imaging applications of biochip.

    PubMed

    Huang, Guoliang; Xu, Shukuan; Zhu, Jiang; Deng, Cheng; Dong, Zhonghua; Yang, Yang; Yang, Xiaoyong; Wang, Xianhua; Jin, Guofan

    2007-01-01

    A novel confocal optical system design and a dual laser confocal scanner have been developed to meet the requirements of highly sensitive detection of biomolecules on microarray chips, which is characterized by a long working distance (wd>3.0 mm), high numerical aperture (NA=0.72), and only 3 materials and 7 lenses used. This confocal optical system has a high scanning resolution, an excellent contrast and signal-to-noise ratio, and an efficiency of collected fluorescence of more than 2-fold better than that of other commercial confocal biochip scanners. The scanner is as equally good for the molecular imaging detection of enclosed biochips as for the detection of biological samples on a slide surface covered with a cover-slip glass. Some applications of gene and protein imagings using the dual laser confocal scanner are described. PMID:18256735

  6. Fiber optic confocal reflectance microscopy: a new real-time technique to view nuclear morphology in cervical squamous epithelium in vivo

    NASA Astrophysics Data System (ADS)

    Sung, Kung-Bin; Richards-Kortum, Rebecca; Follen, Michele; Malpica, Anais; Liang, Chen; Descour, Michael R.

    2003-12-01

    We present a fiber optic confocal reflectance microscope (FCRM) which can be used to image epithelial tissue with sub-cellular resolution in vivo. Confocal images of normal and abnormal appearing cervical tissue were obtained in vivo from eighteen patients undergoing colposcopic examination of the cervix; biopsy specimens were taken from imaged sites. The measured lateral and axial resolutions of the system were 1.6 µm and 3 µm, respectively. Morphologic features, including nuclear size and nuclear-to-cytoplasmic ratio, were extracted from confocal images obtained at various depths beneath the epithelial surface. Image features extracted from confocal images compared well with features extracted from confocal images obtained in vitro and from previous histopathologic studies. This study shows that fiber optic confocal reflectance microscopy can be used to visualize the morphology of cervical epithelium in vivo.

  7. Fast intracellular motion in the living cell by video rate reflection confocal laser scanning microscopy

    PubMed Central

    VESELY, PAVEL; BOYDE, ALAN

    2001-01-01

    Fast intracellular motion (FIM) was first revealed by back scattered light (BSL) imaging in video rate confocal scanning laser microscopy (VRCSLM), beyond the limits of spatial and temporal resolution obtainable with conventional optical microscopy. BSL imaging enabled visualisation of intra and extracellular motion with resolution in space down to 0.2 ?m and in time to 1/25th of a second. Mapping the cell space at 0.2 ?m0.2 ?m (XY = in instantaneous best focal plane)0.5 ?m (Z = height/depth, optic axis direction) volume steps revealed a communication layer above the known contact layer and an integrated dynamic spatial network (IDSN) towards the cell centre. FIM was originally observed as localised quasichaotic dancing (dithering) or reflecting patches/spots in the cell centre, faster in the darker nuclear space. Later, a second type of FIM was recognised which differed by the presence of a varied proportion of centrifugal and centripetal directional movements and/or jumping of patches/spots in the cell centre and outside the nuclear space. The first type is characteristic for cells in slightly adverse conditions while the second type has so far only been found in eutrophic cells. Temporal speeding up and coarsening of FIM, followed by slowing and eventually cessation at cell death, was found on exposure to strong stressors. It was concluded that the state of FIM provides instantaneous information about individual cell reactions to actual treatment and about cell survival. A putative switch between the first and second type FIM could be considered as an indicator of timing of cellular processes. The significance of FIM for the biology of the cell is seen in the rapid assessment of the condition of an individual live cell investigated by combination of various methods. Requirements for further development of this approach are outlined. PMID:11465857

  8. Clinical applications of in vivo fluorescence confocal laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Oh, Chilhwan; Park, Sangyong; Kim, Junhyung; Ha, Seunghan; Park, Gyuman; Lee, Gunwoo; Lee, Onseok; Chun, Byungseon; Gweon, Daegab

    2008-02-01

    Living skin for basic and clinical research can be evaluated by Confocal Laser Scanning Microscope (CLSM) non-invasively. CLSM imaging system can achieve skin image its native state either "in vivo" or "fresh biopsy (ex vivo)" without fixation, sectioning and staining that is necessary for routine histology. This study examines the potential fluorescent CLSM with a various exogenous fluorescent contrast agent, to provide with more resolution images in skin. In addition, in vivo fluorescent CLSM researchers will be extended a range of potential clinical application. The prototype of our CLSM system has been developed by Prof. Gweon's group. The operating parameters are composed of some units, such as illuminated wavelength 488 nm, argon illumination power up to 20mW on the skin, objective lens, 0.9NA oil immersion, axial resolution 1.0μm, field of view 200μm x 100μm (lateral resolution , 0.3μm). In human volunteer, fluorescein sodium was administrated topically and intradermally. Animal studies were done in GFP transgenic mouse, IRC mouse and pig skin. For imaging of animal skin, fluorescein sodium, acridine orange, and curcumine were used for fluorescein contrast agent. We also used the GFP transgenic mouse for fluorescein CLSM imaging. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. Curcumin is a yellow food dye that has similar fluorescent properties to fluorescein sodium. Acridin Orange can be highlight nuclei in viable keratinocyte. In vivo CLSM of transgenic GFP mouse enable on in vivo, high resolution view of GFP expressing skin tissue. GFP signals are brightest in corneocyte, kertinocyte, hair and eccrine gland. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. In papillary dermis, fluorescein distribution is more homogeneous. Curcumin is a yellow food dye that has similar fluorescent properties to fluorescein sodium. In vivo CLSM of transgenic GFP mouse enable on in vivo, high resolution view of GFP expressing skin tissue. GFP signals are brightest in corneocyte, kertinocyte, skin appendage and blood vessels. In conclusion, this study demonstrates the usefulness of CLSM as technique for imaging skin in vivo. In addition, CLSM is non-invasive, the same tissue site may be imaged over a period of time to monitor the various change such as wound healing, severity of skin diseases and effect of therapeutic management.

  9. Current application of confocal endomicroscopy in gastrointestinal disorders.

    PubMed

    Nguyen, Nam Q; Leong, Rupert W L

    2008-10-01

    Confocal endomicroscopy (CEM) is a recent advancement in imaging technology that incorporates a confocal laser microscope into the tip of a flexible endoscope. The 1000-fold magnification and high resolution allows for real time in vivo histology or "virtual biopsies" of the gastrointestinal tract mucosa. CEM has the capability to instantaneously diagnose intra-epithelial neoplasia during endoscopy, alone or in combination with a "red-flag" technique, such as chromoendoscopy. Therefore, there is clinical utility in the surveillance or diagnosis of Barrett's esophagus, gastric intestinal metaplasia and cancer, longstanding ulcerative colitis, and colonic neoplasia. Furthermore, CEM also appears to be useful in the evaluation of coeliac disease, microscopic colitis, and in diagnosing Helicobacter pylori chronic gastritis. This review examines the current available data on the utility of this new technology in clinical gastroenterology and its potential impact in the future. PMID:18761561

  10. Confocal device and application strategies for endoluminal optical coherence microscopy

    NASA Astrophysics Data System (ADS)

    George, Markus; Schnieder, Ludger; Buess, Gerhard F.

    2003-10-01

    While endoscopic optical coherence tomography has been established successfully in vivo ,implementation of endoluminal optical coherence microscopy remains demanding,s suitable confocal probe is lacking. A miniaturized confocal laser scanning microscope is presented,which fulfills the requirements for endoluminal optical coherence microscopy. First,imaging experience gained for optical coherence microscopy of nimal gastrointestinal tissue samples is described. For this purpose,laboratory scale optical coherence microscope with an image acquisition time of 1min 30 s was employed. Cellular membranes can be identified throughout the gastrointestinal organs. Frequency domain image analysis can be used to distinguish columnar from squamous epithelium. Profilometric information on sample surfaces can be obtained directly as isophase lines. Second, the miniaturized confocal laser scanning microscope is characterized. Having an effective diameter of 25 mm, it houses single-mode optical fiber,scanning mirror and an objective lens. The micro-electro-mechanical mirror with gimballed suspension allows two dimensional scanning without introducing an optical path difference. The sinusoidal movement of both axes has to be considered to approximate cartesian image coordinates. Field geometry is illustrated s function of excitation amplitude and frequency. Acceptable image quality is chieved for frame rate of 0.5 Hz. A strategy to position the focal plane axially within the sample volume is discussed.

  11. [Application of Three Dimensional Confocal Micro X-Ray Fluorescence Technology Based on Polycapillary X-Ray Lens in Analysis of Rock and Mineral Samples].

    PubMed

    Li, Fang-zuo; Liu, Zhi-guo; Sun, Tian-xi; Yi, Long-tao; Zhao, Wei-gang; He, Jia-lin; Peng, Song; Wang, Li-li; Zhao, Guang-cui; Ding, Xun-liang

    2015-09-01

    Confocal three dimensional (3D) micro X-ray fluorescence (XRF) spectrometer based on a polycapillary focusing X-ray lens (PFXRL) in the excitation channel and a polycapillary parallel X-ray lens (PPXRL) in the detection channel was developed. The PFXRL and PPXRL were placed in a confocal configuration. This was helpful in improving the signal-to-noise ratio of the XRF spectra, and accordingly lowered the detection limitation of the XRF technology. The confocal configuration ensured that only the XRF signal from the confocal micro-volume overlapped by the output focal spot of the PFXRL and the input focal spot of the PPXRL could be detected by the detector. Therefore, the point-to-point information of XRF for samples could be obtained non-destructively by moving the sample located at the confocal position. The magnitude of the gain in power density of the PFXRL was 10(3). This let the low power conventional X-ray source be used in this confocal XRF, and, accordingly, decreased the requirement of high power X-ray source for the confocal XRF based on polycapillary X-ray optics. In this paper, we used the confocal 3D micro X-ray fluorescence spectrometer to non-destructively analyzed mineral samples and to carry out a 3D point-to-point elemental mapping scanning, which demonstrated the capabilities of confocal 3D micro XRF technology for non-destructive analysis elements composition and distribution for mineral samples. For one mineral sample, the experimental results showed that the area with high density of element of iron had high density of copper. To some extent, this reflected the growth mechanisms of the mineral sample. The confocal 3D micro XRF technology has potential applications in such fields like the analysis identification of ore, jade, lithoid utensils, "gamble stone" and lithoid flooring. PMID:26669153

  12. A simple method for overcoming some problems when observing thick reflective biological samples with a confocal scanning laser microscope.

    PubMed

    Rumio, C; Morini, M; Miani, A; Barajon, I; Castano, P

    1995-01-01

    A simple device is described, which allows the range of depth of scanning to be reduced when observing thick reflecting biological samples with a confocal scanning laser microscope (CSLM). Thick histological sections of human skin and rat brain stem were mounted between two coverslips ('sandwich' style) and the optical tomography was performed from both sides by turning the 'sandwich' upside-down. The samples were impregnated using standard Golgi-Cox, 'rapid Golgi' or other silver methods. The ability to turn the 'sandwich' upside-down is particularly useful when the reflective structure inspected is deep inside the section, i.e., near the lower surface of the specimen, or when it is opaque to the laser beam or excessively reflective. PMID:7897649

  13. Detection of living Sarcoptes scabiei larvae by reflectance mode confocal microscopy in the skin of a patient with crusted scabies

    NASA Astrophysics Data System (ADS)

    Levi, Assi; Mumcuoglu, Kosta Y.; Ingber, Arieh; Enk, Claes D.

    2012-06-01

    Scabies is an intensely pruritic disorder induced by a delayed type hypersensitivity reaction to infestation of the skin by the mite Sarcoptes scabiei. The diagnosis of scabies is established clinically and confirmed by identifying mites or eggs by microscopic examination of scrapings from the skin or by surface microscopy using a dermatoscope. Reflectance-mode confocal microscopy is a novel technique used for noninvasive imaging of skin structures and lesions at a resolution compatible to that of conventional histology. Recently, the technique was employed for the confirmation of the clinical diagnosis of scabies. We demonstrate the first ever documentation of a larva moving freely inside the skin of a patient infected with scabies.

  14. Combined reflectance confocal microscopy-optical coherence tomography for delineation of basal cell carcinoma margins: an ex vivo study

    NASA Astrophysics Data System (ADS)

    Iftimia, Nicusor; Peterson, Gary; Chang, Ernest W.; Maguluri, Gopi; Fox, William; Rajadhyaksha, Milind

    2016-01-01

    We present a combined reflectance confocal microscopy (RCM) and optical coherence tomography (OCT) approach, integrated within a single optical layout, for diagnosis of basal cell carcinomas (BCCs) and delineation of margins. While RCM imaging detects BCC presence (diagnoses) and its lateral spreading (margins) with measured resolution of ˜1 μm, OCT imaging delineates BCC depth spreading (margins) with resolution of ˜7 μm. When delineating margins in 20 specimens of superficial and nodular BCCs, depth could be reliably determined down to ˜600 μm, and agreement with histology was within about ±50 μm.

  15. [Application of confocal simultaneous scanner unit in the study of forebrain neurodevelopment in zebrafish].

    PubMed

    Shi, Ying; Gao, Jing-Xia; Peng, Gang; Jiang, Min

    2013-02-25

    With the application of the photoconversion technology of genetically expressed fluorescent proteins in biologic field, more powerful confocal imaging ability was demanded. The aim of the present study was to establish an experimental model employing confocal simultaneous scanner unit for simultaneous laser stimulation and imaging, taking study of forebrain neurodevelopment in zebrafish as an example. In the present study, 36-48-hour-old Tg(lhx5:kaede) zebrafish embryos were mounted with 1.2% low melting temperature agarose. The forebrain neurons marked with kaede were observed using the simultaneous scanner unit of confocal microscopy. The 405 nm laser was used to stimulate the region of interest (ROI), while 488 and 559 nm lasers were used to acquire images at the same time. The photoconversion state of kaede protein was then reviewed, and the projecting pattern of neurons stimulated by the ultraviolet laser was examined. The results showed that, the fluorescence of stimulated kaede turned from green to red, and the photoconversion of kaede demonstrated anterior dorsal telencephalon (ADt) neurons projected axons ventrally into the anterior commissure (AC) and supraoptic tract (SOT). These results suggest the confocal simultaneous scanner unit meets the demand of the photoconversion experiment. The application of confocal simultaneous scanning technology in examining Tg(lhx5:kaede) zebrafish embryos affords an ideal experimental model in neurodevelopment study. PMID:23426517

  16. Label-free in vivo imaging of myelinated axons in health and disease with spectral confocal reflectance microscopy

    PubMed Central

    Schain, Aaron J.; Hill, Robert A.; Grutzendler, Jaime

    2013-01-01

    We report a new technique for high-resolution in vivo imaging of myelinated axons in the brain, spinal cord and peripheral nerve that requires no fluorescent labeling. This method, based on spectral confocal reflectance microscopy (SCoRe), uses a conventional laser scanning confocal system to generate images by merging the simultaneously reflected signals from multiple lasers of different wavelengths. Striking color patterns unique to individual myelinated fibers are generated that facilitate their tracing in dense axonal areas. These patterns highlight nodes of Ranvier and Schmidt-Lanterman incisures and can be used to detect various myelin pathologies. Using SCoRe we performed chronic brain imaging up to 400 ?m deep, capturing for the first time de novo myelination of mouse cortical axons in vivo. We also established the feasibility of imaging myelinated axons in the human cerebral cortex. SCoRe adds a powerful component to the evolving toolbox for imaging myelination in living animals and potentially in humans. PMID:24681598

  17. Confocal reflectance and two-photon microscopy studies of a songbird skull for preparation of transcranial imaging

    NASA Astrophysics Data System (ADS)

    Abi-Haidar, Darine; Oliver, Thomas

    2009-05-01

    We present experiments and analyses of confocal reflectance and two-photon microscopy studies of zebra finch skull samples. The thin and hollow structure of these birds' skulls is quite translucent, which can allow in vivo transcranial two-photon imaging for brain activation monitoring. However, the skull structure is also quite complex, with high refractive index changes on a macroscopic scale. These studies aim at exploring the geometrical and scattering properties of these skull samples with the use of several confocal microscopy contrasts. Moreover, the study of the axial reflectance exponential decay is used to estimate the scattering coefficients of the bone. Finally, two-photon imaging experiments of a fluorescent object located beneath the skull are carried out. It reveals that two-photon fluorescence can be collected through the skull with a strong signal. It also reveals that the spatial resolution loss is quite high and cannot be fully explained by the bulk scattering properties of the bone, but also by the presence of the high refractive index inhomogeneity of this pneumatic skull structure. Even if the optical properties of the skull are different during in vivo experiments, these preliminary studies are aimed at preparing and optimizing transcranial brain activation monitoring experiments on songbirds.

  18. Pipette SNOM/AFM probe applicable to any wavelength and combination with confocal microscope

    NASA Astrophysics Data System (ADS)

    Muramatsu, Hiroshi; Chiba, Norio; Homma, Katsunori; Yamamoto, Noritaka; Egawa, Akira

    1999-06-01

    A novel pipette SNOM/AFM probe has been developed for its simple fabrication and applicability to wide wavelength range. The pipette probe is simply fabricated by a successive process of pulling, bending with a CO2 laser, making a hole and coating with a metal layer. The hole is made on the tube at the back side of the tip for applying a light. The pipette probe is mounted on the SNOM/AFM system which includes a confocal microscope. The light is introduced to the hole directly by focusing from a confocal microscope or through an optical fiber. The probe provided clear topographic and optical images for a sample of a patterned chromium layer on a glass substrate and fluorescence beads. A confocal images was also obtained in a wide area of the same samples.

  19. HIV detection by in-situ hybridization based on confocal reflected light microscopy

    NASA Astrophysics Data System (ADS)

    Smith, Louis C.; Jericevic, Zeljko; Cuellar, Roland; Paddock, Stephen W.; Lewis, Dorothy E.

    1991-05-01

    Elucidation of the pathogenesis of AIDS is confounded by the finding that few actively infected CD4+ cells (1 in 104-105) can be detected in the peripheral blood, even though there is dramatic depletion (often >90%) of CD4+ cells as the disease progresses. A sensitive, 35S-based human immunodeficiency virus (HIV) mRNA in situ hybridization technique was coupled with a new detection method, confocal laser scanning microscopy, to examine transcriptionally active HIV-infected cells from individuals at different disease stages. An algorithm for image segmentation and analysis has been developed to determine the proportion of HIV-positive cells. Data obtained using this improved detection method suggest that there are more HIV mRNA-producing cells in HIV-infected individuals than previously thought, based on other detection methods.

  20. Application of confocal laser endomicroscopy in the diagnosis and management of Barretts esophagus

    PubMed Central

    Leggett, Cadman L.; Gorospe, Emmanuel C.

    2014-01-01

    Confocal laser endomicroscopy is an advanced endoscopic imaging modality that can be used for the diagnosis of early mucosal dysplasia in various gastrointestinal conditions. It provides histology-like images at 1000-fold magnification. The technology offers potential advantages in the diagnosis of Barretts esophagus and early esophageal cancer due to the low yield of the current practice of surveillance endoscopy with biopsies. Confocal laser endomicroscopy has the potential to eliminate the need for biopsy, establish diagnosis and facilitate application of endoscopic therapy during the time of actual endoscopy. There are several studies that have demonstrated reasonable diagnostic accuracy in patients undergoing surveillance for Barretts esophagus from tertiary academic medical centers. However, the application of confocal laser endomicroscopy in routine clinical endoscopy is still in the process of refinement. Its role in the diagnosis and treatment of Barretts-associated dysplasia will continue to evolve with improvement in technology, criteria for diagnosis and experience among endoscopists in interpreting confocal imaging. PMID:24976007

  1. In vivo confocal microscopy in dermatology: from research to clinical application

    NASA Astrophysics Data System (ADS)

    Ulrich, Martina; Lange-Asschenfeldt, Susanne

    2013-06-01

    Confocal laser scanning microscopy (CLSM) represents an emerging technique for the noninvasive histomorphological analysis of skin in vivo and has shown its applicability for dermatological research as well as its value as an adjunct tool in the clinical management of skin cancer patients. Herein, we aim to give an overview on the current clinical indications for CLSM in dermatology and also highlight the diverse applications of CLSM in dermatological research.

  2. Analysis of the efficiency of hair removal by different optical methods: comparison of Trichoscan, reflectance confocal microscopy, and optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Kuck, Monika; Schanzer, Sabine; Ulrich, Martina; Bartels, Natalie Garcia; Meinke, Martina C.; Fluhr, Joachim; Krah, Martin; Blume-Peytavi, Ulrike; Stockfleth, Eggert; Lademann, Jürgen

    2012-10-01

    Noninvasive diagnostic tools, such as Trichoscan, reflectance confocal microscopy (RCM), and optical coherence tomography (OCT), are efficient methods of hair shaft and growth evaluation. The aim of this study was to carry out a comparative assessment of these three medical procedures by measuring the hair shaft and hair growth after hair removal for a defined period of five days. The application of these techniques was demonstrated by measuring hair growth on the lower leg of six female volunteers. After removal of the hair shaft with a shaving system, the hair follicle infundibula and the length of the growing hairs were measured with the Trichoscan, RCM, and OCT method. All three methods are reliable hair measuring tools after hair removal. Trichoscan is best suited in the implementation of hair growth measurement and RCM in the analysis of hair follicles, whereas the OCT system can be consulted as an additional measurement for the evaluation of the hair follicle and length.

  3. Evaluation through in vivo reflectance confocal microscopy of the cutaneous neurogenic inflammatory reaction induced by capsaicin in human subjects

    NASA Astrophysics Data System (ADS)

    C?runtu, Constantin; Boda, Daniel

    2012-08-01

    We perform an in vivo analysis of the effects of capsaicin on cutaneous microvascularization. A total of 29 healthy subjects are administered a solution of capsaicin (CAP group) or a vehicle solution (nonCAP group) on the dorsal side of the nondominant hand. The evaluation is performed using in vivo reflectance confocal microscopy (RCM). Ten minutes after administration, the area of the section, the perimeter, and the Feret's diameter of the capillaries in the dermal papillae become significantly larger in the CAP group as against the nonCAP group, and this difference is maintained until the conclusion of the experiment. In vivo RCM allows the investigation of cutaneous vascular reactions induced by capsaicin. As such, this method may constitute an useful technique both for research and clinical practice.

  4. Dual-mode reflectance and fluorescence near-video-rate confocal microscope for architectural, morphological and molecular imaging of tissue.

    PubMed

    Carlson, Alicia L; Coghlan, Lezlee G; Gillenwater, Ann M; Richards-Kortum, Rebecca R

    2007-10-01

    We have developed a near-video-rate dual-mode reflectance and fluorescence confocal microscope for the purpose of imaging ex vivo human specimens and in vivo animal models. The dual-mode confocal microscope (DCM) has light sources at 488, 664 and 784 nm, a frame rate of 15 frames per second, a maximum field of view of 300 x 250 mum and a resolution limit of 0.31 mum laterally and 1.37 mum axially. The DCM can image tissue architecture and cellular morphology, as well as molecular properties of tissue, using reflective and fluorescent molecular-specific optical contrast agents. Images acquired with the DCM demonstrate that the system has the sub-cellular resolution needed to visualize the morphological and molecular changes associated with cancer progression and has the capability to image animal models of disease in vivo. In the hamster cheek pouch model of oral carcinogenesis, the DCM was used to image the epithelium and stroma of the cheek pouch; blood flow was visible and areas of dysplasia could be distinguished from normal epithelium using 6% acetic acid contrast. In human oral cavity tissue slices, DCM reflectance images showed an increase in the nuclear-to-cytoplasmic ratio and density of nuclei in neoplastic tissues as compared to normal tissue. After labelling tissue slices with fluorescent contrast agents targeting the epidermal growth factor receptor, an increase in epidermal growth factor receptor expression was detected in cancerous tissue as compared to normal tissue. The combination of reflectance and fluorescence imaging in a single system allowed imaging of two different parameters involved in neoplastic progression, providing information about both the morphological and molecular expression changes that occur with cancer progression. The dual-mode imaging capabilities of the DCM allow investigation of both morphological changes as well as molecular changes that occur in disease processes. Analyzing both factors simultaneously may be advantageous when trying to detect and diagnose disease. The DCM's high resolution and near-video-rate image acquisition and the growing inventory of molecular-specific contrast agents and disease-specific molecular markers holds significant promise for in vivo studies of disease processes such as carcinogenesis. PMID:17910693

  5. Application of confocal laser microscopy for monitoring mesh implants in herniology

    SciTech Connect

    Zakharov, V P; Belokonev, V I; Bratchenko, I A; Timchenko, P E; Vavilov, A V; Volova, L T

    2011-04-30

    The state of the surface of mesh implants and their encapsulation region in herniology is investigated by laser confocal microscopy. A correlation between the probability of developing relapses and the size and density of implant microdefects is experimentally shown. The applicability limits of differential reverse scattering for monitoring the post-operation state of implant and adjacent tissues are established based on model numerical experiments. (optical technologies in biophysics and medicine)

  6. Optical biopsy of early gastroesophageal cancer by catheter-based reflectance-type laser-scanning confocal microscopy.

    PubMed

    Nakao, Madoka; Yoshida, Shigeto; Tanaka, Shinji; Takemura, Yoshito; Oka, Shiro; Yoshihara, Masaharu; Chayama, Kazuaki

    2008-01-01

    Magnified endoscopic observation of the gastrointestinal tract has become possible. However, such observation at the cellular level remains difficult. Laser-scanning confocal microscopy (LCM) is a novel, noninvasive optical imaging method that provides instant microscopic images of untreated tissue under endoscopy. We compare prototype catheter-based reflectance-type LCM images in vivo and histologic images of early gastroesophageal cancer to assess the usefulness of LCM in diagnosing such cancer. 20 sites in the esophagus and 40 sites in the stomach are examined by LCM under endoscopy prior to endoscopic or surgical resection. A prototype catheter LCM system, equipped with a semiconductor laser that oscillates at 685 nm and analyzes reflected light (Mauna Kea Technologies, Paris, France; Fujinon, Saitama, Japan) is used in vivo without fluorescent agent. In all normal esophageal mucosa and esophageal cancers, the nuclei are visualized. In nine of the ten normal esophageal mucosa, cell membranes are visualized, and in five of the ten esophageal cancers, cell membranes are visualized. In all normal gastric mucosa, nuclei and cell membranes are not visualized, but in ten of the 20 gastric cancers, nuclei are visualized. This novel method will aid in immediate diagnosis under endoscopy without the need for biopsy. PMID:19021423

  7. A novel technique using potassium permanganate and reflectance confocal microscopy to image biofilm extracellular polymeric matrix reveals non-eDNA networks in Pseudomonas aeruginosa biofilms.

    PubMed

    Swearingen, Matthew C; Mehta, Ajeet; Mehta, Amar; Nistico, Laura; Hill, Preston J; Falzarano, Anthony R; Wozniak, Daniel J; Hall-Stoodley, Luanne; Stoodley, Paul

    2016-02-01

    Biofilms are etiologically important in the development of chronic medical and dental infections. The biofilm extracellular polymeric substance (EPS) determines biofilm structure and allows bacteria in biofilms to adapt to changes in mechanical loads such as fluid shear. However, EPS components are difficult to visualize microscopically because of their low density and molecular complexity. Here, we tested potassium permanganate, KMnO4, for use as a non-specific EPS contrast-enhancing stain using confocal laser scanning microscopy in reflectance mode. We demonstrate that KMnO4 reacted with EPS components of various strains of Pseudomonas, Staphylococcus and Streptococcus, yielding brown MnO2 precipitate deposition on the EPS, which was quantifiable using data from the laser reflection detector. Furthermore, the MnO2 signal could be quantified in combination with fluorescent nucleic acid staining. COMSTAT image analysis indicated that KMnO4 staining increased the estimated biovolume over that determined by nucleic acid staining alone for all strains tested, and revealed non-eDNA EPS networks in Pseudomonas aeruginosa biofilm. In vitro and in vivo testing indicated that KMnO4 reacted with poly-N-acetylglucosamine and Pseudomonas Pel polysaccharide, but did not react strongly with DNA or alginate. KMnO4 staining may have application as a research tool and for diagnostic potential for biofilms in clinical samples. PMID:26536894

  8. Pilot study of semiautomated localization of the dermal∕epidermal junction in reflectance confocal microscopy images of skin

    PubMed Central

    Kurugol, Sila; Dy, Jennifer G.; Brooks, Dana H.; Rajadhyaksha, Milind

    2011-01-01

    Reflectance confocal microscopy (RCM) continues to be translated toward the detection of skin cancers in vivo. Automated image analysis may help clinicians and accelerate clinical acceptance of RCM. For screening and diagnosis of cancer, the dermal∕epidermal junction (DEJ), at which melanomas and basal cell carcinomas originate, is an important feature in skin. In RCM images, the DEJ is marked by optically subtle changes and features and is difficult to detect purely by visual examination. Challenges for automation of DEJ detection include heterogeneity of skin tissue, high inter-, intra-subject variability, and low optical contrast. To cope with these challenges, we propose a semiautomated hybrid sequence segmentation∕classification algorithm that partitions z-stacks of tiles into homogeneous segments by fitting a model of skin layer dynamics and then classifies tile segments as epidermis, dermis, or transitional DEJ region using texture features. We evaluate two different training scenarios: 1. training and testing on portions of the same stack; 2. training on one labeled stack and testing on one from a different subject with similar skin type. Initial results demonstrate the detectability of the DEJ in both scenarios with epidermis∕dermis misclassification rates smaller than 10% and average distance from the expert labeled boundaries around 8.5 μm. PMID:21456869

  9. Comparison of reflectance confocal microscopy and two-photon second harmonic generation microscopy in fungal keratitis rabbit model ex vivo

    PubMed Central

    Lee, Jun Ho; Lee, Seunghun; Yoon, Calvin J.; Park, Jin Hyoung; Tchah, Hungwon; Kim, Myoung Joon; Kim, Ki Hean

    2016-01-01

    Fungal keratitis is an infection of the cornea by fungal pathogens. Diagnosis methods based on optical microscopy could be beneficial over the conventional microbiology method by allowing rapid and non-invasive examination. Reflectance confocal microscopy (RCM) and two-photon second harmonic generation microscopy (TPSHGM) have been applied to pre-clinical or clinical studies of fungal keratitis. In this report, RCM and TPSHGM were characterized and compared in the imaging of a fungal keratitis rabbit model ex vivo. Fungal infection was induced by using two strains of fungi: aspergillus fumigatus and candida albicans. The infected corneas were imaged in fresh condition by both modalities sequentially and their images were analyzed. Both RCM and TPSHGM could detect both fungal strains within the cornea based on morphology: aspergillus fumigatus had distinctive filamentous structures, and candida albicans had round structures superficially and elongated structures in the corneal stroma. These imaging results were confirmed by histology. Comparison between RCM and TPSHGM showed several characteristics. Although RCM and TPSHGM images had good correlation each other, their images were slightly different due to difference in contrast mechanism. RCM had relatively low image contrast with the infected turbid corneas due to high background signal. TPSHGM visualized cells and collagen in the cornea clearly compared to RCM, but used higher laser power to compensate low autofluorescence. Since these two modalities provide complementary information, combination of RCM and TPSHGM would be useful for fungal keratitis detection by compensating their weaknesses each other. PMID:26977371

  10. Reflectance Confocal Microscopy as an Aid to Dermoscopy to Improve Diagnosis on Equivocal Lesions: Evaluation of Three Bluish Nodules

    PubMed Central

    Bassoli, Sara; Seidenari, Stefania; Pellacani, Giovanni; Longo, Caterina; Cesinaro, Anna Maria

    2010-01-01

    Nodular lesions can be difficult to diagnose under dermoscopy alone, since they often lack specific diagnostic features. Confocal microscopy can be used as an aid to dermoscopy, to increase the diagnostic accuracy on equivocal skin lesions. We report three cases of bluish nodular lesions, difficult to diagnose under dermoscopy alone. Confocal features were very useful in these cases to lead us to the correct diagnosis, recognizing benign versus malignant entities. Histopathology is also reported, with high correspondence compared to the confocal imaging. PMID:20886008

  11. Application of confocal X-ray fluorescence micro-spectroscopy to the investigation of paint layers.

    PubMed

    Sun, Tianxi; Liu, Zhiguo; Wang, Guangfu; Ma, Yongzhong; Peng, Song; Sun, Weiyuan; Li, Fangzuo; Sun, Xuepeng; Ding, Xunliang

    2014-12-01

    A confocal micro X-ray fluorescence (MXRF) spectrometer based on polycapillary X-ray optics was used for the identification of paint layers. The performance of the confocal MXRF was studied. Multilayered paint fragments of a car were analyzed nondestructively to demonstrate that this confocal MXRF instrument could be used in the discrimination of the various layers in multilayer paint systems. PMID:25151613

  12. Reflective coatings for solar applications

    SciTech Connect

    Jorgensen, G.

    1993-05-01

    Many applications of solar energy require large mirrors to provide high levels of concentrated sunlight. The success of such conversion systems hinges on the optical durability and economic viability of the reflector materials. A major effort at the National Renewable Energy Laboratory (NREL) has been to improve the existing reflector materials technology and to identify candidates that retain optical performance and durability criteria and offer potential for reduced cost. To attain the goals, it is desirable to maintain and increase the involvement of industrial organizations in reflective materials R D related to the conversion of solar resources to useful energy. Toward this end, NREL has recently initiated several collaborative efforts with industry to develop advanced reflector materials.

  13. Reflective coatings for solar applications

    SciTech Connect

    Jorgensen, G.

    1993-05-01

    Many applications of solar energy require large mirrors to provide high levels of concentrated sunlight. The success of such conversion systems hinges on the optical durability and economic viability of the reflector materials. A major effort at the National Renewable Energy Laboratory (NREL) has been to improve the existing reflector materials technology and to identify candidates that retain optical performance and durability criteria and offer potential for reduced cost. To attain the goals, it is desirable to maintain and increase the involvement of industrial organizations in reflective materials R&D related to the conversion of solar resources to useful energy. Toward this end, NREL has recently initiated several collaborative efforts with industry to develop advanced reflector materials.

  14. Reflectance confocal microscope for imaging oral tissues in vivo, potentially with line scanning as a low-cost approach for clinical use

    NASA Astrophysics Data System (ADS)

    Peterson, Gary; Abeytunge, Sanjeewa; Eastman, Zachary; Rajadhyaksha, Milind

    2012-02-01

    Reflectance confocal microscopy with a line scanning approach potentially offers a smaller, simpler and less expensive approach than traditional methods of point scanning for imaging in living tissues. With one moving mechanical element (galvanometric scanner), a linear array detector and off-the-shelf optics, we designed a compact (102x102x76mm) line scanning confocal reflectance microscope (LSCRM) for imaging human tissues in vivo in a clinical setting. Custom-designed electronics, based on field programmable gate array (FPGA) logic has been developed. With 405 nm illumination and a custom objective lens of numerical aperture 0.5, lateral resolution was measured to be 0.8 um (calculated 0.64 um). The calculated optical sectioning is 3.2 um. Preliminary imaging shows nuclear and cellular detail in human skin and oral epithelium in vivo. Blood flow is also visualized in the deeper connective tissue (lamina propria) in oral mucosa. Since a line is confocal only in one dimension (parallel) but not in the other, the detection is more sensitive to multiply scattered out of focus background noise than in the traditional point scanning configuration. Based on the results of our translational studies thus far, a simpler, smaller and lower-cost approach based on a LSCRM appears to be promising for clinical imaging.

  15. Applicability of confocal laser scanning microscopy for evaluation and monitoring of cutaneous wound healing

    NASA Astrophysics Data System (ADS)

    Lange-Asschenfeldt, Susanne; Bob, Adrienne; Terhorst, Dorothea; Ulrich, Martina; Fluhr, Joachim; Mendez, Gil; Roewert-Huber, Hans-Joachim; Stockfleth, Eggert; Lange-Asschenfeldt, Bernhard

    2012-07-01

    There is a high demand for noninvasive imaging techniques for wound assessment. In vivo reflectance confocal laser scanning microscopy (CLSM) represents an innovative optical technique for noninvasive evaluation of normal and diseased skin in vivo at near cellular resolution. This study was designed to test the feasibility of CLSM for noninvasive analysis of cutaneous wound healing in 15 patients (7 male/8 female), including acute and chronic, superficial and deep dermal skin wounds. A commercially available CLSM system was used for the assessment of wound bed and wound margins in order to obtain descriptive cellular and morphological parameters of cutaneous wound repair noninvasively and over time. CLSM was able to visualize features of cutaneous wound repair in epidermal and superficial dermal wounds, including aspects of inflammation, neovascularisation, and tissue remodelling in vivo. Limitations include the lack of mechanic fixation of the optical system on moist surfaces restricting the analysis of chronic skin wounds to the wound margins, as well as a limited optical resolution in areas of significant slough formation. By describing CLSM features of cutaneous inflammation, vascularisation, and epithelialisation, the findings of this study support the role of CLSM in modern wound research and management.

  16. Neurosurgical confocal endomicroscopy: A review of contrast agents, confocal systems, and future imaging modalities

    PubMed Central

    Zehri, Aqib H.; Ramey, Wyatt; Georges, Joseph F.; Mooney, Michael A.; Martirosyan, Nikolay L.; Preul, Mark C.; Nakaji, Peter

    2014-01-01

    Background: The clinical application of fluorescent contrast agents (fluorescein, indocyanine green, and aminolevulinic acid) with intraoperative microscopy has led to advances in intraoperative brain tumor imaging. Their properties, mechanism of action, history of use, and safety are analyzed in this report along with a review of current laser scanning confocal endomicroscopy systems. Additional imaging modalities with potential neurosurgical utility are also analyzed. Methods: A comprehensive literature search was performed utilizing PubMed and key words: In vivo confocal microscopy, confocal endomicroscopy, fluorescence imaging, in vivo diagnostics/neoplasm, in vivo molecular imaging, and optical imaging. Articles were reviewed that discussed clinically available fluorophores in neurosurgery, confocal endomicroscopy instrumentation, confocal microscopy systems, and intraoperative cancer diagnostics. Results: Current clinically available fluorescent contrast agents have specific properties that provide microscopic delineation of tumors when imaged with laser scanning confocal endomicroscopes. Other imaging modalities such as coherent anti-Stokes Raman scattering (CARS) microscopy, confocal reflectance microscopy, fluorescent lifetime imaging (FLIM), two-photon microscopy, and second harmonic generation may also have potential in neurosurgical applications. Conclusion: In addition to guiding tumor resection, intraoperative fluorescence and microscopy have the potential to facilitate tumor identification and complement frozen section analysis during surgery by providing real-time histological assessment. Further research, including clinical trials, is necessary to test the efficacy of fluorescent contrast agents and optical imaging instrumentation in order to establish their role in neurosurgery. PMID:24872922

  17. 3D Imaging of Porous Media Using Laser Scanning Confocal Microscopy with Application to Microscale Transport Processes

    SciTech Connect

    Fredrich, J.T.

    1999-02-10

    We present advances in the application of laser scanning confocal microscopy (LSCM) to image, reconstruct, and characterize statistically the microgeometry of porous geologic and engineering materials. We discuss technical and practical aspects of this imaging technique, including both its advantages and limitations. Confocal imaging can be used to optically section a material, with sub-micron resolution possible in the lateral and axial planes. The resultant volumetric image data, consisting of fluorescence intensities for typically {approximately}50 million voxels in XYZ space, can be used to reconstruct the three-dimensional structure of the two-phase medium. We present several examples of this application, including studying pore geometry in sandstone, characterizing brittle failure processes in low-porosity rock deformed under triaxial loading conditions in the laboratory, and analyzing the microstructure of porous ceramic insulations. We then describe approaches to extract statistical microgeometric descriptions from volumetric image data, and present results derived from confocal volumetric data sets. Finally, we develop the use of confocal image data to automatically generate a three-dimensional mesh for numerical pore-scale flow simulations.

  18. Treatment of Basal Cell Carcinoma Using a One-Stop-Shop With Reflectance Confocal Microscopy: Study Design and Protocol of a Randomized Controlled Multicenter Trial

    PubMed Central

    Wolkerstorfer, Albert; Elshot, Yannick; Zupan-Kajcovski, Biljana; Crijns, Marianne B; Starink, Markus V; Bekkenk, Marcel W; van der Wal, Allard C; Spuls, Phyllis I; de Rie, Menno A

    2015-01-01

    Background Basal cell carcinoma (BCC) is the most common cancer diagnosed in white populations worldwide. The rising incidence of BCC is becoming a major worldwide public health problem. Therefore, there is a need for more efficient management. Objective The aim of this research is to assess the efficacy and safety of a one-stop-shop (OSS) concept, using real-time in vivo reflectance confocal microscopy (RCM) (Vivascope 1500; Lucid Technologies, Henrietta, NY, USA) as a diagnostic tool, prior to surgical management of new primary BCCs. Methods This is a prospective non-inferiority multi-center RCT designed to compare the “OSS concept using RCM” to current standards of care in diagnosing and treating clinically suspected BCC. Patients ≥ 18 years attending our outpatient clinic at the Department of Dermatology, Academic Medical Center, University of Amsterdam, and the Department of Dermatology, the Netherlands Cancer Institute-Antoni van Leeuwenhoek Hospital (Amsterdam, The Netherlands) with a clinically suspected new primary BCC lesion will be considered for enrollment using predefined inclusion and exclusion criteria, and will be randomly allocated to the experimental or control group. The main outcome parameter is the assessment of incomplete surgical excision margins on the final pathology report of confirmed BCC lesions (either by punch biopsy or RCM imaging). Other outcome measures include diagnostic accuracy (sensitivity and specificity) of RCM for diagnosing BCC and dividing between subtypes, and throughput time. Patient satisfaction data will be collected postoperatively after 3 months during routine follow-up. Results This research is investigator-initiated and received ethics approval. Patient recruitment started in February 2015, and we expect all study-related activities to be completed by fall 2015. Conclusions This RCT is the first to examine an OSS concept using RCM for diagnosing and treating clinically suspected BCC lesions. Results of this research are expected to have applications in evidence-based practice for the increasing number of patients suffering from BCC and possibly lead to a more efficient disease management strategy. Trial Registration ClinicalTrials.gov: NCT02285790; https://clinicaltrial.gov/ct2/show/NCT02285790 (Archived by WebCite at http://www.webcitation.org/6b2LfDKWu). PMID:26362616

  19. Comparative confocal microscopy of internal genitalia of phytoptine mites (Eriophyoidea, Phytoptidae): new generic diagnoses reflecting host-plant associations.

    PubMed

    Chetverikov, Philipp E

    2014-02-01

    Confocal laser scanning microscopy (CLSM) methods are still rarely used by acarologists although they are very appropriate for studying minute arthropod pests, especially eriophyoid mites. In this paper, the female reproductive system of phytoptines, including the bud mite Phytoptus avellanae, the well-known pest of hazelnut, was studied using CLSM and resulted in new interpretations of the functioning anatomy of phytoptid genitalia. Comparison of cuticle-lined internal genitalia, based on novel CLSM-based 2D and 3D imaging, and multivariate analysis of morphometric measurements, show that two basic types of internal genitalia can be found within Phytoptinae: one type in phytoptines associated with monocotyledoneous hosts (especially Cyperaceae and Asparagaceae), and another one in those associated with various dicotyledoneous hosts. Phytoptines from monocots (genera Oziella and Acathrix and Phytoptus "caricis" sp. group) possess a spherical distal part of the spermathecal tube and a semitriangular transverse genital apodeme, whereas phytoptines from dicots (genus Phytoptus "avellenae" sp. group) possess an elongate distal part of the tube and a trapezoidal apodeme. These differences in the internal genitalic anatomy were used for modifying the diagnosis of phytoptine genera (Phytoptus, Oziella and Acathrix), and reorganizing the Phytoptinae, resulting in new synonymies: 11 species were transferred from genus Phytoptus "caricis" sp. group to the genus Oziella. PMID:24037537

  20. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: FOUNDATIONS FOR QUANTIFYING CYTOMETRIC APPLICATIONS WITH SPECTROSCOPIC INSTRUMENTS

    EPA Science Inventory

    The confocal laser-scanning microscopy (CLSM) has enormous potential in many biological fields. The goal of a CLSM is to acquire and quantify fluorescence and in some instruments acquire spectral characterization of the emitted signal. The accuracy of these measurements demands t...

  1. Digital holographic confocal microscope

    NASA Astrophysics Data System (ADS)

    Goy, Alexandre S.; Psaltis, Demetri

    2013-02-01

    We demonstrate experimentally a scanning confocal microscopy technique based on digital holographic recording of the scanned spot. The data collected in this way contains all the information to produce three-dimensional images. Several methods to treat the data are presented, such as the dynamic placement of the pinhole. Examples of reflection and transmission images of epithelial cells and mouse brain tissue are shown. The computations can be performed in real time, the speed being limited only by the frame rate of the camera. This method enables a convenient implementation of confocal microscopy, especially in transmission as no de-scan device is required.

  2. Application of regularized RichardsonLucy algorithm for deconvolution of confocal microscopy images

    PubMed Central

    Laasmaa, M; Vendelin, M; Peterson, P

    2011-01-01

    Although confocal microscopes have considerably smaller contribution of out-of-focus light than widefield microscopes, the confocal images can still be enhanced mathematically if the optical and data acquisition effects are accounted for. For that, several deconvolution algorithms have been proposed. As a practical solution, maximum-likelihood algorithms with regularization have been used. However, the choice of regularization parameters is often unknown although it has considerable effect on the result of deconvolution process. The aims of this work were: to find good estimates of deconvolution parameters; and to develop an open source software package that would allow testing different deconvolution algorithms and that would be easy to use in practice. Here, RichardsonLucy algorithm has been implemented together with the total variation regularization in an open source software package IOCBio Microscope. The influence of total variation regularization on deconvolution process is determined by one parameter. We derived a formula to estimate this regularization parameter automatically from the images as the algorithm progresses. To assess the effectiveness of this algorithm, synthetic images were composed on the basis of confocal images of rat cardiomyocytes. From the analysis of deconvolved results, we have determined under which conditions our estimation of total variation regularization parameter gives good results. The estimated total variation regularization parameter can be monitored during deconvolution process and used as a stopping criterion. An inverse relation between the optimal regularization parameter and the peak signal-to-noise ratio of an image is shown. Finally, we demonstrate the use of the developed software by deconvolving images of rat cardiomyocytes with stained mitochondria and sarcolemma obtained by confocal and widefield microscopes. PMID:21323670

  3. Application of real-time confocal laser scanning microscopy to observe living cells in tissue specimens.

    PubMed

    Saino, Tomoyuki; Satoh, Yoh-ichi

    2004-01-01

    Fluorescence microscopy imaging has developed into an important tool for the study of cell structure and function in cell biology. This non-invasive technique permits the characterization, localization and qualitative quantification of free ions, messengers, pH, voltage and other molecules in living cells. The regulation of cytosolic Ca2+ homeostasis is essential for cells. However, most investigations have used cultured or isolated cells as an experimental model and, consequently, provide only limited insight into the mechanisms that operate in tissue in situ. More useful information could be obtained by studying intact tissue specimens. The calcium dynamics of some tissue specimens, such as arteriole smooth muscle cells, supra cervical ganglia and peripheral nerve bundles, were analysed in this study. Real-time confocal microscopy revealed that individual cells exhibited different [Ca2+]i dynamics and the responses to transmitters/modulators were heterogeneous. It is important that the confocal microscopes have good detection performances, due to the reduction of stray light. We conclude that real-time confocal microscopy is a useful tool for investigating structural and functional changes of cells in living tissues, although suitable tissue-preparation is important for these measurements. PMID:15077899

  4. Modern confocal microscopy.

    PubMed

    Rajwa, Bartek

    2005-02-01

    This unit re-examines confocal microscopy from a current perspective. It outlines many of the most modern applications of confocal microscopy and the issues surrounding them. The expanding applications of confocal microscopy demand both minor and major modifications of the technology to enhance imaging capabilities for a growing variety of samples. Techniques of interest such as FRET, FLIP, FRAP, and IFRAP are described. The unit includes a discussion on multispectral imaging, potentially the most exciting innovation in the field of biological imaging. The other area that is beginning to have considerable impact on biology, medicine, and pharmacology comprises high-content screening (HCS) and high-throughput screening (HTS). This unit also introduces programmable array microscopes (PAMs), which are based upon a technique in which spatial modulators are placed in the imaging plane of a microscope and used to generate patterns of illumination and/or detection. PMID:18770814

  5. Scanning computed confocal imager

    DOEpatents

    George, John S. (Los Alamos, NM)

    2000-03-14

    There is provided a confocal imager comprising a light source emitting a light, with a light modulator in optical communication with the light source for varying the spatial and temporal pattern of the light. A beam splitter receives the scanned light and direct the scanned light onto a target and pass light reflected from the target to a video capturing device for receiving the reflected light and transferring a digital image of the reflected light to a computer for creating a virtual aperture and outputting the digital image. In a transmissive mode of operation the invention omits the beam splitter means and captures light passed through the target.

  6. Broadband reflectance coatings for vacuum ultraviolet application

    NASA Technical Reports Server (NTRS)

    Herzig, Howard; Fleetwood, C. M., Jr.; Flint, B. K.

    1987-01-01

    An experimental investigation has obtained results indicating that neither LaF3 nor LiYF4 are acceptable alternatives to MgF2 as coatings for vacuum-deposited aluminum mirrors from which high UV reflectance down to 1150 A is required. Nevertheless, LaF3 may prove useful in those specialized applications in which the suppression of lower wavelength emissions, such as the 1216-A hydrogen line, is desirable.

  7. Towards real-time image deconvolution: application to confocal and STED microscopy

    PubMed Central

    Zanella, R.; Zanghirati, G.; Cavicchioli, R.; Zanni, L.; Boccacci, P.; Bertero, M.; Vicidomini, G.

    2013-01-01

    Although deconvolution can improve the quality of any type of microscope, the high computational time required has so far limited its massive spreading. Here we demonstrate the ability of the scaled-gradient-projection (SGP) method to provide accelerated versions of the most used algorithms in microscopy. To achieve further increases in efficiency, we also consider implementations on graphic processing units (GPUs). We test the proposed algorithms both on synthetic and real data of confocal and STED microscopy. Combining the SGP method with the GPU implementation we achieve a speed-up factor from about a factor 25 to 690 (with respect the conventional algorithm). The excellent results obtained on STED microscopy images demonstrate the synergy between super-resolution techniques and image-deconvolution. Further, the real-time processing allows conserving one of the most important property of STED microscopy, i.e the ability to provide fast sub-diffraction resolution recordings. PMID:23982127

  8. Application of Laser Scanning Confocal Microscopy to Heat and Mass Transport Modeling in Porous Microstructures

    NASA Technical Reports Server (NTRS)

    Marshall, Jochen; Milos, Frank; Fredrich, Joanne; Rasky, Daniel J. (Technical Monitor)

    1997-01-01

    Laser Scanning Confocal Microscopy (LSCM) has been used to obtain digital images of the complicated 3-D (three-dimensional) microstructures of rigid, fibrous thermal protection system (TPS) materials. These orthotropic materials are comprised of refractory ceramic fibers with diameters in the range of 1 to 10 microns and have open porosities of 0.8 or more. Algorithms are being constructed to extract quantitative microstructural information from the digital data so that it may be applied to specific heat and mass transport modeling efforts; such information includes, for example, the solid and pore volume fractions, the internal surface area per volume, fiber diameter distributions, and fiber orientation distributions. This type of information is difficult to obtain in general, yet it is directly relevant to many computational efforts which seek to model macroscopic thermophysical phenomena in terms of microscopic mechanisms or interactions. Two such computational efforts for fibrous TPS materials are: i) the calculation of radiative transport properties; ii) the modeling of gas permeabilities.

  9. Rheology and Confocal Reflectance Microscopy as Probes of Mechanical Properties and Structure during Collagen and Collagen/Hyaluronan Self-Assembly

    PubMed Central

    Yang, Ya-li; Kaufman, Laura J.

    2009-01-01

    In this work, the gelation of three-dimensional collagen and collagen/hyaluronan (HA) composites is studied by time sweep rheology and time lapse confocal reflectance microscopy (CRM). To investigate the complementary nature of these techniques, first collagen gel formation is investigated at concentrations of 0.5, 1.0, and 1.5 mg/mL at 37°C and 32°C. The following parameters are used to describe the self-assembly process in all gels: the crossover time (tc), the slope of the growth phase (kg), and the arrest time (ta). The first two measures are determined by rheology, and the third by CRM. A frequency-independent rheological measure of gelation, tg, is also measured at 37°C. However, this quantity cannot be straightforwardly determined for gels formed at 32°C, indicating that percolation theory does not fully capture the dynamics of collagen network formation. The effects of collagen concentration and gelation temperature on kg, tc, and ta as well as on the mechanical properties and structure of these gels both during gelation and at equilibrium are elucidated. Composite collagen/HA gels are also prepared, and their properties are monitored at equilibrium and during gelation at 37°C and 32°C. We show that addition of HA subtly alters mechanical properties and structure of these systems both during the gelation process and at equilibrium. This occurs in a temperature-dependent manner, with the ratio of HA deposited on collagen fibers versus that distributed homogeneously between fibers increasing with decreasing gelation temperature. In addition to providing information on collagen and collagen/HA structure and mechanical properties during gelation, this work shows new ways in which rheology and microscopy can be used complementarily to reveal details of gelation processes. PMID:19217873

  10. Micro- and nanodomain imaging in uniaxial ferroelectrics: Joint application of optical, confocal Raman, and piezoelectric force microscopy

    SciTech Connect

    Shur, V. Ya. Zelenovskiy, P. S.

    2014-08-14

    The application of the most effective methods of the domain visualization in model uniaxial ferroelectrics of lithium niobate (LN) and lithium tantalate (LT) family, and relaxor strontium-barium niobate (SBN) have been reviewed in this paper. We have demonstrated the synergetic effect of joint usage of optical, confocal Raman, and piezoelectric force microscopies which provide extracting of the unique information about formation of the micro- and nanodomain structures. The methods have been applied for investigation of various types of domain structures with increasing complexity: (1) periodical domain structure in LN and LT, (2) nanodomain structures in LN, LT, and SBN, (3) nanodomain structures in LN with modified surface layer, (4) dendrite domain structure in LN. The self-assembled appearance of quasi-regular nanodomain structures in highly non-equilibrium switching conditions has been considered.

  11. FluoRender: An Application of 2D Image Space Methods for 3D and 4D Confocal Microscopy Data Visualization in Neurobiology Research

    PubMed Central

    Wan, Yong; Otsuna, Hideo; Chien, Chi-Bin; Hansen, Charles

    2013-01-01

    2D image space methods are processing methods applied after the volumetric data are projected and rendered into the 2D image space, such as 2D filtering, tone mapping and compositing. In the application domain of volume visualization, most 2D image space methods can be carried out more efficiently than their 3D counterparts. Most importantly, 2D image space methods can be used to enhance volume visualization quality when applied together with volume rendering methods. In this paper, we present and discuss the applications of a series of 2D image space methods as enhancements to confocal microscopy visualizations, including 2D tone mapping, 2D compositing, and 2D color mapping. These methods are easily integrated with our existing confocal visualization tool, FluoRender, and the outcome is a full-featured visualization system that meets neurobiologists’ demands for qualitative analysis of confocal microscopy data. PMID:23584131

  12. Reflection: Its Concepts and Applications in Geometry

    ERIC Educational Resources Information Center

    Man, Yiu Kwong

    2004-01-01

    This paper discusses the basic concepts of reflection and its related concepts in optics. It aims at providing examples on how to apply the principle of reflection in geometry. Explorations of the concepts involved via dynamic geometry software are also included.

  13. Confocal Microscopy Core Facility

    Cancer.gov

    The Confocal Microscopy Core Facility provides "open access" confocal laser scanning microscopy (LSM) services to all CCR investigators. The Facility's equipment includes a: Zeiss LSM 510 META NLO for 2-photon imaging system Zeiss LSM 710 NLO for 2-photon

  14. Confocal Microscopy Core Facility

    Cancer.gov

    The Confocal Microscopy Core Facility is supported by CCR and there is no charge to individual users for confocal time. Collaborations with laboratories outside the CCR are also considered, time permitting. Please refer to our publications list for exampl

  15. Identifying brain neoplasms using dye-enhanced multimodal confocal imaging

    NASA Astrophysics Data System (ADS)

    Wirth, Dennis; Snuderl, Matija; Sheth, Sameer; Kwon, Churl-Su; Frosch, Matthew P.; Curry, William; Yaroslavsky, Anna N.

    2012-02-01

    Brain tumors cause significant morbidity and mortality even when benign. Completeness of resection of brain tumors improves quality of life and survival; however, that is often difficult to accomplish. The goal of this study was to evaluate the feasibility of using multimodal confocal imaging for intraoperative detection of brain neoplasms. We have imaged different types of benign and malignant, primary and metastatic brain tumors. We correlated optical images with histopathology and evaluated the possibility of interpreting confocal images in a manner similar to pathology. Surgical specimens were briefly stained in 0.05 mg/ml aqueous solution of methylene blue (MB) and imaged using a multimodal confocal microscope. Reflectance and fluorescence signals of MB were excited at 642 nm. Fluorescence emission of MB was registered between 670 and 710 nm. After imaging, tissues were processed for hematoxylin and eosin (H&E) histopathology. The results of comparison demonstrate good correlation between fluorescence images and histopathology. Reflectance images provide information about morphology and vascularity of the specimens, complementary to that provided by fluorescence images. Multimodal confocal imaging has the potential to aid in the intraoperative detection of microscopic deposits of brain neoplasms. The application of this technique may improve completeness of resection and increase patient survival.

  16. EVALUATION OF CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: APPLICATIONS FOR IMAGING MORPHOLOGY AND DEATH IN EMBRYOS AND REPRODUCTIVE TISSUE/ORGANS

    EPA Science Inventory

    The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. It is remarkable that procedures to test the performance of these machines are not done routinely by most investigators and thus many of the machines in the field are working at level...

  17. Histopathological confirmation of similar intramucosal distribution of fluorescein in both intravenous administration and local mucosal application for probe-based confocal laser endomicroscopy of the normal stomach.

    PubMed

    Nonaka, Kouichi; Ohata, Ken; Ban, Shinichi; Ichihara, Shin; Takasugi, Rumi; Minato, Yohei; Tashima, Tomoaki; Matsuyama, Yasushi; Takita, Maiko; Matsuhashi, Nobuyuki; Neumann, Helmut

    2015-12-16

    Probe-based confocal laser endomicroscopy (pCLE) is capable of acquiring in vivo magnified cross-section images of the gastric mucosa. Intravenous injection of fluorescein sodium is used for confocal imaging. However, it is still under debate if local administration of the dye to the mucosa is also effective for confocal imaging as it is not yet clear if topical application also reveals the intramucosal distribution of fluorescein. The objective of this study was to evaluate the intramucosal distribution of fluorescein sodium after topical application and to compare the distribution to the conventional intravenous injection used for confocal imaging. pCLE of the stomach uninfected with Helicobacter pylori was performed in a healthy male employing intravenous administration and local mucosal application of fluorescein. The mucosa of the lower gastric body was biopsied 1 min and 5 min after intravenous administration or local mucosal application of fluorescein, and the distribution of fluorescein in the biopsy samples was examined histologically. Green fluorescence was already observed in the cytoplasm of fundic glandular cells in the biopsied deep mucosa 1 min after local mucosal application of fluorescein. It was also observed in the foveolar lumen and inter-foveolar lamina propria, although it was noted at only a few sites. In the tissue biopsied 5 min after the local mucosal application of fluorescein, green fluorescence was more frequently noted in the cytoplasm of fundic glandular cells than in that 1 min after the local mucosal application of fluorescein, although obvious green fluorescence was not identified in the foveolar lumen or inter-foveolar lamina propria. The distribution of intravenously administered fluorescein in the cytoplasm of fundic glandular cells was also clearly observed similarly to that after local mucosal application of fluorescein. Green fluorescence in more cells was observed in many cells 5 min after intravenous administration compared with that after 1 min. The presence of fluorescein in the mucosa was observed within a short time after local mucosal application of fluorescein, suggesting that pCLE images similarly to those after intravenous fluorescein administration can be acquired by local mucosal application of fluorescein. PMID:26677449

  18. Histopathological confirmation of similar intramucosal distribution of fluorescein in both intravenous administration and local mucosal application for probe-based confocal laser endomicroscopy of the normal stomach

    PubMed Central

    Nonaka, Kouichi; Ohata, Ken; Ban, Shinichi; Ichihara, Shin; Takasugi, Rumi; Minato, Yohei; Tashima, Tomoaki; Matsuyama, Yasushi; Takita, Maiko; Matsuhashi, Nobuyuki; Neumann, Helmut

    2015-01-01

    Probe-based confocal laser endomicroscopy (pCLE) is capable of acquiring in vivo magnified cross-section images of the gastric mucosa. Intravenous injection of fluorescein sodium is used for confocal imaging. However, it is still under debate if local administration of the dye to the mucosa is also effective for confocal imaging as it is not yet clear if topical application also reveals the intramucosal distribution of fluorescein. The objective of this study was to evaluate the intramucosal distribution of fluorescein sodium after topical application and to compare the distribution to the conventional intravenous injection used for confocal imaging. pCLE of the stomach uninfected with Helicobacter pylori was performed in a healthy male employing intravenous administration and local mucosal application of fluorescein. The mucosa of the lower gastric body was biopsied 1 min and 5 min after intravenous administration or local mucosal application of fluorescein, and the distribution of fluorescein in the biopsy samples was examined histologically. Green fluorescence was already observed in the cytoplasm of fundic glandular cells in the biopsied deep mucosa 1 min after local mucosal application of fluorescein. It was also observed in the foveolar lumen and inter-foveolar lamina propria, although it was noted at only a few sites. In the tissue biopsied 5 min after the local mucosal application of fluorescein, green fluorescence was more frequently noted in the cytoplasm of fundic glandular cells than in that 1 min after the local mucosal application of fluorescein, although obvious green fluorescence was not identified in the foveolar lumen or inter-foveolar lamina propria. The distribution of intravenously administered fluorescein in the cytoplasm of fundic glandular cells was also clearly observed similarly to that after local mucosal application of fluorescein. Green fluorescence in more cells was observed in many cells 5 min after intravenous administration compared with that after 1 min. The presence of fluorescein in the mucosa was observed within a short time after local mucosal application of fluorescein, suggesting that pCLE images similarly to those after intravenous fluorescein administration can be acquired by local mucosal application of fluorescein. PMID:26677449

  19. Clinical applications of a real-time scanning-slit confocal microscope designed for real-time observations of the in-vivo human cornea

    NASA Astrophysics Data System (ADS)

    Masters, Barry R.

    1995-05-01

    We describe a new, real-time, flying slit confocal microscope, that has unique features and imaging characteristics for in vivo human ocular imaging. In vivo real-time confocal microscopy is currently used to investigate the tear film, renewal of the ocular surface, the role of epithelial innervation in epithelial cell proliferation, wound healing, kinetics of drug penetration, the effects of laser refractive surgery on the keratocyte activation and distribution in the stroma, and the nature of endothelial defects. The following clinical examples will be presented and discussed: confocal microscopy of normal human basal and wing cells in the epithelium, confocal microscopy of lamellar and penetrating corneal grafts, confocal microscopy of corneal ulcer, confocal microscopy of scar formation after herpes keratitis, and confocal microscopy of corneal innervation. The use of scanning slit confocal microscopes has unique advantages over other instrumental systems based on pinhole-containing Nipkow disks (tandem-scanning confocal microscopes) for clinical in vivo confocal microscopy.

  20. Binary phase digital reflection holograms - Fabrication and potential applications

    NASA Technical Reports Server (NTRS)

    Gallagher, N. C., Jr.; Angus, J. C.; Coffield, F. E.; Edwards, R. V.; Mann, J. A., Jr.

    1977-01-01

    A novel technique for the fabrication of binary-phase computer-generated reflection holograms is described. By use of integrated circuit technology, the holographic pattern is etched into a silicon wafer and then aluminum coated to make a reflection hologram. Because these holograms reflect virtually all the incident radiation, they may find application in machining with high-power lasers. A number of possible modifications of the hologram fabrication procedure are discussed.

  1. Forensic applications of microscopical infrared internal reflection spectroscopy

    NASA Astrophysics Data System (ADS)

    Tungol, Mary W.; Bartick, Edward G.; Reffner, John A.

    1994-01-01

    Applications of microscopical infrared internal reflection spectroscopy in forensic science are discussed. Internal reflection spectra of single fibers, hairs, paint chips, vehicle rubber bumpers, photocopy toners, carbon copies, writing ink on paper, lipstick on tissue, black electrical tape, and other types of forensic evidence have been obtained. The technique is convenient, non-destructive, and may permit smeared materials to be analyzed in situ.

  2. Confocal fluorescence microendoscopy of bronchial epithelium.

    PubMed

    Lane, Pierre M; Lam, Stephen; McWilliams, Annette; Leriche, Jean C; Anderson, Marshall W; Macaulay, Calum E

    2009-01-01

    Confocal microendoscopy permits the acquisition of high-resolution real-time confocal images of bronchial mucosa via the instrument channel of an endoscope. We report here on the construction and validation of a confocal fluorescence microendoscope and its use to acquire images of bronchial epithelium in vivo. Our objective is to develop an imaging method that can distinguish preneoplastic lesions from normal epithelium to enable us to study the natural history of these lesions and the efficacy of chemopreventive agents without biopsy removal of the lesion that can introduce a spontaneous regression bias. The instrument employs a laser-scanning engine and bronchoscope-compatible confocal probe consisting of a fiber-optic image guide and a graded-index objective lens. We assessed the potential of topical application of physiological pH cresyl violet (CV) as a fluorescence contrast-enhancing agent for the visualization of tissue morphology. Images acquired ex vivo with the confocal microendoscope were first compared with a bench-top confocal fluorescence microscope and conventional histology. Confocal images from five sites topically stained with CV were then acquired in vivo from high-risk smokers and compared to hematoxylin and eosin stained sections of biopsies taken from the same site. Sufficient contrast in the confocal imagery was obtained to identify cells in the bronchial epithelium. However, further improvements in the miniature objective lens are required to provide sufficient axial resolution for accurate classification of preneoplastic lesions. PMID:19405738

  3. Confocal fluorescence microendoscopy of bronchial epithelium

    NASA Astrophysics Data System (ADS)

    Lane, Pierre M.; Lam, Stephen; McWilliams, Annette; Leriche, Jean C.; Anderson, Marshall W.; Macaulay, Calum E.

    2009-03-01

    Confocal microendoscopy permits the acquisition of high-resolution real-time confocal images of bronchial mucosa via the instrument channel of an endoscope. We report here on the construction and validation of a confocal fluorescence microendoscope and its use to acquire images of bronchial epithelium in vivo. Our objective is to develop an imaging method that can distinguish preneoplastic lesions from normal epithelium to enable us to study the natural history of these lesions and the efficacy of chemopreventive agents without biopsy removal of the lesion that can introduce a spontaneous regression bias. The instrument employs a laser-scanning engine and bronchoscope-compatible confocal probe consisting of a fiber-optic image guide and a graded-index objective lens. We assessed the potential of topical application of physiological pH cresyl violet (CV) as a fluorescence contrast-enhancing agent for the visualization of tissue morphology. Images acquired ex vivo with the confocal microendoscope were first compared with a bench-top confocal fluorescence microscope and conventional histology. Confocal images from five sites topically stained with CV were then acquired in vivo from high-risk smokers and compared to hematoxylin and eosin stained sections of biopsies taken from the same site. Sufficient contrast in the confocal imagery was obtained to identify cells in the bronchial epithelium. However, further improvements in the miniature objective lens are required to provide sufficient axial resolution for accurate classification of preneoplastic lesions.

  4. Liquid level sensing based on laser differential confocal detectors

    NASA Astrophysics Data System (ADS)

    Gao, Haibo; Fan, Chunshi; Zhang, Li; Hu, Yao

    2015-01-01

    Liquid level measurement plays an important part in industry and daily life. Applications include oil tanks, gasoline stations and public water supplies. Traditional electronic sensors cannot satisfy the demands in harsh environments. Recently, optical sensors have been particularly attractive in these applications. We propose a sensing method based on laser differential confocal detectors for discrete or continuous liquid level sensing. No target or supplementary device need to be immersed into the liquid. The sensitivity of the liquid level is about 0.01 mm with current systematic parameters. Measurement experiment of simulated liquid surface with a reflective mirror is carried out to verify the method.

  5. Confocal microscope with enhanced lateral resolution using engineered illumination pupil

    NASA Astrophysics Data System (ADS)

    Boruah, B. R.

    2010-02-01

    The maximum lateral resolution achievable with a confocal microscope is twice that of a wide field microscope. However, the spatial frequency content in the confocal image near the cutoff has very poor signal and is hardly of any practical use. Barring in the fluorescence mode, no technique can provide significant resolution enhancement simultaneously both in the reflection and fluorescence mode of the confocal microscope. This paper describes a technique based on aperture engineering that can significantly enhance the high spatial frequency content in the image of a confocal microscope, in principle, working either in the reflection or the fluorescence mode. Results obtained from numerical simulations and experimental implementation are presented.

  6. How the confocal laser scanning microscope entered biological research.

    PubMed

    Amos, W B; White, J G

    2003-09-01

    A history of the early development of the confocal laser scanning microscope in the MRC Laboratory of Molecular Biology in Cambridge is presented. The rapid uptake of this technology is explained by the wide use of fluorescence in the 80s. The key innovations were the scanning of the light beam over the specimen rather than vice-versa and a high magnification at the level of the detector, allowing the use of a macroscopic iris. These were followed by an achromatic all-reflective relay system, a non-confocal transmission detector and novel software for control and basic image processing. This design was commercialized successfully and has been produced and developed over 17 years, surviving challenges from alternative technologies, including solid-state scanning systems. Lessons are pointed out from the unusual nature of the original funding and research environment. Attention is drawn to the slow adoption of the instrument in diagnostic medicine, despite promising applications. PMID:14519550

  7. Application of confocal Raman microscopy to investigate casein micro-particles in blend casein/pectin films.

    PubMed

    Zhuang, Yu; Sterr, Julia; Kulozik, Ulrich; Gebhardt, Ronald

    2015-03-01

    Pectin triggers formation of casein micro-particles during solution casting. Confocal Raman microscopy revealed their composition and spatial dimension in resulting films. Peaks in the Raman spectra corresponded to those found in films prepared by either casein or pectin. This suggested that no conformational changes occurred after mixing. Raman images revealed incompatibility of both polymers because particles consisted of casein only and the surrounding matrix of pectin. Deformation of micro-particles into an oblate shape took place during film formation. In dried films, an empty space between casein and pectin was found in lateral dimension. In contrast, casein micro-particles overlapped with the pectin matrix in the vertical dimension. PMID:25482531

  8. Expanding Imaging Capabilities for Microfluidics: Applicability of Darkfield Internal Reflection Illumination (DIRI) to Observations in Microfluidics

    PubMed Central

    Kawano, Yoshihiro; Otsuka, Chino; Sanzo, James; Higgins, Christopher; Nirei, Tatsuo; Schilling, Tobias; Ishikawa, Takuji

    2015-01-01

    Microfluidics is used increasingly for engineering and biomedical applications due to recent advances in microfabrication technologies. Visualization of bubbles, tracer particles, and cells in a microfluidic device is important for designing a device and analyzing results. However, with conventional methods, it is difficult to observe the channel geometry and such particles simultaneously. To overcome this limitation, we developed a Darkfield Internal Reflection Illumination (DIRI) system that improved the drawbacks of a conventional darkfield illuminator. This study was performed to investigate its utility in the field of microfluidics. The results showed that the developed system could clearly visualize both microbubbles and the channel wall by utilizing brightfield and DIRI illumination simultaneously. The methodology is useful not only for static phenomena, such as clogging, but also for dynamic phenomena, such as the detection of bubbles flowing in a channel. The system was also applied to simultaneous fluorescence and DIRI imaging. Fluorescent tracer beads and channel walls were observed clearly, which may be an advantage for future microparticle image velocimetry (?PIV) analysis, especially near a wall. Two types of cell stained with different colors, and the channel wall, can be recognized using the combined confocal and DIRI system. Whole-slide imaging was also conducted successfully using this system. The tiling function significantly expands the observing area of microfluidics. The developed system will be useful for a wide variety of engineering and biomedical applications for the growing field of microfluidics. PMID:25748425

  9. Confocal Microscopy in Biopsy Proven Argyrosis

    PubMed Central

    Guven Yilmaz, Suzan; Akalin, Taner; Egrilmez, Sait; Yagci, Ayse

    2013-01-01

    Purpose. To evaluate the confocal microscopy findings of a 46-year-old male with bilateral biopsy proven argyrosis. Materials and Methods. Besides routine ophthalmologic examination, anterior segment photography and confocal microscopy with cornea Rostoch module attached to HRT II (Heidelberg Engineering GmbH, Heidelberg, Germany) were performed. Findings. Squamous metaplastic changes on conjunctival epithelium and intense highly reflective extracellular punctiform deposits in conjunctival substantia propria were detected. Corneal epithelium was normal. Highly reflective punctiform deposits starting from anterior to mid-stroma and increasing through Descemet's membrane were evident. Corneal endothelium could not be evaluated due to intense stromal deposits. Conclusion. Confocal microscopy not only supports diagnosis in ocular argyrosis, but also demonstrates the intensity of the deposition in these patients. PMID:23970986

  10. Fiber optic confocal microscope: In vivo precancer detection

    NASA Astrophysics Data System (ADS)

    Carlson, Kristen Dawn

    Cancer is a significant public health problem worldwide. Many cancers originate as precancerous lesions in the epithelium which, when removed in sufficient time, can prevent progression to cancer. However, current detection techniques are typically time-consuming and expensive, limiting their acceptance and accessibility. Optical techniques, such as confocal microscopy, have significant potential to provide clinicians with real-time, high-resolution images of cells and tissue without tissue removal. These images of cell morphology and tissue architecture can be used to characterize tissue and determine the presence or extent of precancer and cancer. This dissertation explores the instrumentation and application of fiber optic reflectance confocal microscopy for in vivo precancer detection. The first part of the dissertation presents in vivo imaging of suspicious lesions in the human uterine cervix and oral mucosa using a fiber bundle based confocal microscope with a complex glass miniature objective lens. Images are analyzed quantitatively and qualitatively to determine the potential of this technology in vivo. An analysis of nuclear density from images of 30 cervical epithelium sites shows differentiation between normal and precancerous sites. Similarly, images from 20 oral mucosa sites demonstrate changes in nuclear density and tissue architecture indicative of progression of precancer and cancer. In addition to this multi-fiber confocal microscope used with a glass objective lens for the clinical studies, imaging of tissue samples has been accomplished with the same confocal system using an injection molded plastic miniature objective lens demonstrating comparable optical quality for a significantly less expensive optical component. Finally, a benchtop prototype of a single fiber confocal microscope using a gimbaled two-axis MEMS scanner has been designed and constructed. Imaging of a resolution target and cellular samples demonstrates sufficient resolution and field of view for cellular imaging. The results from the imaging studies presented here indicate that in vivo confocal microscopy has the potential to improve early precancer detection in epithelial tissue. Advances in imaging technology will continue to reduce the cost of imaging systems and improve the imaging capability, leading to an inexpensive, real-time, minimally-invasive tool for in vivo imaging.

  11. Multimodal imaging including spectral-domain optical coherence tomography and confocal near-infrared reflectance for characterization of lacquer cracks in highly myopic eyes

    PubMed Central

    Liu, C-F; Liu, L; Lai, C-C; Chou, J CL; Yeh, L-K; Chen, K-J; Chen, Y-P; Wu, W-C; Chuang, L-H; Sun, C-C; Wang, N-K

    2014-01-01

    Purpose To compare multimodal imaging in detecting lacquer cracks in highly myopic eyes, and to correlate these findings with those of spectral-domain optical coherence tomography (SD-OCT). Methods An observational case series study. Patients with a refractive error worse than ?8 diopters and lacquer cracks were recruited. The rates of detection of the lacquer cracks using multimodal imaging including near-infrared reflectance (NIR) imaging, fundus autofluorescence (FAF) imaging, and fluorescence angiography (FA) were compared. The characteristic findings of multimodal imaging were correlated with those of SD-OCT. Results NIR imaging was more sensitive (92.9%) in detecting lacquer cracks than either FAF (12.5%) or FA (67.9%). Lacquer cracks showed hyperreflectance on NIR, and they were consistently associated with a continuous retinal pigment epithelium-Bruch's membrane complex, thinner choroid, and acoustic shadows on SD-OCT. Conclusions NIR imaging is superior to blue laser light (FAF and FA) imaging in detecting lacquer cracks. SD-OCT in combination with NIR located primary pathological lacquer cracks in the intact retinal pigment epithelium-Bruch's membrane complex as well as thinner choroid. These findings indicate that multimodal cSLO and SD-OCT imaging allow for detecting of lacquer cracks in highly myopic eyes. PMID:25233819

  12. Application of a reflectance model to the sensor planning system

    NASA Astrophysics Data System (ADS)

    Koutecký, Tomáś; Paloušek, David; Brandejs, Jan

    2015-05-01

    This study describes a new sensor planning system for the automatic generation of scanning positions based on a computer model of the part for digitization of sheet metal parts. The focus of this paper is in the application of a reflectance model into this sensor planning system. The goal of this sensor planning system and application of this model is to ensure fast, complete and accurate digitization of the parts for their inspection during serial-line production, especially in the automotive industry. A methodology of the sensor planning system consists of positions planning, their simulation for true visibility of the part elements using a reflectance model, and a simulation of the positions for robot reachability. Compared to previous studies, visual properties of the scanned parts' surface can be simulated precisely. The Nayar model is used as a reflectance model. This model is suitable for materials that are characterized by the combination of diffuse and specular reflections and uses three components of reflection: diffuse, specular lobe and specular spike. Results of the scanning that were obtained using an ATOS III Triple Scan fringe projection 3D scanner and a KUKA KR 60 HA industrial robot were compared to the simulation. The comparison based on the correspondence of the polygons area acquired in each sensor position (in simulation and in scanning) shows that in the performed measurements the median of differences between simulation and scanning is around 16%.

  13. Spectral reflectance of selected aqueous solutions for water quality applications

    NASA Technical Reports Server (NTRS)

    Querr, M. R.; Waring, R. C.; Holland, W. E.; Nijm, W.; Hale, G. M.

    1972-01-01

    The relative specular reflectances of individual aqueous solutions having a particular chemical salt content were measured in the 2 to 20 micrometers region of the infrared component or radiant flux. Distilled water was the reflectance standard. The angle of incidence was 70.03 deg plus or minus 0.23 deg. Absolute reflectances of the solutions for the same polarization and angle of incidence were computed by use of the measured relative reflectances, one of the Fresnel equations, and the optical constants of distilled water. Phase shift and phase difference spectra were obtained by respectively applying a Kramers-Kronig dispersion analysis to the absolute and relative reflectance spectra. The optical constants of the solutions were determined by algorithms commonly associated with the Kramers-Kronig analysis. Spectral signatures that qualitatively and quantitatively characterize the solute and that show structure of the infrared bands of water were noted in the phase difference spectra. The relative and absolute reflectances, the phase shift and phase difference spectra and the optical constants are presented in graphical form. Application of these results to remote sensing of the chemical quality of natural waters is discussed briefly.

  14. Reflectance of metallic indium for solar energy applications

    NASA Technical Reports Server (NTRS)

    Bouquet, F. L.; Hasegawa, T.

    1984-01-01

    An investigation has been conducted in order to compile quantitative data on the reflective properties of metallic indium. The fabricated samples were of sufficiently high quality that differences from similar second-surface silvered mirrors were not apparent to the human eye. Three second-surface mirror samples were prepared by means of vacuum deposition techniques, yielding indium thicknesses of approximately 1000 A. Both hemispherical and specular measurements were made. It is concluded that metallic indium possesses a sufficiently high specular reflectance to be potentially useful in many solar energy applications.

  15. High-speed multispectral confocal biomedical imaging

    PubMed Central

    Carver, Gary E.; Locknar, Sarah A.; Morrison, William A.; Krishnan Ramanujan, V.; Farkas, Daniel L.

    2014-01-01

    Abstract. A new approach for generating high-speed multispectral confocal images has been developed. The central concept is that spectra can be acquired for each pixel in a confocal spatial scan by using a fast spectrometer based on optical fiber delay lines. This approach merges fast spectroscopy with standard spatial scanning to create datacubes in real time. The spectrometer is based on a serial array of reflecting spectral elements, delay lines between these elements, and a single element detector. The spatial, spectral, and temporal resolution of the instrument is described and illustrated by multispectral images of laser-induced autofluorescence in biological tissues. PMID:24658777

  16. Stereoscopic images in confocal (tandem scanning) microscopy.

    PubMed

    Boyde, A

    1985-12-13

    Stereoscopic pair images with parallel projection geometry are obtained by through-focusing along two inclined axes while recording two (summed and stacked) images with a microscope with a very shallow depth of field. The two stack images sample the same depth slice of translucent or reflective specimens. The method will work most conveniently with a tandem scanning microscope (a direct-view, confocal scanning optical microscope). This is a direct method for recording stereo images that can be used to the limit of resolution in optical microscopy. It demonstrates a previously unrealized advantage of confocal optical microscopy. PMID:4071051

  17. Reflective Array with Controlled Focusing for Radiotomographic Application

    NASA Astrophysics Data System (ADS)

    Shipilov, S. E.; Eremeev, A. I.; Yakubov, V. P.

    2016-01-01

    It's considered the principle possibility of creation the managed reflectors for formulation of given field distribution in the focus area. Reflectors change the reflect ratio in dependence of the external control. The proposed theoretical modeling of such controlled focused device which provides focuse to a specific point in a given distribution of the reflectors. On the basis of numerical simulation it's considered the application of this approach for the solution of the problem of radiotomography.

  18. Confocal Microscopy Core Facility

    Cancer.gov

    Susan H. Garfield, Facility Head Confocal Microscopy Core Facility, CCR, NCI, NIH Building 37, Room B114 E Tel: 301.435.6187 Fax: 301.496.0734 email: susan_garfield@nih.gov Poonam Mannan, Biologist Building 37, Room B114 F Tel: 301.451.7816 e-mail: mannan

  19. Re-scan confocal microscopy: scanning twice for better resolution

    PubMed Central

    De Luca, Giulia M.R.; Breedijk, Ronald M.P.; Brandt, Rick A.J.; Zeelenberg, Christiaan H.C.; de Jong, Babette E.; Timmermans, Wendy; Azar, Leila Nahidi; Hoebe, Ron A.; Stallinga, Sjoerd; Manders, Erik M.M.

    2013-01-01

    We present a new super-resolution technique, Re-scan Confocal Microscopy (RCM), based on standard confocal microscopy extended with an optical (re-scanning) unit that projects the image directly on a CCD-camera. This new microscope has improved lateral resolution and strongly improved sensitivity while maintaining the sectioning capability of a standard confocal microscope. This simple technology is typically useful for biological applications where the combination high-resolution and high-sensitivity is required. PMID:24298422

  20. Clinical applications of lightguide diffuse reflectance spectrophotometry in vascular disease

    NASA Astrophysics Data System (ADS)

    Harrison, David K.; Delaney, Colin; Brown, Linda; Newton, David J.; McCollum, Peter T.

    1994-02-01

    There is enormous potential for application of lightguide tissue reflectance spectrophotometry in the diagnosis and treatment of peripheral vascular disease. In the present study, measurements were carried out in 10 such pre-amputation patients to compare the use of micro-lightguide spectrophotometry with the macro-lightguide technique. These preliminary results show excellent agreement between the new, non-invasive micro-lightguide technique and the `gold standard' skin blood flow measurements. This technique could thus provide a more functional, non-invasive assessment of healing potential than skin blood flow measurement.

  1. Olivine-metal mixtures: Spectral reflectance properties and application to asteroid reflectance spectra

    NASA Astrophysics Data System (ADS)

    Cloutis, Edward A.; Sanchez, Juan A.; Reddy, Vishnu; Gaffey, Michael J.; Binzel, Richard P.; Burbine, Thomas H.; Hardersen, Paul S.; Hiroi, Takahiro; Lucey, Paul G.; Sunshine, Jessica M.; Tait, Kimberly T.

    2015-05-01

    Olivine-rich asteroids appear to be common in the main asteroid belt as well as present in the near-Earth asteroid population. There are a number of meteorite classes that are dominated by olivine ± metal. To determine whether relationships exist between these asteroids and meteorites, we spectrally characterized a number of olivine + meteoritic metal powder intimate and areal mixtures, pallasite slabs, and olivine powders on a metal slab. Our goal is to understand the spectral characteristics of olivine + metal assemblages and develop spectral metrics that can be used to analyze reflectance spectra of olivine-dominated asteroids. We found that the major olivine absorption band in the 1 μm region is resolvable in intimate mixtures for metal abundances as high as ∼90 wt.%. The wavelength position of the 1 μm region olivine absorption band center is sensitive to Fa content but insensitive to other variables. However, the band minimum position moves to shorter wavelengths with increasing metal abundance due to changes in spectral slope. The full width at half maximum (FWHM) of this band and reflectance at 1.8 μm are both most sensitive to olivine Fa content, metal abundance, and grain size, and much less to the presence of nanophase iron that reddens spectra. Reflectance at 0.56 μm and the 1.8/0.56 μm reflectance ratio are sensitive to these same parameters as well as to nanophase iron-associated spectral reddening. The wavelength position of the local reflectance maximum in the 0.7 μm region moves to longer wavelengths with increasing metal abundance and is most useful for constraining metal abundance in high metal-content mixtures. Pallasite slab spectra differ in a number of respects from powdered assemblages and multiple spectral parameters can be used to discriminate them. The spectra of increasingly fine-grained olivine + metal assemblages and those involving low-Fa olivine show increasing spectral dominance by metal. Systematic application of multiple spectral metrics allows olivine + metal assemblage properties such as Fa content, olivine/metal ratio, and grain size to be quantified or constrained. Analysis of reflectance spectra of 22 presumed olivine ± metal-rich asteroids indicates that most of them possess low- to medium-Fa content olivine (Fa<∼67), with variable abundances of macroscopic metal. A number exhibit visible region absorption bands that are indicative of some fraction of coarser-grained olivine (>45 μm). Most asteroid spectra can be plausibly linked to specific olivine ± metal-bearing meteorite classes. Most of the asteroid spectra examined exhibit some degree of spectral reddening below ∼1.8 μm which is most consistent with the presence of fine-grained nanophase iron, likely produced by space weathering.

  2. Multimodal confocal hyperspectral imaging microscopy with wavelength sweeping source

    NASA Astrophysics Data System (ADS)

    Kim, Young-Duk; Do, Dukho; Yoo, Hongki; Gweon, DaeGab

    2015-02-01

    There exist microscopes that are able to obtain the chemical properties of a sample, because there are some cases in which it is difficult to find out causality of a phenomenon by using only the structural information of a sample. Obtaining the chemical properties of a sample is important in biomedical imaging, because most biological phenomena include changes in the chemical properties of the sample. Hyperspectral imaging (HSI) is one of the popular imaging methods for characterizing materials and biological samples by measuring the reflectance or emission spectrum of the sample. Because all materials have a unique reflectance spectrum, it is possible to analyze material properties and detect changes in the chemical properties of a sample by measuring the spectral changes with respect to the original spectrum. Because of its ability to measure the spectrum of a sample, HSI is widely used in materials identification applications such as aerial reconnaissance and is the subject of various studies in microscopy. Although there are many advantages to using the method, conventional HSI has some limitations because of its complex configuration and slow speed. In this research we propose a new type of multimodal confocal hyperspectral imaging microscopy with fast image acquisition and a simple configuration that is capable of both confocal and HSI microscopies.

  3. Confocal filtering in cathodoluminescence microscopy of nanostructures

    SciTech Connect

    Narvez, Angela C. E-mail: j.p.hoogenboom@tudelft.nl; Weppelman, I. Gerward C.; Moerland, Robert J.; Hoogenboom, Jacob P. E-mail: j.p.hoogenboom@tudelft.nl; Kruit, Pieter

    2014-06-23

    Cathodoluminescence (CL) microscopy allows optical characterization of nanostructures at high spatial resolution. At the nanoscale, a main challenge of the technique is related to the background CL generated within the sample substrate. Here, we implement confocal detection of the CL signal to minimize the background contribution to the measurement. Nano-phosphors were used as point sources to evaluate the filtering capabilities of our confocal CL system, obtaining an axial intensity profile with 2.7??m full width at half maximum for the central peak, in good correspondence with theoretical expectations. Considering the electron interaction volume, we found that the confocal filter becomes effective for electron energies above 20?keV, when using a 25??m pinhole (0.86 Airy units). To illustrate our approach, we present confocal CL imaging of gold nanowires and triangular shaped plates deposited on an indium-tin oxide covered glass substrate, comparing the images with those obtained in standard unfiltered CL detection. The results show that confocal CL microscopy is a valuable tool for the investigation of nanostructures on highly cathodoluminescent substrates, widely used in biological and optical applications.

  4. Advanced microscopy: laser scanning confocal microscopy.

    PubMed

    Hanrahan, Orla; Harris, James; Egan, Chris

    2011-01-01

    Fluorescence microscopy is an important and fundamental tool for biomedical research. Optical microscopy is almost non-invasive and allows highly spatially resolved images of organisms, cells, macromolecular complexes, and biomolecules to be obtained. Generally speaking, the architecture of the observed structures is not significantly modified and the environmental conditions can be kept very close to physiological reality. The development of fluorescence microscopy was revolutionized with the invention of laser scanning confocal microscopy (LSCM). With its unique three-dimensional representation and analysis capabilities, this technology gives us a more real view of the world.This chapter introduces the reader to the methodology of setting up basic experiments for use with a laser scanning confocal microscope. There are practical guidelines about sample preparation for both fixed and living specimens, as well as examples of some of the applications of confocal microscopy. PMID:21898220

  5. Optimal pupil design for confocal microscopy

    NASA Astrophysics Data System (ADS)

    Patel, Yogesh G.; Rajadhyaksha, Milind; DiMarzio, Charles A.

    2010-02-01

    Confocal reflectance microscopy may enable screening and diagnosis of skin cancers noninvasively and in real-time, as an adjunct to biopsy and pathology. Current instruments are large, complex, and expensive. A simpler, confocal line-scanning microscope may accelerate the translation of confocal microscopy in clinical and surgical dermatology. A confocal reflectance microscope may use a beamsplitter, transmitting and detecting through the pupil, or a divided pupil, or theta configuration, with half used for transmission and half for detection. The divided pupil may offer better sectioning and contrast. We present a Fourier optics model and compare the on-axis irradiance of a confocal point-scanning microscope in both pupil configurations, optimizing the profile of a Gaussian beam in a circular or semicircular aperture. We repeat both calculations with a cylindrical lens which focuses the source to a line. The variable parameter is the fillfactor, h, the ratio of the 1/e2 diameter of the Gaussian beam to the diameter of the full aperture. The optimal values of h, for point scanning are 0.90 (full) and 0.66 for the half-aperture. For line-scanning, the fill-factors are 1.02 (full) and 0.52 (half). Additional parameters to consider are the optimal location of the point-source beam in the divided-pupil configuration, the optimal line width for the line-source, and the width of the aperture in the divided-pupil configuration. Additional figures of merit are field-of-view and sectioning. Use of optimal designs is critical in comparing the experimental performance of the different configurations.

  6. Tunable fiber confocal sensor with LED

    NASA Astrophysics Data System (ADS)

    Weng, Chun-Jen; Lan, Tzu-Hsien; Hwang, Chi-Hung; Chu, Nien-Nan; Huang, Chien-Yao; Cheng, Pi-Ying

    2015-07-01

    A novel concept of confocal sensor based on focal lens is proposed to measure the displacement. The light source is a stabilized fiber coupled LED. A 1x2 graded-index multimode fiber optic coupler is used in this sensor. One port is a LED input port via SMA connector, another port is a LED output port connected to a reflective collimator and the other port is a reflective sensor port connected to a photo detector. The focusing sensor head is the cascade of a focal lens and a 20X objective lens. In this confocal displacement sensor, LED passes through a focal lens and an objective lens so that the LED beam focuses at a fixed focal point. A test target is placed after the objective lens. The displacement between the sensor head and a target can be measured quickly by detecting the reflective power according to the confocal principle. The long-term stability of LED is under 0.5%. The effective back focal length is varied from 5.67mm to 6.57mm by 0-290mA current driving so that the measuring range is about 0.9mm. The FWHM resolution of displacement is about 50μm. This sensor has the features of low cost, high stability, high precision and compact.

  7. The sociomoral reflection measure: applicability to Swedish children and adolescents.

    PubMed

    Nilsson, I; Crafoord, J; Hedengren, M; Ekehammar, B

    1991-01-01

    The Sociomoral Reflection Measure (SRM; Gibbs & Widaman, 1982) was developed as a group administerable instrument for measuring developmental stages of moral reasoning. The aim of this study was to examine its reliability and construct validity, employing a sample of 542 Swedish elementary and high school students (aged 8-17), from schools within metropolitan Stockholm. Interrater reliability varied between 0.83 and 0.92, and the internal consistency coefficient was 0.76. Factor analyses of the norms (on which the overall score is based) revealed one factor for the oldest age group (16-17 years), but two factors among the younger subjects (8-15 years), clearly separating the norms related to each of the two moral dilemmas. The Sociomoral Reflection Maturity Score was moderately, but significantly, related to both age and grade. The results suggests that, although used in a non-American context and with self-trained raters, the SRM seems highly applicable in Sweden, at least for research purposes. PMID:2047796

  8. Bidirectional Reflectance Functions for Application to Earth Radiation Budget Studies

    NASA Technical Reports Server (NTRS)

    Manalo-Smith, N.; Tiwari, S. N.; Smith, G. L.

    1997-01-01

    Reflected solar radiative fluxes emerging for the Earth's top of the atmosphere are inferred from satellite broadband radiance measurements by applying bidirectional reflectance functions (BDRFs) to account for the anisotropy of the radiation field. BDRF's are dependent upon the viewing geometry (i.e. solar zenith angle, view zenith angle, and relative azimuth angle), the amount and type of cloud cover, the condition of the intervening atmosphere, and the reflectance characteristics of the underlying surface. A set of operational Earth Radiation Budget Experiment (ERBE) BDRFs is available which was developed from the Nimbus 7 ERB (Earth Radiation Budget) scanner data for a three-angle grid system, An improved set of bidirectional reflectance is required for mission planning and data analysis of future earth radiation budget instruments, such as the Clouds and Earth's Radiant Energy System (CERES), and for the enhancement of existing radiation budget data products. This study presents an analytic expression for BDRFs formulated by applying a fit to the ERBE operational model tabulations. A set of model coefficients applicable to any viewing condition is computed for an overcast and a clear sky scene over four geographical surface types: ocean, land, snow, and desert, and partly cloudy scenes over ocean and land. The models are smooth in terms of the directional angles and adhere to the principle of reciprocity, i.e., they are invariant with respect to the interchange of the incoming and outgoing directional angles. The analytic BDRFs and the radiance standard deviations are compared with the operational ERBE models and validated with ERBE data. The clear ocean model is validated with Dlhopolsky's clear ocean model. Dlhopolsky developed a BDRF of higher angular resolution for clear sky ocean from ERBE radiances. Additionally, the effectiveness of the models accounting for anisotropy for various viewing directions is tested with the ERBE along tract data. An area viewed from nadir and from the side give two different radiance measurements but should yield the same flux when converted by the BDRF. The analytic BDRFs are in very good qualitative agreement with the ERBE models. The overcast scenes exhibit constant retrieved albedo over viewing zenith angles for solar zenith angles less than 60 degrees. The clear ocean model does not produce constant retrieved albedo over viewing zenith angles but gives an improvement over the ERBE operational clear sky ocean BDRF.

  9. Feasibility of digitally stained multimodal confocal mosaics to simulate histopathology

    NASA Astrophysics Data System (ADS)

    Gareau, Daniel S.

    2009-05-01

    Fluorescence confocal mosaicing microscopy of tissue biopsies stained with acridine orange has been shown to accurately identify tumors and with an overall sensitivity of 96.6% and specificity of 89.2%. However, fluorescence shows only nuclear detail similar to hematoxylin in histopathology and does not show collagen or cytoplasm, which may provide necessary negative contrast information similar to eosin used in histopathology. Reflectance mode contrast is sensitive to collagen and cytoplasm without staining. To further improve sensitivity and specificity, digitally stained confocal mosaics combine confocal fluorescence and reflectance images in a multimodal pseudo-color image to mimic the appearance of histopathology with hematoxylin and eosin and facilitate the introduction of confocal microscopy into the clinical realm.

  10. Diagnosis of Acanthamoeba keratitis in vivo with confocal microscopy.

    PubMed

    Winchester, K; Mathers, W D; Sutphin, J E; Daley, T E

    1995-01-01

    We present eight cases of Acanthamoeba keratitis. In each case; the Acanthamoeba organisms were visualized in the epithelium and anterior stroma using tandem scanning confocal microscopy. The organisms were highly reflective, ovoid, and were 10-25 microns in diameter. The Acanthamoeba organisms in the human corneas were identical in size and shape to Acanthamoeba organisms on an agar plate visualized with the same confocal microscope. Confocal microscopy is a useful method for identifying Acanthamoeba organisms in vivo within the corneal epithelium and anterior stroma. PMID:7712728

  11. Frustrated Total Internal Reflection: A Simple Application and Demonstration.

    ERIC Educational Resources Information Center

    Zanella, F. P.; Magalhaes, D. V.; Oliveira, M. M.; Bianchi, R. F.; Misoguti, L.; Mendonca, C. R.

    2003-01-01

    Describes the total internal reflection process that occurs when the internal angle of incidence is equal to or greater than the critical angle. Presents a demonstration of the effect of frustrated total internal reflection (FTIR). (YDS)

  12. Confocal microscopy for healthy and pathological nail.

    PubMed

    Cinotti, E; Fouilloux, B; Perrot, J L; Labeille, B; Douchet, C; Cambazard, F

    2014-07-01

    Nail diseases are often annoying for the patient and diagnostically challenging for dermatologists. New imaging techniques are of high interest in the diagnosis of nail disorders to reduce the number of nail biopsies. Confocal microscopy is a high-resolution emerging imaging technique that can be used to explore the entire body surface, including skin, mucosa, hair and nails. A systematic review of the literature concerning the use of confocal microscopy for the study of either healthy or pathological nail has been performed to evaluate the current use of this technique and possible future applications. Confocal microscopy is particularly suitable for nails because it allows a non-invasive in vivo examination of this sensitive body area, and nail plate transparency permits to image up to the nail bed with an easy identification of corneocytes. Confocal microscopy can play a role in the diagnosis of onychomycosis and melanonichia, and in the study of drug penetration through the nail plate. It could be used in the future as a non-invasive procedure for the investigation of different nail diseases, such as psoriasis and lichen planus. Further application could be the intra-operative ex vivo examination of nail specimens to outline tumour margins to assist surgery. PMID:24320009

  13. Applications of microstructured silicon wafers as internal reflection elements in attenuated total reflection Fourier transform infrared spectroscopy.

    PubMed

    Schumacher, Henrik; Knzelmann, Ulrich; Vasilev, Boris; Eichhorn, Klaus-Jochen; Bartha, Johann W

    2010-09-01

    A novel internal reflection element (IRE) for attenuated total reflection Fourier transform infrared (ATR-FT-IR) spectral acquisition is introduced and applied for several surface-sensitive measurements. It is based on microstructured double-side-polished (100) silicon wafers with v-shaped grooves of {111} facets on their backside. These facets of the so-called "microstructured single-reflection elements" (mSRE) are formed by a crystal-oriented anisotropic wet etching process within a conventional wafer structuring process. They are used to couple infrared radiation into and out of the IRE. In contrast to the application of the commonly used silicon multiple-reflection elements (MRE), the new elements provide single-reflection ATR measurements at the opposite wafer side by using simple reflection accessories without any special collimation. Due to the short light path, the spectral range covers the entire mid-infrared region with a high optical throughput, including the range of silicon lattice vibrations from 300 to 1500 cm(-1). In addition to typical ATR applications, i.e., the measurement of bulk liquids and soft materials, the new reflection elements can be effectively used and customer-specifically designed for in situ and ex situ investigations of aqueous solutions, thin films, and monolayers on Si. Examples presented in this article are in situ etching of native as well as thermal SiO(2) and characterization of polydimethylsiloxane (PDMS) films on Si under various measuring conditions. PMID:20828439

  14. Confocal coded aperture imaging

    DOEpatents

    Tobin, Jr., Kenneth William; Thomas, Jr., Clarence E.

    2001-01-01

    A method for imaging a target volume comprises the steps of: radiating a small bandwidth of energy toward the target volume; focusing the small bandwidth of energy into a beam; moving the target volume through a plurality of positions within the focused beam; collecting a beam of energy scattered from the target volume with a non-diffractive confocal coded aperture; generating a shadow image of said aperture from every point source of radiation in the target volume; and, reconstructing the shadow image into a 3-dimensional image of the every point source by mathematically correlating the shadow image with a digital or analog version of the coded aperture. The method can comprise the step of collecting the beam of energy scattered from the target volume with a Fresnel zone plate.

  15. Confocal microscopy and exfoliative cytology

    PubMed Central

    Reddy, Shyam Prasad; Ramani, Pratibha; Nainani, Purshotam

    2013-01-01

    Context: Early detection of potentially malignant lesions and invasive squamous-cell carcinoma in the oral cavity could be greatly improved through techniques that permit visualization of subtle cellular changes indicative of the neoplastic transformation process. One such technique is confocal microscopy. Combining rapidity with reliability, an innovative idea has been put forward using confocal microscope in exfoliative cytology. Aims: The main objective of this study was to assess confocal microscopy for cytological diagnosis and the results were compared with that of the standard PAP stain. Settings and Design: Confocal microscope, acridine orange (AO) stain, PAP (Papanicolaou) stain. The study was designed to assess confocal microscopy for cytological diagnosis. In the process, smears of patients with (clinically diagnosed and/or suspected) oral squamous cell carcinoma as well as those of controls (normal people) were stained with acridine orange and observed under confocal microscope. The results were compared with those of the standard PAP method. Materials and Methods: Samples of buccal mucosa smears from normal patients and squamous cell carcinoma patients were made, fixed in 100% alcohol, followed by AO staining. The corresponding set of smears was stained with PAP stain using rapid PAP stain kit. The results obtained were compared with those obtained with AO confocal microscopy. Results: The study had shown nuclear changes (malignant cells) in the smears of squamous cell carcinoma patients as increased intensity of fluorescence of the nucleus, when observed under confocal microscope. Acridine orange confocal microscopy showed good amount of sensitivity and specificity (93%) in identifying malignant cells in exfoliative cytological smears. Conclusion: Confocal microscopy was found to have good sensitivity in the identification of cancer (malignant) cells in exfoliative cytology, at par with the PAP method. The rapidity of processing and screening a specimen resulted in saving of time. It added a certain amount of objectivity to the process of arriving at a diagnosis. PMID:24250082

  16. Reflectance spectroscopy: quantitative analysis techniques for remote sensing applications.

    USGS Publications Warehouse

    Clark, R.N.; Roush, T.L.

    1984-01-01

    Several methods for the analysis of remotely sensed reflectance data are compared, including empirical methods and scattering theories, both of which are important for solving remote sensing problems. The concept of the photon mean path length and the implications for use in modeling reflectance spectra are presented.-from Authors

  17. Diffuse reflection imaging at terahertz frequencies for security applications

    NASA Astrophysics Data System (ADS)

    Dean, P.; Khanna, S.; Chakraborty, S.; Lachab, M.; Davies, A. G.; Linfield, E. H.

    2007-10-01

    We report diffuse reflection imaging of concealed powdered samples in atmospheric air using a quantum cascade laser operating at 2.83 THz. The imaging system uses a helium-cooled silicon bolometer for mapping radiation diffusely reflected and scattered from samples, and a room-temperature pyroelectric sensor for simultaneously acquiring a specular image. A range of powders concealed within plastic packaging and standard FedEx envelopes was imaged with a resolution of better than 0.5 mm, and it was possible to detect powdered samples concealed within packaging from which there was a strong component of surface reflection. The feasibility of performing dual-wavelength diffuse reflection imaging for identification of illicit drugs and explosives is discussed.

  18. Illumination and Reflectance Estimation with its Application in Foreground Detection.

    PubMed

    Tu, Gang Jun; Karstoft, Henrik; Pedersen, Lene Juul; Jrgensen, Erik

    2015-01-01

    In this paper, we introduce a novel approach to estimate the illumination and reflectance of an image. The approach is based on illumination-reflectance model and wavelet theory. We use a homomorphic wavelet filter (HWF) and define a wavelet quotient image (WQI) model based on dyadic wavelet transform. The illumination and reflectance components are estimated by using HWF and WQI, respectively. Based on the illumination and reflectance estimation we develop an algorithm to segment sows in grayscale video recordings which are captured in complex farrowing pens. Experimental results demonstrate that the algorithm can be applied to detect the domestic animals in complex environments such as light changes, motionless foreground objects and dynamic background. PMID:26343675

  19. Illumination and Reflectance Estimation with its Application in Foreground Detection

    PubMed Central

    Tu, Gang Jun; Karstoft, Henrik; Pedersen, Lene Juul; Jrgensen, Erik

    2015-01-01

    In this paper, we introduce a novel approach to estimate the illumination and reflectance of an image. The approach is based on illumination-reflectance model and wavelet theory. We use a homomorphic wavelet filter (HWF) and define a wavelet quotient image (WQI) model based on dyadic wavelet transform. The illumination and reflectance components are estimated by using HWF and WQI, respectively. Based on the illumination and reflectance estimation we develop an algorithm to segment sows in grayscale video recordings which are captured in complex farrowing pens. Experimental results demonstrate that the algorithm can be applied to detect the domestic animals in complex environments such as light changes, motionless foreground objects and dynamic background. PMID:26343675

  20. Transmission-reflection tomography: Application to reverse VSP data

    SciTech Connect

    Mao, W.; Stuart, G.W.

    1997-05-01

    A multiphase tomographic algorithm is presented that allows 2-D and 3-D slowness (inverse of velocity) and variable reflector depth to be reconstructed simultaneously from both transmission and reflection traveltimes. The authors analyze the ambiguity in the determination of velocity and depth in transmission and reflection data and realize that depth in transmission and reflection data and realize that depth perturbation is more sensitive to reflection traveltime anomalies than slowness perturbation, whereas the reverse is true of transmission traveltime anomalies. Because of the constraints on velocity and depth provided by the different wave types, this algorithm reduces the ambiguity substantially between velocity and depth prevalent in reflection tomography and also avoids the undetermined problem in transmission tomography. The linearized inversion was undertaken iteratively by decoupling velocity parameters from reflector depths. A rapid 2-D and 3-D ray-tracing algorithm is used to compute transmission and reflection traveltimes and partial derivatives with respect to slowness and reflector depth. Both depth and velocity are parameterized in terms of cubic B-spline functions. The method has been applied to a reverse vertical seismic profile (VSP) data set recorded on the British coal measures along a crossed-linear array.

  1. Spectral characteristics of chromatic confocal imaging systems.

    PubMed

    Hillenbrand, Matthias; Mitschunas, Beate; Brill, Florian; Grewe, Adrian; Sinzinger, Stefan

    2014-11-10

    We present signal-generation models for chromatic confocal imaging systems with illumination and detection pinholes of finite size: a collinear model that considers neither aberrations nor diffraction effects, a geometrical model that accounts for aberrations, and a wave optical model covering both aberrations and diffraction effects. These models are aimed at describing the spectral response of multipoint sensor systems with field-dependent aberrations and vignetting effects. They are suitable for single- and double-pass systems with either diffusely or specularly reflecting surfaces under test. We show experimental results to verify our models. PMID:25402983

  2. Any Way You Slice It—A Comparison of Confocal Microscopy Techniques

    PubMed Central

    Jonkman, James

    2015-01-01

    The confocal fluorescence microscope has become a popular tool for life sciences researchers, primarily because of its ability to remove blur from outside of the focal plane of the image. Several different kinds of confocal microscopes have been developed, each with advantages and disadvantages. This article will cover the grid confocal, classic confocal laser-scanning microscope (CLSM), the resonant scanning-CLSM, and the spinning-disk confocal microscope. The way each microscope technique works, the best applications the technique is suited for, the limitations of the technique, and new developments for each technology will be presented. Researchers who have access to a range of different confocal microscopes (e.g., through a local core facility) should find this paper helpful for choosing the best confocal technology for specific imaging applications. Others with funding to purchase an instrument should find the article helpful in deciding which technology is ideal for their area of research. PMID:25802490

  3. Combined Confocal and Magnetic Resonance Microscopy

    SciTech Connect

    Wind, Robert A.; Majors, Paul D.; Minard, Kevin R.; Ackerman, Eric J.; Daly, Don S.; Holtom, Gary R.; Thrall, Brian D.; Weber, Thomas J.

    2002-05-12

    Confocal and magnetic resonance microscopy are both used to study live cells in a minimally invasive way. Both techniques provide complementary information. Therefore, by examining cells simultaneously with both methodologies, more detailed information is obtained than is possible with each of the microscopes individually. In this paper two configurations of a combined confocal and magnetic resonance microscope described. In both cases the sample compartment is part of a temperature regulated perfusion system. The first configuration is capable of studying large single cells or three-dimensional cell agglomerates, whereas with the second configuration monolayers of mammalian cells can be investigated . Combined images are shown of Xenopus laevis frog oocytes, model JB6 tumor spheroids, and a single layer of Chinese hamster ovary cells. Finally, potential applications of the combined microscope are discussed.

  4. A new multichannel spectral imaging laser scanning confocal microscope.

    PubMed

    Zhang, Yunhai; Hu, Bian; Dai, Yakang; Yang, Haomin; Huang, Wei; Xue, Xiaojun; Li, Fazhi; Zhang, Xin; Jiang, Chenyu; Gao, Fei; Chang, Jian

    2013-01-01

    We have developed a new multichannel spectral imaging laser scanning confocal microscope for effective detection of multiple fluorescent labeling in the research of biological tissues. In this paper, the design and key technologies of the system are introduced. Representative results on confocal imaging, 3-dimensional sectioning imaging, and spectral imaging are demonstrated. The results indicated that the system is applicable to multiple fluorescent labeling in biological experiments. PMID:23585775

  5. Optical tweezers for confocal microscopy

    NASA Astrophysics Data System (ADS)

    Hoffmann, A.; Meyer zu Hrste, G.; Pilarczyk, G.; Monajembashi, S.; Uhl, V.; Greulich, K. O.

    2000-11-01

    In confocal laser scanning microscopes (CLSMs), lasers can be used for image formation as well as tools for the manipulation of microscopic objects. In the latter case, in addition to the imaging lasers, the light of an extra laser has to be focused into the object plane of the CLSM, for example as optical tweezers. Imaging as well as trapping by optical tweezers can be done using the same objective lens. In this case, z-sectioning for 3D imaging shifts the optical tweezers with the focal plane of the objective along the optical axis, so that a trapped object remains positioned in the focal plane. Consequently, 3D imaging of trapped objects is impossible without further measures. We present an experimental set-up keeping the axial trapping position of the optical tweezers at its intended position whilst the focal plane can be axially shifted over a distance of about 15 ?m. It is based on fast-moving correctional optics synchronized with the objective movement. First examples of application are the 3D imaging of chloroplasts of Elodea densa (Canadian waterweed) in a vigorous cytoplasmic streaming and the displacement of zymogen granules in pancreatic cancer cells (AR42 J).

  6. In vivo confocal microscopy of Thygeson's superficial punctate keratitis.

    PubMed

    Cheng, Lulu L; Young, Alvin L; Wong, Angus K K; Law, Ricky W K; Lam, Dennis S C

    2004-06-01

    A 56-year-old Chinese man diagnosed with Thygeson's keratitis by clinical biomicroscopy was examined using a tandem scanning confocal microscope. Among normal superficial epithelial cells, clumps of markedly enlarged epithelial cells were identified. Multiple highly reflective filamentary structures were observed in the deeper layers. Most of these lesions were linear; some demonstrated curled ends and others demonstrated branching lesions with 'sprouts'. No inflammatory cells were evident in the areas of corneal stroma sampled. In vivo confocal microscopy may be helpful in the diagnosis of Thygeson's superficial keratitis. To the best of our knowledge, we report the first in vivo confocal images of focal desquamation of epithelium and intraepithelial hyper-reflective linear lesions in English literature. PMID:15180847

  7. Instrumentation for Reflectance Spectroscopy and Microspectroscopy with Application to Astrobiology

    NASA Technical Reports Server (NTRS)

    Mouroulis, Pantazis; Blaney, Diana L.; Green, Robert O.

    2008-01-01

    We present instrument concepts for in-situ reflectance spectroscopy over a spatial resolution range from several meters to tens of micrometers. These have been adapted to the low mass and power requirements of rover or similar platforms. Described are a miniaturized imaging spectrometer for rover mast, a combined mast and arm point spectrometer, and an imaging microspectrometer for the rover arm.

  8. Laboratory laser reflectance measurement and applications to asteroid surface analysis

    NASA Astrophysics Data System (ADS)

    Shaw, A.; Daly, M. G.; Cloutis, E. A.; Tait, K. T.; Izawa, M. R. M.; Barnouin, O. S.; Hyde, B. C.; Nicklin, I.

    2014-07-01

    Introduction Laboratory reflectance measurement of asteroid analogs is an important tool for interpreting the reflectance of asteroids. One dominant factor affecting how measured reflectance changes as a function of phase angle (180 minus the scattering angle) is surface roughness [1], which is related to grain size. A major goal of this study is to be able to use the angular distributions (phase functions) of scattered light from various regions on an asteroid surface to determine the relative grain size between those regions. Grain size affects the spectral albedo and continuum slopes of surface materials, has implications in terms of understanding geologic processes on asteroids and is also valuable for the planning and operations of upcoming missions to asteroids, such as the New Frontiers OSIRIS-REx sample return mission to the asteroid (101955) Bennu [2]. Information on surface roughness is particularly powerful when combined with other datasets, such as thermal inertia maps (e.g., a smooth, low-backscatter surface of low thermal inertia likely contains fine grains). Approach To better constrain the composition and surface texture of Bennu, we are conducting experiments to investigate the laser return signature of terrestrial and meteorite analogs to Bennu. The objective is to understand the nature of laser returns given possible compositional, grain size and slope distributions on the surface of Bennu to allow surface characterization, particularly surface grain size, which would significantly aid efforts to identify suitable sites for sampling by the OSIRIS-REx mission. Setup A 1064-nm laser is used to determine the reflectance of Bennu analogs and their constituents (1064 nm is the wavelength of many laser altimeters including the one planned to fly on OSIRIS-REx). Samples of interest include serpentinites (greenalite, etc.), magnetite, and shungite. To perform the experiments, a goniometer has been built. This instrument allows reflectance measurements at various illumination and viewing geometries. The goniometer has an an arm and a caddy that travels the length of an arc. Both the caddy and arm can accommodate either a source or detector. The arm rotates in azimuth and elevation, allowing data acquisition over the whole hemisphere. The optical assembly that we mount on the caddy for the first two sets of experiments described below has also been built. Experiments We have determined a series of three sets of experiments for measuring reflectance as a function of grain size while successively broadening the range of illumination and viewing geometries: 1) The first set of experiments involves measuring reflectance of a set of samples, each of a different grain size, at constant viewing and illumination geometry (nadir for both). 2) The second set of experiments involves a similar set of measurements, but this time the incidence angle will be varied, while keeping the phase angle constant (at zero, i.e., the lidar geometry). The results will be important for calibrating OSIRIS-REx Laser Altimeter (OLA) data, including separating the contributions of range, surface roughness, and surface composition. 3) The third set of experiments builds on the previous experiments by also allowing phase angle to vary, resulting in phase function (angular scattering intensity distribution) measurements. These data are particularly useful for the interpretation of OSIRIS-REx Visible and IR Spectrometer (OVIRS) and OSIRIS-REx Camera Suite (OCAMS) data taken at varying illumination and viewing geometries. These datasets can then be analyzed together with the OLA dataset for a more complete picture of surface reflectance characteristics. Conclusion The experiments outlined above and the resulting database are intended to benefit 1) proper interpretation of photometric data to determine surface roughness and 2) generation of albedo maps from laser altimeter measurements of planetary surfaces, such as that of Bennu. We have built a facility to collect this database of reflectance measurements, and the facility has already seen ''fi

  9. 834-A reflective coating for magnetospheric imagery applications

    NASA Astrophysics Data System (ADS)

    Chakrabarti, Supriya; Edelstein, Jerry; Keski-Kuha, Ritva A.; Threat, Felix T.

    1992-06-01

    Imaging upflowing O(superscript +) ions of ionospheric origin and plasmaspheric O(superscript +) can be achieved through solar resonance scattering at 834 angstroms. Unfortunately, several strong background emissions, including the ones at 1025 angstroms and 1216 angstroms due to geocoronal hydrogen atoms, pose serious problems to its implementation. Most common optical coatings have higher reflectivity at 1025 angstroms and 1216 angstroms than at 834 angstroms. We have designed a multiple-layer coating which selectively reflects 834 angstroms radiation and suppresses 1025 angstroms and 1216 angstroms radiation. The structure of the coating material consists of a very thin (50 - 150 angstroms) method (nickel) layer, on top of a semitransparent dielectric material (magnesium fluoride), over an aluminum substrate. Three such coatings were produced at NASA Goddard Space Flight Center using an existing coating facility which is not optimized for thin coatings. In spite of such fabrication difficulties, we have obtained encouraging results.

  10. Application of multispectral reflectance for early detection of tomato disease

    NASA Astrophysics Data System (ADS)

    Xu, Huirong; Zhu, Shengpan; Ying, Yibin; Jiang, Huanyu

    2006-10-01

    Automatic diagnosis of plant disease is important for plant management and environmental preservation in the future. The objective of this study is to use multispectral reflectance measurements to make an early discrimination between the healthy and infected plants by the strain of tobacco mosaic virus (TMV-U1) infection. There were reflectance changes in the visible (VIS) and near infrared spectroscopy (NIR) between the healthy and infected plants. Discriminant models were developed using discriminant partial least squares (DPLS) and Mahalanobis distance (MD). The DPLS models had a root mean square error of calibration (RMSEC) of 0.397 and correlation coefficient (r) of 0.59 and the MD model correctly classified 86.7% healthy plants and up to 91.7% infected plants.

  11. The Application of Reflected GPS Signals to Ocean Remote Sensing

    NASA Technical Reports Server (NTRS)

    Garrison, James L.; Katzberg, Stephen J.

    1998-01-01

    The L-band broadcast signal from the Global Positioning System (GPS) which has reflected off of the sea surface is under study for use as a ocean, coastal and wetlands remote sensing tool. The reflected signal from a given GPS satellite is cross-correlated with the pseudorandom noise code uniquely identifying that satellite. The shape of this cross-correlation, ordinarily a very sharp triangle when tracking a direct line of sight signal, becomes wider and smoother as the mean square slope of the reflecting surface increases. It is proposed that the surface wind speed can be determined by matching the recorded shape of this cross-correlation to that predicted by theoretical models as a function of wind speed and direction. The significance of these effects increases with altitude of the receiver. Experimental data have been collected using a specially modified GPS receiver on aircraft and on a balloon at altitudes of up to 25 km. These data compare favorably with predictions of analytical models and demonstrate the dependence of the waveform shape on surface wind speed and receiver altitude. The advantages that this measurement technique has over conventional scatterometers is the small size, low cost and simplicity of the receiver hardware, no requirement for a transmitter, and the ability to simultaneously collect data from usually 10 or more points (from a low earth orbiting satellite). This number could if the Russian GLONASS (Global Navigation Satellite System) satellites are also considered as additional sources of radiation. Furthermore, the bistatic scattering geometry is complementary to the backscatter used by conventional scatterometers.

  12. An 834 A reflective coating for magnetospheric imagery applications

    NASA Technical Reports Server (NTRS)

    Chakrabarti, Supriya; Edelstein, Jerry; Keski-Kuha, Ritva A. M.; Threat, Feliz T.

    1992-01-01

    We have designed a multiple-layer coating which selectively reflects 834 A radiation and suppresses 1025 A and 1216 A radiation. The structure of the coating material consists of a very thin (50-150 A) metal (nickel) layer, on top of a semitransparent dielectric material (magnesium fluoride), over an aluminum substrate. Three such coatings were produced at NASA Goddard Space Flight Center using an existing coating facility which is not optimized for thin coatings. In spite of such fabrication difficulties, we have obtained encouraging results.

  13. anti-reflective films for solar cell applications

    NASA Astrophysics Data System (ADS)

    Ali, Khuram; Khan, Sohail A.; Jafri, Mohd Zubir Mat

    2014-04-01

    Indium tin oxide (ITO) and titanium dioxide (TiO2) anti-reflective coatings (ARCs) were deposited on a (100) P-type monocrystalline Si substrate by a radio-frequency (RF) magnetron sputtering. Polycrystalline ITO and anatase TiO2 films were obtained at room temperature (RT). The thickness of ITO (60 to 64 nm) and TiO2 (55 to 60 nm) films was optimized, considering the optical response in the 400- to 1,000-nm wavelength range. The deposited films were characterized by X-ray diffraction (XRD), Raman spectroscopy, field emission scanning electron microscopy (FESEM), energy dispersive spectroscopy (EDS), and atomic force microscopy (AFM). The XRD analysis showed preferential orientation along (211) and (222) for ITO and (200) and (211) for TiO2 films. The XRD analysis showed that crystalline ITO/TiO2 films could be formed at RT. The crystallite strain measurements showed compressive strain for ITO and TiO2 films. The measured average optical reflectance was about 12% and 10% for the ITO and TiO2 ARCs, respectively.

  14. Skeleton-migration: Applications in deep crustal reflection seismic profiling

    NASA Astrophysics Data System (ADS)

    Eaton, D. W.; Vasudevan, K.

    2009-12-01

    The reflection geometry of the sub-surface is three-dimensional in character. A 3-D seismic data acquisition and processing would be the ideal modus operandi for true seismic interpretation. However, almost all deep-crustal reflection profiles recorded on land follow quasi-linear geometry, for economic reasons. Although conventional processing of the lines accommodates crooked-line geometry, the migration algorithms used to produce seismic images for interpretation are generally 2-D in nature. Consequently, the effects of 3-D geometry are not usually well-accounted for. For example, the out-of-plane reflections lead to mislocation errors that increase with recording time. The events may be mislocated by 10s of km and show spurious apparent dip after migration. In order to circumvent these problems and to gain insight into 3-D structures, we present an easy-to-implement Skeleton-migration algorithm. The skeleton-migration method follows a two-step procedure. In the first step, we introduce a fast skeletonization of the final pre-processed stack to generate a digital catalogue containing a variety of event attributes including two-way travel times and location information in UTM co-ordinates. In the second step, we apply ray-based migration to the catalogue of events or two-way travel times of the 2-D stack using an appropriate velocity model for the crust and upper mantle. Since often we do not know a priori the strike direction of the reflectors, we have implemented a fast visualization-based optimization procedure to determine the strike. In subsequent steps, we use visualization methods to view and interpret the skeleton-migration results. We illustrate the usefulness of the method with examples from both the synthetic and deep crustal seismic reflection data. For the synthetic examples, we consider physical models corresponding to a point-scatterer, a synform, a fault and a subducting slab. In all these instances, we use an elastic Kirchhoff algorithm with a shooting geometry that mimics a crooked-line in a medium of constant background velocity to generate synthetic seismograms. We apply the present skeleton-migration method for a set of trial strike directions not only to establish the functionality of the method but also affirm the model geometry from the migrated results. For a real data example, we apply the technique to deep crustal reflection seismic profile of SNORCLE (Slave-Northern Cordilleran Lithospheric Evolution Transect) line 1 to understand the Paleoproterozoic geometry of the reflectors interpreted to be a relic of the subducted slab. We use the coherency-filtered stack as the starting point for skeletonization. Despite the noise reduction of the stack profile through coherency-filtering, we introduce procedures to select sections of the data, i.e. ribbon-filtering, for ray-based migration to minimize the effects of residual noise during migration. We demonstrate the usefulness of the skeleton-migration method in understanding the geometry of the Moho and the sub-Moho geometry of the mantle reflectors with the migration velocity corresponding to an average 1-D velocity extracted from refraction surveys conducted in the study area.

  15. Custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of Inositol 1,4,5-trisphosphate-mediated calcium signals

    NASA Astrophysics Data System (ADS)

    Sigaut, Lorena; Barella, Mariano; Espada, Roco; Ponce, Mara Laura; Dawson, Silvina Ponce

    2011-06-01

    The flash photolysis of ``caged'' compounds is a powerful experimental technique for producing rapid changes in concentrations of bioactive signaling molecules. These caged compounds are inactive and become active when illuminated with ultraviolet light. This paper describes an inexpensive adaptation of an Olympus confocal microscope that uses as source of ultraviolet light the mercury lamp that comes with the microscope for conventional fluorescence microscopy. The ultraviolet illumination from the lamp (350 - 400 nm) enters through an optical fiber that is coupled to a nonconventional port of the microscope. The modification allows to perform the photolysis of caged compounds over wide areas (~200 ?m) and obtain confocal fluorescence images simultaneously. By controlling the ultraviolet illumination exposure time and intensity it is possible to regulate the amount of photolyzed compounds. In the paper we characterize the properties of the system and show its capabilities with experiments done in aqueous solution and in Xenopus Laevis oocytes. The latter demonstrate its applicability for the study of Inositol 1,4,5-trisphosphate-mediated intracellular calcium signals.

  16. Effects of Reflection Category and Reflection Quality on Learning Outcomes during Web-Based Portfolio Assessment Process: A Case Study of High School Students in Computer Application Course

    ERIC Educational Resources Information Center

    Chou, Pao-Nan; Chang, Chi-Cheng

    2011-01-01

    This study examines the effects of reflection category and reflection quality on learning outcomes during Web-based portfolio assessment process. Experimental subjects consist of forty-five eight-grade students in a "Computer Application" course. Through the Web-based portfolio assessment system, these students write reflection, and join…

  17. Affine reflection groups for tiling applications: Knot theory and DNA

    NASA Astrophysics Data System (ADS)

    Bodner, M.; Patera, J.; Peterson, M.

    2012-01-01

    We present in this paper some non-conventional applications of affine Weyl groups Waff of rank 2, the symmetry group of the tiling/lattice. We first develop and present the tools for applications requiring tilings of a real Euclidean plane {R}^2. We then elucidate the equivalence of these tilings with 2D projections of knots. The resulting mathematical structure provides a framework within which is encompassed recent work utilizing knot theory for modeling the structure and function of genetic molecules, specifically the action of particular enzymes in altering the topology of DNA in site-specific recombination.

  18. 33 CFR 148.708 - Must the applicant's proposal reflect potential regulations?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ..., DEPARTMENT OF HOMELAND SECURITY (CONTINUED) DEEPWATER PORTS DEEPWATER PORTS: GENERAL Environmental Review Criteria for Deepwater Ports § 148.708 Must the applicant's proposal reflect potential regulations... foreseeable environmental regulations in planning, operating, and decommissioning a deepwater port....

  19. 33 CFR 148.708 - Must the applicant's proposal reflect potential regulations?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ..., DEPARTMENT OF HOMELAND SECURITY (CONTINUED) DEEPWATER PORTS DEEPWATER PORTS: GENERAL Environmental Review Criteria for Deepwater Ports § 148.708 Must the applicant's proposal reflect potential regulations... foreseeable environmental regulations in planning, operating, and decommissioning a deepwater port....

  20. 33 CFR 148.708 - Must the applicant's proposal reflect potential regulations?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., DEPARTMENT OF HOMELAND SECURITY (CONTINUED) DEEPWATER PORTS DEEPWATER PORTS: GENERAL Environmental Review Criteria for Deepwater Ports § 148.708 Must the applicant's proposal reflect potential regulations... foreseeable environmental regulations in planning, operating, and decommissioning a deepwater port....

  1. 33 CFR 148.708 - Must the applicant's proposal reflect potential regulations?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ..., DEPARTMENT OF HOMELAND SECURITY (CONTINUED) DEEPWATER PORTS DEEPWATER PORTS: GENERAL Environmental Review Criteria for Deepwater Ports § 148.708 Must the applicant's proposal reflect potential regulations... foreseeable environmental regulations in planning, operating, and decommissioning a deepwater port....

  2. 33 CFR 148.708 - Must the applicant's proposal reflect potential regulations?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ..., DEPARTMENT OF HOMELAND SECURITY (CONTINUED) DEEPWATER PORTS DEEPWATER PORTS: GENERAL Environmental Review Criteria for Deepwater Ports § 148.708 Must the applicant's proposal reflect potential regulations... foreseeable environmental regulations in planning, operating, and decommissioning a deepwater port....

  3. Hybrid hyperchromats for chromatic confocal sensor systems

    NASA Astrophysics Data System (ADS)

    Hillenbrand, Matthias; Mitschunas, Beate; Wenzel, Christian; Grewe, Adrian; Ma, Xuan; Feßer, Patrick; Bichra, Mohamed; Sinzinger, Stefan

    2012-07-01

    The combination of diffractive and refractive elements in hybrid optical systems allows for precise control of the longitudinal chromatic aberration. We provide comprehensive design strategies for hybrid hyperchromatic lenses that maximise the longitudinal chromatic aberrations. These lenses are mainly used in chromatic confocal sensor systems for efficient non-contact profilometry as well as for measurements of distances and wall thicknesses of transparent materials. Our design approach enables the tailoring of the sensor properties to the specific measurement problem and assists designers in finding optimised solutions for industrial applications. We, for example, demonstrate a hybrid system that significantly exceeds the longitudinal chromatic aberration of purely diffractive elements.

  4. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: SPECTROSCOPY

    EPA Science Inventory

    The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. The goal of a CLSM is to acquire and quantify fluorescence and in some instruments acquire spectral characterization of emitted signals. The accuracy of these measurements demands that...

  5. In vivo confocal microscopy of ocular surface squamous neoplasia

    PubMed Central

    Parrozzani, R; Lazzarini, D; Dario, A; Midena, E

    2011-01-01

    Purpose To analyse in vivostructural and cellular features of ocular surface squamous neoplasia using clinical confocal microscopy. Methods Ten consecutive cases of untreated ocular surface squamous neoplasia were in vivoinvestigated using clinical confocal microscopy (ConfoScan4, Nidek Co. Ltd, Gamagori, Japan) with a 40 surface non-contact objective lens. Confocal microscopy images were compared with cytologic samples obtained by scraping technique. Results Confocal microscopy examination revealed large areas of superficial cells debris and/or keratin debris accompanied by syncytial-like groupings, loss of the normal structure of the conjunctival epithelium andor of the corneal basal epithelium layer, papillomatous organization, large fibrovascular structures, and fine vessels perpendicular to the tumour surface. Sub-epithelial (pre-Bowman) space involvement was documented in four cases (50%). Irregular healthy tissue infiltration at the lateral edge of the lesion was documented in two cases (20%) whereas abrupt demarcation between neoplastic cells and normal epithelium was documented in eight cases (80%). In vivocyto-morphologic study using clinical confocal microscopy showed cellular anisocytosis, pleocytosis, and anisonucleosis, enlarged nuclei with high nuclear to cytoplasmic ratio, high reflective cytoplasm and indistinct cytoplasmic borders in all cases (100%). Conclusion CCM appears to be a promising and non-invasive method for in vivostructural and cellular analysis of OSSN. PMID:21311574

  6. Benefits of metal reflective surfaces for concentrating solar applications

    NASA Astrophysics Data System (ADS)

    Braendle, Stefan

    2010-08-01

    Concentrating photovoltaic (CPV) companies are constantly making gains in efficiency and a lower levelized cost of energy, but continue to face questions of reliability and efficiency at scale remain. New technologies such as highly efficient aluminum mirrors help CPV companies fulfill both of these demands by allowing for performance and reliability gains, while also enabling high volume production for scaled deployment. In testing, metal mirrors have shown to be good matches for concentrating applications while performing at the same level as glass mirrors in accelerated weather tests. When combined with the inherent lighter weight and formability of aluminum, these new mirrors provide CPV solutions with a compelling advantage in the field.

  7. Use of probe-based confocal laser endomicroscopy (pCLE) in gastrointestinal applications. A consensus report based on clinical evidence

    PubMed Central

    Wang, Kenneth K; Carr-Locke, David L; Singh, Satish K; Neumann, Helmut; Bertani, Helga; Arsenescu, Razvan I; Caillol, Fabrice; Chang, Kenneth J; Chaussade, Stanislas; Coron, Emmanuel; Costamagna, Guido; Dlugosz, Aldona; Ian Gan, S; Giovannini, Marc; Gress, Frank G; Haluszka, Oleh; Ho, Khek Y; Kahaleh, Michel; Konda, Vani J; Prat, Frederic; Shah, Raj J; Sharma, Prateek; Slivka, Adam; Wolfsen, Herbert C; Zfass, Alvin

    2015-01-01

    Background Probe-based confocal laser endomicroscopy (pCLE) provides microscopic imaging during an endoscopic procedure. Its introduction as a standard modality in gastroenterology has brought significant progress in management strategies, affecting many aspects of clinical care and requiring standardisation of practice and training. Objective This study aimed to provide guidance on the standardisation of its practice and training in Barrett’s oesophagus, biliary strictures, colorectal lesions and inflammatory bowel diseases. Methods Initial statements were developed by five group leaders, based on the available clinical evidence. These statements were then voted and edited by the 26 participants, using a modified Delphi approach. After two rounds of votes, statements were validated if the threshold of agreement was higher than 75%. Results Twenty-six experts participated and, among a total of 77 statements, 61 were adopted (79%) and 16 were rejected (21%). The adoption of each statement was justified by the grade of evidence. Conclusion pCLE should be used to enhance the diagnostic arsenal in the evaluation of these indications, by providing microscopic information which improves the diagnostic performance of the physician. In order actually to implement this technology in the clinical routine, and to ensure good practice, standardised initial and continuing institutional training programmes should be established. PMID:26137298

  8. Novel efficient methods for measuring mesophyll anatomical characteristics from fresh thick sections using stereology and confocal microscopy: application on acid rain-treated Norway spruce needles.

    PubMed

    Albrechtov, Jana; Jancek, Jir; Lhotkov, Zuzana; Radochov, Barbora; Kubnov, Lucie

    2007-01-01

    Recent design-based stereological methods that can be applied to thick sections cut in an arbitrary direction are presented and their implementation for measuring mesophyll anatomical characteristics is introduced. These methods use software-randomized virtual 3D probes, such as disector and fakir test probes, in stacks of optical sections acquired using confocal microscopy. They enable unbiased estimations of the mean mesophyll cell volume, mesophyll cell number in a needle, and for the first time an internal surface area of needles or other narrow leaves directly from the fresh tissue cross-sections cut using a hand microtome. Therefore, reliable results can be obtained much faster than when using a standard microtechnical preparation. The proposed methods were tested on Norway spruce needles affected for 1 year by acid rain treatment. The effect of acid rain resulted in changes of mesophyll parameters: the ratio of intercellular spaces per mesophyll cell volume increased, while needle internal surface area, total number of mesophyll cells, and number of mesophyll cells per unit volume of a needle decreased in the treated needles. PMID:17322549

  9. Developments of new force reflecting control schemes and an application to a teleoperation training simulator

    NASA Technical Reports Server (NTRS)

    Kim, Won S.

    1992-01-01

    Two schemes of force reflecting control, position-error based force reflection and low-pass-filtered force reflection, both combined with shared compliance control, were developed for dissimilar master-slave arms. These schemes enabled high force reflection gains, which were not possible with a conventional scheme when the slave arm was much stiffer than the master arm. The experimental results with a peg-in-hole task indicated that the newly force reflecting control schemes combined with compliance control resulted in best task performances. As a related application, a simulated force reflection/shared compliance control teleoperation trainer was developed that provided the operator with the feel of kinesthetic force virtual reality.

  10. Optical Electronic Bragg Reflection Sensor System with Hydrodynamic Flow Applications

    NASA Technical Reports Server (NTRS)

    Lyons, D. R.

    2003-01-01

    This project, as described in the following report, involved design and fabrication of fiber optic sensors for the detection and measurement of dynamic fluid density variations. These devices are created using UV (ultraviolet) ablation and generally modified transverse holographic fiber grating techniques. The resulting phase gratings created on or immediately underneath the flat portion of D-shaped optical waveguides are characterized as evanescent field sensing devices. The primary applications include the sensor portion of a real-time localized or distributed measurement system for hydrodynamic flow, fluid density measurements, and phase change phenomena. Several design modifications were implemented in an attempt to accomplish the tasks specified in our original proposal. In addition, we have established key collaborative relationships with numerous people and institutions.

  11. Automated cellular pathology in noninvasive confocal microscopy

    NASA Astrophysics Data System (ADS)

    Ting, Monica; Krueger, James; Gareau, Daniel

    2014-03-01

    A computer algorithm was developed to automatically identify and count melanocytes and keratinocytes in 3D reflectance confocal microscopy (RCM) images of the skin. Computerized pathology increases our understanding and enables prevention of superficial spreading melanoma (SSM). Machine learning involved looking at the images to measure the size of cells through a 2-D Fourier transform and developing an appropriate mask with the erf() function to model the cells. Implementation involved processing the images to identify cells whose image segments provided the least difference when subtracted from the mask. With further simplification of the algorithm, the program may be directly implemented on the RCM images to indicate the presence of keratinocytes in seconds and to quantify the keratinocytes size in the en face plane as a function of depth. Using this system, the algorithm can identify any irregularities in maturation and differentiation of keratinocytes, thereby signaling the possible presence of cancer.

  12. Confocal in vivo microscopy and confocal laser-scanning fluorescence microscopy in keratoconus.

    PubMed

    Somodi, S; Hahnel, C; Slowik, C; Richter, A; Weiss, D G; Guthoff, R

    1996-11-01

    The purpose of this study was the determination of morphological changes in the corneal epithelium and the keratocyte network in keratoconus. In all, 33 eyes of 19 patients were examined in vivo using the confocal slit-scanning microscope Microphthal. After penetrating keratoplasty, recipients' trephanates were stained with the Live/Dead kit and examined using the confocal laser-scanning fluorescence microscope Diaphot 300/Odyssey. The fluorescence images were reconstructed three-dimensionally. All findings were compared with data from healthy corneas. Morphological alterations were found only in the area of the corneal apex; obviously elongated superficial epithelial cells arranged in a whorl-like fashion were found. Near Bowman's membrane, highly reflective changes and fold-like structures were visible. The anterior stroma also showed an increased reflectivity. In the posterior stroma, typical findings were Vogt's striae and keratocytes with extremely long processes arranged nearly in parallel. In scarred stroma the keratocytes were spindle-shaped and arranged irregularly. The spatial organization of the living keratocyte network could be demonstrated through three-dimensional reconstructions. PMID:9479549

  13. Confocal microscopy in a case of crystalline keratopathy in a patient with smouldering multiple myeloma.

    PubMed

    Mazzotta, Cosimo; Caragiuli, Stefano; Caporossi, Aldo

    2014-06-01

    We report the clinical and confocal microscopic findings of the cornea in a patient with smouldering multiple myeloma (SMM) using in vivo scanning laser confocal microscopy. A 72-year-old female underwent a complete ophthalmological examination including slit-lamp biomicroscopy with digital photography, HRT II laser scanning in vivo confocal microscopy and haematological laboratory assessment. Corneal biomicroscopy revealed the presence of bilateral diffuse microgranular tiny grey opacities. In vivo confocal microscopy showed randomly oriented hyper-reflective needle-shaped crystals throughout all levels of the stroma, sparing epithelium and endothelium. In vivo confocal microscopy was very helpful in the differential diagnosis by allowing the nature of the corneal deposits to be established, revealing the typical aspect of the crystals, and excluding granular dystrophy, leading to a suspected diagnosis of SMM. Crystalline corneal deposits may easily be confused as crumb-like opacities typical of granular dystrophy on slit-lamp examination even by experienced ophthalmologists. PMID:23928943

  14. Simple method for modeling radar reflections in a homogeneous halfspace, with applications

    NASA Astrophysics Data System (ADS)

    Greenfield, Roy J.; Moran, Mark L.; Davis, J. L.

    2000-04-01

    We have developed a method to rapidly compute synthetic radar records from complex reflecting surfaces. The approach is a 3- D time domain Hemholtz-Kirchhoff (HK) representation, similar to Hilterman (1981), that includes the radiation characteristics of GPR dipoles on the surface of a uniform dielectric halfspace. Validity is established by making comparisons with published model results and by comparisons with field data. Comparison to the ray theory results of Zeng et al. (1997) show excellent agreement in reflection arrival times for pipes of various diameters. We also reproduce the non-specular reflection results of Schleicher et al. (1991), which show that large amplitude reflections can originate from the inflection points of curved surfaces. Our comparisons with field data use reflection records taken at a test site in Borden, Ontario, over horizontally oriented buried metal drums. The H-plane reflection data were collected using shielded 700-MHz dipoles. Our raw synthetic amplitude trends show reasonable agreement to the field data, but are not perfect. Using a small diameter synthetic dipole array, we show that the mismatch is most likely caused by antenna shielding effects. The versatility of the HK method is demonstrated by giving results for a number of interesting applications. These include synthetic records for crisscrossing pipes buried at various depths, reflection synthetics from a truncated cone representing the slag heaps in Daniels and Brower (1998), and reflections from a rough surface. The slag heap models demonstrate the effect of antenna polarization on reflections from sloping surfaces. Analysis of synthetic reflections from rough surfaces shows that the coda following the first impulsive arrival can be used to estimate the surface roughness. This is of interest for interpreting reflections from glacier data. Our results demonstrate that the HK method is useful in interpreting data, as well as for developing field survey strategies.

  15. Cross-polarization confocal imaging of subsurface flaws in silicon nitride.

    SciTech Connect

    Liu, Z.; Sun, J. G.; Pei, Z.

    2011-03-01

    A cross-polarization confocal microscopy (CPCM) method was developed to image subsurface flaws in optically translucent silicon nitride (Si{sub 3}N{sub 4}) ceramics. Unlike conventional confocal microscopy, which measures reflected light so is applicable only to transparent and semi-transparent materials, CPCM detects scattered light from subsurface while filtering out the reflected light from ceramic surface. For subsurface imaging, the refractive-index mismatch between imaging (air) and imaged (ceramic) medium may cause image distortion and reduce resolution in the depth direction. This effect, characterized by an axial scaling factor (ASF), was analyzed and experimentally determined for glass and Si{sub 3}N{sub 4} materials. The experimental CPCM system was used to image Hertzian C-cracks generated by various indentation loads in the subsurface of a Si{sub 3}N{sub 4} specimen. It was demonstrated that CPCM may provide detailed information of subsurface cracks, such as crack angle and path, and subsurface microstructural variations.

  16. Confocal fluorescence microendoscopy using a digital micro-mirror device

    NASA Astrophysics Data System (ADS)

    Feng, Zhifeng; Wang, Liqiang; Duan, Huilong

    2010-11-01

    A design of confocal fluorescence microendoscopy utilizing a digital micro-mirror device (DMD) is described. Laser beams of the microendoscope are coupled into the body through a telescopic optics system, rather than through fibers or fiber bundles which are widely used in existing microendoscopes. Each micro-mirror of the DMD is used as a confocal pinhole. The DMD not only couples the laser beams into the body by a random time-varying speckle pattern and performs the scanning mechanism of the body tissue with different positions, but also couples the fluorescent signal emitted from the markers out to the CCD camera. Because of the CCD's integration feature and DMD's rapid parallel scanning feature, a complete predetermined depth tomography image accumulated by different scanning patterns of DMD can be acquired through only one CCD exposure procedure. The objective lens to realize high resolution and high sensitivity fluorescence imaging is the other function of the telescopic optics, with a numerical aperture of 0.35. The resolution of confocal microendoscope is superior to 228 lp/mm determined by 1951USAF resolution test target. Images of a tendon specimen are also shown to demonstrate practical application of the design. The confocal microendoscope using a DMD permits the acquisition of high-resolution real-time confocal images of epithelial tissue in vivo organ and realizes the aim of non-invasive diagnosis and treatment.

  17. Confocal Raman microspectroscopy of the skin.

    PubMed

    Förster, Matthias; Bolzinger, Marie-Alexandrine; Montagnac, Gilles; Briançon, Stéphanie

    2011-01-01

    Confocal Raman spectroscopy is a technique with considerable potential for the non-invasive study of biological tissues and skin samples in vitro or in vivo. It can be used to study skin physiology and possible pathological conditions and to obtain data about molecular composition and the structure of skin, for example, water content, moisturization and changes in the skin barrier function can all be observed. In-depth measurements also allow biopharmaceutical studies, such as analyzing the rate of penetration of a drug and the biochemical changes that may be induced by an applied formulation. Confocal Raman microspectroscopy is now at such a stage of refinement that it opens up new vistas. The big leap forward in its ease of use enables this technology to be used as an analytical method by more and more non-specialist laboratories. This review gives an overview of the state of the art of this technology by presenting an update on the principles of Raman spectroscopy and then by looking at examples of new developments in in vivo and in vitro applications. PMID:21914580

  18. Study of liquid jet instability by confocal microscopy

    NASA Astrophysics Data System (ADS)

    Yang, Lisong; Adamson, Leanne J.; Bain, Colin D.

    2012-07-01

    The instability of a liquid microjet was used to measure the dynamic surface tension of liquids at the surface ages of ?1 ms using confocal microscopy. The reflected light from a laser beam at normal incidence to the jet surface is linear in the displacement of the surface near the confocal position, leading to a radial resolution of 4 nm and a dynamic range of 4 ?m in the surface position, thus permitting the measurement of amplitude of oscillation at the very early stage of jet instability. For larger oscillations outside the linear region of the confocal response, the swell and neck position of the jet can be located separately and the amplitude of oscillation determined with an accuracy of 0.2 ?m. The growth rate of periodically perturbed water and ethanol/water mixture jets with a 100-?m diameter nozzle and mean velocity of 5.7 m s-1 has been measured. The dynamic surface tension was determined from the growth rate of the instability with a linear, axisymmetric, constant property model. Synchronisation of the confocal imaging system with the perturbation applied to the jet permitted a detailed study of the temporal evolution of the neck into a ligament and eventually into a satellite drop.

  19. Intravital Confocal and Two-photon Imaging of Dual-color Cells and Extracellular Matrix Mimics

    PubMed Central

    Bal, Ufuk; Andresen, Volker; Baggett, Brenda; Utzinger, Urs

    2013-01-01

    To optimize imaging of cells in three dimensional culture we studied confocal backscattering, Second Harmonic Generation (SHG) and autofluorescence as source of contrast in extracellular matrix (ECM) mimics and evaluated the attenuation as well as bleaching of endogenous cellular fluorescence signals. All common ECM mimics exhibit contrast observable with confocal reflectance microscopy. SHG imaging on collagen I based hydrogels provides high contrast and good optical penetration depth. Agarose is a useful embedding medium because it allows for large optical penetration and exhibits minimal autofluorescence while still providing good reflectance to detect voids in the embedding medium. We labeled breast cancer cells outline with DsRed2 and nucleus with eGFP. DsRed2 can be excited with confocal imaging at 568nm, and with two photon excitation (TPE) in the red and longer NIR. eGFP was excited at 488nm for confocal and in the NIR for TPE. While there is small difference in the bleaching rate for eGFP between confocal and TPE we observed significant difference for DsRed2 where bleaching is strongest during TPE in the red wavelengths and smallest during confocal imaging. After a few hundred microns depth in a collagen I hydrogel, TPE fluorescence becomes twice as strong compared to confocal imaging. PMID:23380006

  20. Combined confocal Raman and quantitative phase microscopy system for biomedical diagnosis

    PubMed Central

    Kang, Jeon Woong; Lue, Niyom; Kong, Chae-Ryon; Barman, Ishan; Dingari, Narahara C.; Goldfless, Stephen J.; Niles, Jacquin C.; Dasari, Ramachandra R.; Feld, Michael S.

    2011-01-01

    We have developed a novel multimodal microscopy system that incorporates confocal Raman, confocal reflectance, and quantitative phase microscopy (QPM) into a single imaging entity. Confocal Raman microscopy provides detailed chemical information from the sample, while confocal reflectance and quantitative phase microscopy show detailed morphology. Combining these intrinsic contrast imaging modalities makes it possible to obtain quantitative morphological and chemical information without exogenous staining. For validation and characterization, we have used this multi-modal system to investigate healthy and diseased blood samples. We first show that the thickness of a healthy red blood cell (RBC) shows good correlation with its hemoglobin distribution. Further, in malaria infected RBCs, we successfully image the distribution of hemozoin (malaria pigment) inside the cell. Our observations lead us to propose morphological screening by QPM and subsequent chemical imaging by Raman for investigating blood disorders. This new approach allows monitoring cell development and cell-drug interactions with minimal perturbation of the biological system of interest. PMID:21991542

  1. Applications of high power lasers. [using reflection holograms for machining and surface treatment

    NASA Technical Reports Server (NTRS)

    Angus, J. C.

    1979-01-01

    The use of computer generated, reflection holograms in conjunction with high power lasers for precision machining of metals and ceramics was investigated. The Reflection holograms which were developed and made to work at both optical wavelength (He-Ne, 6328 A) and infrared (CO2, 10.6) meet the primary practical requirement of ruggedness and are relatively economical and simple to fabricate. The technology is sufficiently advanced now so that reflection holography could indeed be used as a practical manufacturing device in certain applications requiring low power densities. However, the present holograms are energy inefficient and much of the laser power is lost in the zero order spot and higher diffraction orders. Improvements of laser machining over conventional methods are discussed and addition applications are listed. Possible uses in the electronics industry include drilling holes in printed circuit boards making soldered connections, and resistor trimming.

  2. TOTAL INTERNAL REFLECTION WITH FLUORESCENCE CORRELATION SPECTROSCOPY: APPLICATIONS TO SUBSTRATE-SUPPORTED PLANAR MEMBRANES

    PubMed Central

    Thompson, Nancy L.; Wang, Xiang; Navaratnarajah, Punya

    2009-01-01

    In this review paper, the conceptual basis and experimental design of total internal reflection with fluorescence correlation spectroscopy (TIR-FCS) is described. The few applications to date of TIR-FCS to supported membranes are discussed, in addition to a variety of applications not directly involving supported membranes. Methods related, but not technically equivalent, to TIR-FCS are also summarized. Future directions for TIR-FCS are outlined. PMID:19269331

  3. "Knowing Is Not Enough; We Must Apply": Reflections on a Failed Action Learning Application

    ERIC Educational Resources Information Center

    Reese, Simon

    2015-01-01

    This paper reflects upon a sub-optimal action learning application with a strategic business re-design project. The objective of the project was to improve the long-term business performance of a subsidiary business and build the strategic plan. Action learning was introduced to aid the group in expanding their view of the real problems…

  4. Promoting Reflective Thinking Skills by Using Web 2.0 Application

    ERIC Educational Resources Information Center

    Abdullah, Mohamed

    2015-01-01

    The study aims to investigate are using Web 2.0 applications promoting reflective thinking skills for higher education student in faculty for education. Although the literature reveals that technology integration is a trend in higher education and researchers and educators have increasingly shared their ideas and examples of implementations of Web…

  5. "Knowing Is Not Enough; We Must Apply": Reflections on a Failed Action Learning Application

    ERIC Educational Resources Information Center

    Reese, Simon

    2015-01-01

    This paper reflects upon a sub-optimal action learning application with a strategic business re-design project. The objective of the project was to improve the long-term business performance of a subsidiary business and build the strategic plan. Action learning was introduced to aid the group in expanding their view of the real problems

  6. Total Reflection X-Ray Microscopy in a SEM: 2. Application to surfaces and interfaces

    NASA Astrophysics Data System (ADS)

    Jibaoui, H.; Erre, D.; Cazaux, J.

    2000-05-01

    Some applications of Total Reflection X-Ray Microscopy (TRXRM) are given. It is demonstrated that this new imaging technique permits to acquire rapidly (a few seconds) digital images related to the topography of the irradiated surfaces. An important illustration is the direct imaging of slightly buried solid/solid interfaces.

  7. Immunofluorescence and Confocal Microscopy of Neutrophils

    PubMed Central

    Allen, Lee-Ann H.

    2015-01-01

    Rapid recruitment of neutrophils to sites of infection and their ability to phagocytose and kill microbes is an important aspect of the innate immune response. Challenges associated with imaging of these cells include their short lifespan and small size and the fact that unstimulated cells are nonadherent. In addition, although cytoplasmic granules are plentiful, the abundance of many other organelles is diminished. Here we reprise methods for analysis of resting and activated cells using immunofluorescence and confocal microscopy, including kinetic analysis of phagosome maturation and degranulation, and detection of intraphagosomal superoxide accumulation. We describe approaches for rapid cell fixation and permeabilization that maximize antigen detection and discuss other variables that also affect data interpretation and image quality (such as cell spreading, degranulation, and phagocytosis). Finally, we show that these methods are also applicable to studies of neutrophil interactions with the extracellular matrix. PMID:24504957

  8. EVALUATION OF CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: PRETTY PICTURES OR CONFOCAL QA

    EPA Science Inventory

    Evaluation of confocal microscopy system performance: Pretty pictures or confocal QA?

    Robert M. Zucker

    Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, N...

  9. Diagrammatic expansion of the Casimir energy in multiple reflections: Theory and applications

    SciTech Connect

    Maghrebi, Mohammad F.

    2011-02-15

    We develop a diagrammatic representation of the Casimir energy of a multibody configuration. The diagrams represent multiple reflections between the objects and can be organized by a few simple rules. The lowest-order diagrams (or reflections) give the main contribution to the Casimir interaction which proves the usefulness of this expansion. Among some applications of this, we find analytical formulae describing the interaction between edges, i.e. semi-infinite plates, where we also give a first example of blocking in the context of the Casimir energy. We also find the interaction of edges with a needle and describe analytically a recent model of the repulsion due to the Casimir interaction.

  10. Z-polarized confocal microscopy.

    PubMed

    Huse, N; Schnle, A; Hell, S W

    2001-10-01

    In light microscopy the transverse nature of the electromagnetic field precludes a strongly focused longitudinal field component, thus confining polarization spectroscopy and imaging to two dimensions (x,y). Here we describe a simple confocal microscopy arrangement that optimizes for signal from molecules with transition dipoles oriented parallel to the optic axis. In the proposed arrangement, we not only generate a predominant longitudinally (z) polarized focal field, but also engineer the detection scheme in such a way that in a bulk of randomly oriented molecules, the microscope's effective point-spread function is dominated by the contribution of those molecules that are oriented along the optic axis. Our arrangement not only implicitly allows for the determination of the orientation of transition dipoles of single molecules in three dimensions, but also highlights the contribution of z-oriented molecules in three-dimensional imaging. PMID:11806348

  11. Z-polarized confocal microscopy.

    PubMed

    Huse, N; Schnle, A; Hell, S W

    2001-07-01

    In light microscopy the transverse nature of the electromagnetic field precludes a strongly focused longitudinal field component, thus confining polarization spectroscopy and imaging to two dimensions (x,y). Here we describe a simple confocal microscopy arrangement that optimizes for signal from molecules with transition dipoles oriented parallel to the optic axis. In the proposed arrangement, we not only generate a predominant longitudinally (z) polarized focal field, but also engineer the detection scheme in such a way that in a bulk of randomly oriented molecules, the microscope's effective point-spread function is dominated by the contribution of those molecules that are oriented along the optic axis. Our arrangement not only implicitly allows for the determination of the orientation of transition dipoles of single molecules in three dimensions, but also highlights the contribution of z-oriented molecules in three-dimensional imaging. PMID:11516316

  12. Confocal endomicroscopy of the larynx

    NASA Astrophysics Data System (ADS)

    Just, T.; Wiechmann, T.; Stachs, O.; Stave, J.; Guthoff, R.; Hüttmann, G.; Pau, H. W.

    2012-02-01

    Beside the good image quality with the confocal laser scanning microscope (HRTII) and the Rostock Cornea Module (RCM), this technology can not be used to investigate the human larynx in vivo. To accomplish this, a rigid custom-made endoscope (KARL STORZ GmbH & Co. KG; Tuttlingen Germany) was developed. A connector was developed to connect the scanner head of the HRTII to the rigid endoscope. With the connector, the starting plane can be set manually. To achieve optical sectioning of the laryngeal tissue (80 μm per volume scan), the scanning mechanism of the HRTII needs to be activated using a foot switch. The devices consisting of the endoscope, HRTII, and the connector supply images of 400 x 400 μm and reach average penetration depths of 100-300 μm (λ/4 plate of the scanner head of the HRTII was removed). The lateral and axial resolutions are about 1-2 μm and 2 μm, respectively. In vivo rigid confocal endoscopy is demonstrated with an acquisition time for a volume scan of 6 s. The aim of this study was to differentiate pre-malignant laryngeal lesions from micro-invasive carcinoma of the larynx. 22 patients with suspicious lesions of the true vocal cords were included. This pilot study clearly demonstrates the possibility to detect dysplastic cells close to the basal cell layer and within the subepithelial space in lesions with small leukoplakia (thin keratin layer). These findings may have an impact on microlaryngoscopy to improve the precision for biopsy and on microlaryngoscopic laser surgery of the larynx to identify the margins of the pre-malignant lesion.

  13. Multimodal confocal mosaicing microscopy: an emphasis on squamous cell carcinoma

    NASA Astrophysics Data System (ADS)

    Chen, Nathaniel W.; Sensibaugh, Jordan; Ardeshiri, Ardaland; Blanchard, Adam; Jacques, Steven; Gareau, Daniel

    2010-02-01

    Our previous study reported a sensitivity of 96.6% and a specificity of 89.2% in rapidly detecting Basal Cell Carcinomas (BCCs) when nuclei were stained with acridine orange. Squamous Cell Carcinomas (SCCs) and infiltrative BCCs remain difficult to detect. More complete screening can be achieved utilizing both acridine orange for nuclei staining and eosin for cytoplasmic contrast, using two lasers to excite the two stains independently. Nuclear fluorescence is achieved by staining with acridine orange (0.5mM, 60 s), and cytoplasmic fluorescence is achieved by staining with eosin working solution (30 s). This work shows good morphological contrast of SCC and infiltrative BCC with eosin, acridine orange, and reflectance, and presents a means for rapid SCC and infiltrative BCC detection in fresh skin excisions using multimodal confocal microscopy. In addition, digital staining is shown to effectively simulate hematoxylin and eosin (H&E) histology with confocal mosaics.

  14. Confocal Microscope Alignment of Nanocrystals for Coherent Diffraction Imaging

    SciTech Connect

    Beitra, Loren; Watari, Moyu; Matsuura, Takashi; Shimamoto, Naonobu; Harder, Ross; Robinson, Ian

    2010-06-23

    We have installed and tested an Olympus LEXT confocal microscope at the 34-ID-C beamline of the Advanced Photon Source (APS). The beamline is for Coherent X-ray Diffraction (CXD) experiments in which a nanometre-sized crystal is aligned inside a focussed X-ray beam. The microscope was required for three-dimensional (3D) sample alignment to get around sphere-of-confusion issues when locating Bragg peaks in reciprocal space. In this way, and by use of strategic sample preparations, we have succeeded in measuring six Bragg peaks from a single 200 nm gold crystal and obtained six projections of its internal displacement field. This enables the clear identification of stacking-fault bands within the crystal. The confocal alignment method will allow a full determination of the strain tensor provided three or more Bragg reflections from the same crystal are found.

  15. Clinical feasibility of rapid confocal melanoma feature detection

    NASA Astrophysics Data System (ADS)

    Hennessy, Ricky; Jacques, Steve; Pellacani, Giovanni; Gareau, Daniel

    2010-02-01

    In vivo reflectance confocal microscopy shows promise for the early detection of malignant melanoma. One diagnostic trait of malignancy is the presence of pagetoid melanocytes in the epidermis. For automated detection of MM, this feature must be identified quantitatively through software. Beginning with in vivo, noninvasive confocal images from 10 unequivocal MMs and benign nevi, we developed a pattern recognition algorithm that automatically identified pagetoid melanocytes in all four MMs and identified none in five benign nevi. One data set was discarded due to artifacts caused by patient movement. With future work to bring the performance of this pattern recognition technique to the level of the clinicians on difficult lesions, melanoma diagnosis could be brought to primary care facilities and save many lives by improving early diagnosis.

  16. Needle-based confocal laser endomicroscopy

    PubMed Central

    Giovannini, Marc

    2015-01-01

    New applications of confocal laser endomicroscopy were developed as pCLE in the bile duct and nCLE for pancreatic cystic tumors, pancreatic masses and lymph nodes. The aim of this paper would be to give you an update in this new technology and to try to define its place in the diagnosis of cystic and solid pancreatic masses. The material used was a 19G EUS-needle in which the stylet was replaced by the Confocal mini-probe. The mini-probe (0.632 mm of diameter) is pre-loaded and screwed by a locking device in the EUS-Needle and guided endosonographically in the target. Regarding pancreatic cystic lesion, the presence of epithelial villous structures based on nCLE was associated with pancreatic cystic neoplasm (IPMN) (P = 0.004) and provided a sensitivity of 59%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 50%. A superficial vascular network pattern visualized on nCLE was identified in serous cystadenomas. It corresponded on pathological specimen to a dense and subepithelial capillary vascularization. The accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of this sign for the diagnosis of SCA were 87%, 69%, 100%, 100%, and 82%, respectively. In pancreatic adenocarcinomas, nCLE found vascular leakage with irregular vessels with leakage of fluorescein into the tumor, large dark clumps which correspond to humps of malignant cells. These criteria correlate with the histological structure of those tumors which are characterized by tumoral glands, surrounded by fibrosis in case of fibrous stroma tumor. Neuroendocrine tumors showed a dense network of small vessels on a dark background, which fits with the histological structure based on cord of cells surrounded by vessels and by fibrosis. nCLE is feasible during a EUS examination; these preliminary results are very encouraging and may be used in the future in case of inconclusive EUS-FNA. PMID:26643694

  17. EVALUATION OF CONFOCAL MICROSCOPY SYSTEM PERFORMANCE

    EPA Science Inventory

    BACKGROUND. The confocal laser scanning microscope (CLSM) has enormous potential in many biological fields. Currently there is a subjective nature in the assessment of a confocal microscope's performance by primarily evaluating the system with a specific test slide provided by ea...

  18. Measured energy savings from the application of reflective roofsin 2 small non-residential buildings

    SciTech Connect

    Akbari, Hashem

    2003-01-14

    Energy use and environmental parameters were monitored in two small (14.9 m{sup 2}) non-residential buildings during the summer of 2000. The buildings were initially monitored for about 1 1/2 months to establish a base condition. The roofs of the buildings were then painted with a white coating and the monitoring was continued. The original solar reflectivities of the roofs were about 26%; after the application of roof coatings the reflectivities increased to about 72%. The monitored electricity savings were about 0.5kWh per day (33 Wh/m2 per day). The estimated annual savings are about 125kWh per year (8.4 kWh/m2); at a cost of $0.1/kWh, savings are about $0.86/m2 per year. Obviously, it costs significantly more than this amount to coat the roofs with reflective coating, particularly because of the remote locations of these buildings. However, since the prefabricated roofs are already painted green at the factory, painting them a white (reflective) color would bring no additional cost. Hence, a reflective roof saves energy at no incremental cost.

  19. Multiplanar OCT/confocal ophthalmoscope in the clinic

    NASA Astrophysics Data System (ADS)

    Rosen, Richard B.; Podoleanu, Adrian Gh.; Rogers, John A.; Dunne, Shane; Dobre, George M.; Cucu, Radu G.; Jackson, David A.; Garcia, Patricia; Orlock, Dennis A.; Yannuzzi, Lawrence A.; Fisher, Yale; Nieto, Juan; Gentile, Ronald C.; Rosenthal, Jeanne L.; Muldoona, Thomas O.; Walsh, Joseph B.; Costa, Danielle; Huang, Sheau; Slakter, Jason; Spaide, Richard

    2003-07-01

    This paper demonstrates the clinical application of a multiplanar imaging system, which simultaneously acquires en-face (C-scan) OCT and corresponding confocal ophthalmoscopic images along with cross-sectional (B-scan) OCT at cursor designated locations on the confocal image. Advantages of the simultaneous OCT/confocal acquisition as well as the challenges of interpreting the C-scan OCT images are discussed. Variations in tissue inclination with respect to th coherence wave surface alters the sampling of structures within the depth in the retina, producing novel slice orientations which are often challenging to interpret. We evaluate for the first time the utility of C-scan OCT for a variety of pathologies including exudative ARMD, macular hole, central serous retinopathy, diabetic retinopathy, polypoidal choroidal vasculopathy and macular pucker. Several remarkable observations of new aspects of clinical anatomy were noted. The versatility of selective capture of C-scan OCT images and B-scan OCT images at precise points on the confocal image affords the clinician a more complete and interactive tool for 3D imaging of retinal pathology.

  20. Design and demonstration of multimodal optical scanning microscopy for confocal and two-photon imaging.

    PubMed

    Chun, Wanhee; Do, Dukho; Gweon, Dae-Gab

    2013-01-01

    We developed a multimodal microscopy based on an optical scanning system in order to obtain diverse optical information of the same area of a sample. Multimodal imaging researches have mostly depended on a commercial microscope platform, easy to use but restrictive to extend imaging modalities. In this work, the beam scanning optics, especially including a relay lens, was customized to transfer broadband (400-1000 nm) lights to a sample without any optical error or loss. The customized scanning optics guarantees the best performances of imaging techniques utilizing the lights within the design wavelength. Confocal reflection, confocal fluorescence, and two-photon excitation fluorescence images were obtained, through respective implemented imaging channels, to demonstrate imaging feasibility for near-UV, visible, near-IR continuous light, and pulsed light in the scanning optics. The imaging performances for spatial resolution and image contrast were verified experimentally; the results were satisfactory in comparison with theoretical results. The advantages of customization, containing low cost, outstanding combining ability and diverse applications, will contribute to vitalize multimodal imaging researches. PMID:23387653

  1. Confocal MicroscopyGuided Laser Ablation for Superficial and Early Nodular Basal Cell Carcinoma

    PubMed Central

    Chen, Chih-Shan Jason; Sierra, Heidy; Cordova, Miguel; Rajadhyaksha, Milind

    2014-01-01

    Importance Laser ablation is a rapid and minimally invasive approach for the treatment of superficial skin cancers, but efficacy and reliability vary owing to lack of histologic margin control. High-resolution reflectance confocal microscopy (RCM) may offer a means for examining margins directly on the patient. Observations We report successful elimination of superficial and early nodular basal cell carcinoma (BCC) in 2 cases-, using RCM imaging to guide Er-:YAG laser ablation. Three-dimensional (3-D) mapping is feasible with RCM-, to delineate the lateral border and thickness of the tumor. Thus, the surgeon may deliver laser fluence and passes with localized controlie, by varying the ablation parameters in sub-lesional areas with specificity that is governed by the 3-D topography of the BCC. We further demonstrate intra-operative detection of residual BCC after initial laser ablation and complete removal of remaining tumor by additional passes. Both RCM imaging and histologic sections confirm the final clearance of BCC. Conclusions and Relevance Confocal microscopy may enhance the efficacy and reliability of laser tumor ablation. This report represents a new translational application for RCM imaging, which, when combined with an ablative laser, may one day provide an efficient and cost-effective treatment for BCC. PMID:24827701

  2. ARES: a new reflective/emissive imaging spectrometer for terrestrial applications

    NASA Astrophysics Data System (ADS)

    Mueller, Andreas A.; Richter, Rolf; Habermeyer, Martin; Mehl, Harald; Dech, Stefan; Kaufmann, Hermann J.; Segl, Karl; Strobl, Peter; Haschberger, Peter; Bamler, Richard

    2003-04-01

    A new airborne imaging spectrometer introduced: the ARES (Airborne Reflective Emissive Spectrometer) to be built by Integrated Spectronics, Sydney, Australia, financed by DLR German Aerospace Center and the GFZ GeoResearch Center Potsdam, Germany, and will be available to the scientific community from 2003/2004 on. The ARES sensor will provide 160 channels in the solar reflective region (0.45-2.45 μm) and the thermal region (8-13 μm). It will consists of two separate coregistered optical systems for the reflective and thermal part of the spectrum. The spectral resolution is intended to be between 12 and 15 nm in the solar wavelength range and should reach 150nm in the thermal. ARES will be used mainly for environmental applications in terrestrial ecosystems. The thematic focus is thought to be on soil sciences, geology, agriculture and forestry. Limnologic applications should be possible but will not play a key role in the thematic applications. For all above mentioned key application scenarios the spectral response of soils, rocks, and vegetation as well as their mixtures contain the valuable information to be extracted and quantified. The radiometric requirements for the instrument have been modelled based on realistic application scenarios and account for the most demanding requirements of the three application fields: a spectral bandwidth of 15 nm in the 0.45-1.8 μm region, and 12 nm in the 2 - 2.45 μm region. The required noise equivalent radiance is 0.005, 0.003, and 0.003 mWcm-2sr-1μm-1 for the spectral regions 0.45-1 μm, 1 - 1.8 μm, and 2 - 2.45 μm, respectively.

  3. Diagnosis of microsporidial keratitis with in vivo confocal microscopy.

    PubMed

    Hsiao, Ya-Chuan; Tsai, I-Lun; Kuo, Chin-Tzu; Yang, Tsung-Lin

    2013-01-01

    As a rare cause of microbial keratitis, microsporidial keratitis (MK) is first described in a patient with acquired immunodeficiency syndrome. As increased use of topical steroid creates a localized immunosuppressive environment of the eyes, MK occurs more commonly than expected in immunocompetent patients nowadays. Owing to initial insidious growth of pathogens and nonspecific ocular symptoms of infected patients, its frequent misdiagnosis has posed a major clinical challenge in recent decades. Without appropriate treatments, MK can progress deeply into corneal stroma, anterior and posterior segments, subsequently deteriorating vision severely and ultimately requiring corneal transplant. Related risk factors for the occurrence of MK in immunocompetent individuals include contact lens wear, topical steroid use, previous corneal trauma, and a history of laser refractive surgery. The conventional standard of MK diagnosis is based on a tissue biopsy by superficial corneal scrapping. In vivo confocal laser scanning microscopy can obtain images through the cornea in a plane paralleling to the vertical axis. This approach provides an effective method of identifying tissue layers that correspond to corneal histologic structures. This current study investigates the efficacy of \\textit{in vivo} confocal laser scanning microscopy in diagnosing MK in immunocompetent patients. The clinical presentations of enrolled patients, including features of slit lamp biomicroscopy and the histopathological results of corneal scrapping, were described. In these patients, the confocal microscopy identified multiple small intracellular hyper-reflective dots in the cytoplasm of corneal epithelial cells and stromal keratocytes. Additionally, the confocal microscopic images clearly revealed the enhanced cytoplasm of cell with intracellular round hyper-reflective dots. The size and morphology of hyper-reflective dots were compatible with the spores of microsporidia found in corneal tissue. Moreover, vision recovered after topical use of antimicrobial medicine. This observation suggests that in vivo confocal laser scanning microscopy provides a rapid, non-invasive, and high resolution scheme for diagnosing MK. In addition to diminishing the risk of secondary infection from epithelial defect created by superficial debridement, this approach facilitates early diagnosis and appropriate treatments. Furthermore, from a series of images taken during the clinical courses, this method is highly promising for use in monitoring treatment effects and identifying the recurrence of MK. PMID:23507856

  4. Submillimeter Confocal Imaging Active Module

    NASA Technical Reports Server (NTRS)

    Hong, John; Mehdi, Imran; Siegel, Peter; Chattopadhyay, Goutam; Cwik, Thomas; Rowell, Mark; Hacker, John

    2009-01-01

    The term submillimeter confocal imaging active module (SCIAM) denotes a proposed airborne coherent imaging radar system that would be suitable for use in reconnaissance, surveillance, and navigation. The development of the SCIAM would include utilization and extension of recent achievements in monolithic microwave integrated circuits capable of operating at frequencies up to and beyond a nominal radio frequency of 340 GHz. Because the SCIAM would be primarily down-looking (in contradistinction to primarily side-looking), it could be useful for imaging shorter objects located between taller ones (for example, objects on streets between buildings). The SCIAM would utilize a confocal geometry to obtain high cross-track resolution, and would be amenable to synthetic-aperture processing of its output to obtain high along-track resolution. The SCIAM (see figure) would include multiple (two in the initial version) antenna apertures, separated from each other by a cross-track baseline of suitable length (e.g., 1.6 m). These apertures would both transmit the illuminating radar pulses and receive the returns. A common reference oscillator would generate a signal at a controllable frequency of (340 GHz + (Delta)f)/N, where (Delta)f is an instantaneous swept frequency difference and N is an integer. The output of this oscillator would be fed to a frequency- multiplier-and-power-amplifier module to obtain a signal, at 340 GHz + (Delta)f, that would serve as both the carrier signal for generating the transmitted pulses and a local-oscillator (LO) signal for a receiver associated with each antenna aperture. Because duplexers in the form of circulators or transmit/receive (T/R) switches would be lossy and extremely difficult to implement, the antenna apertures would be designed according to a spatial-diplexing scheme, in which signals would be coupled in and out via separate, adjacent transmitting and receiving feed horns. This scheme would cause the transmitted and received beams to be aimed in slightly different directions, and, hence, to not overlap fully on the targets on the ground. However, a preliminary analysis has shown that the loss of overlap would be small enough that the resulting loss in signal-to-noise ratio (SNR) would be much less than the SNR loss associated with the use of a 340-GHz T/R switch.

  5. Measured Energy Savings from the Application of Reflective Roofs in 3 AT and T Regeneration Buildings

    SciTech Connect

    Akbari, Hashen; Rainer, Leo

    2000-11-01

    Energy use and environmental parameters were monitored in three AT and T regeneration buildings during the summer of 2000. These buildings are constructed with concrete and are about 14.9 m2 (160 f2; 10x16 ft)in size. The buildings were initially monitored for about 1 1/2 months to establish a base condition. Then, the roofs of the buildings were painted with a white coating and the monitoring was continued. The original roof reflectances were about 26 percent; after the application of roof coatings the reflectivities increased to about 72 percent. In two of these buildings, we monitored savings of about 0.5kWh per day (8.6 kWh/m2 [0.8 kWh/ft2]). The third building showed a reduction in air-conditioning energy use of about 13kWh per day. These savings probably resulted from the differences in the performance (EER) of the two dissimilar AC units in this building. The estimated annual savings for two of the buildings are about 125kWh per year; at a cost of dollar 0.1/kWh, savings are about dollar 12.5 per year. Obviously, it costs significantly more than this amount to coat the roofs with reflective coating, particularly because of the remote location of the buildings. However, since the prefabricated roofs are already painted green at the factory, painting them with white (reflective) color would bring no additional cost. Hence the payback time for having reflective roofs is nil, and the reflective roofs save an accumulated 370kWh over 30 years of the life of the roof.

  6. Study and Development of near-Infrared Reflective and Absorptive Materials for Energy Saving Application

    NASA Astrophysics Data System (ADS)

    Cui, Yu Xing

    Near-Infrared (NIR) materials find applications in the field of energy saving. Both NIR reflective and absorptive materials can be used as energy saving materials with different working principles. The reflective materials can reflect the NIR light preventing it from being transmitted. Silver thin films are the best option as reflective films based on its reflectivity and cost. On the other hand, NIR absorptive materials can effectively convert the absorbed NIR light from sunlight to heat or electric energy. The first part of this research explored methods of preparing silver thin films that could be processed at low cost. The second part involved the design, synthesis and characterization of nickel coordination polymers as NIR absorptive materials. In part 1, different solution based methods of preparing silver thin films were studied. A silver nanoparticles solution was used to make thin film by a spray-pyrolysis process. Another method involved the surface activation with a fluoro-compound or silver nanoparticles followed by electroless silver plating on different substrates. Both methods could be processed at low cost. The obtained silver films showed NIR reflection of 5090% with transmission of 15-28% in the visible region. In part 2, two Nickel coordination polymers were explored. Tetraamino compounds were used as bridging ligands to increase the scope of electronic delocalization and metal-ligand orbital overlap which would reduce the energy gap to the NIR region. As a result, both polymers showed broad NIR absorption with maximum of 835 and 880 nm, respectively. In addition, the polymer showed NIR halochromism. This ground study pointed out both Ni coordination polymers as NIR absorptive materials with NIR halochromism.

  7. Integrated photoacoustic, confocal, and two-photon microscope.

    PubMed

    Rao, Bin; Soto, Florentina; Kerschensteiner, Daniel; Wang, Lihong V

    2014-03-01

    The invention of green fluorescent protein and other molecular fluorescent probes has promoted applications of confocal and two-photon fluorescence microscopy in biology and medicine. However, exogenous fluorescence contrast agents may affect cellular structure and function, and fluorescence microscopy cannot image nonfluorescent chromophores. We overcome this limitation by integrating optical-resolution photoacoustic microscopy into a modern Olympus IX81 confocal, two-photon, fluorescence microscope setup to provide complementary, label-free, optical absorption contrast. Automatically coregistered images can be generated from the same sample. Imaging applications in ophthalmology, developmental biology, and plant science are demonstrated. For the first time, in a familiar microscopic fluorescence imaging setting, this trimodality microscope provides a platform for future biological and medical discoveries. PMID:24589986

  8. In vivo 783-channel diffuse reflectance imaging system and its application

    NASA Astrophysics Data System (ADS)

    Yang, Joon-Mo; Han, Yong-Hui; Yoon, Gilwon; Ahn, Byung Soo; Lee, Byung-Cheon; Soh, Kwang-Sup

    2007-08-01

    A fiber-based reflectance imaging system was constructed to produce in vivo absorption spectroscopic images of biological tissues with diffuse light in the cw domain. The principal part of this system is the 783-channel fiber probe, composed of 253 illumination fibers and 530 detection fibers distributed in a 2020 mm square region. During illumination with the 253 illumination fibers, diffuse reflected lights are collected by the 530 detection fibers and recorded simultaneously as an image with an electron multiplying CCD camera for fast data acquisition. After signal acquisition, a diffuse reflectance image was reconstructed by applying the spectral normalization method we devised. To test the applicability of the spectral normalization, we conducted two phantom experiments with chicken breast tissue and white Delrin resin by using animal blood as an optical inhomogeneity. In the Delrin phantom experiment, we present images produced by two methods, spectral normalization and reference signal normalization, along with a comparison of the two. To show the feasibility of our system for biomedical applications, we took images of a human vein in vivo with the spectral normalization method.

  9. Confocal Fluorescence Microscopy of Mung Beanleaves

    NASA Astrophysics Data System (ADS)

    Chen, Zhiwei; Liu, Dongwu

    Recently, confocal microscope has become a routine technique and indispensable tool for cell biological studies and molecular investigations. The light emitted from the point out-of-focus is blocked by the pinhole and can not reach the detector, which is one of the critical features of the confocal microscope. In present studies, the probes acridine orange (AO) and rhodamine-123 were used to research stoma and mitochondria of mung bean leaves, respectively. The results indicated that the stomatal guard cells and mitochondria were clearly seen in epidermic tissue of mung bean leaves. Taken together, it is a good method to research plant cells with confocal microscope and fluorescence probes.

  10. 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.

  11. High laser damage threshold surface relief micro-structures for anti-reflection applications

    NASA Astrophysics Data System (ADS)

    Hobbs, Douglas S.; MacLeod, Bruce D.

    2008-01-01

    Microstructures built into the surfaces of an optic or window, are an effective replacement for thin-film coatings in anti-reflection (AR) and narrow-band filter applications. AR microstructures exhibit particularly noteworthy performance where an average reflection loss of less than 0.2% over a four-octave range (400-1800nm) has been demonstrated, and a loss of less than 0.03% is routinely achieved for narrow-band applications. Because AR micro-textures provide a gradual change in the refractive index at a material boundary, it is expected that light can propagate through the boundary without material damage at energy levels that are much higher than that found with thin-film interference coatings. Recently, it was shown that the laser induced damage threshold (LIDT) of an inexpensive borosilicate glass window containing AR microstructures was nearly 57 J/cm2 at 1064nm (20ns pulse). This LIDT is two to three times greater than the damage threshold of single-layer sol-gel AR coatings on fused silica often reported in the literature. The development of surface relief AR textures for use in high-energy laser applications is presented. Data from scanning electron microscope (SEM) analysis, reflection measurements, and LIDT testing, is shown for high performance AR microstructures fabricated in fused silica, and borosilicate glass. Results of LIDT testing at wavelengths ranging from the near ultraviolet through the near infrared confirm the initial result that AR microstructures can operate at pulsed laser power levels at least two times higher than thin-film coatings. For near infrared applications such as laser weapons and fiber optic communications requiring high performance AR, LIDT levels for AR microstructures in fused silica are found to be at least five times greater than conventional multi-layer thin film coatings. An initial surface absorption test at 1064nm shows that AR microstructures may also exhibit improved lifetimes within continuous wave laser systems.

  12. Big five personality traits reflected in job applicants' social media postings.

    PubMed

    Stoughton, J William; Thompson, Lori Foster; Meade, Adam W

    2013-11-01

    Job applicants and incumbents often use social media for personal communications allowing for direct observation of their social communications "unfiltered" for employer consumption. As such, these data offer a glimpse of employees in settings free from the impression management pressures present during evaluations conducted for applicant screening and research purposes. This study investigated whether job applicants' (N=175) personality characteristics are reflected in the content of their social media postings. Participant self-reported social media content related to (a) photos and text-based references to alcohol and drug use and (b) criticisms of superiors and peers (so-called "badmouthing" behavior) were compared to traditional personality assessments. Results indicated that extraverted candidates were prone to postings related to alcohol and drugs. Those low in agreeableness were particularly likely to engage in online badmouthing behaviors. Evidence concerning the relationships between conscientiousness and the outcomes of interest was mixed. PMID:23790360

  13. Large Area Microencapsulated Reflective Guest-Host Liquid Crystal Displays and Their Applications

    NASA Astrophysics Data System (ADS)

    Nakai, Yutaka; Tanaka, Masao; Enomoto, Shintaro; Iwanaga, Hiroki; Hotta, Aira; Kobayashi, Hitoshi; Oka, Toshiyuki; Kizaki, Yukio; Kidzu, Yuko; Naito, Katsuyuki

    2002-07-01

    We have developed reflective liquid crystal displays using microencapsulated guest-host liquid crystals, whose size was sufficiently large for viewing documents. A high-brightness image can be realized because there is no need for polarizers. Easy fabrication processes, consisting of screen-printing of microencapsulated liquid crystal and film adhesion, have enabled the realization of thinner and lighter cell structures. It has been confirmed that the display is tolerant of the pressures to which it would be subject in actual use. The optimization of fabrication processes has enabled the realization of reflectance uniformity in the display area and reduction of the driving voltage. Our developed display is suitable for portable information systems, such as electronic book applications.

  14. Detection of forests using mid-IR reflectance: An application for aerosol studies

    SciTech Connect

    Kaufman, Y.J. . Goddard Space Flight Center); Remer, L.A. )

    1994-05-01

    The detection of dark, dense vegetation is an important step in the remote sensing of aerosol loading. Current methods that employ the red (0.64 [mu]m) and the near-IR (0.84 [mu]m) regions are unsatisfactory in that the presence of aerosols in the scene distorts the apparent reflectance in the visible and near-IR ranges of the spectrum. The mid-IR spectral region is also sensitive to vegetation due to the absorption of liquid water in the foliage, but is not sensitive to the presence of most aerosols (except for dust). Therefore, mid-IR channels on the AVHRR and EOS-MODIS (e.g., the 3.75 [mu]m or the 3.95 [mu]m channels) have a unique potential for the remote sensing of dark, dense vegetation, particularly in the presence of biomass burning smoke or industrial/urban haze. The reflective part of the 3.75 [mu]m channel ([rho][sub 3.75]) is applied to images of the AVHRR over the eastern US. This channel was found to be correlated to reflectance at 0.64 [mu]m ([rho][sub 0.64]), less sensitive to haze than the visible channel and superior to both the 0.64 [mu]m reflectance and the normalized difference vegetation index (NDVI) to determine forest pixels in an image. However, its application to monitor the seasonal evolution of vegetation is presently questionable. For the purpose of the remote sensing of aerosol over dark, dense vegetation, it is proposed that the dark, dense vegetation be determined from [rho][sub 3.75] < 0.025. These findings may have further implications for other specific applications of the remote sensing of vegetation in hazy atmospheres.

  15. Confocal unstable-resonator semiconductor laser.

    PubMed

    Salzman, J; Lang, R; Larson, A; Yariv, A

    1986-08-01

    GaAs/GaAlAs heterostructure lasers with a monolithic confocal unstable resonator were demonstrated. The curved mirrors satisfying the confocal condition were fabricated by etching. Close to threshold, the lasers operate in a single lateral mode with a nearly collimated output beam. A single-lobe far-field intensity distribution as narrow as 1.9 degrees full width at half maximum was measured. PMID:19738671

  16. Confocal unstable-resonator semiconductor laser

    SciTech Connect

    Salzman, J.; Lang, R.; Larson, A.; Yariv

    1986-08-01

    GaAs/GaA1As heterostructure lasers with a monolithic confocal unstable resonator were demonstrated. The curved mirrors satisfying the confocal condition were fabricated by etching. Close to threshold, the lasers operate in a single lateral mode with a nearly collimated out beam. A single-lobe far-field intensity distribution as narrow as 1.9 deg full width at half maximum was measured.

  17. Confocal unstable-resonator semiconductor laser

    SciTech Connect

    Salzman, J.; Lang, R.; Larson, A.; Yariv, A.

    1986-08-01

    GaAs/GaAlAs heterostructure lasers with a monolithic confocal unstable resonator were demonstrated. The curved mirrors satisfying the confocal condition were fabricated by etching. Close to threshold, the lasers operate in a single lateral mode with a nearly collimated output beam. A single-lobe far-field intensity distribution as narrow as 1.9/sup 0/ full width at half maximum was measured.

  18. A novel photodiode for reflectance pulse oximetry in low-power applications.

    PubMed

    Haahr, Rasmus G; Duun, Sune; Birkelund, Karen; Raahauge, Palle; Petersen, Peter; Dam, Henrik; Nrgaard, Lars; Thomsen, Erik V

    2007-01-01

    The amount of light collected is crucial for low-power applications of pulse oximetry. In this work a novel ring-shaped backside photodiode has been developed for a wearable reflectance pulse oximeter. The photodiode is proven to work with a dual LED with wavelengths of 660 nm and 940 nm. For the purpose of continuously monitoring vital signs of a human, a temperature sensor is integrated onto the chip containing the photodiode. This biomedical multisensor chip is made for integration into "the Electronic Patch", an autonomous monitoring system for humans. PMID:18002464

  19. Confocal Raman Microspectroscopy of Oral Streptococci

    NASA Astrophysics Data System (ADS)

    Beier, Brooke D.

    Raman spectroscopy has been used in a variety of applications throughout the field of biomedical optics. It has the ability to acquire chemically-specific information in a non-invasive manner, without the need for exogenous markers. This makes it useful in the identification of bacterial species, as well as in the study of tissues and other cells. In this work, a species identification model has been created in order to discriminate between the oral bacterial species Streptococcus sanguinis and Streptococcus mutans. These are two of the most prevalent species within the human mouth and their relative concentrations can be an indicator of a patient's oral health and risk of tooth decay. They are predominantly found within plaque on the tooth's surface. To study a simplified model for dental plaque, we have examined S. sanguinis and S. mutans grown in biofilm forms. Raman spectroscopy has been implemented here through a confocal microscope. The optical system has been equipped with computationally controlled stages to allow for automated scanning, including autofocusing to probe a consistent depth within a sample. A spectrum has been acquired from each position within a scan and sent for spectral preprocessing before being submitted for species identification. This preprocessing includes an algorithm that has been developed to remove fluorescence features from known contaminants within the confocal volume, to include signal from a fluorescent substrate. Species classification has been accomplished using a principal component score-fed logistic regression model constructed from a variety of biofilm samples that have been transferred and allowed to dry, as might occur with the study of plaque samples. This binary classification model has been validated on other samples with identical preparations. The model has also been transferred to determine the species of hydrated biofilms studied in situ. Artificially mixed biofilms have been examined to test the spatial capabilities of our species identification model. The work included in this thesis has been focused on the study of S. sanguinis and S. mutans, though the principles could easily be applied to the study of other biofilms.

  20. Confocal microlaparoscope for imaging the fallopian tube

    NASA Astrophysics Data System (ADS)

    Wu, Tzu-Yu; Schafer, Rachel; Rouse, Andrew R.; Gmitro, Arthur F.

    2012-02-01

    Recent evidence suggests that epithelial ovarian cancer may originate in the fimbriated end of the fallopian tube1. Unlike many other cancers, poor access to the ovary and fallopian tubes has limited the ability to study the progression of this deadly disease and to diagnosis it during the early stage when it is most amenable to therapy. We have previously reported on a rigid confocal microlaparoscope system that is currently undergoing a clinical trial to image the epithelial surface of the ovary2. In order to gain in vivo access to the fallopian tubes we have developed a new confocal microlaparoscope with an articulating distal tip. The new instrument builds upon the technology developed for the existing confocal microlaparoscope. It has an ergonomic handle fabricated by a rapid prototyping printer. While maintaining compatibility with a 5 mm trocar, the articulating distal tip of the instrument consists of a 2.2 mm diameter bare fiber bundle catheter with automated dye delivery for fluorescence imaging. This small and flexible catheter design should enable the confocal microlaparoscope to image early stage ovarian cancer arising inside the fallopian tube. Early ex vivo mages of human fallopian tube and in vivo imaging results from recent open surgeries using the rigid confocal microlaparoscope system are presented. Ex vivo images from animal models using the new articulating bare fiber system are also presented. These high quality images collected by the new flexible system are similar in quality to those obtained from the epithelial surface of ovaries with the rigid clinical confocal microlaparoscope.

  1. Confocal microscopy of skin cancers: Translational advances toward clinical utility

    PubMed Central

    Rajadhyaksha, Milind

    2014-01-01

    Recent advances in translational research in and technology for confocal microscopy of skin cancers, toward clinical applications, are described. Advances in translational research are in diagnosis of melanoma in vivo, pre-operative mapping of lentigo maligna melanoma margins to guide surgery and intra-operative imaging of residual basal cell carcinomas to guide shave-biopsy. Advances in technology include mosaicing microscopy for detection of basal cell carcinomas in large areas of excised tissue, toward rapid pathology-at-the-bedside, and development of small, simple and low-cost line-scanning confocal microscopes for worldwide use in diverse primary healthcare settings. Current limitations and future opportunities and challenges for both clinicians and technologists are discussed. PMID:19964286

  2. Chromatic confocal microscopy using staircase diffractive surface.

    PubMed

    Rayer, Mathieu; Mansfield, Daniel

    2014-08-10

    A chromatic confocal microscope (CCM) is a high-dynamic-range noncontact distance measurement sensor; it is based on a hyperchromatic lens. The vast majority of commercial CCMs use refractive-based chromatic dispersion to chromatically code the optical axis. This approach significantly limits the range of applications and performance of the CCM. In order to be a suitable alternative to a laser triangulation gauge and laser encoder, the performance of the CCM must be improved. In this paper, it is shown how hybrid aspheric diffractive (HAD) lenses can bring the CCM to its full potential by increasing the dynamic range by a factor of 2 and the resolution by a factor of 5 while passively athermizing and increasing the light throughput efficiency of the optical head [M. Rayer, U.S. patent 1122052.2 (2011)]. The only commercially suitable manufacturing process is single-point diamond turning. However, the optical power carried by the diffractive side of a hybrid aspheric diffractive lens is limited by the manufacturing process. A theoretical study of manufacturing losses has revealed that the HAD configuration with the highest diffraction efficiency is for a staircase diffractive surface (SDS). SDS lenses have the potential to reduce light losses associated with manufacturing limits by a factor of 5 without increasing surface roughness, allowing scalar diffraction-limited optical design with a diffractive element. PMID:25320920

  3. Confocal photothermal flow cytometry in vivo

    NASA Astrophysics Data System (ADS)

    Zharov, Vladimir P.; Galanzha, Ekaterina I.; Ferguson, Scott; Tuchin, Valery V.

    2005-04-01

    The new experimental design of an integrated flow cytometry (FC) is presented, combining high-resolution transmission digital microscopy (TDM) with photothermal (PT), photoacoustic (PA), and fluorescence techniques. We used phantom in vitro to verify this concept with moving living cells, and micro- and nanoparticles. The transistion in vivo study was realized by using unique rat mesentery model for real-time detection of circulating red and white blood cells. The adaptation of confocal schematics to PT microscopy to provide 3-D measurement is discussed. We demonstrated that simulataneous transmission, PT and fluorescent imaging provide the basis for nanodiagnostics and nanotherpeutics in vivo with gold nanoparticles as PT probes and sensitizers as well as identification cells with specific absorbing endogenous and exogenous structures. First attempt to use in parallel PA methods with detection PA signals from single live cells are presented. Potential applications of integrated FC are discussed, including identification of selected cells with different natural absorptive properties, characterization of bioflow (e.g., velocity profile), and PT nanotherapeutics and nanodiagnostics of metastatic cells with gold nanoparticles.

  4. Precision measurement of acoustic reflectivity for a scanning acoustic microscope that measures amplitude and phase: applicability in biology

    NASA Astrophysics Data System (ADS)

    Ngwa, Wilfred; Knauth, Stefan; Laforsch, Christian; Grill, Wolfgang

    2004-07-01

    Sundry image processing schemes for a high-frequency (1.2 GHz) scanning acoustic microscope that measures amplitude and phase are presented. Particular emphasis is paid to the acquisition of precise in-focus information, like the acoustic reflectivity, of three-dimensional microscopic objects. The brightness of a surface element of any object under observation depends not solely on the reflectivity. It is also affected by the tilt angle of the surface with respect to the axis of the microscope. Vector microscopy with synchronous observation of the phase and amplitude has been employed to determine the tilt from the image in phase contrast and correct the observed brightness in the image in amplitude contrast accordingly. Additionally three-dimensional scanning has been used to determine the maximum intensity obtained for confocal positioning of any surface element. The relevance of such schemes for truly quantitative measurements in biology is demonstrated, with results that have led to the ascertainment of phenotypic plasticity in daphnia (waterfleas) species.

  5. [Application of near-infrared reflectance spectroscopy in grass breeding with space flight mutagenesis].

    PubMed

    Ren, Wei-Bo; Han, Jian-Guo; Zhang, Yun-Wei; Guo, Hui-Qin

    2008-02-01

    Near infrared reflectance spectroscopy is a new fast and efficient analysis method. It has been wildly used in many areas such as evaluation of feedstuff, assessment of soil fertilizer and so on. In the present paper, the principle, technique method and merits of NIRS were introduced. The potential application of NIRS in grass breeding with space flight mutagenesis was discussed in areas such as analysis of grass nutrition, estimate of secondary metabolism compounds, forecast of disease and insects resistance, and evaluation of abiotic stress. The conclusion is that application of NIRS in grass breeding with space mutagenesis is significant in both academic and technical areas because it not only improves the efficiency of mutation selection but helps uncover the mechanism of space mutation breeding. PMID:18479009

  6. Ghost reflections of Gaussian beams in anamorphic optical systems with an application to Michelson interferometer.

    PubMed

    Abd El-Maksoud, Rania H

    2016-02-20

    In this paper, a methodology is developed to model and analyze the effect of undesired (ghost) reflections of Gaussian beams that are produced by anamorphic optical systems. The superposition of these beams with the nominal beam modulates the nominal power distribution at the recording plane. This modulation may cause contrast reduction, veiling parts of the nominal image, and/or the formation of spurious interference fringes. The developed methodology is based on synthesizing the beam optical paths into nominal and ghost optical beam paths. Similar to the nominal beam, we present the concept that each ghost beam is characterized by a beam size, wavefront radius of curvature, and Gouy phase in the paraxial regime. The nominal and ghost beams are sequentially traced through the system and formulas for estimating the electric field magnitude and phase of each ghost beam at the recording plane are presented. The effective electric field is the addition of the individual nominal and ghost electric fields. Formulas for estimating Gouy phase, the shape of the interference fringes, and the central interference order are introduced. As an application, the theory of the formation of the interference fringes by Michelson interferometer is presented. This theory takes into consideration the ghost reflections that are formed by the beam splitter. To illustrate the theory and to show its wide applicability, simulation examples that include a Mangin mirror, a Michelson interferometer, and a black box optical system are provided. PMID:26906582

  7. Multidepth imaging by chromatic dispersion confocal microscopy

    NASA Astrophysics Data System (ADS)

    Olsovsky, Cory A.; Shelton, Ryan L.; Saldua, Meagan A.; Carrasco-Zevallos, Oscar; Applegate, Brian E.; Maitland, Kristen C.

    2012-03-01

    Confocal microscopy has shown potential as an imaging technique to detect precancer. Imaging cellular features throughout the depth of epithelial tissue may provide useful information for diagnosis. However, the current in vivo axial scanning techniques for confocal microscopy are cumbersome, time-consuming, and restrictive when attempting to reconstruct volumetric images acquired in breathing patients. Chromatic dispersion confocal microscopy (CDCM) exploits severe longitudinal chromatic aberration in the system to axially disperse light from a broadband source and, ultimately, spectrally encode high resolution images along the depth of the object. Hyperchromat lenses are designed to have severe and linear longitudinal chromatic aberration, but have not yet been used in confocal microscopy. We use a hyperchromat lens in a stage scanning confocal microscope to demonstrate the capability to simultaneously capture information at multiple depths without mechanical scanning. A photonic crystal fiber pumped with a 830nm wavelength Ti:Sapphire laser was used as a supercontinuum source, and a spectrometer was used as the detector. The chromatic aberration and magnification in the system give a focal shift of 140?m after the objective lens and an axial resolution of 5.2-7.6?m over the wavelength range from 585nm to 830nm. A 400x400x140?m3 volume of pig cheek epithelium was imaged in a single X-Y scan. Nuclei can be seen at several depths within the epithelium. The capability of this technique to achieve simultaneous high resolution confocal imaging at multiple depths may reduce imaging time and motion artifacts and enable volumetric reconstruction of in vivo confocal images of the epithelium.

  8. An addressable confocal microscope for functional imaging of neuronal activity

    NASA Astrophysics Data System (ADS)

    Bansal, Vivek

    2005-07-01

    The study of computation occurring in single neurons and small networks of interconnected neurons is often limited by (1) the number of sites that can be simultaneously probed with electrophysiology tools such as patch pipettes and (2) the recording speed of fluorescence imaging tools such as confocal or multiphoton microscopy. Even in the line scan mode of galvanometer-based scanners, where one scan dimension is sacrificed to gain overall speed, the effective frame rate is limited to less than 1 kHz with no flexibility in site selection. To overcome these limitations and allow the study of many sites throughout the dendritic arbor, we have developed an addressable confocal laser-scanning microscope that permits recording from user-selected sites-of-interest at high frame rates, in addition to conventional full frame imaging. Our system utilizes acousto-optic deflectors (AODs) in the illumination pathway to allow for rapid user-defined positioning of a focused laser spot. However, since AODs rely on diffraction to steer a laser beam, they cannot effectively descan the fluorescence emission spectrum as done in mirror-based systems which utilize reflection; this prevents the use of a stationary pinhole as a spatial filter. Instead, we implement an addressable spatial filter using a digital micromirror device (DMD) in conjunction with the AODs to achieve confocality. A registration algorithm synchronizes the AODs and DMD such that point illumination and point detection are always colocalized in conjugate image planes. The current version of the confocal system has a spatial resolution of 1 mum. Furthermore, by letting the user tailor which sites are visited, we have shown that recordings can be made at an aggregate frame rate of 40 kHz. We have successfully demonstrated that the system is capable of optical sectioning and thus exhibits the main advantage of a confocal microscope for light-scattering biological tissue. This property was used to create three-dimensional reconstructions of fluorescently labeled test specimens. Additionally, we have used the system to record intracellular calcium transients using the fluorescent calcium indicator Oregon Green BAPTA-1. The transients were a result of back-propagating action potentials elicited via 1 nA current injections in cultured hippocampal neurons from wild-type mice.

  9. Dye-Enhanced Multimodal Confocal Imaging of Brain Cancers

    NASA Astrophysics Data System (ADS)

    Wirth, Dennis; Snuderl, Matija; Sheth, Sameer; Curry, William; Yaroslavsky, Anna

    2011-04-01

    Background and Significance: Accurate high resolution intraoperative detection of brain tumors may result in improved patient survival and better quality of life. The goal of this study was to evaluate dye enhanced multimodal confocal imaging for discriminating normal and cancerous brain tissue. Materials and Methods: Fresh thick brain specimens were obtained from the surgeries. Normal and cancer tissues were investigated. Samples were stained in methylene blue and imaged. Reflectance and fluorescence signals were excited at 658nm. Fluorescence emission and polarization were registered from 670 nm to 710 nm. The system provided lateral resolution of 0.6 μm and axial resolution of 7 μm. Normal and cancer specimens exhibited distinctively different characteristics. H&E histopathology was processed from each imaged sample. Results and Conclusions: The analysis of normal and cancerous tissues indicated clear differences in appearance in both the reflectance and fluorescence responses. These results confirm the feasibility of multimodal confocal imaging for intraoperative detection of small cancer nests and cells.

  10. ARES: a new reflective/emissive imaging spectrometer for terrestrial applications

    NASA Astrophysics Data System (ADS)

    Mueller, Andreas; Richter, Rolf; Habermeyer, Martin; Mehl, Harald; Dech, Stefan; Kaufmann, Hermann J.; Segl, Karl; Strobl, Peter; Haschberger, Peter; Bamler, Richard

    2004-10-01

    Airborne imaging spectrometers have a history of about 20 years starting with the operation of AIS in 1982. During the following years, many other instruments were built and successfully operated, e.g., AVIRIS, CASI, DAIS-7915, and HyMap. Since imaging spectrometers cover a spectral region with a large number of narrow contiguous bands they are able to retrieve the spectral reflectance signature of the earth allowing tasks such as mineral identification and abundance mapping, monitoring of vegetation properties, and assessment of water constituents. An essential prerequisite for the evaluation of imaging spectrometer data is a stable spectral and radiometric calibration. Although a considerable progress has been achieved in this respect over the last two decades, this issue is still technically challenging today, especially for low-to-medium cost instruments. This paper introduces a new airborne imaging spectrometer, the ARES (Airborne Reflective Emissive Spectrometer) to be built by Integrated Spectronics, Sydney, Australia, and co-financed by DLR German Aerospace Center and the GFZ GeoResearch Center Potsdam, Germany. The instrument shall feature a high performance over the entire optical wavelength range and will be available to the scientific community from 2006 on. The ARES sensor will provide 150 channels in the solar reflective region (0.47-2.42 μm) and the thermal region (8.1-12.1 μm). It will consist of two co-registered optical systems for the reflective and thermal part of the spectrum. The spectral resolution is intended to be between 12 and 16 nm in the solar wavelength range and should reach 150 nm in the thermal range. ARES will be used mainly for environmental applications in terrestrial ecosystems. The thematic focus is thought to be on soil sciences, geology, agriculture and forestry. Limnologic applications should be possible but will not play a key role in the thematic applications. For all above mentioned key application scenarios, the spectral response of soils, rocks, and vegetation as well as their mixtures contain the valuable information to be extracted and quantified. The radiometric requirements for the instrument have been modeled based on realistic application scenarios and account for the most demanding requirements of the three application fields: a spectral bandwidth of 16 nm in the 0.47-1.8 μm region, and 12 nm in the 2.02 - 2.42 μm region. The required noise equivalent radiance is 0.05, 0.03, and 0.02 Wm-2sr-1μm-1 for the spectral regions 0.47- 0.89 μm, 0.89 - 1.8 μm, and 2.02 - 2.42 μm, respectively. In the thermal region similar simulations have been carried out. Results suggest a required noise equivalent temperature of 0.05 K for the retrieval of emissivity spectra in the desired accuracy. Nevertheless, due to system restrictions these requirements might have to be reduced to 0.1 K in the wavelength range between 8.1 and 10 μm and 0.1-0.2 K from 10 to 12.1 μm.

  11. Optical characterization and confocal fluorescence imaging of mechanochromic acrylate polymers

    NASA Astrophysics Data System (ADS)

    van Horn, M.; Smith, P.; Mason, B. P.; Hemmer, J. R.; Read de Alaniz, J.; Hooper, J. P.; Osswald, S.

    2015-01-01

    The development of mechanochromic molecules has opened new pathways for the study of localized stress and failure in polymers. Their application as stress or temperature diagnostics, however, requires suitable measurement techniques capable of detecting the force- and temperature-sensitive chemical species with high spatial resolution. Confocal imaging techniques offer excellent spatial resolution but the energy input during these measurements can itself affect the activation state of the mechanochromic species. Here, we present a systematic study of the effects of laser-based imaging on the activation and fluorescence behavior of mechanochromic spiropyran (SP) integrated into poly(methyl acrylate) (PMA) and poly(methyl methacrylate) matrices using a confocal Raman microspectrometer. Localized stress and temperature activation were studied by means of high-rate compressive loading and dynamic fracture. Laser illumination of SP in PMA revealed a strong excitation wavelength- and power-dependence. Suitable correction functions were established and used to account for the observed laser effects. The presented study demonstrates that confocal imaging using conventional Raman spectrometers is a powerful characterization tool for localized stress analysis in mechanochromic polymers, offering quantifiable information on the activation state with high spatial resolution. However, laser-mechanophore interactions must be well understood and effects of laser excitation and exposure times must be taken into consideration when interpreting the obtained results.

  12. 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. PMID:25717185

  13. Ultrafast superresolution fluorescence imaging with spinning disk confocal microscope optics

    PubMed Central

    Hayashi, Shinichi; Okada, Yasushi

    2015-01-01

    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 microtubules, mitochondria, lysosomes, and endosomes were observed with temporal resolutions of 30100 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. PMID:25717185

  14. Multi-spectral confocal microendoscope for in-vivo imaging

    NASA Astrophysics Data System (ADS)

    Rouse, Andrew Robert

    The concept of in-vivo multi-spectral confocal microscopy is introduced. A slit-scanning multi-spectral confocal microendoscope (MCME) was built to demonstrate the technique. The MCME employs a flexible fiber-optic catheter coupled to a custom built slit-scan confocal microscope fitted with a custom built imaging spectrometer. The catheter consists of a fiber-optic imaging bundle linked to a miniature objective and focus assembly. The design and performance of the miniature objective and focus assembly are discussed. The 3mm diameter catheter may be used on its own or routed though the instrument channel of a commercial endoscope. The confocal nature of the system provides optical sectioning with 3mum lateral resolution and 30mum axial resolution. The prism based multi-spectral detection assembly is typically configured to collect 30 spectral samples over the visible chromatic range. The spectral sampling rate varies from 4nm/pixel at 490nm to 8nm/pixel at 660nm and the minimum resolvable wavelength difference varies from 7nm to 18nm over the same spectral range. Each of these characteristics are primarily dictated by the dispersive power of the prism. The MCME is designed to examine cellular structures during optical biopsy and to exploit the diagnostic information contained within the spectral domain. The primary applications for the system include diagnosis of disease in the gastro-intestinal tract and female reproductive system. Recent data from the grayscale imaging mode are presented. Preliminary multi-spectral results from phantoms, cell cultures, and excised human tissue are presented to demonstrate the potential of in-vivo multi-spectral imaging.

  15. Confocal microlaparoscope for imaging the fallopian tube

    NASA Astrophysics Data System (ADS)

    Wu, Tzu-Yu; Rouse, Andrew R.; Chambers, Setsuko K.; Hatch, Kenneth D.; Gmitro, Arthur F.

    2014-11-01

    Recent evidence suggests that ovarian cancer can originate in the fallopian tube. Unlike many other cancers, poor access to the ovary and fallopian tubes has limited the ability to study the progression of this deadly disease and to diagnosis it during the early stage when it is most amenable to therapy. A rigid confocal microlaparoscope system designed to image the epithelial surface of the ovary in vivo was previously reported. A new confocal microlaparoscope with an articulating distal tip has been developed to enable in vivo access to human fallopian tubes. The new microlaparoscope is compatible with 5-mm trocars and includes a 2.2-mm-diameter articulating distal tip consisting of a bare fiber bundle and an automated dye delivery system for fluorescence confocal imaging. This small articulating device should enable the confocal microlaparoscope to image early stage ovarian cancer arising inside the fallopian tube. Ex vivo images of animal tissue and human fallopian tube using the new articulating device are presented along with in vivo imaging results using the rigid confocal microlaparoscope system.

  16. Performance of "Moth Eye" Anti-Reflective Coatings for Solar Cell Applications

    SciTech Connect

    Clark, E.; Kane, M.; Jiang, P.

    2011-03-14

    An inexpensive, effective anti-reflective coating (ARC) has been developed at the University of Florida to significantly enhance the absorption of light by silicon in solar cells. This coating has nano-scale features, and its microstructure mimics that of various night active insects (e.g. a moth's eye). It is a square array of pillars, each about 700 nm high and having a diameter of about 300 nm. Samples of silicon having this coating were exposed either to various combinations of either elevated temperature and humidity or to gamma irradiation ({sup 60}Co) at the Savannah River National Laboratory, or to a broad spectrum ultraviolet light and to a 532 nm laser light at the University of Florida. The anti-reflective properties of the coatings were unaffected by any of these environmental stresses, and the microstructure of the coating was also unaffected. In fact, the reflectivity of the gamma irradiated ARC became lower (advantageous for solar cell applications) at wavelengths between 400 and 1000 nm. These results show that this coating is robust and should be tested in actual systems exposed to either weather or a space environment. Structural details of the ARCs were studied to optimize their performance. Square arrays performed better than hexagonal arrays - the natural moth-eye coating is indeed a square array. The optimal depth of the templated nanopillars in the ARC was investigated. A wet etching technology for ARC formation was developed that would be less expensive and much faster than dry etching. Theoretical modeling revealed that dimple arrays should perform better than nipple arrays. A method of fabricating both dimple and nipple arrays having the same length was developed, and the dimple arrays performed better than the nipple arrays, in agreement with the modeling. The commercial viability of the technology is quite feasible, since the technology is scalable and inexpensive. This technology is also compatible with current industrial fabrication of solar cells.

  17. [Quantitative measurement of induced skin reddening using optical reflection spectroscopy--methodology and clinical application].

    PubMed

    Smesny, S; Riemann, S; Riehemann, S; Bellemann, M E; Sauer, H

    2001-10-01

    Optical reflection spectroscopy is a simple and quick method for the quantification of colour intensity, and is thus suitable for the determination of changes in skin reddening (erythema) due to local vasodilatation. To quantify the time course of this erythema, the oxyhaemoglobin absorption double peak with maxima at 542 and 577 nm is an appropriate parameter. A compact handheld optical spectrometer makes the technique applicable to clinical use, an example being the niacin patch test described herein. This noninvasive test provides information about the cell membrane metabolism via the skin flush induced by niacin (vitamin B3) and mediated by prostaglandin. The aim of this study was to adapt optical reflection spectroscopy to the requirements of the clinical niacin patch test. To that end, we investigated 60 healthy volunteers. Analysis of the spectroscopic data with regard to physiological covariables of niacin sensitivity revealed faster and more intense erythema in females--a gender effect that to our knowledge has not previously been reported. In the light of these results, the findings of other researchers based on semi-quantitative test methods should be reassessed, with consideration given to the gender effect. PMID:11721583

  18. Radius measurement by laser confocal technology.

    PubMed

    Yang, Jiamiao; Qiu, Lirong; Zhao, Weiqian; Zhang, Xin; Wang, Xu

    2014-05-01

    A laser confocal radius measurement (LCRM) method is proposed for high-accuracy measurement of the radius of curvature (ROC). The LCRM uses the peak points of confocal response curves to identify the cat eye and confocal positions precisely. It then accurately measures the distance between these two positions to determine the ROC. The LCRM also uses conic fitting, which significantly enhances measurement accuracy by restraining the influences of environmental disturbance and system noise on the measurement results. The experimental results indicate that LCRM has a relative expanded uncertainty of less than 10 ppm for both convex and concave spheres. Thus, LCRM is a feasible method for ROC measurements with high accuracy and concise structures. PMID:24921872

  19. Confocal multiview light-sheet microscopy

    PubMed Central

    Medeiros, Gustavo de; Norlin, Nils; Gunther, Stefan; Albert, Marvin; Panavaite, Laura; Fiuza, Ulla-Maj; Peri, Francesca; Hiiragi, Takashi; Krzic, Uros; Hufnagel, Lars

    2015-01-01

    Selective-plane illumination microscopy has proven to be a powerful imaging technique due to its unsurpassed acquisition speed and gentle optical sectioning. However, even in the case of multiview imaging techniques that illuminate and image the sample from multiple directions, light scattering inside tissues often severely impairs image contrast. Here we combine multiview light-sheet imaging with electronic confocal slit detection implemented on modern camera sensors. In addition to improved imaging quality, the electronic confocal slit detection doubles the acquisition speed in multiview setups with two opposing illumination directions allowing simultaneous dual-sided illumination. Confocal multiview light-sheet microscopy eliminates the need for specimen-specific data fusion algorithms, streamlines image post-processing, easing data handling and storage. PMID:26602977

  20. Confocal multiview light-sheet microscopy

    NASA Astrophysics Data System (ADS)

    Medeiros, Gustavo De; Norlin, Nils; Gunther, Stefan; Albert, Marvin; Panavaite, Laura; Fiuza, Ulla-Maj; Peri, Francesca; Hiiragi, Takashi; Krzic, Uros; Hufnagel, Lars

    2015-11-01

    Selective-plane illumination microscopy has proven to be a powerful imaging technique due to its unsurpassed acquisition speed and gentle optical sectioning. However, even in the case of multiview imaging techniques that illuminate and image the sample from multiple directions, light scattering inside tissues often severely impairs image contrast. Here we combine multiview light-sheet imaging with electronic confocal slit detection implemented on modern camera sensors. In addition to improved imaging quality, the electronic confocal slit detection doubles the acquisition speed in multiview setups with two opposing illumination directions allowing simultaneous dual-sided illumination. Confocal multiview light-sheet microscopy eliminates the need for specimen-specific data fusion algorithms, streamlines image post-processing, easing data handling and storage.

  1. Total-Internal-Reflection Platforms for Chemical and Biological Sensing Applications

    NASA Astrophysics Data System (ADS)

    Sapsford, Kim E.

    Sensing platforms based on the principle of total internal reflection (TIR) represent a fairly mature yet still expanding and exciting field of research. Sensor development has mainly been driven by the need for rapid, stand-alone, automated devices for application in the fields of clinical diagnosis and screening, food and water safety, environmental monitoring, and chemical and biological warfare agent detection. The technologies highlighted in this chapter are continually evolving, taking advantage of emerging advances in microfabrication, lab-on-a-chip, excitation, and detection techniques. This chapter describes many of the underlying principles of TIR-based sensing platforms and additionally focusses on planar TIR fluorescence (TIRF)-based chemical and biological sensors.

  2. Application of Neutron Reflectivity for Studies of Biomolecular Structures and Functions at Interfaces

    SciTech Connect

    Johs, Alexander; Liang, Liyuan; Gu, Baohua; Ankner, John Francis; Wang, Wei

    2009-01-01

    Structures and functions of cell membranes are of central importance in understanding processes such as cell signaling, chemotaxis, redox transformation, biofilm formation, and mineralization occurring at interfaces. This chapter provides an overview of the application of neutron reflectivity (NR) as a unique tool for probing biomolecular structures and mechanisms as a first step toward understanding protein protein, protein lipid, and protein mineral interactions at the membrane substrate interfaces. Emphasis is given to the review of existing literature on the assembly of biomimetic membrane systems, such as supported membranes for NR studies, and demonstration of model calculations showing the potential of NR to elucidate molecular fundamentals of microbial cell mineral interactions and structure functional relationships of electron transport pathways. The increased neutron flux afforded by current and upcoming neutron sources holds promise for elucidating detailed processes such as phase separation, formation of microdomains, and membrane interactions with proteins and peptides in biological systems.

  3. Confocal acoustic radiation force optical coherence elastography using a ring ultrasonic transducer

    PubMed Central

    Qi, Wenjuan; Li, Rui; Ma, Teng; Kirk Shung, K.; Zhou, Qifa; Chen, Zhongping

    2014-01-01

    We designed and developed a confocal acoustic radiation force optical coherence elastography system. A ring ultrasound transducer was used to achieve reflection mode excitation and generate an oscillating acoustic radiation force in order to generate displacements within the tissue, which were detected using the phase-resolved optical coherence elastography method. Both phantom and human tissue tests indicate that this system is able to sense the stiffness difference of samples and quantitatively map the elastic property of materials. Our confocal setup promises a great potential for point by point elastic imaging in vivo and differentiation of diseased tissues from normal tissue. PMID:24737920

  4. Confocal acoustic radiation force optical coherence elastography using a ring ultrasonic transducer

    SciTech Connect

    Qi, Wenjuan; Li, Rui; Ma, Teng; Kirk Shung, K.; Zhou, Qifa; Chen, Zhongping

    2014-03-24

    We designed and developed a confocal acoustic radiation force optical coherence elastography system. A ring ultrasound transducer was used to achieve reflection mode excitation and generate an oscillating acoustic radiation force in order to generate displacements within the tissue, which were detected using the phase-resolved optical coherence elastography method. Both phantom and human tissue tests indicate that this system is able to sense the stiffness difference of samples and quantitatively map the elastic property of materials. Our confocal setup promises a great potential for point by point elastic imaging in vivo and differentiation of diseased tissues from normal tissue.

  5. Power and limits of laser scanning confocal microscopy.

    PubMed

    Laurent, M; Johannin, G; Gilbert, N; Lucas, L; Cassio, D; Petit, P X; Fleury, A

    1994-01-01

    In confocal microscopy, the object is illuminated and observed so as to rid the resulting image of the light from out-of-focus planes. Imaging may be performed in the reflective or in the fluorescence mode. Confocal microscopy allows accurate and non-destructive optical sectioning in a plane perpendicular or parallel to the optical axis of the microscope. Further digital three-dimensional treatments of the data may be performed so as to visualize the specimen from a variety of angles. Several examples illustrating each of these possibilities are given. Three-dimensional reconstitution of nuclear components using a cubic representation and a ray-tracing based method are also given. Instrumental and experimental factors can introduce some bias into the acquisition of the 3-D data set: self-shadowing effects of thick specimens, spherical aberrations due to the sub-optimum use of the objective lenses and photo-bleaching processes. This last phenomenon is the one that most heavily hampers the quantitative analysis needed for a 3-D reconstruction. We delineate each of these problems and indicate to what extent they can be solved. Some tips are given for the practice of confocal microscope and image recovery: how to determine empirically the thickness of the optical slices, how to deal with extreme contrasts in an image, how to prevent artificial flattening of the specimens. Finally, future prospects in the field are outlined. Particular mention of the use of pulsed lasers is made as they may be an alternative to UV-lasers and a possible means to attenuate photodamage to biological specimens. PMID:8087072

  6. Confocal Microscopy for Modeling Electron Microbeam Irradiation of Skin

    SciTech Connect

    Miller, John H.; Chrisler, William B.; Wang, Xihai; Sowa, Marianne B.

    2011-08-01

    For radiation exposures employing targeted sources such as particle microbeams, the deposition of energy and dose will depend on the spatial heterogeneity of the spample. Although cell structural variations are relatively minor for two-dimensional cell cultures, they can vary significantly for fully differential tissues. Employing high-resolution confocal microscopy, we have determined the spatial distribution, size, and shape of epidermal kerantinocyte nuclei for the full-thickness EpiDerm skin model (MatTek, Ashland, VA). Application of these data to claculate the microdosimetry and microdistribution of energy deposition by an electron microbeam is discussed.

  7. Atherosclerotic plaque detection by confocal Brillouin and Raman microscopies

    NASA Astrophysics Data System (ADS)

    Meng, Zhaokai; Basagaoglu, Berkay; Yakovlev, Vladislav V.

    2015-02-01

    Atherosclerosis, the development of intraluminal plaque, is a fundamental pathology of cardiovascular system and remains the leading cause of morbidity and mortality worldwide. Biomechanical in nature, plaque rupture occurs when the mechanical properties of the plaque, related to the morphology and viscoelastic properties, are compromised, resulting in intraluminal thrombosis and reduction of coronary blood flow. In this report, we describe the first simultaneous application of confocal Brillouin and Raman microscopies to ex-vivo aortic wall samples. Such a non-invasive, high specific approach allows revealing a direct relationship between the biochemical and mechanical properties of atherosclerotic tissue.

  8. [Near infrared reflectance spectroscopy (NIRS) and its application in the determination for the quality of animal feed and products].

    PubMed

    Wang, Li; Meng, Qing-Xiang; Ren, Li-Ping; Yang, Jian-Song

    2010-06-01

    Near-infrared reflectance spectroscopy (NIRS) has been the most rapidly developing and noticeable spectrographic analytical technique in recent years. The determining principle and progresses of near-infrared reflectance spectroscopy are presented briefly. It mainly includes the progresses in pre-processing technique and analyzing model of near-infrared reflectance spectroscopy. Two pre-processing techniques, including differential coefficient-dealt with technique, the signal-smoothing technique, and four analyzing models of near-infrared spectroscopy, including the multiplied lined regression (MLR), principal component analysis (PCA), partial least squares (PLS), and artificial nerve network (ANN). The application of near-infrared reflectance spectroscopy to the first time. The investigation of reviewed papers shows that the near-infrared reflectance spectroscopy is widely applied in feed analysis and animal products analysis because of its rapidness, non-destruction and non-pollution. The near infrared reflectance spectroscopy has been used to determine the feed common ingredient, such as dry matter, crude protein, crude fiber, crude fat and so on, micro-components including amino acid, vitamin, and noxious components, and to determine the physical and chemical properties of animal products which including egg, mutton, beef and pork. Details of the analytical characteristics of feed and animal products described in the reviewed papers are given. New trends and limits to the application of near-infrared reflectance spectroscopy in these fields are also discussed. PMID:20707134

  9. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: AXIAL RESOLUTION

    EPA Science Inventory

    Abstract

    Confocal Microscopy System Performance: Axial resolution.
    Robert M. Zucker, PhD

    Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research and Development, U.S. Environmental Protection Agency, Re...

  10. Confocal microscopy imaging of solid tissue

    EPA Science Inventory

    Confocal laser scanning microscopy (CLSM) is a technique that is capable of generating serial sections of whole-mount tissue and then reassembling the computer acquired images as a virtual 3-dimensional structure. In many ways CLSM offers an alternative to traditional sectioning ...

  11. Active confocal imaging for visual prostheses.

    PubMed

    Jung, Jae-Hyun; Aloni, Doron; Yitzhaky, Yitzhak; Peli, Eli

    2015-06-01

    There are encouraging advances in prosthetic vision for the blind, including retinal and cortical implants, and other "sensory substitution devices" that use tactile or electrical stimulation. However, they all have low resolution, limited visual field, and can display only few gray levels (limited dynamic range), severely restricting their utility. To overcome these limitations, image processing or the imaging system could emphasize objects of interest and suppress the background clutter. We propose an active confocal imaging system based on light-field technology that will enable a blind user of any visual prosthesis to efficiently scan, focus on, and "see" only an object of interest while suppressing interference from background clutter. The system captures three-dimensional scene information using a light-field sensor and displays only an in-focused plane with objects in it. After capturing a confocal image, a de-cluttering process removes the clutter based on blur difference. In preliminary experiments we verified the positive impact of confocal-based background clutter removal on recognition of objects in low resolution and limited dynamic range simulated phosphene images. Using a custom-made multiple-camera system based on light-field imaging, we confirmed that the concept of a confocal de-cluttered image can be realized effectively. PMID:25448710

  12. Vibrometry using a chromatic confocal sensor

    NASA Astrophysics Data System (ADS)

    Berkovic, G.; Zilberman, S.; Shafir, E.; Cohen-Sabban, J.

    2014-05-01

    We demonstrate vibrometry using a chromatic confocal sensor which measures displacements with 0.1 μm resolution at a rate of 10 kHz. This technique was used to study the vibration of a musical tuning fork with a resonance at 523 Hz. Other examples presented include vibration of water waves and multiple point vibrometry of a vibrating steel rod.

  13. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: LASER POWER MEASUREMENTS

    EPA Science Inventory

    Laser power abstract
    The reliability of the confocal laser-scanning microscope (CLSM) to obtain intensity measurements and quantify fluorescence data is dependent on using a correctly aligned machine that contains a stable laser power. The laser power test appears to be one ...

  14. Mapping of dendritic lesions in patients with herpes simplex keratitis using in vivo confocal microscopy

    PubMed Central

    Yokogawa, Hideaki; Kobayashi, Akira; Mori, Natsuko; Sugiyama, Kazuhisa

    2015-01-01

    Purpose To produce a two-dimensional reconstruction map of dendritic lesions in patients with herpes simplex keratitis (HSK) using in vivo confocal microscopy. Methods Four eyes of four patients (mean 65.8 years) with HSK presenting with a dendritic lesion were enrolled. Slit-lamp biomicroscopy and in vivo laser confocal microscopy were performed. Acquired confocal images at the level of the epithelium were arranged and mapped into subconfluent montages. Changes in the shape and degree of light reflection of abnormal cells and deposits around dendritic lesions as well as other corneal layers were qualitatively evaluated. Results Mapping of dendritic lesion was successful in all cases, and the subconfluent montages clearly showed the larger image of dendritic lesion. In all cases, the dendritic lesion consisted of hyperreflective irregular epithelial cells, and was surrounded by distorted and elongated epithelial cells. In three cases, hyperreflective deposits were noted at the midline of the lesion. The corneal stroma showed a hyperreflective honeycomb pattern. In two cases, inflammatory cells were observed at the level of endothelial cell layer. Conclusion Mapping of dendritic lesions in patients with HSK was successful in all patients using in vivo confocal microscopy. Cellular level observation of dendritic lesion at a relatively larger magnification may help understand the in vivo morphological change of HSK. Further study in more patients with HSK and nonherpetic dendritic lesion is needed to utilize confocal microscopy images in differential diagnosis and follow-up of the epithelial lesions with dendrite. PMID:26445524

  15. Confocal mosaicing microscopy of basal-cell carcinomas ex vivo: progress in digital staining to simulate histology-like appearance

    NASA Astrophysics Data System (ADS)

    Bini, Jason; Spain, James; Nehal, Kishwer; Hazelwood, Vikki; DiMarzio, Charles; Rajadhyaksha, Milind

    2011-03-01

    Confocal mosaicing microscopy enables rapid imaging of large areas of fresh tissue, without the processing that is necessary for conventional histology. Using acridine orange (1 milliMolar, 20 seconds) to stain nuclei, basal cell carcinomas were detected in fluorescence confocal mosaics of Mohs surgical excisions with sensitivity of 96.6% and specificity of 89.2%. A possible barrier toward clinical acceptance is that confocal mosaics are based on a single mode of contrast and appear in grayscale, whereas histology is based on two (hematoxylin for nuclei, eosin for cellular cytoplasm and dermis) and appears purple-and-pink. Toward addressing this barrier, we report progress in developing a multispectral analytical model for digital staining: fluorescence confocal mosaics, which show only nuclei, are digitally stained purple and overlaid on reflectance confocal mosaics, which show only cellular cytoplasm and dermis, and digitally stained pink, to mimic the appearance of histology. Comparison of digitally stained confocal mosaics by our Mohs surgeon to the corresponding Mohs histology shows good correlation for normal and tumor detail. Digitally stained confocal mosaicing microscopy may allow direct examination of freshly excised tissue and serve as an adjunct for rapid pathology at-the-bedside.

  16. Miniature objective lens with variable focus for confocal endomicroscopy

    PubMed Central

    Kim, Minkyu; Kang, DongKyun; Wu, Tao; Tabatabaei, Nima; Carruth, Robert W.; Martinez, Ramses V; Whitesides, George M.; Nakajima, Yoshikazu; Tearney, Guillermo J.

    2014-01-01

    Spectrally encoded confocal microscopy (SECM) is a reflectance confocal microscopy technology that can rapidly image large areas of luminal organs at microscopic resolution. One of the main challenges for large-area SECM imaging in vivo is maintaining the same imaging depth within the tissue when patient motion and tissue surface irregularity are present. In this paper, we report the development of a miniature vari-focal objective lens that can be used in an SECM endoscopic probe to conduct adaptive focusing and to maintain the same imaging depth during in vivo imaging. The vari-focal objective lens is composed of an aspheric singlet with an NA of 0.5, a miniature water chamber, and a thin elastic membrane. The water volume within the chamber was changed to control curvature of the elastic membrane, which subsequently altered the position of the SECM focus. The vari-focal objective lens has a diameter of 5 mm and thickness of 4 mm. A vari-focal range of 240 μm was achieved while maintaining lateral resolution better than 2.6 μm and axial resolution better than 26 μm. Volumetric SECM images of swine esophageal tissues were obtained over the vari-focal range of 260 μm. SECM images clearly visualized cellular features of the swine esophagus at all focal depths, including basal cell nuclei, papillae, and lamina propria. PMID:25574443

  17. Imaging intracellular protein dynamics by spinning disk confocal microscopy

    PubMed Central

    Stehbens, Samantha; Pemble, Hayley; Murrow, Lindsay; Wittmann, Torsten

    2012-01-01

    The palette of fluorescent proteins has grown exponentially over the last decade, and as a result live imaging of cells expressing fluorescently tagged proteins is becoming more and more main stream. Spinning disk confocal microscopy (SDC) is a high speed optical sectioning technique, and a method of choice to observe and analyze intracellular fluorescent protein dynamics at high spatial and temporal resolution. In an SDC system, a rapidly rotating pinhole disk generates thousands of points of light that scan the specimen simultaneously, which allows direct capture of the confocal image with low noise scientific grade cooled charged-coupled device (CCD) cameras, and can achieve frame rates of up 1000 frames per second. In this chapter we describe important components of a state-of-the-art spinning disk system optimized for live cell microscopy, and provide a rationale for specific design choices. We also give guidelines how other imaging techniques such as total internal reflection (TIRF) microscopy or spatially controlled photoactivation can be coupled with SDC imaging, and provide a short protocol on how to generate cell lines stably expressing fluorescently tagged proteins by lentivirus-mediated transduction. PMID:22264541

  18. Cosmetic assessment of the human hair by confocal microscopy.

    PubMed

    Hadjur, Christophe; Daty, Grard; Madry, Genevive; Corcuff, Pierre

    2002-01-01

    The optical sectioning property of the confocal microscope offers a breakthrough from the classic observation of the hair in a scanning electron microscope (SEM). Confocal microscopy requires minimal sampling preparation, and the hair can be observed in its natural environment with less damage than by other microscopic methods such as SEM. While used in the reflection mode, the true morphology of the cuticle and the various exogenous deposits at the surface can be identified and quantified. This relatively noninvasive, nondestructive technique is routinely used by us to monitor the efficiency of cleansing shampoos, to assess the homogeneity of layering polymers, and to evaluate the changes they induce in the optical properties of the hair surface in terms of opacity, transparency, and brilliancy. A second important field of investigation uses the fluorescence channel which reveals the internal structure of the hair. Fluorescent probes (rhodamine and its derivatives) demonstrate the routes of penetration and outline the geometry of cortical cells and of the medulla according to their lipophilic or hydrophilic properties. A volume rendering of a hair cylinder provides a better understanding of the interrelationships between cuticle cells, cortical cells, and the medullar channel. This recent technology is becoming an invaluable tool for the cosmetic assessment of the hair. PMID:11998902

  19. Statistical characterization of engineered tissues using confocal mosaic technology

    NASA Astrophysics Data System (ADS)

    Levitz, David; Ardeshiri, Ardalan; Ahmed, Jabeer; Gareau, Daniel S.; Jacques, Steven L.

    2010-02-01

    Characterization of engineered tissues using optical methods often involves tradeoff between the fraction of total volume that is imaged and the spatial resolution. The limitation is not technological but rather practical, having more to do with effective probe designs and computer memory storage for large datasets. In this paper, we propose using confocal mosaicing, a technique used to characterize large volumes of excisioned biopsies from Mohs surgeries, to characterizing collagen gels. This technique stitches together high-resolution 3D images of a series adjacent millimeter sized regions that collectively make up areas that are ~cm2. Image acquisition time is approximately 5 min. The resulting high-resolution images closely resemble hematoxylin and eosin histological sections, only obtained without the time-consuming embedding and sectioning steps. Disk-shaped collagen gels that are 1 ml volume and ~1.5 cm diameter were prepared with smooth muscle cells and imaged at days 1 and 5. Using the digital staining technique, we were able to survey the spatial distribution of cells in the hydrogel and assess spatial heterogeneity in 3D from the fluorescence data. The reflectance data provided information on collagen fibril structure and matrix remodeling by the cells. Digital staining presented the data in a way that is easily interpreted by tissue engineers. Altogether, we believe confocal mosaicing and digital staining represents an important technological novelty that significantly advances nondestructive optical evaluation of engineered tissues.

  20. Characterization of lased enamel organic matrix using confocal microscopy

    NASA Astrophysics Data System (ADS)

    Hsu, Chin-Ying S.; Girija, Veerappan

    2001-10-01

    In the past decade several studies have demonstrated the increased acid resistance in enamel demineralization after laser irradiation. However, the exact mechanism of action to this effect still remains a speculation. Recently, the role of organic matrix was revealed to be significant in the laser-induced inhibition of enamel demineralization. The aim of the present study was to characterize the lipid component of organic matrix in mature lazed enamel and unlazed enamel histochemically using a hydrophobic fluorescent probe with Confocal Laser Scanning Microscope (CLSM). Partial decalcification of thin enamel sections was carried out using 0.5 M of EDTA in a stainless steel grid for 5 hours, following fixation with 3.5% paraformaldehyde. Thereafter the sections were stained with Nile red coupled with CLSM. The intensity of the light reflection was analyzed under the same conditions for all specimens, ruling out the autofluorescence in the control sections. Confocal imaging revealed a diffuse and increased fluorescence of the lipid stain in the lazed areas suggesting that the swelling and coating of organic matrix on the surface of enamel crystals in the peri and interprismatic spaces is rendering the increased acid resistance. These findings will substantiate the proposed organic blocking theory in partially explaining the laser-induced prevention of enamel demineralization.

  1. Applicability of a Diffuse Reflectance Infrared Fourier Transform handheld spectrometer to perform in situ analyses on Cultural Heritage materials

    NASA Astrophysics Data System (ADS)

    Arrizabalaga, Iker; Gómez-Laserna, Olivia; Aramendia, Julene; Arana, Gorka; Madariaga, Juan Manuel

    2014-08-01

    This work studies the applicability of a Diffuse Reflectance Infrared Fourier Transform handheld device to perform in situ analyses on Cultural Heritage assets. This portable diffuse reflectance spectrometer has been used to characterise and diagnose the conservation state of (a) building materials of the Guevara Palace (15th century, Segura, Basque Country, Spain) and (b) different 19th century wallpapers manufactured by the Santa Isabel factory (Vitoria-Gasteiz, Basque Country, Spain) and by the well known Dufour and Leroy manufacturers (Paris, France), all of them belonging to the Torre de los Varona Castle (Villanañe, Basque Country, Spain). In all cases, in situ measurements were carried out and also a few samples were collected and measured in the laboratory by diffuse reflectance spectroscopy (DRIFT) in order to validate the information obtained by the handheld instrument. In the analyses performed in situ, distortions in the diffuse reflectance spectra can be observed due to the presence of specular reflection, showing the inverted bands caused by the Reststrahlen effect, in particular on those IR bands with the highest absorption coefficients. This paper concludes that the results obtained in situ by a diffuse reflectance handheld device are comparable to those obtained with laboratory diffuse reflectance spectroscopy equipment and proposes a few guidelines to acquire good spectra in the field, minimising the influence caused by the specular reflection.

  2. Applicability of a Diffuse Reflectance Infrared Fourier Transform handheld spectrometer to perform in situ analyses on Cultural Heritage materials.

    PubMed

    Arrizabalaga, Iker; Gmez-Laserna, Olivia; Aramendia, Julene; Arana, Gorka; Madariaga, Juan Manuel

    2014-08-14

    This work studies the applicability of a Diffuse Reflectance Infrared Fourier Transform handheld device to perform in situ analyses on Cultural Heritage assets. This portable diffuse reflectance spectrometer has been used to characterise and diagnose the conservation state of (a) building materials of the Guevara Palace (15th century, Segura, Basque Country, Spain) and (b) different 19th century wallpapers manufactured by the Santa Isabel factory (Vitoria-Gasteiz, Basque Country, Spain) and by the well known Dufour and Leroy manufacturers (Paris, France), all of them belonging to the Torre de los Varona Castle (Villanae, Basque Country, Spain). In all cases, in situ measurements were carried out and also a few samples were collected and measured in the laboratory by diffuse reflectance spectroscopy (DRIFT) in order to validate the information obtained by the handheld instrument. In the analyses performed in situ, distortions in the diffuse reflectance spectra can be observed due to the presence of specular reflection, showing the inverted bands caused by the Reststrahlen effect, in particular on those IR bands with the highest absorption coefficients. This paper concludes that the results obtained in situ by a diffuse reflectance handheld device are comparable to those obtained with laboratory diffuse reflectance spectroscopy equipment and proposes a few guidelines to acquire good spectra in the field, minimising the influence caused by the specular reflection. PMID:24747846

  3. Light localization properties of biological cells via confocal imaging

    NASA Astrophysics Data System (ADS)

    Sahay, Peeyush; Ghimire, Hemendra M.; Almabadi, Huda; Pradhan, Prabhakar

    2015-03-01

    Detection and characterization of the spatial refractive index fluctuations of very weakly disordered optical dielectric media has ample applications in various fields ranging from soft condensed matter to biological research. We report a study of the submicron scale degree of the structural disorder of heterogeneous weakly disordered optical dielectric media, such as biological cells, by quantifying their submicron scale light-localization properties. Confocal microscopy is used to construct disordered optical lattices of these dielectric media. Light-localization properties are studied by the statistical analysis of the inverse participation ratio (IPR) of the localized eigenfunctions of these optical lattices at the submicron scales. The method is described and its importance is highlighted. As one of the applications, we demonstrate that using this method, different types of normal and cancerous cells can be distinguished by quantifying the structural disorder inside the cells via their confocal micrographs. Other potential applications of the technique to characterize weakly disordered media, as well as biological cells, in particular cancer detection, are also discussed. NIH and University of Memphis.

  4. Improving the optical contrast of backscattering signal in reflectance-based imaging with gold nanoshells

    NASA Astrophysics Data System (ADS)

    Kah, James C. Y.; Chow, Tzu-Hao; Olivo, Malini C.; Ng, Beng-Koon; Gulam, Razul S.; Sheppard, Colin J. R.

    2007-07-01

    The application of gold nanoparticles as a contrast agent in optical bioimaging is well appreciated, but limited to a narrow excitation range due to its rather invariable optical resonance typically at 520 nm. Compared to gold nanoparticles, the optical response of gold nanoshells can be tuned to match the higher excitation wavelength of many promising clinical reflectance-based imaging modalities such as the optical coherence tomography (OCT). In this study, we demonstrate the tunability of gold nanoshells to improve the optical contrast of backscattering signal under confocal reflectance microscopy and OCT. The gold nanoshells were synthesized and conjugated to antibodies for in vitro demonstration of their selective optical contrast in cancer cells over normal cells under the confocal reflectance microscopy. The OCT signals from these gold nanoshells were compared to that from bare silica cores and intrinsic tissue scattering using 1% Intralipid. We have shown that gold nanoshells are able to elicit an optical contrast to discriminate between cancerous and normal cells under the confocal reflectance microscopy based on differences in molecular markers expression. Compared to bare silica core, the presence of the gold shell is able to effect a higher backscattered OCT signal with an apparent contrast over 1% Intralipid. This contrast can be made to be dependent on the molecular marker expression with antibody specificity.

  5. Application of a wide-field phantom eye for optical coherence tomography and reflectance imaging

    PubMed Central

    Corcoran, Anthony; Muyo, Gonzalo; van Hemert, Jano; Gorman, Alistair; Harvey, Andrew R.

    2015-01-01

    Optical coherence tomography (OCT) and reflectance imaging are used in clinical practice to measure the thickness and transverse dimensions of retinal features. The recent trend towards increasing the field of view (FOV) of these devices has led to an increasing significance of the optical aberrations of both the human eye and the device. We report the design, manufacture and application of the first phantom eye that reproduces the off-axis optical characteristics of the human eye, and allows the performance assessment of wide-field ophthalmic devices. We base our design and manufacture on the wide-field schematic eye, [Navarro, R. J. Opt. Soc. Am. A, 1985, 2.] as an accurate proxy to the human eye and enable assessment of ophthalmic imaging performance for a ±70∘ external FOV. We used multi-material 3D-printed retinal targets to assess imaging performance of the following ophthalmic instruments: the Optos 200Tx, Heidelberg Spectralis, Zeiss FF4 fundus camera and Optos OCT SLO and use the phantom to provide an insight into some of the challenges of wide-field OCT. PMID:26740737

  6. The application of visible wavelength reflectance hyperspectral imaging for the detection and identification of blood stains.

    PubMed

    Li, Bo; Beveridge, Peter; O'Hare, William T; Islam, Meez

    2014-12-01

    Current methods of detection and identification of blood stains rely largely on visual examination followed by presumptive tests such as Kastle-Meyer, Leuco-malachite green or luminol. Although these tests are useful, they can produce false positives and can also have a negative impact on subsequent DNA tests. A novel application of visible wavelength reflectance hyperspectral imaging has been used for the detection and positive identification of blood stains in a non contact and non destructive manner on a range of coloured substrates. The identification of blood staining was based on the unique visible absorption spectrum of haemoglobin between 400 and 500 nm. Images illustrating successful discrimination of blood stains from nine red substances are included. It has also been possible to distinguish between blood and approximately 40 other reddish stains. The technique was also successfully used to detect latent blood stains deposited on white filter paper at dilutions of up to 1 in 512 folds and on red tissue at dilutions of up to 1 in 32 folds. Finally, in a blind trial, the method successfully detected and identified a total of 9 blood stains on a red T-shirt. PMID:25498930

  7. Confocal micro-Raman spectroscopy of black soap films

    NASA Astrophysics Data System (ADS)

    Lecourt, B.; Capelle, F.; Adamietz, F.; Malaplate, A.; Blaudez, D.; Kellay, H.; Turlet, J. M.

    1998-01-01

    Black soap films from aqueous solutions of sodium dodecyl sulphate are studied by micro-Raman confocal spectroscopy. At the end of the draining process films of different thicknesses are obtained depending on the experimental conditions: Working in a closed humidified chamber leads to common black films while, under evaporation or in the presence of electrolyte, Newton black films are observed. From the Raman spectra of these films, quantitative information is deduced about the conformational and lateral order of the aliphatic surfactant chains, as well as the thickness of the residual water layer. More accurate measurements of the thickness of these ultimate films have been carried out by transmission ellipsometry and their effective refractive index measured by Brewster angle reflectivity. The thinner films present higher molecular organization and their aqueous core exhibits unusual spectral features.

  8. Assessing the tissue-imaging performance of confocal microscope architectures via Monte Carlo simulations.

    PubMed

    Chen, Ye; Wang, Danni; Liu, Jonathan T C

    2012-11-01

    Various confocal microscope architectures have been developed for in vivo tissue imaging, including single-axis confocal (SAC) and dual-axis confocal (DAC) configurations utilizing both point-scanning (PS) and line-scanning (LS) approaches. While it is known that these design variations lead to tradeoffs in imaging performance, a quantitative comparison of the imaging performance of these configurations in highly turbid media would be of value. Here, we perform Monte Carlo simulations to evaluate the optical-sectioning capability of these various confocal microscope architectures in reflectance mode. In particular, we investigate the axial and transverse responses of these configurations to reflective targets at various depths within a homogenous scattering medium. We find that the DAC-PS configuration results in superior rejection of multiply scattered background light compared to all other configurations, followed in performance by the SAC-PS, the DAC-LS, and then the SAC-LS. Line scanning with both the DAC and SAC configurations leads to photon crosstalk between pixels. However, at shallow depths, the axial and transverse resolution of all configurations is maintained in a homogeneous scattering medium. PMID:23114341

  9. [Confocal laser scanning microscopy: a deep look into the cell].

    PubMed

    Roderfeld, M; Matern, S; Roeb, E

    2003-11-28

    Confocal laser scanning microscopy (CLSM) is a powerful technology for assaying biomolecular distribution and dynamics in cells and tissues. Innovations in CLSM-techniques, coupled with the development of new dyes and genetically encoded indicators, have increased both in vitro and in vivo imaging approaches. CLSM has had wide application in basic science, but little impact so far on medical investigations. As a "cutting edge" technology CLSM has proved to be a valuable tool in some areas within medical applications including pathology, dermatology, ophthalmology and research in various other fields of medicine. This paper gives an overview about the wide range of CLSM-applications and shows the enormous potential of this technology in medical research and development. PMID:14648437

  10. Bidirectional Reflectance of icy Samples: Application to water ice detection on the Moon and Mercury

    NASA Astrophysics Data System (ADS)

    Yoldi Martnez de Mandojana, Z.; Pommerol, A.; Jost, B.; Poch, O.; Gouman, J.; Thomas, N.

    2015-10-01

    The reflectance of water ice and lunar regolith simulant (JSC-1AF) mixtures has been measured under different geometries. We have found that considerable amounts of water ice can be mixed within the soil without producing any noticeable photometric signature, as the relation between the reflectance and the amount of ice in the sample is strongly non-linear. Some reflectance models have been tested to try to reproduce this non-linearity.

  11. Probe based confocal laser endomicroscopy of the pancreatobiliary system

    PubMed Central

    Almadi, Majid A; Neumann, Helmut

    2015-01-01

    AIM: To review applications of confocal laser endomicroscopy (CLE) in pancreatobiliary lesions and studies that assessed training and interpretation of images. METHODS: A computerized literature search was performed using OVID MEDLINE, EMBASE, Cochrane library, and the ISI Web of Knowledge from 1980 to October 2014. We also searched abstracts from major meetings that included the Digestive Disease Week, Canadian Digestive Disease Week and the United European Gastroenterology Week using a combination of controlled vocabulary and text words related to pCLE, confocal, endomicroscopy, probe-based confocal laser endomicroscopy, and bile duct to identify reports of trials. In addition, recursive searches and cross-referencing was performed, and manual searches of articles identified after the initial search was also completed. We included fully published articles and those in abstract form. Given the relatively recent introduction of CLE we included randomized trials and cohort studies. RESULTS: In the evaluation of indeterminate pancreatobiliary strictures CLE with ERCP compared to ERCP alone can increase the detection of cancerous strictures with a sensitivity of (98% vs 45%) and has a negative predictive value (97% vs 69%), but decreased the specificity (67% vs 100%) and the positive predictive value (71% vs 100%) when compared to index pathology. Modifications in the classification systems in indeterminate biliary strictures have increased the specificity of pCLE from 67% to 73%. In pancreatic cystic lesions there is a need to develop similar systems to interpret and characterize lesions based on CLE images obtained. The presence of superficial vascular network predicts serous cystadenomas accurately. Also training in acquiring and interpretation of images is feasible in those without any prior knowledge in CLE in a relatively simple manner and computer-aided diagnosis software is a promising innovation. CONCLUSION: The role of pCLE in the evaluation of pancreatobiliary disorders might be better suited for those with an intermediate and low probability. PMID:26640347

  12. In-vivo multi-spectral confocal microscopy

    NASA Astrophysics Data System (ADS)

    Rouse, Andrew R.; Udovich, Joshua A.; Gmitro, Arthur F.

    2005-03-01

    A multi-spectral confocal microendoscope (MCME) for in-vivo imaging has been developed. The MCME employs a flexible fiber-optic catheter coupled to a slit-scan confocal microscope with an imaging spectrometer. The catheter consists of a fiber-optic imaging bundle linked to a miniature objective and focus assembly. The focus mechanism allows for imaging to a maximum tissue depth of 200 microns. The 3mm diameter catheter may be used on its own or routed though the instrument channel of a commercial endoscope. The confocal nature of the system provides optical sectioning with 3 micron lateral resolution and 30 micron axial resolution. The system incorporates two laser sources and is therefore capable of simultaneous acquisition of spectra from multiple dyes using dual excitation. The prism based multi-spectral detection assembly is typically configured to collect 30 spectral samples over the visible range. The spectral sampling rate varies from 4nm/pixel at 490nm to 8nm/pixel at 660nm and the minimum resolvable wavelength difference varies from 8nm to 16nm over the same spectral range. Each of these characteristics are primarily dictated by the dispersion characteristics of the prism. The MCME is designed to examine cellular structures during optical biopsy and to exploit the diagnostic information contained within the spectral domain. The primary applications for the system include diagnosis of disease in the gastro-intestinal tract and female reproductive system. In-vitro, and ex-vivo multi-spectral results are presented.

  13. Laser differential confocal paraboloidal vertex radius measurement.

    PubMed

    Yang, Jiamiao; Qiu, Lirong; Zhao, Weiqian; Shen, Yang; Jiang, Hongwei

    2014-02-15

    This Letter proposes a laser differential confocal paraboloidal vertex radius measurement (DCPRM) method for the high-accuracy measurement of the paraboloidal vertex radius of curvature. DCPRM constructs an autocollimation vertex radius measurement light path for the paraboloid by placing a reflector in the incidence light path. This technique is based on the principle that a paraboloid can aim a parallel beam at its focus without aberration and uses differential confocal positioning technology to identify the paraboloid focus and vertex accurately. Measurement of the precise distance between these two positions is achieved to determine the paraboloid vertex radius. Preliminary experimental results indicate that DCPRM has a relative expanded uncertainty of less than 0.001%. PMID:24562218

  14. Technology and New Directions in Professional Development: Applications of Digital Video, Peer Review, and Self-Reflection

    ERIC Educational Resources Information Center

    Collins, James L.; Cook-Cottone, Catherine P.; Robinson, Judith Schick; Sullivan, Roberta R.

    2004-01-01

    Pedagogical applications of technology--including digital video, course management systems, online discussion forums, and CD-ROM com-pilations--for development of professional skills were tested in three distinct professional graduate programs. Role-playing, peer review, and self-reflection instructional methods were technologically enhanced by

  15. MEMS-Based Dual Axes Confocal Microendoscopy

    PubMed Central

    Piyawattanametha, Wibool; Wang, Thomas D.

    2011-01-01

    We demonstrate a miniature, near-infrared microscope (λ = 785 nm) that uses a novel dual axes confocal architecture. Scalability is achieved with post-objective scanning, and a MEMS mirror provides real time (>4 Hz) in vivo imaging. This instrument can achieve sub-cellular resolution with deep tissue penetration and large field of view. An endoscope-compatible version can image digestive tract epithelium to guide tissue biopsy and monitor therapy. PMID:22190845

  16. Quasi-simultaneous OCT/confocal imaging

    NASA Astrophysics Data System (ADS)

    Trifanov, Irina; Hughes, Michael; Rosen, Richard B.; Podoleanu, Adrian

    2008-04-01

    A new approach of acquiring quasi-simultaneous OCT and confocal images is presented. The two images are generated using different principles, optical coherence tomography (OCT) and confocal microscopy (CM). When the system is used to image the retina, the two images have depth resolutions, at present, of less than 20 ?m and approximately 1 mm respectively. The acquisition and display of en-face OCT and confocal images are quasi-simultaneous, without the need of a beam splitter. By using a chopper to periodically obstruct the reference beam in the OCT interferometer, synchronized with the XY-transversal scanner, much higher acquisition speed is obtained than in a previous report where we flipped an opaque screen in the reference arm of the interferometer. Successful operation of the novel configuration was achieved by: (1) stable synchronization of the chopper's movement with the horizontal line scanner and (2) fast self-adjusting of the gain value of avalanche photodiodes depending on the optical power. Images from coin, leafs and retina in vivo have been collected to demonstrate the functionality of the system.

  17. Characterization and Application of a Grazing Angle Objective for Quantitative Infrared Reflection Microspectroscopy

    NASA Technical Reports Server (NTRS)

    Pepper, Stephen V.

    1995-01-01

    A grazing angle objective on an infrared microspectrometer is studied for quantitative spectroscopy by considering the angular dependence of the incident intensity within the objective's angular aperture. The assumption that there is no angular dependence is tested by comparing the experimental reflectance of Si and KBr surfaces with the reflectance calculated by integrating the Fresnel reflection coefficient over the angular aperture under this assumption. Good agreement was found, indicating that the specular reflectance of surfaces can straight-forwardly be quantitatively integrated over the angular aperture without considering non-uniform incident intensity. This quantitative approach is applied to the thickness determination of dipcoated Krytox on gold. The infrared optical constants of both materials are known, allowing the integration to be carried out. The thickness obtained is in fair agreement with the value determined by ellipsometry in the visible. Therefore, this paper illustrates a method for more quantitative use of a grazing angle objective for infrared reflectance microspectroscopy.

  18. Analysis for Mar Vel Black and acetylene soot low reflectivity surfaces for star tracker sunshade applications

    NASA Technical Reports Server (NTRS)

    Yung, E.

    1974-01-01

    Mar Vel Black is a revolutionary new extremely low reflectivity anodized coating developed by Martin Marietta of Denver. It is of great interest in optics in general, and in star trackers specifically because it can reduce extraneous light reflections. A sample of Mar Vel Black was evaluated. Mar Vel Black looks much like a super black surface with many small peaks and very steep sides so that any light incident upon the surface will tend to reflect many times before exiting that surface. Even a high reflectivity surface would thus appear to have a very low reflectivity under such conditions. Conversely, acetylene soot does not have the magnified surface appearance of a super black surface. Its performance is, however, predictable from the surface structure, considering the known configuration of virtually pure carbon.

  19. High harmonic terahertz confocal gyrotron with nonuniform electron beam

    NASA Astrophysics Data System (ADS)

    Fu, Wenjie; Guan, Xiaotong; Yan, Yang

    2016-01-01

    The harmonic confocal gyrotron with nonuniform electron beam is proposed in this paper in order to develop compact and high power terahertz radiation source. A 0.56 THz third harmonic confocal gyrotron with a dual arc section nonuniform electron beam has been designed and investigated. The studies show that confocal cavity has extremely low mode density, and has great advantage to operate at high harmonic. Nonuniform electron beam is an approach to improve output power and interaction efficiency of confocal gyrotron. A dual arc beam magnetron injection gun for designed confocal gyrotron has been developed and presented in this paper.

  20. Study of neutron noise from reflected, metal assemblies with criticality safety applications in mind

    SciTech Connect

    Barnett, C.S.

    1985-08-20

    The author studied the statistics of detected neutrons that leaked from four subcritical reflected, enriched-uranium assemblies, to explore the feasibility of developing a criticality warning system based on neutron noise analysis. The calculated multiplication factors of the assemblies are 0.59, 0.74, 0.82, and 0.92. The author studied three possible discriminators, i.e., three signatures that might be used to discriminate among assemblies of various multiplications. They are: (1) variance-to-mean ratio of the counts in a time bin (V/M); (2) covariance-to-mean ratio of the counts in a common time bin from two different detectors (C/M); and (3) covariance-to-mean ratio of the counts from a single detector in two adjacent time bins of equal length, which the author calls the serial-covariance-to-mean ratio (SC/M). The performances of the three discriminators were not greatly different, but a hierarchy did emerge: SC/M greater than or equal to V/M greater than or equal to C/M. An example of some results: in the neighborhood of k = 0.6 the ..delta..k required for satisfactory discrimination varies from about 3% to 7% as detector solid angle varies from 19% to 5%. In the neighborhood of k = 0.8 the corresponding ..delta..ks are 1% and 2%. The noise analysis techniques studied performed well enough in deeply subcritical situations to deserve testing in an applications environment. They have a good chance of detecting changes in reactivity that are potentially dangerous. One can expect sharpest results when doing comparisons, i.e., when comparing two records, one taken in the past under circumstances known to be normal and one taken now to search for change.

  1. Confocal Endomicroscopic Examination of Malignant Biliary Strictures and Histologic Correlation With Lymphatics

    PubMed Central

    Loeser, Caroline S.; Robert, Marie E.; Mennone, Albert; Nathanson, Michael H.; Jamidar, Priya

    2013-01-01

    Background and Aims Current methods to diagnose malignant biliary strictures are of low sensitivity. Confocal endomicroscopy is a new approach that may improve the diagnosis of indeterminate biliary strictures. The purpose of this study was to evaluate indeterminate biliary strictures using probe-based confocal laser endomicroscopy and to understand the histologic basis for the confocal images. Methods Fourteen patients with indeterminate biliary strictures underwent endoscopic retrograde cholangiopancreatography with examination of their common bile duct with fluorescein-aided probe-based confocal laser endomicroscopy. Standard brushings and biopsies were performed. In parallel, rat bile ducts were examined either with conventional staining and light microscopy or with multiphoton microscopy. Results Earlier published criteria were used to evaluate possible malignancy in the confocal images obtained in the 14 patients. None of the individual criteria were found to be specific enough for malignancy, but a normal-appearing reticular pattern without other putative markers of malignancy was observed in all normal patients. Multiphoton reconstructions of intact rat bile ducts revealed that the reticular pattern seen in normal tissue was in the same focal plane but was smaller than blood vessels. Special stains identified the smaller structures in this network as lymphatics. Conclusions Our limited series suggests that a negative confocal imaging study of the biliary tree can be used to rule out carcinoma, but there are frequent false positives using individual earlier published criteria. An abnormal reticular network, which may reflect changes in lymphatics, was never seen in benign strictures. Better correlation with known histologic structures may lead to improved accuracy of diagnoses. PMID:21063210

  2. ConfocalCheck - A Software Tool for the Automated Monitoring of Confocal Microscope Performance

    PubMed Central

    Hng, Keng Imm; Dormann, Dirk

    2013-01-01

    Laser scanning confocal microscopy has become an invaluable tool in biomedical research but regular quality testing is vital to maintain the systems performance for diagnostic and research purposes. Although many methods have been devised over the years to characterise specific aspects of a confocal microscope like measuring the optical point spread function or the field illumination, only very few analysis tools are available. Our aim was to develop a comprehensive quality assurance framework ranging from image acquisition to automated analysis and documentation. We created standardised test data to assess the performance of the lasers, the objective lenses and other key components required for optimum confocal operation. The ConfocalCheck software presented here analyses the data fully automatically. It creates numerous visual outputs indicating potential issues requiring further investigation. By storing results in a web browser compatible file format the software greatly simplifies record keeping allowing the operator to quickly compare old and new data and to spot developing trends. We demonstrate that the systematic monitoring of confocal performance is essential in a core facility environment and how the quantitative measurements obtained can be used for the detailed characterisation of system components as well as for comparisons across multiple instruments. PMID:24224017

  3. Crystals and collimators for X-ray spectrometry. [Bragg reflection properties and design for astronomical applications

    NASA Technical Reports Server (NTRS)

    Mckenzie, D. L.; Landecker, P. B.; Underwood, J. H.

    1976-01-01

    Results of the measurement of Bragg reflection properties of crystals suitable for use in X-ray astronomy are presented. Measurements with a double crystal spectrometer were performed on rubidium acid phthalate and thallium acid phthalate to yield values of the integrated reflectivity and diffraction width in the range 8-18 A, and measurements of integrated reflectivity were also performed on ammonium dihydrogen phosphate. The theory and design of an arc-minute range multigrid collimator to be flown on a rocket for solar X-ray studies are also described, along with a method for determining the collimator's X-ray axis.

  4. Single-layered PDLC films for electrically variable laser light reflection application

    NASA Astrophysics Data System (ADS)

    Marinov, Yordan G.; Hadjichristov, Georgi B.; Petrov, Alexander G.

    2010-12-01

    Electrically controllable two-beam reflection of coherent light double-passed through the microscale polymer-dispersed liquid crystal (PDLC) single layer is reported. By PDLC medium, the intensities of two reflected beams can be reciprocally changed by applying a suitable voltage in the range 0-100 V. The optical phase shift electrically induced for coherent light propagating in two passes through the single-layered PDLC film, as well as the additional phase shift externally introduced by the substrate of the PDLC cell, are responsible for the electrically controllable reflection in a pair of two beams with complementary voltage-dependent intensity behaviors.

  5. Materials and corrosion characterization using the confocal resonator

    SciTech Connect

    Tigges, C.P.; Sorensen, N.R.; Hietala, V.M.; Plut, T.A.

    1997-05-01

    Improved characterization and process control is important to many Sandia and DOE programs related to manufacturing. Many processes/structures are currently under-characterized including thin film growth, corrosion and semiconductor structures, such as implant profiles. A sensitive tool is required that is able to provide lateral and vertical imaging of the electromagnetic properties of a sample. The confocal resonator is able to characterize the surface and near-surface impedance of materials. This device may be applied to a broad range of applications including in situ evaluation of thin film processes, physical defect detection/characterization, the characterization of semiconductor devices and corrosion studies. In all of these cases, the technology should work as a real-time process diagnostic or as a feedback mechanism regarding the quality of a manufacturing process. This report summarizes the development and exploration of several diagnostic applications.

  6. Site-specific confocal fluorescence imaging of biological microstructures in a turbid medium

    NASA Astrophysics Data System (ADS)

    Saloma, Caesar; Palmes-Saloma, Cynthia; Kondoh, Hisato

    1998-06-01

    Normally transparent biological structures in a turbid medium are imaged using a laser confocal microscope and multiwavelength site-specific fluorescence labelling. The spatial filtering capability of the detector pinhole in the confocal microscope limits the number of scattered fluorescent photons that reach the photodetector. Simultaneous application of different fluorescent markers on the same sample site minimizes photobleaching by reducing the excitation time for each marker. A high-contrast grey-level image is also produced by summing confocal images of the same site taken at different fluorescence wavelengths. Monte Carlo simulations are performed to obtain the quantitative behaviour of confocal fluorescence imaging in turbid media. Confocal images of the following samples were also obtained: (i) 15 m diameter fluorescent spheres placed 1.16 mm deep beneath an aqueous suspension of 0.0823 m diameter polystyrene latex spheres, and (ii) hindbrain of a whole-mount mouse embryo (age 10 days) that was stained to fluoresce at 515 nm and 580 nm peak wavelengths. Expression of RNA transcripts of a gene within the embryo hindbrain was detected by a fluorescence-based whole-mount in situ hybridization procedure that we recently tested.

  7. Variational attenuation correction in two-view confocal microscopy

    PubMed Central

    2013-01-01

    Background Absorption and refraction induced signal attenuation can seriously hinder the extraction of quantitative information from confocal microscopic data. This signal attenuation can be estimated and corrected by algorithms that use physical image formation models. Especially in thick heterogeneous samples, current single view based models are unable to solve the underdetermined problem of estimating the attenuation-free intensities. Results We present a variational approach to estimate both, the real intensities and the spatially variant attenuation from two views of the same sample from opposite sides. Assuming noise-free measurements throughout the whole volume and pure absorption, this would in theory allow a perfect reconstruction without further assumptions. To cope with real world data, our approach respects photon noise, estimates apparent bleaching between the two recordings, and constrains the attenuation field to be smooth and sparse to avoid spurious attenuation estimates in regions lacking valid measurements. Conclusions We quantify the reconstruction quality on simulated data and compare it to the state-of-the art two-view approach and commonly used one-factor-per-slice approaches like the exponential decay model. Additionally we show its real-world applicability on model organisms from zoology (zebrafish) and botany (Arabidopsis). The results from these experiments show that the proposed approach improves the quantification of confocal microscopic data of thick specimen. PMID:24350574

  8. Estimation of anisotropic blur for the restoration of confocal images

    NASA Astrophysics Data System (ADS)

    Rooms, Filip; Philips, Wilfried; Van Oostveldt, Patrick

    2004-02-01

    We present a novel method for joint estimation of the degradation and restoration of photon-limited images. Our method will be demonstrated on confocal microscope images, since confocal microscopy is an important tool in many academic (fundamental biology, . . . ) and industrial (material science, pharmaceutical industry, . . . ) applications. However, the observed images are usually degraded, which hinders analysis and interpretation of the images. Degradation in this kind of images is due to two sources: first, we have blurring due to the bandlimited nature of the optical system; second, Poisson noise contaminates the observations due to the discrete nature of the photon detection process. The proposed method iterates noise reduction and blur estimation using the steerable pyramid transform (which is a variant of the wavelet transform) and deconvolution in the signal domain. These steps are applied in two phases, a training phase and a restoration phase. In the first phase, these three steps are iterated until the blur estimation converges. The second phase is the actual restoration phase. During the iterations the blur estimation serves as a sharpness measure for the restored image, and is used to controls the number of iterations. So, our integrated method provides a completely automatic algorithm where no prior information about the image degradation is required. Our integrated technique was compared with other common restoration techniques for these kind of images, and provided the best restoration results, with least artifacts.

  9. A multi-axis confocal rheoscope for studying shear flow of structured fluids.

    PubMed

    Lin, Neil Y C; McCoy, Jonathan H; Cheng, Xiang; Leahy, Brian; Israelachvili, Jacob N; Cohen, Itai

    2014-03-01

    We present a new design for a confocal rheoscope that enables uniform uniaxial or biaxial shear. The design consists of two precisely positioned parallel plates with a gap that can be adjusted down to 2 0.1 ?m, allowing for the exploration of confinement effects. By using our shear cell in conjunction with a biaxial force measurement device and a high-speed confocal microscope, we are able to measure the real-time biaxial stress while simultaneously imaging the material three-dimensional structure. We illustrate the importance of the instrument capabilities by discussing the applications of this instrument in current and future research topics in colloidal suspensions. PMID:24689598

  10. Optical clearing assisted confocal microscopy of ex vivo transgenic mouse skin

    NASA Astrophysics Data System (ADS)

    Song, Eunjoo; Ahn, YoonJoon; Ahn, Jinhyo; Ahn, Soyeon; Kim, Changhwan; Choi, Sanghoon; Boutilier, Richard Martin; Lee, Yongjoong; Kim, Pilhan; Lee, Ho

    2015-10-01

    We examined the optical clearing assisted confocal microscopy of the transgenic mouse skin. The pinna and dorsal skin were imaged with a confocal microscope after the application of glycerol and FocusClear. In case of the glycerol-treated pinna, the clearing was minimal due to the inefficient permeability. However, the imaging depth was improved when the pinna was treated with FocusClear. In case of dorsal skin, we were able to image deeply to the subcutaneous connective tissue with both agents. Various skin structures such as the vessel, epithelium cells, cartilage, dermal cells, and hair follicles were clearly imaged.

  11. Reflectance spectroscopy - Quantitative analysis techniques for remote sensing applications. [in planetary surface geology

    NASA Technical Reports Server (NTRS)

    Clark, R. N.; Roush, T. L.

    1984-01-01

    The empirical methods and scattering theories that are important for solving remote sensing problems are among the methods for remotely sensed reflectance data analysis presently compared. In the case of the photon mean optical path length concept's implications for reflectance spectra modeling, it is shown that the mean optical path length in a particulate surface is in roughly inverse proportion to the square root of the absorption coefficient. Absorption bands, which are Gaussian in shape when plotted as true absorptance vs photon energy, are also Gaussians in apparent absorptance, although they have a smaller intensity. An apparent continuum in a reflectance spectrum is modeled as a mathematical function that is used to isolate a particular absorption feature for analysis, and it is noted that this continuum should be removed by dividing it into the reflectance spectrum.

  12. Non-reflecting boundary conditions applicable to general purpose CFD simulators

    NASA Astrophysics Data System (ADS)

    Salvesen, Hans-Christen; Teigland, Rune

    1998-09-01

    In simulations of propagating blast waves the effects of artificial reflections at open boundaries can seriously degrade the accuracy of the computations. In this paper, a boundary condition based on a local approximation by a plane traveling wave is presented. The method yields small artificial reflections at open boundaries. The derivation and the theory behind these so-called plane-wave boundary conditions are presented. The method is conceptually simple and is easy to implement in two and three dimensions. These non-reflecting boundary conditions are employed in the three-dimensional computational fluid dynamics (CFD) solver FLACS, capable of simulating gas explosions and blast-wave propagation in complex geometries. Several examples involving propagating waves in one and two dimensions, shock tube and an example of a simulation of a propagating blast wave generated by an explosion in a compressor module are shown. The numerical simulations show that artificial reflections due to the boundary conditions employed are negligible.

  13. Fibre optic confocal imaging (FOCI) for subsurface microscopy of the colon in vivo.

    PubMed

    Delaney, P M; King, R G; Lambert, J R; Harris, M R

    1994-02-01

    Fibre optic confocal imaging (FOCI) is a new type of microscopy which has been recently developed (Delaney et al. 1993). In contrast to conventional light microscopy, FOCI and other confocal techniques allow clear imaging of subsurface structures within translucent objects. However, unlike conventional confocal microscopes which are bulky (because of a need for accurate alignment of large components) FOCI allows the imaging end to be miniaturised and relatively mobile. FOCI is thus particularly suited for clear subsurface imaging of structures within living animals or subjects. The aim of the present study was to assess the suitability of using FOCI for imaging of subsurface structures within the colon, both in vitro (human and rat biopsies) and in vivo (in rats). Images were obtained in fluorescence mode (excitation 488 nm, detection above 515 nm) following topical application of fluorescein. By this technique the glandular structure of the colon was imaged. FOCI is thus suitable for subsurface imaging of the colon in vivo. PMID:8157487

  14. Corneal confocal microscopy: Recent progress in the evaluation of diabetic neuropathy

    PubMed Central

    Papanas, Nikolaos; Ziegler, Dan

    2015-01-01

    The present brief review discusses recent progress with corneal confocal microscopy for the evaluation of diabetic sensorimotor polyneuropathy. Corneal confocal microscopy is a new, non-invasive and reproducible diagnostic modality, and it can also be easily applied for patient follow up. It enables new perspectives of studying the natural history of diabetic sensorimotor polyneuropathy, severity of nerve fiber pathology and documenting early nerve fiber regeneration after therapeutic intervention. It shows moderate to high sensitivity and specificity for the timely diagnosis of diabetic sensorimotor polyneuropathy. Currently, corneal confocal microscopy is mainly used in specialized centers, but deserves more widespread application for the assessment of diabetic sensorimotor polyneuropathy. Finally, further progress is required in terms of technical improvements for automated nerve fiber quantification and for analysis of larger images. PMID:26221515

  15. Design and Demonstration of a Miniature Catheter for a Confocal Microendoscope

    NASA Astrophysics Data System (ADS)

    Rouse, Andrew R.; Kano, Angelique; Udovich, Joshua A.; Kroto, Shona M.; Gmitro, Arthur F.

    2004-11-01

    The fluorescence confocal microendoscope provides high-resolution, in vivo imaging of cellular pathology during optical biopsy. The confocal microendoscope employs a flexible fiber-optic catheter coupled to a custom-built slit-scan confocal microscope. The catheter consists of a fiber-optic imaging bundle linked to a miniature objective and focus assembly. The 3-mm-diameter catheter may be used on its own or routed though the instrument channel of a commercial endoscope, adding microscopic imaging capability to conventional endoscopy. The design and performance of the miniature objective and focus assembly are discussed. Primary applications of the system include diagnosis of disease in the gastrointestinal tract and female reproductive system.

  16. Applications of shallow high-resolution seismic reflection to various environmental problems

    USGS Publications Warehouse

    Miller, R.D.; Steeples, D.W.

    1994-01-01

    Shallow seismic reflection has been successfully applied to environmental problems in a variety of geologic settings. Increased dynamic range of recording equipment and decreased cost of processing hardware and software have made seismic reflection a cost-effective means of imaging shallow geologic targets. Seismic data possess sufficient resolution in many areas to detect faulting with displacement of less than 3 m and beds as thin as 1 m. We have detected reflections from depths as shallow as 2 m. Subsurface voids associated with abandoned coal mines at depths of less than 20 m can be detected and mapped. Seismic reflection has been successful in mapping disturbed subsurface associated with dissolution mining of salt. A graben detected and traced by seismic reflection was shown to be a preferential pathway for leachate leaking from a chemical storage pond. As shown by these case histories, shallow high-resolution seismic reflection has the potential to significantly enhance the economics and efficiency of preventing and/or solving many environmental problems. ?? 1994.

  17. Application of combined experimental and numerical techniques in determining the temperature dependence of reflectivity of semiconductors

    SciTech Connect

    Cerny, R.; Chab, V.; Prikryl, P.

    1996-03-01

    Combined experimental and numerical techniques for determining the temperature dependence of reflectivity of basic semiconductors are analyzed. The method for determination of the reflectivity dependence of liquid semiconductors under pulsed laser irradiation on temperature developed earlier by the authors is modified for the case of solid semiconductors. The results obtained by the time-resolved reflectivity measurement technique together with the known temperature dependencies of the refraction index and the extinction coefficient for the cw probe laser and the room-temperature data for the reflectivity at the frequency of the primary pulsed laser beam are the input parameters of this method. The method itself consists in matching the experimental and computed values of the maximum reflectivity of cw probe laser in dependence on the energy density of the laser pulse and a least-squares fitting procedure. The method is verified on experimental data for the XeCl excimer laser irradiation of Si(100), giving R{sub s} = 0.590 {+-} 0.005 + (4.5 {+-} 0.5) x10{sup -5} (T-293) for the reflectivity of crystalline silicon, which is in good agreement with experimental measurements done by other investigators. In addition, numerical test and error analyses of both the method presented here and the previous method proposed for liquid semiconductors are described and the accuracy and error limits of both methods are discussed.

  18. Hematopoiesis in 3 dimensions: human and murine bone marrow architecture visualized by confocal microscopy

    PubMed Central

    Malide, Daniela; Chen, Jichun; Calado, Rodrigo T.; Kajigaya, Sachiko; Young, Neal S.

    2010-01-01

    In many animals, blood cell production occurs in the bone marrow. Hematopoiesis is complex, requiring self-renewing and pluripotent stem cells, differentiated progenitor and precursor cells, and supportive stroma, adipose tissue, vascular structures, and extracellular matrix. Although imaging is a vital tool in hematology research, the 3-dimensional architecture of the bone marrow tissue in situ remains largely uncharacterized. The major hindrance to imaging the intact marrow is the surrounding bone structures are almost impossible to cut/image through. We have overcome these obstacles and describe a method whereby whole-mounts of bone marrow tissue were immunostained and imaged in 3 dimensions by confocal fluorescence and reflection microscopy. We have successfully mapped by multicolor immunofluorescence the localization pattern of as many as 4 cell features simultaneously over large tiled views and to depths of approximately 150 ?m. Three-dimensional images can be assessed qualitatively and quantitatively to appreciate the distribution of cell types and their interrelationships, with minimal perturbations of the tissue. We demonstrate its application to normal mouse and human marrow, to murine models of marrow failure, and to patients with aplastic anemia, myeloid, and lymphoid cell malignancies. The technique should be generally adaptable for basic laboratory investigation and for clinical diagnosis of hematologic diseases. PMID:20647571

  19. Reflections on current and future applications of multiangle imaging to aerosol and cloud remote sensing

    NASA Astrophysics Data System (ADS)

    Diner, David

    2010-05-01

    The Multi-angle Imaging SpectroRadiometer (MISR) instrument has been collecting global Earth data from NASA's Terra satellite since February 2000. With its 9 along-track view angles, 4 spectral bands, intrinsic spatial resolution of 275 m, and stable radiometric and geometric calibration, no instrument that combines MISR's attributes has previously flown in space, nor is there is a similar capability currently available on any other satellite platform. Multiangle imaging offers several tools for remote sensing of aerosol and cloud properties, including bidirectional reflectance and scattering measurements, stereoscopic pattern matching, time lapse sequencing, and potentially, optical tomography. Current data products from MISR employ several of these techniques. Observations of the intensity of scattered light as a function of view angle and wavelength provide accurate measures of aerosol optical depths (AOD) over land, including bright desert and urban source regions. Partitioning of AOD according to retrieved particle classification and incorporation of height information improves the relationship between AOD and surface PM2.5 (fine particulate matter, a regulated air pollutant), constituting an important step toward a satellite-based particulate pollution monitoring system. Stereoscopic cloud-top heights provide a unique metric for detecting interannual variability of clouds and exceptionally high quality and sensitivity for detection and height retrieval for low-level clouds. Using the several-minute time interval between camera views, MISR has enabled a pole-to-pole, height-resolved atmospheric wind measurement system. Stereo imagery also makes possible global measurement of the injection heights and advection speeds of smoke plumes, volcanic plumes, and dust clouds, for which a large database is now available. To build upon what has been learned during the first decade of MISR observations, we are evaluating algorithm updates that not only refine retrieval accuracies but also include enhancements (e.g., finer spatial resolution) that would have been computationally prohibitive just ten years ago. In addition, we are developing technological building blocks for future sensors that enable broader spectral coverage, wider swath, and incorporation of high-accuracy polarimetric imaging. Prototype cameras incorporating photoelastic modulators have been constructed. To fully capitalize on the rich information content of the current and next-generation of multiangle imagers, several algorithmic paradigms currently employed need to be re-examined, e.g., the use of aerosol look-up tables, neglect of 3-D effects, and binary partitioning of the atmosphere into "cloudy" or "clear" designations. Examples of progress in algorithm and technology developments geared toward advanced application of multiangle imaging to remote sensing of aerosols and clouds will be presented.

  20. Inter-Comparison of ASTER and MODIS Surface Reflectance and Vegetation Index Products for Synergistic Applications to Natural Resource Monitoring

    PubMed Central

    Miura, Tomoaki; Yoshioka, Hiroki; Fujiwara, Kayo; Yamamoto, Hirokazu

    2008-01-01

    Synergistic applications of multi-resolution satellite data have been of a great interest among user communities for the development of an improved and more effective operational monitoring system of natural resources, including vegetation and soil. In this study, we conducted an inter-comparison of two remote sensing products, namely, visible/near-infrared surface reflectances and spectral vegetation indices (VIs), from the high resolution Advanced Thermal Emission and Reflection Radiometer (ASTER) (15 m) and lower resolution Moderate Resolution Imaging Spectroradiometer (MODIS) (250 m 500 m) sensors onboard the Terra platform. Our analysis was aimed at understanding the degree of radiometric compatibility between the two sensors' products due to sensor spectral bandpasses and product generation algorithms. Multiple pairs of ASTER and MODIS standard surface reflectance products were obtained at randomly-selected, globally-distributed locations, from which two types of VIs were computed: the normalized difference vegetation index and the enhanced vegetation indices with and without a blue band. Our results showed that these surface reflectance products and the derived VIs compared well between the two sensors at a global scale, but subject to systematic differences, of which magnitudes varied among scene pairs. An independent assessment of the accuracy of ASTER and MODIS standard products, in which in-house surface reflectances were obtained using in situ Aeronet atmospheric data for comparison, suggested that the performance of the ASTER atmospheric correction algorithm may be variable, reducing overall quality of its standard reflectance product. Atmospheric aerosols, which were not corrected for in the ASTER algorithm, were found not to impact the quality of the derived reflectances. Further investigation is needed to identify the sources of inconsistent atmospheric correction results associated with the ASTER algorithm, including additional quality assessments of the ASTER and MODIS products with other atmospheric radiative transfer codes.

  1. Confocal laser endomicroscopy for gastrointestinal diseases.

    PubMed

    Kiesslich, Ralf; Goetz, Martin; Neurath, Markus F

    2008-07-01

    Confocal laser endomicroscopy enables in vivo microscopy of the mucosal layer of the gastrointestinal tract with subcellular resolution during ongoing endoscopy. Endomicroscopy opens the door to immediate tissue and vessel analysis. Different types of diseases can be diagnosed with optical surface and subsurface analysis. Analysis of the in vivo microarchitecture can be used for targeting biopsies to relevant areas, and subsurface imaging can unmask microscopic diseases or bacterial infection. Molecular imaging is becoming feasible, which will enable new indications in gastrointestinal endoscopy. This article reviews the current and rapidly expanding clinical data on endomicroscopy and gives a look into future research. PMID:18674696

  2. Diffusion of photoacid generators by laser scanning confocal microscopy

    NASA Astrophysics Data System (ADS)

    Zhang, Ping L.; Webber, Stephen E.; Mendenhall, J.; Byers, Jeffrey D.; Chao, Keith K.

    1998-06-01

    Diffusion of the photogenerated acid during the period of time between exposure and development can cause contrast loss and ultimately loss of the latent image. This is especially relevant for chemically amplified photoresists that require a post-exposure baking step, which in turn facilitates acid diffusion due to the high temperature normally employed. It is thus important to develop techniques with good spatial resolution to monitor the photogeneration of acid. More precisely, we need techniques that provide two distinct types of information: spatial resolution on various length scales within the surface layer and also sufficient depth resolution so that one can observe the transition from very surface layer to bulk structure in the polymer blend coated on silicon substrate. Herein laser scanning confocal microscopy is used to evaluate the resist for the first time. We report the use of the confocal microscopy to map the pag/dye distribution in PHS matrices, with both reflectance images and fluorescence images. A laser beam is focused onto a small 3D volume element, termed a voxel. It is typically 200 nm X 200 nm laterally and 800 nm axially. The illuminated voxel is viewed such that only signals emanating from this voxel are detected, i.e., signal from outside the probed voxel is not detected. By adjusting the vertical position of the laser focal point, the voxel can be moved to the designated lateral plane to produce an image. Contrast caused by topology difference between the exposed and unexposed area can be eliminated. Bis-p-butylphenyl iodonium triflat (7% of polyhydroxystyrene) is used as photoacid generators. 5% - 18% (by weight, PHS Mn equals 13 k) resist in PGMEA solution is spin cast onto the treated quartz disk with thickness of 1.4 micrometers , 5 micrometers space/10 micrometers pitch chrome mask is used to generate the pattern with mercury DUV illumination. Fluoresceinamine, the pH-sensitive dye, is also used to enhance the contrast of fluorescence image. The typical PEB temperature is 90 degree(s)C for 90 seconds. 488 nm is used as the excitation wavelength. Both reflectance and fluorescence images (> 510 nm) are processed by using Adobe Photoshop. It was found that the reflectance is more sensitive to the change of the refractive index of the resist while the fluorescence is more sensitive to the distribution of the PAG/dye. The NIH Image software is used for acid exchange rate calculation. Second Fick's Law is applied to analyze the image change. The diffusion coefficient for this PAG in PHS during PEB is smaller than 8.8 X 10-13 cm2/s.

  3. Reflections on Reflections.

    ERIC Educational Resources Information Center

    Merifield, A.

    1990-01-01

    Geometric and algebraic solutions to problems involving reflections of balls on a pool table are presented. The question of whether the ball must eventually enter a pocket is explored. A determination of the number of reflections is discussed. (CW)

  4. Particular features of the application of IR reflection spectroscopy methods in studies in archeology and paleontology

    NASA Astrophysics Data System (ADS)

    Zolotarev, V. M.; Khlopachev, G. A.

    2013-06-01

    We have considered an optical model of a porous rough surface with optical properties of objects (bone, flint) that are typical of archeology and paleontology. We have formulated an approach that makes it possible to perform mathematical processing of the IR reflection spectra of objects of this kind using standard algorithms and determine criteria that ensure obtaining reliable information on objects with a rough surface in the course of interpretation of frequencies in their IR reflection spectra. The potential of the approach has been demonstrated using as an example an investigation by the IR Fourier-transform reflection spectroscopy of mineralization processes of mammoth tusks from two paleolithic sites (14000 and 16000 BCE) located by the town of Yudinovo, Bryansk oblast, Russia.

  5. Critical reflectance derived from MODIS: Application for the retrieval of aerosol absorption over desert regions

    NASA Astrophysics Data System (ADS)

    Wells, Kelley C.; Martins, J. Vanderlei; Remer, Lorraine A.; Kreidenweis, Sonia M.; Stephens, Graeme L.

    2012-02-01

    The determination of aerosol direct radiative forcing over desert regions requires accurate information about the aerosol single-scattering albedo (SSA); however, the brightness of desert surfaces in the visible and near-IR range complicates the retrieval of aerosol optical properties using passive space-based measurements. Here we use the critical reflectance method to retrieve spectral aerosol absorption from space over North Africa, a desert region that is predominantly impacted by absorbing dust and biomass burning aerosol. We examine the sensitivity of the critical reflectance parameter to aerosol physical and optical properties that are representative of the region, and we find that the critical reflectance has low sensitivity to assumptions of aerosol size and refractive index for dust-like particles, except at scattering angles near 180, which should be avoided with this method. We use our findings to retrieve spectral SSA from critical reflectance derived from Moderate Resolution Imaging Spectroradiometer (MODIS) reflectances in the vicinity of two Aerosol Robotic Network (AERONET) stations: Tamanrasset, in the Algerian Sahara, and Banizoumbou, in the Sahel. We retrieve lower aerosol SSAs at Banizoumbou, which is often impacted by dust-smoke mixtures, and higher SSAs at Tamanrasset, where pure desert dust is the dominant aerosol. Our results generally fall within the AERONET uncertainty envelopes, although at Banizoumbou we retrieve a spectral dependence different from that of AERONET. On the basis of our analysis, we expect to be able to retrieve SSA from critical reflectance for pure dust with an uncertainty of 0.02 and to provide spatial and spectral SSA information that will help reduce current uncertainties in the aerosol radiative forcing over desert regions.

  6. [Application of near infrared reflectance spectroscopy to predict meat chemical compositions: a review].

    PubMed

    Tao, Lin-Li; Yang, Xiu-Juan; Deng, Jun-Ming; Zhang, Xi

    2013-11-01

    In contrast to conventional methods for the determination of meat chemical composition, near infrared reflectance spectroscopy enables rapid, simple, secure and simultaneous assessment of numerous meat properties. The present review focuses on the use of near infrared reflectance spectroscopy to predict meat chemical compositions. The potential of near infrared reflectance spectroscopy to predict crude protein, intramuscular fat, fatty acid, moisture, ash, myoglobin and collagen of beef, pork, chicken and lamb is reviewed. This paper discusses existing questions and reasons in the current research. According to the published results, although published results vary considerably, they suggest that near-infrared reflectance spectroscopy shows a great potential to replace the expensive and time-consuming chemical analysis of meat composition. In particular, under commercial conditions where simultaneous measurements of different chemical components are required, near infrared reflectance spectroscopy is expected to be the method of choice. The majority of studies selected feature-related wavelengths using principal components regression, developed the calibration model using partial least squares and modified partial least squares, and estimated the prediction accuracy by means of cross-validation using the same sample set previously used for the calibration. Meat fatty acid composition predicted by near-infrared spectroscopy and non-destructive prediction and visualization of chemical composition in meat using near-infrared hyperspectral imaging and multivariate regression are the hot studying field now. On the other hand, near infrared reflectance spectroscopy shows great difference for predicting different attributes of meat quality which are closely related to the selection of calibration sample set, preprocessing of near-infrared spectroscopy and modeling approach. Sample preparation also has an important effect on the reliability of NIR prediction; in particular, lack of homogeneity of the meat samples influenced the accuracy of estimation of chemical components. In general the predicting results of intramuscular fat, fatty acid and moisture are best, the predicting results of crude protein and myoglobin are better, while the predicting results of ash and collagen are less accurate. PMID:24555369

  7. Imaging System With Confocally Self-Detecting Laser.

    DOEpatents

    Webb, Robert H.; Rogomentich, Fran J.

    1996-10-08

    The invention relates to a confocal laser imaging system and method. The system includes a laser source, a beam splitter, focusing elements, and a photosensitive detector. The laser source projects a laser beam along a first optical path at an object to be imaged, and modulates the intensity of the projected laser beam in response to light reflected from the object. A beam splitter directs a portion of the projected laser beam onto a photodetector. The photodetector monitors the intensity of laser output. The laser source can be an electrically scannable array, with a lens or objective assembly for focusing light generated by the array onto the object of interest. As the array is energized, its laser beams scan over the object, and light reflected at each point is returned by the lens to the element of the array from which it originated. A single photosensitive detector element can generate an intensity-representative signal for all lasers of the array. The intensity-representative signal from the photosensitive detector can be processed to provide an image of the object of interest.

  8. [Application of near-infrared diffuse reflectance spectroscopy to the detection and identification of transgenic corn].

    PubMed

    Rui, Yu-kui; Luo, Yun-bo; Huang, Kun-lun; Wang, Wei-min; Zhang, Lu-da

    2005-10-01

    With the rapid development of the GMO, more and more GMO food has been pouring into the market. Much attention has been paid to GMO labeling under the controversy of GMO safety. Transgenic corns and their parents were scanned by continuous wave of near infrared diffuse reflectance spectroscopy range of 12000-4000 cm(-1); the resolution was 4 cm(-1); scanning was carried out for 64 times; BP algorithm was applied for data processing. The GMO food was easily resolved. Near-infrared diffuse reflectance spectroscopy is unpolluted and inexpensive compared with PCR and ELISA, so it is a very promising detection method for GMO food. PMID:16395887

  9. Reflectance spectra of mafic silicate-opaque assemblages with applications to meteorite spectra

    NASA Technical Reports Server (NTRS)

    Cloutis, Edward A.; Smith, Dorian G. W.; Lambert, Richard St. J.; Gaffey, Michael J.

    1990-01-01

    The addition of fine-grained magnetite to mafic silicate spectra can impart not only an overall blue slope, but also lower overall reflectance and band intensities. The reflectance spectra of the CO and CV magnetite-bearing carbonaceous chondrites are noted to exhibit many of these features; the low band depths of these meteorites suggest that an additional dark, neutral phase, such as ordered carbon, is present. Carbon + mafic silicate spectra possess a red overall slope at low amorphous carbon concentrations. The parent bodies of some of the darkest meteorites should exhibit spectral features attributable to mafic silicates.

  10. Reflecting Reflective Practice

    ERIC Educational Resources Information Center

    Galea, Simone

    2012-01-01

    This paper demystifies reflective practice on teaching by focusing on the idea of reflection itself and how it has been conceived by two philosophers, Plato and Irigaray. It argues that reflective practice has become a standardized method of defining the teacher in teacher education and teacher accreditation systems. It explores how practices of…

  11. Reflecting Reflective Practice

    ERIC Educational Resources Information Center

    Galea, Simone

    2012-01-01

    This paper demystifies reflective practice on teaching by focusing on the idea of reflection itself and how it has been conceived by two philosophers, Plato and Irigaray. It argues that reflective practice has become a standardized method of defining the teacher in teacher education and teacher accreditation systems. It explores how practices of

  12. Numerical study of a confocal ultrasonic setup for creation of cavitation

    NASA Astrophysics Data System (ADS)

    Lafond, Maxime; Chavrier, Franoise; Prieur, Fabrice; Mestas, Jean-Louis; Lafon, Cyril

    2015-10-01

    Acoustic cavitation is used for various therapeutic applications such as local enhancement of drug delivery, histotripsy or hyperthermia. One of the utmost important parameter for cavitation creation is the rarefaction pressure. The typical magnitude of the rarefaction pressure required to initiate cavitation from gas dissolved in tissue is beyond the range of the megapascal. Because nonlinear effects need to be taken into account, a numerical simulator based on the Westervelt equation was used to study the pressure waveform and the acoustic field generated by a setup for creation of cavitation consisting of two high intensity focused ultrasound transducers mounted confocally. At constant acoustic power, simulations with only one and both transducers from the confocal setup showed that the distortion of the pressure waveform due to the combined effects of nonlinearity and diffraction is less pronounced when both confocal transducers are used. Consequently, the confocal setup generates a greater peak negative pressure at focus which is more favorable for cavitation initiation. Comparison between the confocal setup and a single transducer with the same total emitting surface puts in evidence the role of the spatial separation of the two beams. Furthermore, it has been previously shown that the location of the peak negative pressure created by a single transducer shifts from focus towards the transducers in the presence of nonlinear effects. The simulator was used to study a configuration where the acoustical axes of transducers intersect on the peak negative pressure instead of the geometrical focus. For a representative confocal setup, namely moderate nonlinear effects, a 2% increase of the peak negative pressure and 8% decrease of the peak positive pressure resulted from this configuration. These differences tend to increase by increasing nonlinear effects. Although the optimal position of the transducers varies with the nonlinear regimen, the intersection point remains the location of the peak negative pressure in any case. Thus, unlike the location of the peak negative pressure for a single transducer can shift by a few millimeters, the focal point of a confocal device is independent of the power. This point is particularly important for therapeutic applications, frequently requiring high spatial accuracy. An experiment conducted shows that cavitation creation can be achieved easier with confocal ultrasound.

  13. Confocal Raman imaging for cancer cell classification

    NASA Astrophysics Data System (ADS)

    Mathieu, Evelien; Van Dorpe, Pol; Stakenborg, Tim; Liu, Chengxun; Lagae, Liesbet

    2014-05-01

    We propose confocal Raman imaging as a label-free single cell characterization method that can be used as an alternative for conventional cell identification techniques that typically require labels, long incubation times and complex sample preparation. In this study it is investigated whether cancer and blood cells can be distinguished based on their Raman spectra. 2D Raman scans are recorded of 114 single cells, i.e. 60 breast (MCF-7), 5 cervix (HeLa) and 39 prostate (LNCaP) cancer cells and 10 monocytes (from healthy donors). For each cell an average spectrum is calculated and principal component analysis is performed on all average cell spectra. The main features of these principal components indicate that the information for cell identification based on Raman spectra mainly comes from the fatty acid composition in the cell. Based on the second and third principal component, blood cells could be distinguished from cancer cells; and prostate cancer cells could be distinguished from breast and cervix cancer cells. However, it was not possible to distinguish breast and cervix cancer cells. The results obtained in this study, demonstrate the potential of confocal Raman imaging for cell type classification and identification purposes.

  14. Being In-Between: Reflecting on Time, Space and Career during the Tenure Application Process

    ERIC Educational Resources Information Center

    Eichler, Mathew

    2015-01-01

    Part of the process of becoming a tenured faculty member is applying for tenure. This reflective essay reports on the period after the submission of tenure materials for review but before the review process for tenure is completed. This is an "in-between" space, where the race of the tenure track is no longer present, but the role of

  15. Comparison of Three Canopy Reflectance Sensors for Variable-Rate Nitrogen Application in Corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In recent years, canopy reflectance sensing has been investigated for in-season assessment of crop nitrogen (N) health and subsequent control of N fertilization. The several sensor systems that are now commercially available have design and operational differences. One difference is the sensed wavel...

  16. Nadir and oblique canopy reflectance sensing for N application in corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Canopy reflectance sensing can be used to assess in-season crop nitrogen (N) health for subsequent control of N fertilization. The several sensor systems that are now commercially available have design and operational differences, including sensed wavelengths, size of the sensed area, and nadir vs. ...

  17. Design Study of a Visible/Infrared Periscope for Intense Radiation Applications using Reflective Optics

    SciTech Connect

    Medley, S.S.

    1998-05-01

    In magnetically confined fusion devices employing deuterium-tritium (D-T) operation, refractive optical components exposed to neutron and gamma radiation can be subject to degradation of the transmission characteristics, induced luminescence, and altered mechanical properties including dimensional changes. Although radiation resistant refractive optics functioned well for the Tokamak Fusion Test Reactor (TFTR) periscope system during D-T operation, this design approach is unpromising in the much more hostile radiation environment of future D-T devices such as the International Thermonumclear Experimental Reactor (ITER). Under contract to the Princeton Plasma Physics Laboratory, Ball Aerospace of Colorado carried out a periscope design study based on the use of reflective optics. In this design, beryllium reflective input optics supported by a fused silica optical bench were interfaced to a Cassegrain relay system to transfer plasma images to remotely located cameras. This system is also capable of measuring first-wall surface temperatures in the range of 300 - 2,000 degrees C even under projected heating of the reflective optics themselves to several hundred degrees Celsius. Tests of beryllium mirror samples, however, revealed that operation at temperatures above 700 degrees C leads to a loss of specular reflectivity, thus placing an upper limit on the acceptable thermal environment. The main results of this periscope study are presented in this paper.

  18. Practical Application of Polarization and Light Control for Reduction of Reflected Glare.

    ERIC Educational Resources Information Center

    Crouch, C. L.; Kaufman, J. E.

    1963-01-01

    The role of reflected glare and visual viewing angles in near task performance is discussed, and following statements are reported--(1) a worker at a desk normally assumes a position in which his eyes traverse an area of work extending from a point approximately vertically below his eyes to a point not more than 40 degrees from the vertical, (2) a

  19. Practical Application of Polarization and Light Control for Reduction of Reflected Glare.

    ERIC Educational Resources Information Center

    Crouch, C. L.; Kaufman, J. E.

    1963-01-01

    The role of reflected glare and visual viewing angles in near task performance is discussed, and following statements are reported--(1) a worker at a desk normally assumes a position in which his eyes traverse an area of work extending from a point approximately vertically below his eyes to a point not more than 40 degrees from the vertical, (2) a…

  20. Quantitative single-molecule imaging by confocal laser scanning microscopy

    PubMed Central

    Vukojevi?, Vladana; Heidkamp, Marcus; Ming, Yu; Johansson, Bjrn; Terenius, Lars; Rigler, Rudolf

    2008-01-01

    A new approach to quantitative single-molecule imaging by confocal laser scanning microscopy (CLSM) is presented. It relies on fluorescence intensity distribution to analyze the molecular occurrence statistics captured by digital imaging and enables direct determination of the number of fluorescent molecules and their diffusion rates without resorting to temporal or spatial autocorrelation analyses. Digital images of fluorescent molecules were recorded by using fast scanning and avalanche photodiode detectors. In this way the signal-to-background ratio was significantly improved, enabling direct quantitative imaging by CLSM. The potential of the proposed approach is demonstrated by using standard solutions of fluorescent dyes, fluorescently labeled DNA molecules, quantum dots, and the Enhanced Green Fluorescent Protein in solution and in live cells. The method was verified by using fluorescence correlation spectroscopy. The relevance for biological applications, in particular, for live cell imaging, is discussed. PMID:19011092

  1. Signal and noise modeling in confocal laser scanning fluorescence microscopy.

    PubMed

    Herberich, Gerlind; Windoffer, Reinhard; Leube, Rudolf E; Aach, Til

    2012-01-01

    Fluorescence confocal laser scanning microscopy (CLSM) has revolutionized imaging of subcellular structures in biomedical research by enabling the acquisition of 3D time-series of fluorescently-tagged proteins in living cells, hence forming the basis for an automated quantification of their morphological and dynamic characteristics. Due to the inherently weak fluorescence, CLSM images exhibit a low SNR. We present a novel model for the transfer of signal and noise in CLSM that is both theoretically sound as well as corroborated by a rigorous analysis of the pixel intensity statistics via measurement of the 3D noise power spectra, signal-dependence and distribution. Our model provides a better fit to the data than previously proposed models. Further, it forms the basis for (i) the simulation of the CLSM imaging process indispensable for the quantitative evaluation of CLSM image analysis algorithms, (ii) the application of Poisson denoising algorithms and (iii) the reconstruction of the fluorescence signal. PMID:23285574

  2. In vivo confocal microscopy for the oral cavity: Current state of the field and future potential.

    PubMed

    Maher, N G; Collgros, H; Uribe, P; Ch'ng, S; Rajadhyaksha, M; Guitera, P

    2016-03-01

    Confocal microscopy (CM) has been shown to correlate with oral mucosal histopathology in vivo. The purposes of this review are to summarize what we know so far about in vivo CM applications for oral mucosal pathologies, to highlight some current developments with CM devices relevant for oral applications, and to formulate where in vivo CM could hold further application for oral mucosal diagnosis and management. Ovid Medline and/or Google searches were performed using the terms 'microscopy, confocal', 'mouth neoplasms', 'mouth mucosa', 'leukoplakia, oral', 'oral lichen planus', 'gingiva', 'cheilitis', 'taste', 'inflammatory oral confocal', 'mucosal confocal' and 'confocal squamous cell oral'. In summary, inclusion criteria were in vivo use of any type of CM for the human oral mucosa and studies on normal or pathological oral mucosa. Experimental studies attempting to identify proteins of interest and microorganisms were excluded. In total 25 relevant articles were found, covering 8 main topics, including normal oral mucosal features (n=15), oral dysplasia or neoplasia (n=7), inflamed oral mucosa (n=3), taste impairment (n=3), oral autoimmune conditions (n=2), pigmented oral pathology/melanoma (n=1), delayed type hypersensitivity (n=1), and cheilitis glandularis (n=1). The evidence for using in vivo CM in these conditions is poor, as it is limited to mainly small descriptive studies. Current device developments for oral CM include improved probe design. The authors propose that future applications for in vivo oral CM may include burning mouth syndrome, intra-operative mapping for cancer surgery, and monitoring and targeted biopsies within field cancerization. PMID:26786962

  3. Confocal imaging of benign and malignant proliferative skin lesions in vivo

    NASA Astrophysics Data System (ADS)

    Gonzalez, Salvador; Rajadhyaksha, Milind M.; Anderson, R. Rox

    1999-06-01

    Near-infrared confocal reflectance microscopy (CM) provides non- invasive real-time images of thin en-face tissue sections with high resolution and contrast. Imaging of cells, nuclei, other organelles, microvessels, and hair follicles has been possible at resolution comparable to standard histology, to a maximum depth of 250-300 μm in human skin in vivo. We have characterized psoriasis as a prototype of benign proliferative skin conditions, and non-pigmented skin malignancies in vivo based on their unstained, native histologic features using CM. Our data shows that reflectance CM may potentially diagnose and morphometrically evaluate proliferative skin lesions in vivo.

  4. Metrological characterization of optical confocal sensors measurements (20 and 350 travel ranges)

    NASA Astrophysics Data System (ADS)

    Nouira, H.; El-Hayek, N.; Yuan, X.; Anwer, N.; Salgado, J.

    2014-03-01

    Confocal sensors are usually used in dimensional metrology applications, like roughness, form, thickness and surface profile measurements. With the progress of technologies, metrological applications require measurements with nanometer-level of accuracy by using ultra-high precision machines, which should present a minimum and stable metrology loop. The loop is equipped with sensors with nanometer-level of resolution and linear residual. The study presented here, is mainly focused on the characterization of Confocal sensors in order to identify their performance practically. Such information is useful to establish a correction model in the digital signal processing (DSP) software. In this context, LNE developed an ultra-high-precision machine, dedicated to the roughness measurement with an uncertainty of a few nanometres (< 30 nm) by using a tactile sensor. In order to match this machine to Confocal sensors, an experiment has been recently developed to characterize the behaviour of two commercial Confocal sensors with the measuring range of 20 ?m and 350 ? m. The experiment permits the evaluation of the major error sources: axial and radial motion errors as-well-as the deviation/tilt of the sensors.

  5. Application of Extended Inverse Scatter Correction to Mid-Infrared Reflectance Spectra of Soil

    SciTech Connect

    Gallagher, Neal B.; Blake, Thomas A.; Gassman, Paul L.

    2005-05-01

    Scattering artifacts adversely affect infrared reflectance measurements of powders and soils, and extended inverse scatter correction (EISC) is a flexible method useful for correcting for these artifacts. EISC was used to correct mid-infrared reflectance spectra of two different soils coated with dibutyl phosphate and the results were examined using regression analysis. To determine the correction, EISC fits a measured spectrum to a reference spectrum. However, if measured spectra contain features not included in the reference spectrum the fit can be biased resulting in poor correction. Weighted and robust least squares were used to account for these potential biases. Additionally, the present work demonstrates how analyte-free samples can be used to determine basis functions for an extended mixture model used in the correction. Corrected spectra resulted in partial least squares models that performed at least as well as 2nd derivative spectra and were more interpretable.

  6. Near-infrared reflectance spectra-applications to problems in asteroid-meteorite relationships

    NASA Technical Reports Server (NTRS)

    Mcfadden, Lucy A.; Chamberlin, Alan; Vilas, Faith

    1991-01-01

    Near-infrared spectral reflectance data were collected at the Infrared Telescope Facility (IRTF) at Mauna Kea Observatories in 1985 and 1986 for the purpose of searching the region near the 3:1 Kirkwood gap for asteroids with the spectral signatures of ordinary chondrite parent bodies. Twelve reflectance spectra are observed. The presence of ordinary chondrite parent bodies among this specific set of observed asteroids is not obvious, though the sample is biased towards the larger asteroids in the region due to limitations imposed by detector sensitivity. The data set, which was acquired with the same instrumentation used for the 52-color asteroid survey (Bell et al., 1987), also presents some additional findings. The range of spectral characteristics that exist among asteroids of the same taxonomic type is noted. Conclusions based on the findings are discussed.

  7. Application of the Total Reflection X-ray Fluorescence technique to trace elements determination in tobacco

    NASA Astrophysics Data System (ADS)

    Martinez, T.; Lartigue, J.; Zarazua, G.; Avila-Perez, P.; Navarrete, M.; Tejeda, S.

    2008-12-01

    Many studies have identified an important number of toxic elements along with organic carcinogen molecules and radioactive isotopes in tobacco. In this work we have analyzed by Total Reflection X-Ray Fluorescence 9 brands of cigarettes being manufactured and distributed in the Mexican market. Two National Institute of Standards and Technology standards and a blank were equally treated at the same time. Results show the presence of some toxic elements such as Pb and Ni. These results are compared with available data for some foreign brands, while their implications for health are discussed. It can be confirmed that the Total Reflection X-Ray Fluorescence method provides precise (reproducible) and accuracy (trueness) data for 15 elements concentration in tobacco samples.

  8. Optimization and Application of Reflective LSPR Optical Fiber Biosensors Based on Silver Nanoparticles

    PubMed Central

    Chen, Jiangping; Shi, Se; Su, Rongxin; Qi, Wei; Huang, Renliang; Wang, Mengfan; Wang, Libing; He, Zhimin

    2015-01-01

    In this study, we developed a reflective localized surface plasmon resonance (LSPR) optical fiber sensor, based on silver nanoparticles (Ag NPs). To enhance the sensitivity of the LSPR optical sensor, two key parameters were optimized, the length of the sensing area and the coating time of the Ag NPs. A sensing length of 1.5 cm and a 1-h coating time proved to be suitable conditions to produce highly sensitive sensors for biosensing. The optimized sensor has a high refractive index sensitivity of 387 nm/RIU, which is much higher than that of other reported individual silver nanoparticles in solutions. Moreover, the sensor was further modified with antigen to act as a biosensor. Distinctive wavelength shifts were found after each surface modification step. In addition, the reflective LSPR optical fiber sensor has high reproducibility and stability. PMID:26016910

  9. Research and Development Data to Define the Thermal Performance of Reflective Materials Used to Conserve Energy in Building Applications

    SciTech Connect

    Eisenberg, J

    2001-04-09

    A comprehensive experimental laboratory study has been conducted on the thermal performance of reflective insulation systems. The goal of this study was to develop test and evaluation protocols and to obtain thermal performance data on a selected number of idealized and commercial systems containing reflective airspaces for use in analytical models. Steady-state thermal resistance has been measured on 17 different test panels using two guarded hot boxes. Additional instrumentation was installed to measure the temperature of critical locations inside the test panels. The test parameters which have been studied are heat flow direction (horizontal, up, and down), number of airspaces comprising the cavity, airspace effective emittance, airspace aspect ratio, airspace mean temperature and temperature difference, and the thermal resistance of the stud material. Tests have also been performed on similar constructions with mass insulation. Two one-dimensional calculation techniques (ASHRAE and proposed ASTM) have been employed to determine the cavity thermal resistance from the measured test panel results. The measured cavity thermal resistance is compared with literature data which is commonly employed to calculate the thermal resistance of reflective airspace assemblies. A consumer-oriented handbook pertaining to reflective insulation for building and commercial applications has also been prepared as part of this study.

  10. Bidirectional Reflectance of Flat, Optically Thick Particulate Layers: An Efficient Radiative Transfer Solution and Applications to Snow and Soil Surfaces

    NASA Technical Reports Server (NTRS)

    Mishchenko, Michael I.; Dlugach, Janna M.; Yanovitsku, Edgard G.; Zakharova, Nadia T.

    1999-01-01

    We describe a simple and highly efficient and accurate radiative transfer technique for computing bidirectional reflectance of a macroscopically flat scattering layer composed of nonabsorbing or weakly absorbing, arbitrarily shaped, randomly oriented and randomly distributed particles. The layer is assumed to be homogeneous and optically semi-infinite, and the bidirectional reflection function (BRF) is found by a simple iterative solution of the Ambartsumian's nonlinear integral equation. As an exact Solution of the radiative transfer equation, the reflection function thus obtained fully obeys the fundamental physical laws of energy conservation and reciprocity. Since this technique bypasses the computation of the internal radiation field, it is by far the fastest numerical approach available and can be used as an ideal input for Monte Carlo procedures calculating BRFs of scattering layers with macroscopically rough surfaces. Although the effects of packing density and coherent backscattering are currently neglected, they can also be incorporated. The FORTRAN implementation of the technique is available on the World Wide Web at http://ww,,v.giss.nasa.gov/-crmim/brf.html and can be applied to a wide range of remote sensing, engineering, and biophysical problems. We also examine the potential effect of ice crystal shape on the bidirectional reflectance of flat snow surfaces and the applicability of the Henyey-Greenstein phase function and the 6-Eddington approximation in calculations for soil surfaces.

  11. Application of Diffuse Reflectance FT-IR Spectroscopy for the Surface Study of Kevlar Fibers

    NASA Astrophysics Data System (ADS)

    Chatzi, E. G.; Ishida, H.; Koenig, J. L.

    1985-12-01

    The surfaces of Kevlar-49 aramid fibers, being used in high-performance composite materials, have been characterized by diffuse reflectance Fourier transform infrared (FT-IR) spectroscopy. Enhancement of the surface selectivity of the technique has been achieved using KBr overlayers. The water absorbed by both the skin and the core of the fibers has been characterized by using this technique and the accessibility of the fiber functional groups has been evaluated.

  12. Spectral reflectance of carbonate sediments and application to remote sensing classification of benthic habitats

    NASA Astrophysics Data System (ADS)

    Louchard, Eric Michael

    Remote sensing is a valuable tool in marine research that has advanced to the point that images from shallow waters can be used to identify different seafloor types and create maps of benthic habitats. A major goal of this dissertation is to examine differences in spectral reflectance and create new methods of analyzing shallow water remote sensing data to identify different seafloor types quickly and accurately. Carbonate sediments were used as a model system as they presented a relatively uniform, smooth surface for measurement and are a major bottom type in tropical coral reef systems. Experimental results found that sediment reflectance varied in shape and magnitude depending on pigment content, but only varied in magnitude with variations in grain size and shape. Derivative analysis of the reflectance spectra identified wavelength regions that correlate to chlorophyll a and chlorophyllide a as well as accessory pigments, indicating differences in microbial community structure. Derivative peak height also correlated to pigment content in the sediments. In remote sensing data, chlorophyll a, chlorophyllide a, and some xanthophylls were identified in derivative spectra and could be quantified from second derivative peak height. Most accessory pigments were attenuated by the water column, however, and could not be used to quantify pigments in sediments from remote sensing images. Radiative transfer modeling of remote sensing reflectance showed that there was sufficient spectral variation to separate major sediment types, such as ooid shoals and sediment with microbial layers, from different densities of seagrass and pavement bottom communities. Both supervised classification with a spectral library and unsupervised classification with principal component analysis were used to create maps of seafloor type. The results of the experiments were promising; classified seafloor types correlated with ground truth observations taken from underwater video and were comparable to existing maps of seafloor type. Creation of accurate seafloor type maps is an important step in constructing maps of benthic habitats.

  13. Multivariate Data Analysis on Tissue Diffuse Reflectance Spectra for Diagnostic Applications

    NASA Astrophysics Data System (ADS)

    Prince, Shanthi; Malarvizhi, S.

    2011-10-01

    Currently, clinical diagnosis of skin disease is generally accomplished by visual inspection under white light illumination. Aside from physical examination, the diagnosis of most of these lesions is invasive, time-consuming, and costly, often requiring surgical excision or biopsy followed by pathological investigations. Several approaches have been tried to improve dermatological diagnosis. Optical means of characterizing tissues have gained importance due to its noninvasive nature. Diffuse reflectance spectra are unique for normal and diseased tissues. Spectral characteristics of the tissue spectra provide useful information to identify various chromophores present in them, because different chromophores have different spectroscopic responses to electromagnetic waves of certain energy bands. An optical fiber spectrometer is set up for collection of diffuse reflectance data from different skin conditions. The method involves exposure of skin surface to white light produced by an incandescent source. These back scattered photons emerging from various layers of tissue are detected by spectrometer resulting in diffuse reflectance data. PCA can be considered as "the mother of all methods in multivariate data analysis". PCA is performed for data reduction and to obtain specific signature from the spectra to differentiate normal and the diseased skin. The proposed principal component analysis method is able to enhance the peculiar characteristics of the diseased diffuse reflectance spectra. Principal component analysis shows that the spectra from normal and diseased tissues are distinct from each other. PCA is recommended as an exploratory tool to uncover unknown trends in the data. A preliminary study, using PCA on the reparability of the spectra of normal and diseased tissue within each patient shows promise that this method is sensitive to changes in tissue brought upon by the onset of disease.

  14. Application of total internal reflection microscopy for laser damage studies on fused silica

    NASA Astrophysics Data System (ADS)

    Sheehan, Lynn M.; Kozlowski, Mark R.; Camp, David W.

    1998-04-01

    Damage studies show that the majority of damage on UV grade fused silica initiates at the front or rear surface. The grinding and polishing processes used to produce the optical surfaces of transparent optics play a key role in the development of defects which can ultimately initiate damage. These defects can be on or breaking through the surface or can be sub-surface and surface defects in transparent materials. Images taken which compare both total internal reflection microscopy and atomic force microscopy show that the observed defects can be less than one micron in size. Total internal reflection microscopy has the added benefit of being able to observe large areas with sub-micron detection. Both off-line and in- situ systems have been applied in the Lawrence Livermore National Laboratory's damage laboratory in order to understand defects in the surface and subsurface of polished fused silica. There is a preliminary indication that TIRM quality can be related to the damage resistance. The in-situ microscope is coupled into a 355 nm, 7.5 ns, 10 Hz Nd:YAG laser system in order to study damage occurring at localized scatter sites revealed with the total internal reflection microscopy method. The tests indicate damage initiating at observed artifacts which have many different morphologies and damage behaviors. Some of the scatter sites and damage morphologies revealed have been related back to the finishing process.

  15. A microfabricated scanning confocal optical microscope for in situ imaging

    NASA Astrophysics Data System (ADS)

    Dickensheets, David Lee

    Scanning confocal optical microscopes are well suited for imaging living tissue because of their ability to 'cross section' intact tissue. They are not, however, well suited for imaging tissues in situ. This dissertation describes a new, miniature, mirror scanned, high resolution confocal optical microscope that operates in real time. It is small enough to fit into an endoscope, and may eventually be incorporated into a hypodermic needle. Such a device would provide immediate in-situ tissue assessment at the cellular level and may enable, for example, biopsy without tissue removal. Non-medical applications may include process monitoring and endoscopic inspection. The microfabricated confocal optical scanning microscope, or μCOSM, incorporates single mode optical fiber illumination, silicon torsional scan mirrors, and an off- axis micro diffractive lens. The prototype device is monochromatic, at 633 nm, with a 1.1 mm working distance and 0.25 NA. It achieves a line response of 0.98 μm FWHM, and an axial response of 11.1 μm FWHM. The first part of the dissertation describes the opto- mechanical design of the microscope, which was chosen to be compatible with the microfabrication technologies used for its construction. Then the imaging properties of the off-axis diffractive objective lens are developed, including the aberrations of second and third order which constrain its use. The lens is a surface relief phase grating, and a rigorous electromagnetic analysis is employed to specify the pupil function of the microscope. Then the image forming properties of the μCOSM are derived and compared to experimental results. The second part of the dissertation describes the fabrication of the individual elements of the μCOSM, and their assembly into an imaging instrument. The lens is constructed using electron beam lithography and reactive ion etching of a fused silica substrate. The scanning mirrors for the microscope, which comprise a single crystal silicon plate suspended by silicon nitride hinges, are constructed using wafer bonding and surface micromachining techniques. A spacer element is fabricated using bulk silicon micromachining. A package is described which places the μCOSM imaging head, complete with focus control, inside a 3.4 mm OD hypodermic tube. Sample images acquired with the microscope are presented.

  16. Fluorescence confocal endomicroscopy in biological imaging

    NASA Astrophysics Data System (ADS)

    Delaney, Peter; Thomas, Steven; Allen, John; McLaren, Wendy; Murr, Elise; Harris, Martin

    2007-02-01

    In vivo fluorescence microscopic imaging of biological systems in human disease states and animal models is possible with high optical resolution and mega pixel point-scanning performance using optimised off-the-shelf turn-key devices. There are however various trade-offs between tissue access and instrument performance when miniaturising in vivo microscopy systems. A miniature confocal scanning technology that was developed for clinical human endoscopy has been configured into a portable device for direct hand-held interrogation of living tissue in whole animal models (Optiscan FIVE-1 system). Scanning probes of 6.3mm diameter with a distal tip diameter of 5.0mm were constructed either in a 150mm length for accessible tissue, or a 300mm probe for laparoscopic interrogation of internal tissues in larger animal models. Both devices collect fluorescence confocal images (excitation 488 nm; emission >505 or >550 nm) comprised of 1024 x 1204 sampling points/image frame, with lateral resolution 0.7um; axial resolution 7um; FOV 475 x 475um. The operator can dynamically control imaging depth from the tissue surface to approx 250um in 4um steps via an internally integrated zaxis actuator. Further miniaturisation is achieved using an imaging contact probe based on scanning the proximal end of a high-density optical fibre bundle (~30,000 fibres) of <1mm diameter to transfer the confocal imaging plane to tissue in intact small animal organs, albeit at lower resolution (30,000 sampling points/image). In rodent models, imaging was performed using various fluorescent staining protocols including fluorescently labelled receptor ligands, labelled antibodies, FITC-dextrans, vital dyes and labelled cells administered topically or intravenously. Abdominal organs of large animals were accessed laparoscopically and contrasted using i.v. fluorescein-sodium. Articular cartilage of sheep and pigs was fluorescently stained with calcein-AM or fluorescein. Surface and sub-surface cellular and sub-cellular details could be readily visualised in vivo at high resolution. In rodent disease models, in vivo endomicroscopy with appropriate fluorescent agents allowed examination of thrombosis formation, tumour microvasculature and liver metastases, diagnosis and staging of ulcerative colitis, liver necrosis and glomerulonephritis. Miniaturised confocal endomicroscopy allows rapid in vivo molecular and subsurface microscopy of normal and pathologic tissue at high resolution in small and large whole animal models. Fluorescein endomicroscopy has recently been introduced into the medical device market as a clinical imaging tool in GI endoscopy and is undergoing clinical evaluation in laparoscopic surgery. This medical usage is encouraging in-situ endomicroscopy as an important pre-clinical research tool to observe microscopic and molecular system biologic events in vivo in animal models for various human diseases.

  17. Digital confocal microscopy through a multimode fiber

    NASA Astrophysics Data System (ADS)

    Loterie, Damien; Farahi, Salma; Papadopoulos, Ioannis; Goy, Alexandre; Psaltis, Demetri; Moser, Christophe

    2015-09-01

    Confocal laser-scanning microscopy is a well-known optical imaging method where a pinhole is used in the illumination and detection pathways of a normal microscope, in order to selectively excite and detect a particular focal volume. The advantage of this method is a significant increase in contrast, due to the rejection of background contributions to the signal. Here, we propose to apply this method in the context of multimode fiber endoscopy. Due to modal scrambling, it is not possible to use a physical pinhole to filter light signals that have travel through multimode fibers. Instead, we use a transmission matrix approach to characterize the propagation of light through the fiber, and we apply the filtering operation in the digital domain.

  18. Three-dimensional scanning confocal laser microscope

    DOEpatents

    Anderson, R. Rox (Lexington, MA); Webb, Robert H. (Lincoln, MA); Rajadhyaksha, Milind (Charlestown, MA)

    1999-01-01

    A confocal microscope for generating an image of a sample includes a first scanning element for scanning a light beam along a first axis, and a second scanning element for scanning the light beam at a predetermined amplitude along a second axis perpendicular to the first axis. A third scanning element scans the light beam at a predetermined amplitude along a third axis perpendicular to an imaging plane defined by the first and second axes. The second and third scanning element are synchronized to scan at the same frequency. The second and third predetermined amplitudes are percentages of their maximum amplitudes. A selector determines the second and third predetermined amplitudes such that the sum of the percentages is equal to one-hundred percent.

  19. Total reflection X-ray fluorescence trace mercury determination by trapping complexation: Application in advanced oxidation technologies

    NASA Astrophysics Data System (ADS)

    Custo, Graciela; Litter, Marta I.; Rodrguez, Diana; Vzquez, Cristina

    2006-11-01

    It is well known that Hg species cause high noxious effects on the health of living organisms even at very low levels (5 ?g/L). Quantification of this element is an analytical challenge due to the peculiar physicochemical properties of all Hg species. The regulation of the maximal allowable Hg concentration led to search for sensitive methods for its determination. Total reflection X-ray fluorescence is a proved instrumental analytical tool for the determination of trace elements. In this work, the use of total reflection X-ray fluorescence for Hg quantification is investigated. However, experimental determination by total reflection X-ray fluorescence requires depositing a small volume of sample on the reflector and evaporation of the solvent until dryness to form a thin film. Because of volatilization of several Hg forms, a procedure to capture these volatile species in liquid samples by using complexing agents is proposed. Acetate, oxalic acid, ethylenediaminetetracetic acid and ammonium pyrrolidine-dithiocarbamate were assayed for trapping the analytes into the solution during the preparation of the sample and onto the reflector during total reflection X-ray fluorescence measurements. The proposed method was applied to evaluate Hg concentration during TiO 2-heterogeneous photocatalysis, one of the most known advanced oxidation technologies. Advanced oxidation technologies are processes for the treatment of effluents in waters and air that involve the generation of very active oxidative and reductive species. In heterogeneous photocatalysis, Hg is transformed to several species under ultraviolet illumination in the presence of titanium dioxide. Total reflection X-ray fluorescence was demonstrated to be applicable in following the extent of the heterogeneous photocatalysis reaction by determining non-transformed Hg in the remaining solution.

  20. Video-rate confocal scanning laser microscope for imaging human tissues {ital in vivo}

    SciTech Connect

    Rajadhyaksha, M.; Anderson, R.R.; Webb, R.H.

    1999-04-01

    We have built a video-rate confocal scanning laser microscope for reflectance imaging of human skin and oral mucosa {ital in vivo}. Design and imaging parameters were determined for optimum resolution and contrast. Mechanical skin-holding fixtures and oral tissue clamps were made for stable objective lens-to-tissue contact such that gross tissue motion relative to the microscope was minimized. Confocal imaging was possible to maximum depths of 350 {mu}m in human skin and 450 {mu}m in oral mucosa, with measured lateral resolution of 0.5{endash}1 {mu}m and axial resolution (section thickness) of 3{endash}5 {mu}m at the 1064-nm wavelength. This resolution is comparable with that of conventional microscopy of excised biopsies (histology). Normal and abnormal tissue morphology and dynamic processes were observed. {copyright} 1999 Optical Society of America

  1. Gold nanoparticles sensing with diffusion reflection measurement as a new medical diagnostics application

    NASA Astrophysics Data System (ADS)

    Fixler, Dror

    2014-02-01

    The ability to quantitatively and noninvasively detect nanoparticles in vivo has important implications on their development as optical sensors for medical diagnostics. We suggest a new method for cancer detection based on diffusion reflection (DR) measurements of gold nanorods (GNR). In our talk, the ability to extract optical properties of phantoms and their GNR concentrations from DR measurements will demonstrate. We will report, for the first time, GNR detection through upper tissue-like phantom layers, as well as the detection of a tumor presented as highly concentrated GNR placed deep within a phantom.

  2. MEMS-BASED 3D CONFOCAL SCANNING MICROENDOSCOPE USING MEMS SCANNERS FOR BOTH LATERAL AND AXIAL SCAN

    PubMed Central

    Liu, Lin; Wang, Erkang; Zhang, Xiaoyang; Liang, Wenxuan; Li, Xingde; Xie, Huikai

    2014-01-01

    A fiber-optic 3D confocal scanning microendoscope employing MEMS scanners for both lateral and axial scan was designed and constructed. The MEMS 3D scan engine achieved a lateral scan range of over 26 with a 2D MEMS scanning micromirror and a depth scan of over 400 ?m with a 1D MEMS tunable microlens. The lateral resolution and axial resolution of this system were experimentally measured as 1.0 ?m and 7.0 ?m, respectively. 2D and 3D confocal reflectance images of micro-patterns, micro-particles, onion skins and acute rat brain tissue were obtained by this MEMS-based 3D confocal scanning microendoscope. PMID:25013304

  3. Evaluation and purchase of confocal microscopes: Numerous factors to consider

    EPA Science Inventory

    The purchase of a confocal microscope can be a complex and difficult decision for an individual scientist, group or evaluation committee. This is true even for scientists that have used confocal technology for many years. The task of reaching the optimal decision becomes almost i...

  4. CONFOCAL LASER SCANNING MICROSCOPY OF RAT FOLLICLE DEVELOPMENT

    EPA Science Inventory

    This study used confocal laser scanning microscopy (CLSM) to study follicular development in millimeter pieces of rat ovary. To use this technology, it is essential to stain the tissue before laser excitation with the confocal microscope. Various fluorescent stains (Yo-Pro, Bo-Pr...

  5. Three-dimensional reconstruction from multiple reflected views within a realist painting: an application to Scott Fraser's "Three way vanitas"

    NASA Astrophysics Data System (ADS)

    Smith, Brandon M.; Stork, David G.; Zhang, Li

    2009-01-01

    The problem of reconstructing a three-dimensional scene from single or multiple views has been thoroughly studied in the computer vision literature, and recently has been applied to problems in the history of art. Criminisi pioneered the application of single-view metrology to reconstructing the fictive spaces in Renaissance paintings, such as the vault in Masaccio's Trinit and the plaza in Piero della Francesca's Flagellazione. While the vast majority of realist paintings provide but a single view, some provide multiple views, through mirrors depicted within their tableaus. The contemporary American realist Scott Fraser's Three way vanitas is a highly realistic still-life containing three mirrors; each mirror provides a new view of the objects in the tableau. We applied multiple-view reconstruction methods to the direct image and the images reflected by these mirrors to reconstruct the three-dimensional tableau. Our methods estimate virtual viewpoints for each view using the geometric constraints provided by the direct view of the mirror frames, along with the reflected images themselves. Moreover, our methods automatically discover inconsistencies between the different views, including ones that might elude careful scrutiny by eye, for example the fact that the height of the water in the glass differs between the direct view and that in the mirror at the right. We believe our work provides the first application of multiple-view reconstruction to a single painting and will have application to other paintings and questions in the history of art.

  6. Numerical studies on a modified negative-branch confocal unstable resonator (MNBUR)

    NASA Astrophysics Data System (ADS)

    Hall, Thomas; Grnewald, Karin M.; Handke, Jrgen

    2007-05-01

    A modified unstable resonator, suitable for a laser with large gain medium cross section and a small or median output coupling, is presented. The spherical mirrors of this resonator form an off-axis negative-branch confocal unstable resonator. The special shaping of the scraper together with the negative-branch confocal unstable resonator configuration yields a laser output beam in the shape of a half-ring. In contrast to the conventional resonator with a ring-shaped output beam, the cross section of the half-ring is more compact and generates a lower structured far-field distribution, but the rotational symmetry disappears. For a rectangular medium cross section, the modified resonator is applicable, too. Some properties of such a rectangular modified negative-branch confocal unstable resonator (MNBUR) are investigated numerically. The theory is based on the integral equation of the Fresnel-Kirchhoff formulation of Huygens' Principle. The theoretical study pays attention to inaccuracies that may occur in mirror manufacturing. With respect to deviations from the specified mirror radius of curvature, the MNBUR does not show important differences to the performance of a conventional unstable resonator. The tilt of a resonator mirror affects the total coupling loss. The sensitivity of the MNBUR to output mirror misalignment is smaller than that of the common negativebranch confocal unstable resonator. A further important improvement achieved by resonator modification is an increased beam quality.

  7. A white light confocal microscope for spectrally resolved multidimensional imaging.

    PubMed

    Frank, J H; Elder, A D; Swartling, J; Venkitaraman, A R; Jeyasekharan, A D; Kaminski, C F

    2007-09-01

    Spectrofluorometric imaging microscopy is demonstrated in a confocal microscope using a supercontinuum laser as an excitation source and a custom-built prism spectrometer for detection. This microscope system provides confocal imaging with spectrally resolved fluorescence excitation and detection from 450 to 700 nm. The supercontinuum laser provides a broad spectrum light source and is coupled with an acousto-optic tunable filter to provide continuously tunable fluorescence excitation with a 1-nm bandwidth. Eight different excitation wavelengths can be simultaneously selected. The prism spectrometer provides spectrally resolved detection with sensitivity comparable to a standard confocal system. This new microscope system enables optimal access to a multitude of fluorophores and provides fluorescence excitation and emission spectra for each location in a 3D confocal image. The speed of the spectral scans is suitable for spectrofluorometric imaging of live cells. Effects of chromatic aberration are modest and do not significantly limit the spatial resolution of the confocal measurements. PMID:17760615

  8. Reflective and refractive optical materials for earth and space applications; Proceedings of the Meeting, Orlando, FL, Apr. 4, 5, 1991

    NASA Astrophysics Data System (ADS)

    Riedl, Max J.; Hale, Robert R.; Parsonage, Thomas B.

    The present conference discusses beryllium mirror design and fabrication, production of aspheric beryllium optical surfaces by HIP consolidation, the control of thermally induced porosity for the fabrication of beryllium optics, fine-grained beryllium optical coatings, light-absorbing beryllium baffle materials, and advanced broadband baffle materials. Also discussed are radiation-resistant optical glasses, a catalog of IR and cryooptical properties of selected materials, durable metal-dielectric mirror coatings, the optical stability of diffuse reflectance materials, and optical filters for space applications.

  9. Fabrication of highly transparent diamond-like carbon anti-reflecting coating for Si solar cell application

    SciTech Connect

    Banerjee, Amit Das, Debajyoti

    2014-04-24

    ARC grade highly transparent unhydrogenated diamond-like carbon (DLC) films were produced, directly from a-C target, using RF magnetron sputtering deposition technique, for optoelectronic applications. Optical band gap, transmittance, reflectance, sp{sup 3} fraction, I{sub D}/I{sub G}, density, and refractive index of the films have been estimated with the help of optical tools like Uv-vis spectrophotometer, ellipsometer and micro-Raman. Optimum ARC-qualities have been identified in low-temperature grown DLC films at an Ar pressure of 4 mTorr in the reactor, accomplishing its key requirements for use in silicon solar cells.

  10. Usage of cornea and sclera back reflected images captured in security cameras for forensic and card games applications

    NASA Astrophysics Data System (ADS)

    Zalevsky, Zeev; Ilovitsh, Asaf; Beiderman, Yevgeny

    2013-10-01

    We present an approach allowing seeing objects that are hidden and that are not positioned in direct line of sight with security inspection cameras. The approach is based on inspecting the back reflections obtained from the cornea and the sclera of the eyes of people attending the inspected scene and which are positioned in front of the hidden objects we aim to image after performing proper calibration with point light source (e.g. a LED). The scene can be a forensic scene or for instance a casino in which the application is to see the cards of poker players seating in front of you.

  11. Reflectors with directional-mixed reflection properties for application in luminaries with high-power LED diodes

    NASA Astrophysics Data System (ADS)

    Zaremba, Krzysztof

    2008-06-01

    Application of directional-mixed reflectors results in a luminance decrease of the apparent image of light emitting diodes (LEDs), which is advantageous as far as glare reduction is concerned. On the other hand, reflectors have a negative impact on luminous intensity curves of the luminaries. This work analyzes an impact of surfaces with directional-mixed reflection properties in a mirror reflector designed for a luminary equipped with high-power LEDs. We present an algorithm used to determine the shape of the reflector of the surface with small scattering, where the axis twist angle for a parabolic reflector varies in a predefined range and follows a power function.

  12. Fabrication of highly transparent diamond-like carbon anti-reflecting coating for Si solar cell application

    NASA Astrophysics Data System (ADS)

    Banerjee, Amit; Das, Debajyoti

    2014-04-01

    ARC grade highly transparent unhydrogenated diamond-like carbon (DLC) films were produced, directly from a-C target, using RF magnetron sputtering deposition technique, for optoelectronic applications. Optical band gap, transmittance, reflectance, sp3 fraction, ID/IG, density, and refractive index of the films have been estimated with the help of optical tools like Uv-vis spectrophotometer, ellipsometer and micro-Raman. Optimum ARC-qualities have been identified in low-temperature grown DLC films at an Ar pressure of 4 mTorr in the reactor, accomplishing its key requirements for use in silicon solar cells.

  13. Critical Reflectance Derived from MODIS: Application for the Retrieval of Aerosol Absorption over Desert Regions

    NASA Technical Reports Server (NTRS)

    Wells, Kelley C.; Martins, J. Vanderlei; Remer, Lorraine A.; Kreidenweis, Sonia M.; Stephens, Graeme L.

    2012-01-01

    Aerosols are tiny suspended particles in the atmosphere that scatter and absorb sunlight. Smoke particles are aerosols, as are sea salt, particulate pollution and airborne dust. When you look down at the earth from space sometimes you can see vast palls of whitish smoke or brownish dust being transported by winds. The reason that you can see these aerosols is because they are reflecting incoming sunlight back to the view in space. The reason for the difference in color between the different types of aerosol is that the particles arc also absorbing sunlight at different wavelengths. Dust appears brownish or reddish because it absorbs light in the blue wavelengths and scatters more reddish light to space, Knowing how much light is scattered versus how much is absorbed, and knowin that as a function of wavelength is essential to being able to quantify the role aerosols play in the energy balance of the earth and in climate change. It is not easy measuring the absorption properties of aerosols when they are suspended in the atmosphere. People have been doing this one substance at a time in the laboratory, but substances mix when they are in the atmosphere and the net absorption effect of all the particles in a column of air is a goal of remote sensing that has not yet been completely successful. In this paper we use a technique based on observing the point at which aerosols change from brightening the surface beneath to darkening it. If aerosols brighten a surface. they must scatter more light to space. If they darken the surface. they must be absorbing more. That cross over point is called the critical reflectance and in this paper we show that critical reflectance is a monotonic function of the intrinsic absorption properties of the particles. This parameter we call the single scattering albedo. We apply the technique to MODIS imagery over the Sahara and Sahel regions to retrieve the single scattering albedo in seven wavelengths, compare these retrievals to ground-based retrievals from AERONET instruments and compute error bars on each retrieval. The results show that we can retrieve single scattering albedo for pure dust to within +/-0.02 and mixtures of dust and smoke to within +/-0.03. No other space based instrument has achieved a retrieval of single scattering albedo that spans the spectrum from 0.47 microns to 2.13 microns and produces regional maps of aerosol absorption showing gradients and changes. Applied in a more operational fashion, such information will narrow uncertainties in estimating aerosol forcing on climate.

  14. In Vivo Confocal Microscopy after Corneal Collagen Crosslinking.

    PubMed

    Mazzotta, Cosimo; Hafezi, Farhad; Kymionis, George; Caragiuli, Stefano; Jacob, Soosan; Traversi, Claudio; Barabino, Stefano; Randleman, J Bradley

    2015-10-01

    In vivo confocal microscopy (IVCM) findings of 84 patients who had undergone conventional epithelium-off corneal collagen crosslinking (CXL) and accelerated CXL (ACXL) were retrospectively reviewed. Analysis confirmed that despite a significant decrease in the mean density of anterior keratocytes in the first 6 postoperative months, cell density after CXL and ACXL returned to baseline values at 12 months. The demarcation lines observed after treatments represent an expression of light-scattering (reflectivity changes) through different tissue densities. Temporary haze of the anterior-mid stroma after conventional CXL represents an indirect sign of CXL-induced stromal collagen compaction and remodeling. IVCM showed that treatment penetration varies to some extent, but that the endothelium is not damaged and is correlated with CXL biomechanical effects. IVCM of limbal structures shows no evidence of pathological changes. Regeneration of subepithelial and stromal nerves was complete 12 months after the operation with fully restored corneal sensitivity and no neurodystrophic occurrences. IVCM allowed detailed high magnification in vivo micromorphological analysis of corneal layers, enabling the assessment of early and late corneal modifications induced by conventional and accelerated CXL. IVCM confirms that CXL is a safe procedure, which is still undergoing development and protocol adjustments. PMID:26142059

  15. Beamforming with a volumetric array of massless laser spark sources--Application in reflection tracking.

    PubMed

    Eskelinen, Joona; Hæggström, Edward; Delikaris-Manias, Symeon; Bolaños, Javier Gómez; Pulkki, Ville

    2015-06-01

    A volumetric array of laser-induced air breakdown sparks is used to produce a directional and steerable acoustic source. The laser breakdown array element is broadband, point-like, and massless. It produces an impulse-like waveform in midair, thus generating accurate spatio-temporal information for acoustic beamforming. A laser-spark scanning setup and the concept of a massless steerable source are presented and evaluated with a cubic array by using an off-line far field delay-and-sum beamforming method. This virtual acoustic array with minimal source influence can, for instance, produce narrow transmission beams to obtain localized and directional impulse response information by reflection tracking. PMID:26093445

  16. [Quantitative analysis of contents in compound fertilizer and application research using near infrared reflectance spectroscopy].

    PubMed

    Song, Le; Zhang, Hong; Ni, Xiao-Yu; Wu, Lin; Liu, Bin-Mei; Yu, Li-Xiang; Wang, Qi; Wu, Yue-Jin

    2014-01-01

    In the present study, a new approach to fast determining the content of urea, biuret and moisture in compound fertilizer composed of urea, ammonium dihydrogenphosphate and potassium chloride was proposed by using near infrared diffuse reflectance spectroscopy. After preprocessing the original spectrum, partial least squares (PLS) models of urea, biuret and moisture were built with the R2 values of 0.9861, 0.9770 and 0.9713 respectively, the root mean square errors of cross validation were 2.59, 0.38, 0.132 respectively. And the prediction correlation factors were 0.9733, 0.9215 and 0.9679 respectively. The authors detected six kinds of compound fertilizer in market for the model verification, the correlation factors were 0.9237, 0.9786 and 0.9874 respectively. The data implied that the new method can be used for situ quality control in the production process of compound fertilizer. PMID:24783536

  17. Reflections on the development and application of FISH whole chromosome painting.

    PubMed

    Tucker, James D

    2015-01-01

    This review describes my personal reflections on the development of whole chromosome painting using fluorescence in situ hybridization and how my laboratory applied the technology in humans and in animal models. The trials and triumphs of the early years are emphasized, along with some of the scientific surprises that were encountered along the way. Scientific issues that my laboratory addressed using chromosome painting technologies are summarized and related to questions in radiation dosimetry, chemical clastogenesis, translocation persistence, and translocation frequencies in unexposed people. A description is provided of scientific controversies that were encountered and how they were resolved. I hope this paper will encourage young scientists to follow their passions and pursue their scientific dreams even if the task seems daunting and the circumstances appear exceedingly difficult. In my case the journey has been challenging, exciting, and richly rewarding on many levels. PMID:25795112

  18. Total reflection X-ray fluorescence and archaeometry: Application in the Argentinean cultural heritage

    NASA Astrophysics Data System (ADS)

    Vzquez, Cristina; Albornoz, Ana; Hajduk, Adam; Elkin, Dolores; Custo, Graciela; Obrustky, Alba

    2008-12-01

    Archaeometry is an interdisciplinary research area involved in the development and use of scientific methods in order to answer questions concerned with the human history. In this way the knowledge of archaeological objects through advanced chemical and physical analyses permits a better preservation and conservation of the cultural heritage and also reveals materials and technologies used in the past. In this sense, analytical techniques play an important role in order to provide chemical information about cultural objects. Considering the non destructive characteristic of this study, analytical techniques must be adequate in order to prevent any alteration or damage and in addition to allow the conservation of their integrity. Taking into account the irreplaceable character of the archaeological and artistic materials considered in this study, analytical techniques must be adequate in order to prevent any alteration or damage and in addition to allow the conservation of their integrity. Total Reflection X-ray Fluorescence Spectrometry as a geometric variant of conventional X-ray fluorescence is a proved microanalytical technique considering the small amount of sample required for the analysis. A few micrograms are enough in order to reveal valuable information about elemental composition and in this context it is highly recommended for artwork studies. In this paper a case study is presented in which Total Reflection X-Ray Fluorescence Spectrometry has been successfully employed in the archaeometry field. Examples from Argentinean cultural heritage sites related with the determination of pigments in paintings on canvas and in rock sites as well as in underwater archaeology research are shown.

  19. Anti-resonant reflecting guidance in alcohol-filled hollow core photonic crystal fiber for sensing applications.

    PubMed

    Liu, Shuhui; Wang, Ying; Hou, Maoxiang; Guo, Jiangtao; Li, Zhihua; Lu, Peixiang

    2013-12-16

    Mechanism and sensing applications of antiresonant reflecting guidance in an alcohol-filled simplified hollow-core (SHC) photonic crystal fiber (PCF) are demonstrated. By filling one air hole in the air cladding of the PCF with alcohol, anti-resonant reflecting guidance of light can be achieved and energy leakage of the core modes can be induced at resonant wavelengths of the Fabry-Pérot (F-P) resonator formed by the alcohol-filled layer combined with the silica cladding in the cross-section of the PCF. The proposed structure exhibits periodic lossy dips in the transmission spectrum, of which the visibilities are sensitive to the refractive index of surrounding medium due to the reflectivity variation of the F-P resonator. Water level sensing is experimentally realized with this principle and the lossy dip exhibits a linear decrease against water level with a sensitivity of 1.1 dB/mm. The sensor is also sensitive to environmental temperature and a temperature sensitivity of -0.48 nm/°C is obtained between room temperature and 60 °C. PMID:24514741

  20. Transmissive grating-reflective mirror-based fiber optic accelerometer for stable signal acquisition in industrial applications

    NASA Astrophysics Data System (ADS)

    Lee, Yeon-Gwan; Kim, Dae-Hyun; Kim, Chun-Gon

    2012-05-01

    This paper discusses an applicable fiber-optic accelerometer composed of a transmissive grating panel, a reflection mirror, and two optical fibers with a separation of quarter grating pitch as transceivers that monitor the low-frequency accelerations of civil engineering structures. This sensor structure brings together the advantages of both a simple sensor structure, which leads to simplified cable design by 50% in comparison with the conventional transmission-type fiber optic accelerometer, and a stable reflected signals acquisition with repeatability in comparison to the researched grating-reflection type fiber optic accelerometer. The vibrating displacement and sinusoidal acceleration measured from the proposed fiber optic sensor demonstrated good agreement with those of a commercial laser displacement sensor and a MEMS accelerometer without electromagnetic interference. The developed fiber optic accelerometer can be used in frequency ranges below 4.0 Hz with a margin of error that is less than 5% and a high sensitivity of 5.06 rad/(m/s)2.

  1. Dye-enhanced multimodal confocal microscopy for noninvasive detection of skin cancers in mouse models

    NASA Astrophysics Data System (ADS)

    Park, Jesung; Mroz, Pawel; Hamblin, Michael R.; Yaroslavsky, Anna N.

    2010-03-01

    Skin cancer is the most common form of human cancer. Its early diagnosis and timely treatment is of paramount importance for dermatology and surgical oncology. In this study, we evaluate the use of reflectance and fluorescence confocal microscopy for detecting skin cancers in an in-vivo trial with B16F10 melanoma and SCCVII squamous cell carcinoma in mice. For the experiments, the mice are anesthetized, then the tumors are infiltrated with aqueous solution of methylene blue and imaged. Reflectance images are acquired at 658 nm. Fluorescence is excited at 658 nm and registered in the range between 690 and 710 nm. After imaging, the mice are sacrificed. The tumors are excised and processed for hematoxylin and eosin histopathology, which is compared to the optical images. The results of the study indicate that in-vivo reflectance images provide valuable information on vascularization of the tumor, whereas the fluorescence images mimic the structural features seen in histopathology. Simultaneous dye-enhanced reflectance and fluorescence confocal microscopy shows promise for the detection, demarcation, and noninvasive monitoring of skin cancer development.

  2. Polymer Cholesteric-Liquid-Crystal (PCLC) Flake/Fluid Host Electro-Optical Suspensions and Their Applications in Color Flexible Reflective Displays

    SciTech Connect

    Marshall, K.L.; Trajkovska-Petkoska, A.; Hasman, K.; Leitch, M.; Cox, G.; Kosc, T.Z.; Jacobs, S.D.

    2008-03-13

    Polymer cholesteric-liquid-crystal (PCLC) flake/fluid-host suspensions are a novel particle display technology for full-color reflective display applications on rigid or flexible substrates. These polarizing pigments require no polarizers or color filters, switch rapidly at very low voltages, and produce highly saturated colors with a reflection efficiency approaching 80%.

  3. REFLECTANCE CALIBRATION OF FOCAL PLANE ARRAY HYPERSPECTRAL IMAGING SYSTEM FOR AGRICULTURAL AND FOOD SAFETY APPLICATIONS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A method to calibrate a pushbroom hyperspectral imaging system for "near-field" applications in agricultural and food safety has been demonstrated. The method consists of a modified geometric control point correction applied to a focal plane array to remove smile and keystone distortion from the sy...

  4. Polarization Of Light Reflected From Forest Canopies On Earth With Applications To Earth-like Planets With Realistic Cloud Cover

    NASA Astrophysics Data System (ADS)

    Wolstencroft, Ramon D.; Breon, F.; Tranter, G.

    2007-05-01

    Quasi-specular reflection of sunlight at the waxy surface of a leaf yields appreciably polarized light especially near the Brewster angle. Sunlight entering the leaf is either absorbed by photosynthetic pigments or, after reflection at internal surfaces, emerges as a diffuse component with a non-zero but low linear polarization and a yet smaller level of circular polarization. The net polarization varies with the leaf's surface roughness, internal architecture and health and is difficult to model. The polarization of a forest canopy is complex and depends especially on the leaf orientation distribution. The global polarization and reflectance properties of Planet Earth have been measured by the POLDER satellite: at 443nm atmospheric Rayleigh scattering dominates, but at 865nm the average surface properties of ocean, vegetation, desert and snow can be estimated. For cloud-free surfaces at 865nm and 90 degree phase angle the percentage polarization,p, and reflectance,R, are respectively [55%,9%](ocean), [7%,23%](vegetation), [6%,40%](desert) and [3%,80%](snow). Note that the values for clear and cloudy ocean are very different, viz [55%,9%] and [4%,45%] respectively. Allowing for the fractional global areas of each component and a global cloud cover of 55% yields p=7.3% for a pale-blue-dot Earth. pR is greatest for oceans and least for vegetation and hence the prospects for detecting pR from vegetation on an Earth-like planet are poor unless >50% is covered in vegetation. However prospects of using the phase angle and wavelength dependence of a pale-blue-dot planet to deduce its properties as it rotates and orbits are more encouraging: the main obstacle will be to overcome the difficulty of correcting for the unknown and variable cloud cover. The possible application of a circular polarization signal that is unique to vegetation remains an intriguing possibility for remote sensing that requires further study.

  5. Retrieving the optical parameters of biological tissues using diffuse reflectance spectroscopy and Fourier series expansions. I. theory and application

    PubMed Central

    Muoz Morales, Aarn A.; Vzquez y Montiel, Sergio

    2012-01-01

    The determination of optical parameters of biological tissues is essential for the application of optical techniques in the diagnosis and treatment of diseases. Diffuse Reflection Spectroscopy is a widely used technique to analyze the optical characteristics of biological tissues. In this paper we show that by using diffuse reflectance spectra and a new mathematical model we can retrieve the optical parameters by applying an adjustment of the data with nonlinear least squares. In our model we represent the spectra using a Fourier series expansion finding mathematical relations between the polynomial coefficients and the optical parameters. In this first paper we use spectra generated by the Monte Carlo Multilayered Technique to simulate the propagation of photons in turbid media. Using these spectra we determine the behavior of Fourier series coefficients when varying the optical parameters of the medium under study. With this procedure we find mathematical relations between Fourier series coefficients and optical parameters. Finally, the results show that our method can retrieve the optical parameters of biological tissues with accuracy that is adequate for medical applications. PMID:23082281

  6. Retrieving the optical parameters of biological tissues using diffuse reflectance spectroscopy and Fourier series expansions. I. theory and application.

    PubMed

    Muoz Morales, Aarn A; Vzquez Y Montiel, Sergio

    2012-10-01

    The determination of optical parameters of biological tissues is essential for the application of optical techniques in the diagnosis and treatment of diseases. Diffuse Reflection Spectroscopy is a widely used technique to analyze the optical characteristics of biological tissues. In this paper we show that by using diffuse reflectance spectra and a new mathematical model we can retrieve the optical parameters by applying an adjustment of the data with nonlinear least squares. In our model we represent the spectra using a Fourier series expansion finding mathematical relations between the polynomial coefficients and the optical parameters. In this first paper we use spectra generated by the Monte Carlo Multilayered Technique to simulate the propagation of photons in turbid media. Using these spectra we determine the behavior of Fourier series coefficients when varying the optical parameters of the medium under study. With this procedure we find mathematical relations between Fourier series coefficients and optical parameters. Finally, the results show that our method can retrieve the optical parameters of biological tissues with accuracy that is adequate for medical applications. PMID:23082281

  7. Confocal Raman Microscopy on in-situ Structural Evolution of Polyolefin Blends

    NASA Astrophysics Data System (ADS)

    Chang, Wansoo; Jeon, Byungho; Lee, Jong-Won; Ryu, Chang Y.

    2011-03-01

    Polyolefins account for more than half of world-wide consumption of plastic materials, and are typically blended with fillers and other types of polymers in applications. In particular, understanding the miscibility and phase behaviors of polyolefin blends is important for the advancement of a wide array of new applications in medicine, packaging, and other fields. We have used Confocal Raman Microscopy to take the advantages of it capability to locally probe the transformation of physical states in polymeric materials and to characterize morphology of polyolefin blends in-situ for lateral and in-depth imaging with a micron-scale spatial resolution. Upon distinct changes of Raman spectra associated with the melting of semicrystaline polyolefins, we report the in-situ morphological changes upon heating and cooling of polyethylene-polyethylene and polypropylene-polyethylene using confocal Raman microscopy with heating stage. NSF MRI-0722563.

  8. Reflections on clinical applications of yoga in voice therapy with MTD.

    PubMed

    Moore, Carmelle

    2012-12-01

    This paper explores the application of modified yoga techniques, as an adjunct to voice therapy, by a speech pathologist who is also a yoga teacher. Yoga practices, with effects that may be short-term, are not considered a substitute for comprehensive and integrated somatic retraining systems (such as the Alexander Technique or Feldenkrais ATM). However, when yoga is conducted emphasizing kinaesthetic and proprioceptive awareness, the client may achieve an 'awareness state' that facilitates the learning of vocal remediation techniques (for example, by more easily 'tuning in' to the subtle sensations of supralaryngeal deconstriction). Core yoga elements and clinical applications are identified. The potential benefits and considerations when using yoga as an adjunct to the treatment of muscle tension dysphonia (MTD) are explored. PMID:23137146

  9. Near infrared reflectance spectra: Applications to problems in asteroid-meteorite relationships

    NASA Technical Reports Server (NTRS)

    Mcfadden, Lucy A.; Chamberlin, Alan B.

    1992-01-01

    An observing program designed to search for evidence of ordinary chondrite parent bodies near the 3:1 Kirkwood Gap was carried out in 1985 and 1986. Studies by Wisdom (1985), Wetherill (1985), and subsequent work by Milani et al. (1989) indicate that the 3:1 Kirkwood gap is the most probable source region for the majority of ordinary chondrite meteorites. The diversity of the reflectance spectra among this small data set is surprising. Early work by Gaffey and McCord (1978) showed that the inner region of the main asteroid belt is dominated by high albedo objects with mafic silicate surfaces. One would expect to see mostly spectra with 1- and 2-micron absorption bands based on this earlier work. Only 5 (of 12) spectra have these expected features. The distribution of taxonomic types presented by Gradie and Tedesco (1982) is in most cases a useful simplification of the compositional structure of the asteroid belt. The range of spectral characteristics seen with higher resolution in the near-IR has not been previously reported and is not represented in the standard asteroid taxonomy. Near-IR spectra contain valuable mineralogical information which enhances knowledge of the composition and structure of asteroids.

  10. Applications of reflection seismics to mapping coal-seam structure and discontinuities

    SciTech Connect

    Dobecki, T.L.; Bartel, L.C.

    1981-01-01

    As a means of demonstrating the effectiveness of reflection seismology in determining continuity of coal seams, three US field projects are reviewed. The three projects involve coals of varied thickness (2 to 14 m) and age (Pennsylvanian to Eocene) from three coal areas of the US (Pennsylvania, Wyoming, and Washington). Each projet also employed its own particular seismic technique, recording system, and seismic energy source although all are considered state-of-the-art, high resolution, digital seismic surveys. Project 1 (thin, Pennsylvania coal) sought detection of sand channels using dynamite and standard in-line (2-D) seismic technique. Project 2 (thick, Wyoming underground coal gasification) involved a gas-explosion (Dinoseis) source with areal (3-D) acquisition methods. Project 3 (thick Washington underground coal gasification) employed a shotgun-type source and standard in-line methods. Data processing was handled by different contractors for each project. Each project was successful in accomplishing its own particular objective; however, data quality and interpretation seem to be more a function of thickness of the target seam, complexity of the overburden, and processing contractor than a seismic source, acquisition scheme (2-D versus 3-D), or recording instrumentation.

  11. Confocal examination of nonmelanoma cancers in thick skin excisions to potentially guide mohs micrographic surgery without frozen histopathology.

    PubMed

    Rajadhyaksha, M; Menaker, G; Flotte, T; Dwyer, P J; González, S

    2001-11-01

    Precise removal of nonmelanoma cancers with minimum damage to the surrounding normal skin is guided by the histopathologic examination of each excision during Mohs micrographic surgery. The preparation of frozen histopathology sections typically requires 20-45 min per excision. Real-time confocal reflectance microscopy offers an imaging method potentially to avoid frozen histopathology and prepare noninvasive (optical) sections within 5 min. Skin excisions ( approximately 1 mm thick) from Mohs surgeries were washed with 5% acetic acid and imaged with a confocal cross-polarized microscope. The confocal images were compared with the corresponding histopathology. Acetic acid causes compaction of chromatin that increases light back-scatter and makes the nuclei bright and easily detectable. Crossed-polarization strongly enhances the contrast of the nuclei because the compacted chromatin depolarizes the illumination light whereas the surrounding cytoplasm and normal dermis does not. Fast low-resolution examination of cancer lobules in wide fields of view followed by high-resolution inspection of nuclear morphology in small fields of view is possible; this is similar to the procedure for examining histopathology sections. Both the Mohs surgeon and the patient will potentially save several hours per day in the operating room. Fast confocal reflectance microscopic examination of excisions (of any thickness) may improve the management of surgical pathology and guide microsurgery of any human tissue. PMID:11710924

  12. Deep stroma investigation by confocal microscopy

    NASA Astrophysics Data System (ADS)

    Rossi, Francesca; Tatini, Francesca; Pini, Roberto; Valente, Paola; Ardia, Roberta; Buzzonetti, Luca; Canovetti, Annalisa; Malandrini, Alex; Lenzetti, Ivo; Menabuoni, Luca

    2015-03-01

    Laser assisted keratoplasty is nowadays largely used to perform minimally invasive surgery and partial thickness keratoplasty [1-3]. The use of the femtosecond laser enables to perform a customized surgery, solving the specific problem of the single patient, designing new graft profiles and partial thickness keratoplasty (PTK). The common characteristics of the PTKs and that make them eligible respect to the standard penetrating keratoplasty, are: the preservation of eyeball integrity, a reduced risk of graft rejection, a controlled postoperative astigmatism. On the other hand, the optimal surgical results after these PTKs are related to a correct comprehension of the deep stroma layers morphology, which can help in the identification of the correct cleavage plane during surgeries. In the last years some studies were published, giving new insights about the posterior stroma morphology in adult subjects [4,5]. In this work we present a study performed on two groups of tissues: one group is from 20 adult subjects aged 59 +/- 18 y.o., and the other group is from 15 young subjects, aged 12+/-5 y.o.. The samples were from tissues not suitable for transplant in patients. Confocal microscopy and Environmental Scanning Electron Microscopy (ESEM) were used for the analysis of the deep stroma. The preliminary results of this analysis show the main differences in between young and adult tissues, enabling to improve the knowledge of the morphology and of the biomechanical properties of human cornea, in order to improve the surgical results in partial thickness keratoplasty.

  13. Correlative confocal Raman Imaging for 2D materials

    NASA Astrophysics Data System (ADS)

    Yang, Jianyong; Liu, Wei; Dieing, Thomas; Fischer, Harald; Henrich, Marius; Hollricher, Olaf

    2015-03-01

    Graphene was one of the first two-dimensional materials which soon after its first mono-layer production received much attention by many researchers worldwide. Its properties vastly differ from bulk graphite and its potential for applications ranges from transistors to transparent conducting electrodes and solar cell applications. While Graphene is arguably the most prominent two-dimensional material there are to this date many more that are subject to current research such as MoS2, WS2 or MoSe2. Graphene has been already and still is extensively studied using a variety of characterization techniques. Raman spectroscopy and more importantly still, Raman imaging proved to be of great value due to the clearly different spectra obtained from single, double, triple and multi-layered Graphene. This and more information that can be extracted from Raman spectroscopy and imaging can well be complemented with other techniques such as various forms of atomic force microscopy (AFM), Scanning Nearfield Optical Microscopy (SNOM), and scanning electron microscopy (SEM). In this contribution we illustrate the benefit of correlating said techniques with confocal Raman imaging in order to deepen the understanding of the samples in question.

  14. Electrostatically driven micromirrors for a miniaturized confocal laser scanning microscope

    NASA Astrophysics Data System (ADS)

    Hofmann, Ulrich; Muehlmann, Sascha; Witt, Martin; Doerschel, Klaus; Schuetz, Rijk; Wagner, Bernd

    1999-09-01

    A compact two-mirror microscanner has been fabricated to build the central part of a miniaturized confocal laser scanning microscope. This microscope shall be mounted at the tip of an endoscope to provide high resolution imaging for medical diagnostics. In order to achieve a resolution of 500 X 500 image elements large scan angles and also large mirror dimensions have to be realized within a spatially strong limited housing. While bulk silicon technology on the one hand enables fabrication of micromirrors with nearly ideal elastical behavior, those actuators on the other hand often are too fragile for a lot of applications. This paper describes the design, fabrication and assembling of electrostatically driven torsional micromirrors that meet the requirements of fast two-dimensional scanning with high angular precision over large scan angles, compact design and also high shock resistance. This is achieved with the combination of bulk silicon technology with metal surface micromachining. Besides medical diagnostics these microscanners can be used in a wider range of applications such as displays, two-dimensional barcode scanning, multiplexing of fiber optics, etc.

  15. Tri-modal confocal mosaics detect residual invasive squamous cell carcinoma in Mohs surgical excisions

    NASA Astrophysics Data System (ADS)

    Gareau, Dan; Bar, Anna; Snaveley, Nicholas; Lee, Ken; Chen, Nathaniel; Swanson, Neil; Simpson, Eric; Jacques, Steve

    2012-06-01

    For rapid, intra-operative pathological margin assessment to guide staged cancer excisions, multimodal confocal mosaic scan image wide surgical margins (approximately 1 cm) with sub-cellular resolution and mimic the appearance of conventional hematoxylin and eosin histopathology (H&E). The goal of this work is to combine three confocal imaging modes: acridine orange fluorescence (AO) for labeling nuclei, eosin fluorescence (Eo) for labeling cytoplasm, and endogenous reflectance (R) for marking collagen and keratin. Absorption contrast is achieved by alternating the excitation wavelength: 488 nm (AO fluorescence) and 532 nm (Eo fluorescence). Superposition and false-coloring of these modes mimics H&E, enabling detection of cutaneous squamous cell carcinomas (SCC). The sum of mosaic Eo+R is false-colored pink to mimic the appearance of eosin, while the AO mosaic is false-colored purple to mimic the appearance of hematoxylin in H&E. In this study, mosaics of 10 Mohs surgical excisions containing invasive SCC, and five containing only normal tissue were subdivided for digital presentation equivalent to 4× histology. Of the total 50 SCC and 25 normal sub-mosaics presented, two reviewers made two and three type-2 errors (false positives), respectively. Limitations to precisely mimic H&E included occasional elastin staining by AO. These results suggest that confocal mosaics may effectively guide staged SCC excisions in skin and other tissues.

  16. Improving transverse resolution of confocal microscopy through spatiotemporal modulation

    NASA Astrophysics Data System (ADS)

    Wang, Baokai; Zou, Limin; Zhang, Su; Tan, Jiubin

    2015-11-01

    A new method is proposed in this paper to improve transverse resolution of a confocal microscope. By setting up the model of a confocal microscope system through spatiotemporal modulation with moving gratings or acousto-optical modulation without defocus distance under coherent light illumination and deducing two-dimensional coherent image formula and transfer function, simulation tests are run with or without spatiotemporal modulation to prove the effectiveness of the proposed method. Simulation results indicate the proposed method can be used to improve the transverse resolution of a confocal microscope system.

  17. Personal reflections on the highlights and changes in radiation and radioisotope measurement applications

    NASA Astrophysics Data System (ADS)

    Gardner, Robin P.; Lee, Kyoung O.

    2015-11-01

    This paper describes the recent changes that the authors have perceived in the use of radiation and radioisotope measurement applications. The first change is that due to the increased use of Monte Carlo simulation which has occurred from a normal evolutionary process. This is due in large part to the increased accuracy that is being obtained by the use of detector response functions (DRFs) and the simultaneous increased computational efficiency that has become available with these DRFs, the availability of a greatly improved weight windows variance reduction method, and the availability of inexpensive computer clusters. This first change is a happy one. The other change that is occurring is in response to recent terrorist activities. That change is the replacement or major change in the use of long-lived radioisotopes in radioisotope measurement and other radioisotope source applications. In general this can be done by improving the security of these radioisotope sources or by replacing them altogether by using machine sources of radiation. In either case one would like to preclude altogether or at least minimize the possibility of terrorists being able to obtain radioisotopes and use them for clandestine purposes.

  18. Penetration of resin-based materials into initial erosion lesion: A confocal microscopic study.

    PubMed

    Ionta, Franciny Querobim; Boteon, Ana Paula; Moretto, Marcelo Juliano; Jnior, Odair Bim; Honrio, Heitor Marques; Silva, Thiago Cruvinel; Wang, Linda; Rios, Daniela

    2016-02-01

    The application of resin-based materials is an alternative of treatment for eroded lesions. Nevertheless, there are no studies about the penetration of these materials into eroded lesion, which might affect its adhesion. Therefore, this study evaluated the penetration of four resin-based materials, with and without enamel etching. By using an in vitro protocol, types of treatment were studied at five levels (AdheSE() , Tetric N-Bond() , Single Bond 2() , Helioseal Clear() , Icon() ) and types of enamel etching in two levels (with and without). Materials were stained with 0.02 mg/mL ethanolic solution of tetramethylrhodamine isothiocyanate. Bovine enamel samples (4 4 mm) were immersed in 0.01 M HCl, pH 2.3, for 30 seconds to produce initial eroded lesions. Afterward, the materials were applied on half of sample enamel surface following the manufacturer's instructions. On the other half of sample, the materials were applied without etching the enamel. Materials penetration into the enamel was assessed by Confocal Laser Scanning Microscopy on reflection and fluorescence modes. The penetration depth (PD) was measured using ImageJ software. Data were analyzed by two-way ANOVA and Tukey test (P??0.05). It can be concluded that prior enamel etching increased the materials penetration into eroded enamel and the Icon() -infiltrant presented highest penetration. Microsc. Res. Tech. 79:72-80, 2016. 2015 Wiley Periodicals, Inc. PMID:26626706

  19. Optical oblique-incidence reflectivity difference microscopy: Application to label-free detection of reactions in biomolecular microarrays

    NASA Astrophysics Data System (ADS)

    Landry, James Paul

    2008-04-01

    Biomolecular microarrays have emerged as a leading technology for high-throughput in vitro assays in genomics and proteomics. Microarrays contain 100 to 100,000 distinct biomolecular features immobilized on a substrate at high density, enabling parallel assays of entire biomolecular systems or screens of large biomolecular libraries on a single glass slide. Microarrays are typically detected by reacting immobilized targets with fluorescently-labeled probes. For many biomolecules, particularly structurally and functionally diverse proteins, modification with labeling-agents can alter their function. For this reason, it is important to develop label-free microarray detection technology to complement standard fluorescence-based detection. In this dissertation, I report my research into the development of optical oblique-incidence reflectivity difference (OI-RD) microscopy for application to high-throughput and label-free detection of biomolecular microarrays in end-point and real-time modalities. OI-RD is a versatile and sensitive form of nulling polarization-modulated ellipsometry. By reflecting light at oblique incidence from a surface, OI-RD measures changes in thickness and dielectric response of ultrathin molecular layers through disproportionate responses of s- and p-polarization reflectivities. In this dissertation I given an account of the engineering and operation of the first OI-RD microscopes and mathematical theory underpinning them. I then report experiments showing label-free OI-RD detection of DNA hybridization and antibody-antigen binding reactions in microarrays fabricated on standard chemically functionalized glass slides. The experiments demonstrate that: (1) The OI-RD signal quantifies biomolecular film properties, in particular, surface mass density, coverage, and orientation of biomolecules in the films. (2) The properties of targets, probes, and other biomolecular entities within the microarray can be measured throughout the microarray usage cycle. (3) A wide variety of biochemical reactions can be detected with a sensitivity and limit of detection comparable to or better than other label-free optical surface biosensors. (4) Microarrays of thousands of features can be end-point detected for screening applications or microarrays of hundreds of features can be detected in real-time for high-throughput biochemical kinetic analysis, with the potential to increase both of these capacities by at least an order of magnitude. (5) OI-RD is compatible with existing microarray fabrication materials and protocols because it is applicable to any optically flat surface.

  20. Bidirectional Reflectance of a Macroscopically Flat, High-Albedo Particulate Surface: An Efficient Radiative Transfer Solution and Applications to Regoliths

    NASA Technical Reports Server (NTRS)

    Mishchenko, Michael I.; Zakharova, Nadia T.

    1999-01-01

    Many remote sensing applications rely on accurate knowledge of the bidirectional reflection function (BRF) of surfaces composed of discrete, randomly positioned scattering particles. Theoretical computations of BRFs for plane-parallel particulate layers are usually reduced to solving the radiative transfer equation (RTE) using one of existing exact or approximate techniques. Since semi-empirical approximate approaches are notorious for their low accuracy, violation of the energy conservation law, and ability to produce unphysical results, the use of numerically exact solutions of RTE has gained justified popularity. For example, the computation of BRFs for macroscopically flat particulate surfaces in many geophysical publications is based on the adding-doubling (AD) and discrete ordinate (DO) methods. A further saving of computer resources can be achieved by using a more efficient technique to solve the plane-parallel RTE than the AD and DO methods. Since many natural particulate surfaces can be well represented by the model of an optically semi-infinite, homogeneous scattering layer, one can find the BRF directly by solving the Ambartsumian's nonlinear integral equation using a simple iterative technique. In this way, the computation of the internal radiation field is avoided and the computer code becomes highly efficient and very accurate and compact. Furthermore, the BRF thus obtained fully obeys the fundamental physical laws of energy conservation and reciprocity. In this paper, we discuss numerical aspects and the computer implementation of this technique, examine the applicability of the Henyey-Greenstein phase function and the sigma-Eddington approximation in BRF and flux calculations, and describe sample applications demonstrating the potential effect of particle shape on the bidirectional reflectance of flat regolith surfaces. Although the effects of packing density and coherent backscattering are currently neglected, they can also be incorporated. The FORTRAN implementation of the technique is available on the World Wide Web, and can be applied to a wide range of remote sensing problems. BRF computations for undulated (macroscopically rough) surfaces are more complicated and often rely on time consuming Monte Carlo procedures. This approach is especially inefficient for optically thick, weakly absorbing media (e.g., snow and desert surfaces at visible wavelengths since a photon may undergo many internal scattering events before it exists the medium or is absorbed. However, undulated surfaces can often be represented as collections of locally flat tilted facets characterized by the BRF found from the traditional plane parallel RTE. In this way the MOnte Carlo procedure could be used only to evaluate the effects of surface shadowing and multiple surface reflections, thereby bypassing the time-consuming ray tracing inside the medium and providing a great savings of CPU time.

  1. In Situ Confocal Raman Mapping Study of a Single Ti-Assisted ZnO Nanowire.

    PubMed

    Gandhi, Ashish C; Hung, Hsuan-Jung; Shih, Po-Hsun; Cheng, Chia-Liang; Ma, Yuan-Ron; Wu, Shengyun

    2009-01-01

    In this work, we succeeded in preparing in-plane zinc oxide nanowires using a Ti-grid assisted by the chemical vapor deposition method. Optical spatial mapping of the Confocal Raman spectra was used to investigate the phonon and geometric properties of a single ZnO nanowire. The local optical results reveal a red shift in the non-polar E2 high frequency mode and width broadening along the growth direction, reflecting quantum-confinement in the radial direction. PMID:20672140

  2. Three-dimensional in vivo imaging by a handheld dual-axes confocal microscope

    PubMed Central

    Ra, Hyejun; Piyawattanametha, Wibool; Mandella, Michael J.; Hsiung, Pei-Lin; Hardy, Jonathan; Wang, Thomas D.; Contag, Christopher H.; Kino, Gordon S.; Solgaard, Olav

    2008-01-01

    We present a handheld dual-axes confocal microscope that is based on a two-dimensional microelectromechanical systems (MEMS) scanner. It performs reflectance and fluorescence imaging at 488 nm wavelength, with three-dimensional imaging capability. The fully packaged microscope has a diameter of 10 mm and acquires images at 4 Hz frame rate with a maximum field of view of 400 ?m 260 ?m. The transverse and axial resolutions of the handheld probe are 1.7 ?m and 5.8 ?m, respectively. Capability to perform real time small animal imaging is demonstrated in vivo in transgenic mice. PMID:18545427

  3. Spectrally encoded slit confocal microscopy using a wavelength-swept laser

    NASA Astrophysics Data System (ADS)

    Kim, Soocheol; Hwang, Jaehyun; Heo, Jung; Ryu, Suho; Lee, Donghak; Kim, Sang-Hoon; Oh, Seung Jae; Joo, Chulmin

    2015-03-01

    We present an implementation of spectrally encoded slit confocal microscopy. The method employs a rapid wavelength-swept laser as the light source and illuminates a specimen with a line focus that scans through the specimen as the wavelength sweeps. The reflected light from the specimen is imaged with a stationary line scan camera, in which the finite pixel height serves as a slit aperture. This scanner-free operation enables a simple and cost-effective implementation in a small form factor, while allowing for the three-dimensional imaging of biological samples.

  4. Development of optical fiber Bragg grating force-reflection sensor system of medical application for safe minimally invasive robotic surgery

    NASA Astrophysics Data System (ADS)

    Song, Hoseok; Kim, Kiyoung; Lee, Jungju

    2011-07-01

    Force feedback plays a very important role in medical surgery. In minimally invasive surgery (MIS), however, the very long and stiff bars of surgical instruments greatly diminish force feedback for the surgeon. In the case of minimally invasive robotic surgery (MIRS), force feedback is totally eliminated. Previous researchers have reported that the absence of force feedback increased the average force magnitude applied to the tissue by at least 50%, and increased the peak force magnitude by at least a factor of two. Therefore, it is very important to provide force information in MIRS. Recently, many sensors are being developed for MIS and MIRS, but some obstacles to their application in actual medical surgery must be surmounted. The most critical problems are size limit and sterilizability. Optical fiber sensors are among the most suitable sensors for the surgical environment. The optical fiber Bragg grating (FBG) sensor, in particular, offers an important additional advantage over other optical fiber sensors in that it is not influenced by the intensity of the light source. In this paper, we present the initial results of a study on the application of a FBG sensor to measure reflected forces in MIRS environments and suggest the possibility of successful application to MIRS systems.

  5. Solar Confocal Interferometers for Sub-Picometer-Resolution Spectral Filters

    NASA Technical Reports Server (NTRS)

    Gary, G. Allen; Pietraszewski, Chris; West, Edward A.; Dines, Terence C.

    2006-01-01

    The confocal Fabry-Perot interferometer allows sub-picometer spectral resolution of Fraunhofer line profiles. Such high spectral resolution is needed to keep pace with the higher spatial resolution of the new set of large-aperture solar telescopes. The line-of-sight spatial resolution derived for line profile inversions would then track the improvements of the transverse spatial scale provided by the larger apertures. The confocal interferometer's unique properties allow a simultaneous increase in both etendue and spectral power. Methods: We have constructed and tested two confocal interferometers. Conclusions: In this paper we compare the confocal interferometer with other spectral imaging filters, provide initial design parameters, show construction details for two designs, and report on the laboratory test results for these interferometers, and propose a multiple etalon system for future testing of these units and to obtain sub-picometer spectral resolution information on the photosphere in both the visible and near-infrared.

  6. CONFOCAL LASER SCANNING MICROSCOPY OF APOPTOSIS IN WHOLE MOUSE OVARIES

    EPA Science Inventory

    Confocal Laser Scanning Microscopy of Apoptosis in Whole Mouse Ovaries. Robert M. Zucker Susan C. Jeffay and Sally D. Perreault Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle...

  7. WAVELENGTH AND ALIGNMENT TESTS FOR CONFOCAL SPECTRAL IMAGING SYSTEMS

    EPA Science Inventory

    Confocal spectral imaging (CSI) microscope systems now on the market delineate multiple fluorescent proteins, labels, or dyes within biological specimens by performing spectral characterizations. However, we find that some CSI present inconsistent spectral profiles of reference s...

  8. CALIBRATION AND VALIDATION OF CONFOCAL SPECTRAL IMAGING SYSTEMS

    EPA Science Inventory

    Confocal spectral imaging (CSI) microscope systems now on the market can perform spectral characterization of biological specimens containing fluorescent proteins, labels or dyes. Some CSI have been found to present inconsistent spectral characterizations within a particular syst...

  9. Bipolar absolute differential confocal approach to higher spatial resolution.

    PubMed

    Zhao, Weiqian; Tan, Jiubin; Qiu, Lirong

    2004-10-18

    By use of a superresolution pupil filtering technique to achieve a lateral optical superresolution and a differential confocal microscopy technique to achieve an axial resolution at the nanometer level, we propose a high spatial resolution bipolar absolute differential confocal approach for the ultraprecision measurement of three-dimensional microstructures. The feasibility of the proposed approach has been proved by use of a shaped annular beam differential confocal microscopy system. The experimental results indicate that the lateral and axial resolutions of the shaped annular beam differential confocal system are better than 0.2 mum and 2 nm, respectively, when lambda=632.8 nm, epsilon=0.5, uM=6.95, and with a 0.85 numerical aperture. PMID:19484057

  10. Application of Hapke photometric model to three geologic surfaces using PARABOLA bidirectional reflection data

    NASA Technical Reports Server (NTRS)

    Shepard, Michael K.; Arvidson, Raymond E.; Deering, Donald W.

    1991-01-01

    The Geologic Remote Sensing Field Experiment (GRSFE) was conducted in July and September of 1989 to collect data with both ground and airborne instrumentation. A major objective of GRSFE was to collect data which could be used to test radiative transfer models for the extraction of composition and textural surface properties from remotely acquired data. Reported here are the initial results from an application of the Hapke photometric model, using data from the Portable Apparatus for Remote Acquisition of Bidirectional Observations of Land and Atmosphere (PARABOLA), a ground based radiometer with three spectral channels. PARABOLA data was collected in the Lunar Crater Volcanic Field in Nevada, specifically from the region of Lunar Lake, a playa. The Hapke model was found to be inadequate for three relatively common geologic surfaces (a clay-rich, hard packed surface with decimeter sized mudcracks; a cobble site, similar to a playa site, but strewn with basaltic cobbles and pebbles; and a surface mantled basalt lava flow). The model is not at fault; rather, the complexity of most geologic surfaces is not accounted for in the initial assumptions.

  11. Variable-rate nitrogen application algorithm based on canopy reflected spectrum and its influence on wheat

    NASA Astrophysics Data System (ADS)

    Liang, Hongxia; Zhao, Chunjiang; Huang, Wenjiang; Liu, Liangyun; Wang, Jihua; Ma, Youhua

    2005-01-01

    This study was to develop the time-specific and time-critical method to overcome the limitations of traditional field sampling methods for variable rate fertilization. Farmers, agricultural managers and grain processing enterprises are interested in measuring and assessing soil and crop status in order to apply adequate fertilizer quantities to crop growth. This paper focused on studying the relationship between vegetation index (OSAVI) and nitrogen content to determine the amount of nitrogen fertilizer recommended for variable rate management in precision agriculture. The traditional even rate fertilizer management was chosen as the CK. The grain yield, ear numbers, 1000-grain weight and grain protein content were measured among the CK, uniform treatments and variable rate fertilizer treatments. It indicated that variable rate fertilization reduced the variability of wheat yield, ear numbers and dry biomass, but it didn't increased crop yield and grain protein content significantly and did not decrease the variety of 1000-grain weight, compared to traditional rate application. The nitrogen fertilizer use efficiency was improved, for this purpose, the variable rate technology based on vegetation index could be used to prevent under ground water pollution and environmental deterioration.

  12. Combined In Vivo Confocal Raman Spectroscopy and Confocal Microscopy of Human Skin

    PubMed Central

    Caspers, P. J.; Lucassen, G. W.; Puppels, G. J.

    2003-01-01

    In vivo confocal Raman spectroscopy is a noninvasive optical method to obtain detailed information about the molecular composition of the skin with high spatial resolution. In vivo confocal scanning laser microscopy is an imaging modality that provides optical sections of the skin without physically dissecting the tissue. A combination of both techniques in a single instrument is described. This combination allows the skin morphology to be visualized and (subsurface) structures in the skin to be targeted for Raman measurements. Novel results are presented that show detailed in vivo concentration profiles of water and of natural moisturizing factor for the stratum corneum that are directly related to the skin architecture by in vivo cross-sectional images of the skin. Targeting of skin structures is demonstrated by recording in vivo Raman spectra of sweat ducts and sebaceous glands in situ. In vivo measurements on dermal capillaries yielded high-quality Raman spectra of blood in a completely noninvasive manner. From the results of this exploratory study we conclude that the technique presented has great potential for fundamental skin research, pharmacology (percutaneous transport), clinical dermatology, and cosmetic research, as well as for noninvasive analysis of blood analytes, including glucose. PMID:12829511

  13. Automatic morphing using image registration: Application to continuous tracking of radar reflectivity and rain fields

    NASA Astrophysics Data System (ADS)

    Vongsaard, Jearanai

    Rainfall is one of the most important natural phenomenon that influences human life. Accurate rainfall estimation and prediction are crucial for flood forecasting, flood control, climate diagnostics, and water resource management. Rain data may be collected from numerous sources. Conventional rain gauge networks or meteorological radars provide continuous coverage in time. Satellite observations provide snap-shots of precipitation fields at poor temporal resolution. While a number of spaceborne platforms have been deployed for rain observation, the development of continuous space/time rainfall remains a major challenge. This dissertation seeks alternative techniques to automatically generate continuous data streams of rainfall data from sparse or intermittent observations. In order to avoid human intervention in the process, an automatic procedure is needed for real-time operations. For this purpose, Automatic Morphing Using Image Registration (AMIR) model is developed by integrating automatic image registration and image morphing algorithm. The new AMIR technique uses automatic image registration as the basis for finding control points for the morphing process. In the study of data assimilation for weather forecasting, there is a need to generate continuous streams of rainfall data to alleviate the so-called "spin up" problem, or the inability to provide short-term forecasts [Road90]. The proposed algorithm has been tested using remote sensing images from Next Generation Weather Radars (NEXRAD) and Tropical Rainfall Measuring Mission (TRMM). Three cases of rainfall data have been used. These include the passage of a storm in Florida, hurricane Floyd, and scattered rain in the southwestern of the United States for the same period using NEXRAD radar data as surrogate for spaceborne observations. These cases have drastically different spatial and temporal characteristics and hence provide tests on the applicability of the AMIR method. Comparative experimental results have shown that AMIR advance the current state of the art as it is comparable to manual morphing and outperforms automatic morphing without control points proposed in literature.

  14. Correction of terrestrial LiDAR intensity channel using Oren-Nayar reflectance model: An application to lithological differentiation

    NASA Astrophysics Data System (ADS)

    Carrea, Dario; Abellan, Antonio; Humair, Florian; Matasci, Battista; Derron, Marc-Henri; Jaboyedoff, Michel

    2016-03-01

    Ground-based LiDAR has been traditionally used for surveying purposes via 3D point clouds. In addition to XYZ coordinates, an intensity value is also recorded by LiDAR devices. The intensity of the backscattered signal can be a significant source of information for various applications in geosciences. Previous attempts to account for the scattering of the laser signal are usually modelled using a perfect diffuse reflection. Nevertheless, experience on natural outcrops shows that rock surfaces do not behave as perfect diffuse reflectors. The geometry (or relief) of the scanned surfaces plays a major role in the recorded intensity values. Our study proposes a new terrestrial LiDAR intensity correction, which takes into consideration the range, the incidence angle and the geometry of the scanned surfaces. The proposed correction equation combines the classical radar equation for LiDAR with the bidirectional reflectance distribution function of the Oren-Nayar model. It is based on the idea that the surface geometry can be modelled by a relief of multiple micro-facets. This model is constrained by only one tuning parameter: the standard deviation of the slope angle distribution (σslope) of micro-facets. Firstly, a series of tests have been carried out in laboratory conditions on a 2 m2 board covered by black/white matte paper (perfect diffuse reflector) and scanned at different ranges and incidence angles. Secondly, other tests were carried out on rock blocks of different lithologies and surface conditions. Those tests demonstrated that the non-perfect diffuse reflectance of rock surfaces can be practically handled by the proposed correction method. Finally, the intensity correction method was applied to a real case study, with two scans of the carbonate rock outcrop of the Dents-du-Midi (Swiss Alps), to improve the lithological identification for geological mapping purposes. After correction, the intensity values are proportional to the intrinsic material reflectance and are independent from range, incidence angle and scanned surface geometry. The corrected intensity values significantly improve the material differentiation.

  15. CCDiode: an optimal detector for laser confocal microscopes

    NASA Astrophysics Data System (ADS)

    Pawley, James B.; Blouke, Morley M.; Janesick, James R.

    1996-04-01

    The laser confocal microscope (LCM) is now an established research tool in biology and materials science. In biological applications, it is usually employed to detect the location of fluorescent market molecules and, under these conditions, signal levels from bright areas are often < 20 photons/pixel (from the specimen, assuming a standard 512 X 768, 1 sec. scan). Although this data rate limits the speed at which information can be derived from the specimen, saturation of the fluorophor, photobleaching of the dye, and phototoxicity prevent it being increased. Currently, most LCMs use photomultiplier tubes (PMT, QE equals 1 - 30% 400 - 900 nm). By contrast, rear-illuminated, scientific charge-coupled devices (CCD) now routinely readout the signal from square sensors approximately 30 micrometers on a side with a QE of 80 - 90%, a noise of only +/- 3 e-/pix and with no multiplicative noise. For this reason, in 1989, one of us (JJ) developed a rear-illuminated, single-channel Si sensor, called the Turbodiode, employing some of the sophisticated readout techniques used to measure charge in a scientific CCD. We are now extending this work to a device in which a single 36 X 36 micrometers sensor is read out through a low-noise FET charge amplifier with a reset circuit and then passed to a correlated, double-sampling digitizer. To maintain the desired +/- 3 e noise level at the relatively high data rate of 1 MHz, our new device utilizes 64 separate readout amplifier/digitizer systems, operating in sequence. The resulting detector is more compact, efficient and reliable than the PMT it replaces but as its sensitive area is smaller than that of a PMT, it will require auxiliary optics when used with any LCM having a large (mm) pinhole. As the signal light is parallel, a simple lens mounted axially and with the CCDiode at its focus would suffice. Future versions may use 3 X 3 or 5 X 5 arrays of sensors to `track' the confocal spot as it is deflected by inhomogeneities of the specimen, change its effective size or shape or detect system misalignment.

  16. Confocal fluorescence microscopy for detection of cervical preneoplastic lesions

    NASA Astrophysics Data System (ADS)

    Sheikhzadeh, Fahime; Ward, Rabab K.; Carraro, Anita; Chen, Zhaoyang; van Niekerk, Dirk; MacAulay, Calum; Follen, Michele; Lane, Pierre; Guillaud, Martial

    2015-03-01

    We examined and established the potential of ex-vivo confocal fluorescence microscopy for differentiating between normal cervical tissue, low grade Cervical Intraepithelial Neoplasia (CIN1), and high grade CIN (CIN2 and CIN3). Our objectives were to i) use Quantitative Tissue Phenotype (QTP) analysis to quantify nuclear and cellular morphology and tissue architecture in confocal microscopic images of fresh cervical biopsies and ii) determine the accuracy of high grade CIN detection via confocal microscopy. Cervical biopsy specimens of colposcopically normal and abnormal tissues obtained from 15 patients were evaluated by confocal fluorescence microscopy. Confocal images were analyzed and about 200 morphological and architectural features were calculated at the nuclear, cellular, and tissue level. For the purpose of this study, we used four features to delineate disease grade including nuclear size, cell density, estimated nuclear-cytoplasmic (ENC) ratio, and the average of three nearest Delaunay neighbors distance (3NDND). Our preliminary results showed ENC ratio and 3NDND correlated well with histopathological diagnosis. The Spearman correlation coefficient between each of these two features and the histopathological diagnosis was higher than the correlation coefficient between colposcopic appearance and histopathological diagnosis. Sensitivity and specificity of ENC ratio for detecting high grade CIN were both equal to 100%. QTP analysis of fluorescence confocal images shows the potential to discriminate high grade CIN from low grade CIN and normal tissues. This approach could be used to help clinicians identify HGSILs in clinical settings.

  17. Confocal laser endomicroscopy for diagnosing lung cancer in vivo.

    PubMed

    Fuchs, Florian S; Zirlik, Sabine; Hildner, Kai; Schubert, Juergen; Vieth, Michael; Neurath, Markus F

    2013-06-01

    Confocal laser endomicroscopy is a novel endoscopic technique that may allow imaging of living cells in lung tissue in vivo. We assessed the potential of this technique for the detection of histology during screening bronchoscopy for lung cancer. 32 patients with suspected malignancies underwent bronchoscopy with endomicroscopy using acriflavine hydrochloride. Standardised areas and localised lesions were analysed by in vivo confocal imaging during bronchoscopy and biopsies were taken. Confocal images were graded and correlated prospectively with conventional histology from biopsies. Acriflavine hydrochloride yielded high-quality confocal images and strongly labelled airway epithelial cells. No side-effects were noted. 75,522 confocal images from 56 different locations were compared prospectively with histological data from biopsy specimens. Endomicroscopy allowed subsurface imaging with detailed analysis of cellular and subcellular structures. Neoplastic changes could be predicted with high accuracy (sensitivity 96.0%, specificity 87.1%, accuracy 91.0%). Confocal laser endomicroscopy with acriflavine is a novel diagnostic tool for the analysis of living cells during bronchoscopy and permits virtual histology of neoplastic changes in the airways with high accuracy. This technique may enable the rapid diagnosis of neoplasia during ongoing endoscopy in patients with suspected lung cancer. PMID:22997220

  18. Design of a confocal microfluidic particle sorter using fluorescent photon burst detection

    NASA Astrophysics Data System (ADS)

    Kunst, Beno H.; Schots, Arjen; Visser, Antonie J. W. G.

    2004-09-01

    An instrumental system is described for detecting and sorting single fluorescent particles such as microspheres, bacteria, viruses, or even smaller macromolecules in a flowing liquid. The system consists of microfluidic chips (biochips), computer controlled high voltage power supplies, and a fluorescence microscope with confocal optics. The confocal observation volume and detection electro-optics allow measurements of single flowing fluorescent particles. The output of the avalanche photodiode (single photon detector) is coupled to a real-time photon-burst detection device, which output can address the control of high voltage power supplies for sorting purposes. Liquid propulsion systems like electro-osmotic flow and plain electric fields to direct the particles through the observation volume have been tested and evaluated. The detection and real-time sorting of fluorescent microspheres are demonstrated. Applications of these biochips for screening of bacteriophages-type biolibraries are briefly discussed.

  19. Methods for Acquisition of Quantitative Data from Confocal Images of Gene Expression in situ

    PubMed Central

    Surkova, S. Yu.; Myasnikova, E. M.; Kozlov, K. N.; Samsonova, A. A.; Reinitz, J.; Samsonova, M. G.

    2009-01-01

    In this review, we summarize original methods for the extraction of quantitative information from confocal images of gene-expression patterns. These methods include image segmentation, the extraction of quantitative numerical data on gene expression, and the removal of background signal and spatial registration. Finally, it is possible to construct a spatiotemporal atlas of gene expression from individual images recorded at each developmental stage. Initially all methods were developed to extract quantitative numerical information from confocal images of segmentation gene expression in Drosophila melanogaster. The application of these methods to Drosophila images makes it possible to reveal new mechanisms in the formation of segmentation gene expression domains, as well as to construct a quantitative atlas of segmentation gene expression. Most image processing procedures can be easily adapted to process a wide range of biological images. PMID:19343098

  20. Confocal Raman microscopy to monitor extracellular matrix during dental pulp stem cells differentiation

    NASA Astrophysics Data System (ADS)

    Salehi, Hamideh; Collart-Dutilleul, Pierre-Yves; Gergely, Csilla; Cuisinier, Frdric J. G.

    2015-07-01

    Regenerative medicine brings promising applications for mesenchymal stem cells, such as dental pulp stem cells (DPSCs). Confocal Raman microscopy, a noninvasive technique, is used to study osteogenic differentiation of DPSCs. Integrated Raman intensities in the 2800 to 3000 cm-1 region (C-H stretching) and the 960 cm-1 peak (?1 PO43-) were collected (to image cells and phosphate, respectively), and the ratio of two peaks 1660 over 1690 cm-1 (amide I bands) to measure the collagen cross-linking has been calculated. Raman spectra of DPSCs after 21 days differentiation reveal several phosphate peaks: ?1 (first stretching mode) at 960 cm-1, ?2 at 430 cm-1, and ?4 at 585 cm-1 and collagen cross-linking can also be calculated. Confocal Raman microscopy enables monitoring osteogenic differentiation in vitro and can be a credible tool for clinical stem cell based research.

  1. Dental pulp stem cells (DPSCs) differentiation study by confocal Raman microscopy

    NASA Astrophysics Data System (ADS)

    Salehi, H.; Collart-Dutilleul, P.-Y.; Gergely, C.; Cuisinier, F. J. G.

    2014-03-01

    Regenerative medicine brings a huge application for Mesenchymal stem cells such as Dental Pulp Stem Cells (DPSCs). Confocal Raman microscopy, a non-invasive, label free , real time and high spatial resolution imaging technique is used to study osteogenic differentiation of DPSCs. Integrated Raman intensities in the 2800-3000 cm-1 region (C-H stretching) and 960 cm-1 peak (phosphate PO4 3-) were collected. In Dental Pulp Stem Cells 21st day differentiated in buffer solution, phosphate peaks ?1 PO4 3- (first vibrational mode) at 960cm-1 and ?2 PO4 3- at 430cm-1 and ?4 PO4 3- at 585cm-1 are obviously present. Confocal Raman microscopy enables the detection of cell differentiation and it can be used to investigate clinical stem cell research.

  2. Evaluation of human sclera after femtosecond laser ablation using two photon and confocal microscopy

    NASA Astrophysics Data System (ADS)

    Sun, Hui; Kurtz, Ronald; Juhasz, Tibor

    2012-08-01

    Glaucoma is the second-leading cause of blindness worldwide and is often associated with elevated intraocular pressure (IOP). Partial thickness intrascleral channels can be created with a femtosecond laser operating at a wavelength of 1700 nm. Such channels have the potential to increase outflow facility and reduce elevated IOP. Analysis of the dimensions and location of these channels is important in understanding their effects. We describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in human cadaver eyes. High-resolution images, hundreds of microns deep in the sclera, were obtained to allow determination of the shape and dimension of such channels. This demonstrates that concept of integrating femtosecond laser surgery, and two-photon and confocal imaging has the future potential for image-guided high-precision surgery in transparent and translucent tissue.

  3. Single molecule confocal fluorescence lifetime correlation spectroscopy for accurate nanoparticle size determination.

    PubMed

    Chon, Bonghwan; Briggman, Kimberly; Hwang, Jeeseong

    2014-07-14

    We report on an experimental procedure in confocal single molecule fluorescence lifetime correlation spectroscopy (FLCS) to determine the range of excitation power and molecular or particulate concentration in solution under which the application of an unmodified model autocorrelation function is justified. This procedure enables fitting of the autocorrelation to an accurate model to measure diffusion length (r) and diffusion time (?D) of single molecules in solution. We also report on the pinhole size dependency of r and ?D in a confocal FLCS platform. This procedure determines a set of experimental parameters with which the Stokes-Einstein (S-E) equation accurately measures the hydrodynamic radii of spherical nanoparticles, enabling the determination of the particle size range for which the hydrodynamic radius by the S-E equation measures the real particle radius. PMID:24879354

  4. Confocal Brillouin microscopy for three-dimensional mechanical imaging

    PubMed Central

    Scarcelli, Giuliano; Yun, Seok Hyun

    2009-01-01

    Acoustically induced inelastic light scattering, first reported in 1922 by Brillouin1, allows non-contact, direct readout of the viscoelastic properties of a material and has widely been investigated for material characterization2, structural monitoring3 and environmental sensing4. Extending the Brillouin technique from point sampling spectroscopy to imaging modality5 would open up new possibilities for mechanical imaging, but has been challenging because rapid spectrum acquisition is required. Here, we demonstrate a confocal Brillouin microscope based on a fully parallel spectrometera virtually imaged phased arraythat improves the detection efficiency by nearly 100-fold over previous approaches. Using the system, we show the first cross-sectional Brillouin imaging based on elastic properties as the contrast mechanism and monitor fast dynamic changes in elastic modulus during polymer crosslinking. Furthermore, we report the first in situ biomechanical measurement of the crystalline lens in a mouse eye. These results suggest multiple applications of Brillouin microscopy in biomedical and biomaterial science. PMID:19812712

  5. Confocal fluorescence microscopy for minimal-invasive tumor diagnosis

    NASA Astrophysics Data System (ADS)

    Zenzinger, M.; Gtz, M. H.; Fischer, S.; Bille, J.

    The goal of the project ``stereotactic laser-neurosurgery'' is the development of a system for careful and minimal-invasive resection of brain tumors with ultrashort laser pulses through a thin probe. A confocal laser-scanning-microscope is integrated in the probe. In this paper, the simulation of its optical properties by a laboratory setup and the expansion by the ability for fluorescence microscopy are reported. For a valuation of the imaging properties, the point-spread-function in three dimensions and the axial depth-transfer-function were measured and thus, among other things, the resolving power and the capacity for depth discrimination were analysed. The microscope will enable intra-operative detection of tumor cells by the method of immunofluorescence. As a first model of the application in the brain, cell cultures, that fluorescein-labelled antibodies were bound to specifically, were used in this work. Due to the fluorescence signal, it was possible to detect and identify clearly the areas that had been marked in this manner, proving the suitability of the setup for minimal-invasive tumor diagnosis.

  6. IV Neuropathy: An In Vivo Confocal Microscopic Study

    PubMed Central

    Almodovar, Jorge L.; Schifitto, Giovanni; McDermott, Michael P.; Ferguson, Michele; Herrmann, David N.

    2012-01-01

    Several approaches exist for quantitative assessment of human immunodeficiency virus (HIV) associated distal sensory polyneuropathy (DSP). While useful, each has some limitations. This study evaluated non-invasive, in vivo reflectance confocal microscopy (RCM) of Meissner Corpuscles (MCs) as a measure of HIV-DSP. Forty-eight adults (29 HIV-infected, 19 controls) underwent RCM of MC density (MCs/mm2) at the arch, fingertip and thenar eminence (TE), ankle skin biopsy to measure epidermal nerve fiber density (ENFD), electrophysiologic studies, and tactile, vibration and thermal threshold testing. HIV+ subjects were clinically categorized as having DSP signs or no signs. MC densities were lower in HIV+ subjects with DSP signs than in controls (arch, p = 0.0003; fingertip, p < 0.0001; TE, p = 0.0002). Tactile thresholds in the TE and foot were worse in HIV-DSP than in controls, but in this mild DSP cohort, sural amplitudes, ENFD and vibration and thermal thresholds didn't differ significantly from controls. Fingertip MC densities and tactile thresholds at the foot were also lower in HIV+ subjects without DSP signs than in controls. Other sensory measures were not significantly different in HIV+ subjects without DSP signs than in controls. MC density correlated inversely with tactile thresholds at each imaging location. The results suggest that RCM of MC density complements existing sensory DSP measures and discriminates mild HIV-DSP from controls at a stage when sural amplitudes do not. Further studies are required to determine whether RCM of MC density can establish quantitative changes in DSP, in response to treatment or disease progression. PMID:23070817

  7. Reflective Packaging

    NASA Technical Reports Server (NTRS)

    1994-01-01

    The aluminized polymer film used in spacecraft as a radiation barrier to protect both astronauts and delicate instruments has led to a number of spinoff applications. Among them are aluminized shipping bags, food cart covers and medical bags. Radiant Technologies purchases component materials and assembles a barrier made of layers of aluminized foil. The packaging reflects outside heat away from the product inside the container. The company is developing new aluminized lines, express mailers, large shipping bags, gel packs and insulated panels for the building industry.

  8. A comparision between rapid Golgi and Golgi-Cox impregnation methods for 3-D reconstruction of neurons at the confocal scanning laser microscope.

    PubMed

    Castano, P; Gioia, M; Barajon, I; Rumio, C; Miani, A

    1995-01-01

    We utilized two widely used impregnation methods, the silver "rapid Golgi" and the mercuric Golgi-Cox methods, for three-dimensional (3-D) reconstruction of neurons at the confocal scanning laser microscope (CSLM), to determine which of them was more suitable for this application. The Golgi-Cox method is the most consistent arid the cleanest procedure with respect to the "rapid Golgi" one which always produces samples with scattered reflective granules that interfere with the image formation at the CSLM. The interneuronal tissue in the case of Golgi-Cox impregnated specimens (i.e. the non-impregnated tissue among impregnated neurons) contributes less to the decrease of reflected light during z-sectioning than in the case of "rapid Golgi" impregnation, but the mercury impregnated samples reflect less than the silver impregnated ones. Owing to the necessity during deep z-scanning to adjust the sensitivity of the CLSM detector the acquisition of images from the deeper planes of the sample may be difficult. In our opinion the "sandwich" mounting of the specimen between two coverslips is indispensable in order to make it possible to scan it from both sides and, thus reduce the penetration in the sample and the consequent distortion of the image. Neither of the impregnation methods used is completely suitable for CLSM observations due both to their intrinsic limitations and to those imposed by the sample thickness. PMID:11322342

  9. Tethered confocal endomicroscopy capsule for diagnosis and monitoring of eosinophilic esophagitis

    PubMed Central

    Tabatabaei, Nima; Kang, DongKyun; Wu, Tao; Kim, Minkyu; Carruth, Robert W.; Leung, John; Sauk, Jenny S; Shreffler, Wayne; Yuan, Qian; Katz, Aubrey; Nishioka, Norman S; Tearney, Guillermo J.

    2013-01-01

    Eosinophilic esophagitis (EoE) is an allergic condition that is characterized by eosinophils infiltrating the esophageal wall. The treatment of the disease may require multiple follow up sedated endoscopies and biopsies to confirm elimination of eosinophils. These procedures are expensive, time consuming, and may be difficult for patients to tolerate. Here we report on the development of a confocal microscopy capsule for diagnosis and monitoring of EoE. The swallowable capsule implements a high-speed fiber-based reflectance confocal microscopy technique termed Spectrally Encoded Confocal Microscopy (SECM). SECM scans the sample in one dimension without moving parts by using wavelength swept source illumination and a diffraction grating at the back plane of the objective lens. As the wavelength of the source is tuned, the SECM optics within the 7 x 30 mm capsule are rotated using a driveshaft enclosed in a 0.8 mm flexible tether. A single rotation of the optics covered a field of view of 22 mm x 223 m. The lateral and axial resolutions of the device were measured to be 2.1 and 14 m, respectively. Images of Acetic Acid stained swine esophagus obtained with the capsule ex vivo and in vivo clearly showed squamous epithelial nuclei, which are smaller and less reflective than eosinophils. Imaging of esophageal biopsies from EoE patients ex vivo demonstrated the capability of this technology to visualize individual eosinophils. Based on the results of this study, we believe that this capsule will be a simpler and more effective device for diagnosing EoE and monitoring the therapeutic response of this disease. PMID:24466487

  10. Confocal Raman microscopy for investigating synthesis and characterization of individual optically trapped vinyl-polymerized surfactant particles.

    PubMed

    Schaefer, Jonathan J; Crawford, Alexis C; Porter, Marc D; Harris, Joel M

    2014-01-01

    Small polymeric particles are increasingly employed as adsorbent materials, as molecular carriers, as delivery vehicles, and in preconcentration applications. The rational development of these materials requires in situ methods of analysis to characterize their synthesis, structure, and applications. Optical-trapping confocal Raman microscopy is a spectroscopic method capable of acquiring information at several stages of the development of such dispersed particulate materials. In the present study, an example material is developed and tested using confocal Raman microscopy for characterization at each stage of the process. Specifically, the method is used to investigate the synthesis, structure, and applications of individual polymeric surfactant particles produced by the vinyl polymerization of sodium 11-acrylamidoundecanoate (SAAU). The kinetics of polymerization can be monitored over time by measuring the loss of the acrylamide C=C functional groups using confocal Raman microscopy of particles optically trapped by the excitation laser, where, within the limits of detecting the vinyl functional group, the complete polymerization of the SAAU monomer was achieved. The polymerized SAAU particles are spherical, and they exhibit uniform access to water throughout their structure, as tested by the penetration of heavy water (D2O) and collection of spatially resolved Raman spectra from the interior of the particle. These porous particles contain hydrophobic domains that can be used to accumulate molecules for adsorption or carrier applications. This property was tested by using confocal Raman microscopy to measure the accumulation equilibria and kinetics of a model compound, dioxybenzone. The partitioning of this compound into the polymer surfactant could be determined on a quantitative basis using relative scattering cross sections of the SAAU monomer and the adsorbate. The study points out the utility of optical-trapping confocal Raman microscopy for investigating the synthesis, structure, and potential carrier applications of polymeric particle materials. PMID:25014718

  11. A line-scanning semi-confocal multi-photon fluorescence microscope with a simultaneous broadband spectral acquisition and its application to the study of the thylakoid membrane of a cyanobacterium Anabaena PCC7120.

    PubMed

    Kumazaki, Shigeichi; Hasegawa, Makoto; Ghoneim, Mohammad; Shimizu, Yugo; Okamoto, Kenji; Nishiyama, Masayoshi; Oh-Oka, Hirozo; Terazima, Masahide

    2007-11-01

    We describe the construction and characterization of a laser-line-scanning microscope capable of detection of broad fluorescence spectra with a resolution of 1 nm. A near-infrared femtosecond pulse train at 800 nm was illuminated on a line (one lateral axis, denoted as X axis) in a specimen by a resonant scanning mirror oscillating at 7.9 kHz, and total multi-photon-induced fluorescence from the linear region was focused on the slit of an imaging polychromator. An electron-multiplying CCD camera was used to resolve fluorescence of different colours at different horizontal pixels and fluorescence of different spatial positions in a specimen at different vertical pixels. Scanning on the other two axes (Y and Z) was achieved by a closed-loop controlled sample scanning stage and a piezo-driven objective actuator. The full widths at half maximum of the point-spread function of the system were estimated to be 0.39-0.40, 0.33 and 0.56-0.59 mum for the X (lateral axis along the line-scan), Y (the other lateral axis) and Z axes (the axial direction), respectively, at fluorescence wavelengths between 644 and 690 nm. A biological application of this microscope was demonstrated in a study of the sub-cellular fluorescence spectra of thylakoid membranes in a cyanobacterium, Anabaena PCC7120. It was found that the fluorescence intensity ratio between chlorophyll molecules mainly of photosystem II and phycobilin molecules of phycobilisome (chlorophyll/phycobilin), in the thylakoid membranes, became lower as one probed deeper inside the cells. This was attributable not to position dependence of re-absorption or scattering effects, but to an intrinsic change in the local physiological state of the thylakoid membrane, with the help of a transmission spectral measurement of sub-cellular domains. The efficiency of the new line-scanning spectromicroscope was estimated in comparison with our own point-by-point scanning spectromicroscope. Under typical conditions of observing cyanobacterial cells, the total exposure time became shorter by about 50 times for a constant excitation density. The improvement factor was proportional to the length of the line-scanned region, as expected. PMID:17970923

  12. Interfacial shape and contact-angle measurement of transparent samples with confocal interference microscopy.

    PubMed

    Fischer, D G; Ovryn, B

    2000-04-01

    A model has been developed that predicts the effective optical path through a thick, refractive specimen on a reflective substrate, as measured with a scanning confocal interference microscope equipped with a high-numerical-aperture objective. Assuming that the effective pinhole of the confocal microscope has an infinitesimal diameter, only one ray in the illumination bundle (the magic ray) contributes to the differential optical path length (OPL). A pinhole with finite diameter, however, allows rays within a small angular cone centered on the magic ray to contribute to the OPL. The model was incorporated into an iterative algorithm that allows the measured phase to be corrected for refractive errors by use of an a priori estimate of the sample profile. The algorithm was validated with a reflected-light microscope equipped with a phase-shifting laser-feedback interferometer to measure the interface shape and the 68 degrees contact angle of a silicone-oil drop on a coated silicon wafer. PMID:18064085

  13. Reflection Coefficients.

    ERIC Educational Resources Information Center

    Greenslade, Thomas B., Jr.

    1994-01-01

    Discusses and provides an example of reflectivity approximation to determine whether reflection will occur. Provides a method to show thin-film interference on a projection screen. Also applies the reflectivity concepts to electromagnetic wave systems. (MVL)

  14. Fibre optic confocal imaging (FOCI) for subsurface microscopy of the colon in vivo.

    PubMed Central

    Delaney, P M; King, R G; Lambert, J R; Harris, M R

    1994-01-01

    Fibre optic confocal imaging (FOCI) is a new type of microscopy which has been recently developed (Delaney et al. 1993). In contrast to conventional light microscopy, FOCI and other confocal techniques allow clear imaging of subsurface structures within translucent objects. However, unlike conventional confocal microscopes which are bulky (because of a need for accurate alignment of large components) FOCI allows the imaging end to be miniaturised and relatively mobile. FOCI is thus particularly suited for clear subsurface imaging of structures within living animals or subjects. The aim of the present study was to assess the suitability of using FOCI for imaging of subsurface structures within the colon, both in vitro (human and rat biopsies) and in vivo (in rats). Images were obtained in fluorescence mode (excitation 488 nm, detection above 515 nm) following topical application of fluorescein. By this technique the glandular structure of the colon was imaged. FOCI is thus suitable for subsurface imaging of the colon in vivo. Images Fig. 2 Fig. 3 PMID:8157487

  15. In vivo molecular and morphological imaging by real time confocal mini-microscopy

    NASA Astrophysics Data System (ADS)

    Goetz, Martin; Gregor, Sebastian; Fottner, Christian; Garcia-Lazaro, Jose; Schirrmacher, Esther; Kempski, Oliver; Bartenstein, Peter; Weber, Mathias; Biesterfeld, Stefan; Galle, Peter R.; Neurath, Markus F.; Kiesslich, Ralf

    2006-02-01

    We evaluated a newly developed miniaturized confocal laser microscopy probe for real-time in vivo molecular and morphological imaging of normal, inflammatory, and malignant tissue in rodents. In the rigid mini-microscopy probe (diameter 7 mm), a single line laser delivers an excitation wavelength of 488 nm. Optical slice thickness is 7 ?m, lateral resolution 0.7 ?m. The range of the z-axis is 0 - 250 ?m below the tissue surface. Organ systems were examined in vivo in rodent models of human diseases. FITC-labeled Lycopersion esculentum lectin was injected or selected cell populations stained for molecular targeting. Morphological imaging was performed using fluorescein sodium, FITC-labeled dextran, and/or acriflavine hydrochloride. Cellular and subcellular details could be readily visualised in vivo at high resolution. Tissue characteristics of different organs were rendered at real time. Selective blood cell staining allowed observation of blood flow and cell migration. Inflammatory diseases such as hepatitis were diagnosed, and tumors were characterized under microscopic control in vivo. Confocal mini-microscopy allows real time in vivo molecular and morphological histologic imaging at high resolution of normal and diseased tissue. Since confocal microscopy is applicable to humans, this technology will have a high impact on different faculties in medicine.

  16. Super-resolution mapping of glutamate receptors in C. elegans by confocal correlated PALM.

    PubMed

    Vangindertael, Jeroen; Beets, Isabel; Rocha, Susana; Dedecker, Peter; Schoofs, Liliane; Vanhoorelbeeke, Karen; Hofkens, Johan; Mizuno, Hideaki

    2015-01-01

    Photoactivated localization microscopy (PALM) is a super-resolution imaging technique based on the detection and subsequent localization of single fluorescent molecules. PALM is therefore a powerful tool in resolving structures and putative interactions of biomolecules at the ultimate analytical detection limit. However, its limited imaging depth restricts PALM mostly to in vitro applications. Considering the additional need for anatomical context when imaging a multicellular organism, these limitations render the use of PALM in whole animals difficult. Here we integrated PALM with confocal microscopy for correlated imaging of the C. elegans nervous system, a technique we termed confocal correlated PALM (ccPALM). The neurons, lying below several tissue layers, could be visualized up to 10 ?m deep inside the animal. By ccPALM, we visualized ionotropic glutamate receptor distributions in C. elegans with an accuracy of 20 nm, revealing super-resolution structure of receptor clusters that we mapped onto annotated neurons in the animal. Pivotal to our results was the TIRF-independent detection of single molecules, achieved by genetic regulation of labeled receptor expression and localization to effectively reduce the background fluorescence. By correlating PALM with confocal microscopy, this platform enables dissecting biological structures with single molecule resolution in the physiologically relevant context of whole animals. PMID:26323790

  17. Super-resolution mapping of glutamate receptors in C. elegans by confocal correlated PALM

    PubMed Central

    Vangindertael, Jeroen; Beets, Isabel; Rocha, Susana; Dedecker, Peter; Schoofs, Liliane; Vanhoorelbeeke, Karen; Hofkens, Johan; Mizuno, Hideaki

    2015-01-01

    Photoactivated localization microscopy (PALM) is a super-resolution imaging technique based on the detection and subsequent localization of single fluorescent molecules. PALM is therefore a powerful tool in resolving structures and putative interactions of biomolecules at the ultimate analytical detection limit. However, its limited imaging depth restricts PALM mostly to in vitro applications. Considering the additional need for anatomical context when imaging a multicellular organism, these limitations render the use of PALM in whole animals difficult. Here we integrated PALM with confocal microscopy for correlated imaging of the C. elegans nervous system, a technique we termed confocal correlated PALM (ccPALM). The neurons, lying below several tissue layers, could be visualized up to 10??m deep inside the animal. By ccPALM, we visualized ionotropic glutamate receptor distributions in C. elegans with an accuracy of 20?nm, revealing super-resolution structure of receptor clusters that we mapped onto annotated neurons in the animal. Pivotal to our results was the TIRF-independent detection of single molecules, achieved by genetic regulation of labeled receptor expression and localization to effectively reduce the background fluorescence. By correlating PALM with confocal microscopy, this platform enables dissecting biological structures with single molecule resolution in the physiologically relevant context of whole animals. PMID:26323790

  18. Confocal soft X-ray scanning transmission microscopy: setup, alignment procedure and limitations

    PubMed Central

    Späth, Andreas; Raabe, Jörg; Fink, Rainer H.

    2015-01-01

    Zone-plate-based scanning transmission soft X-ray microspectroscopy (STXM) is a well established technique for high-contrast imaging of sufficiently transparent specimens (e.g. ultrathin biological tissues, polymer materials, archaeometric specimens or magnetic thin films) with spatial resolutions in the regime of 20 nm and high spectroscopic or chemical sensitivity. However, due to the relatively large depth of focus of zone plates, the resolution of STXM along the optical axis so far stays unambiguously behind for thicker X-ray transparent specimens. This challenge can be addressed by the implementation of a second zone plate in the detection pathway of the beam, resulting in a confocal arrangement. Within this paper a first proof-of-principle study for a confocal STXM (cSTXM) and an elaborate alignment procedure in transmission and fluorescence geometry are presented. Based on first confocal soft X-ray micrographs of well known specimens, the advantage and limitation of cSTXM as well as further development potentials for future applications are discussed. PMID:25537596

  19. To see the unseeable: confocal miniprobes for routine microscopic imaging during endoscopy

    NASA Astrophysics Data System (ADS)

    Osdoit, A.; Lacombe, F.; Cav, C.; Loiseau, S.; Peltier, E.

    2007-02-01

    Confocal fluorescence high resolution imaging during standard endoscopic procedures has been presented as a very promising tool to enhance patient care and physician practice by providing supplementary diagnostic information in real-time. The purpose of this paper is to show not only potential, but convincing results of endoscopic microscopy using a catheter-based approach. Mauna Kea Technologies' core technology, Cellvizio, delivers dynamic imaging at 12 frames/second using confocal miniprobes inserted through the operating channel of regular endoscopes. Cellvizio is composed of 3 parts including (a) a Laser Scanning Unit, (b) Confocal Miniprobe TM with the following characteristics: 5-15 ?m axial resolution, 2-5 ?m lateral resolution, 15-100 ?m depth of penetration, field of view of 600x500 ?m and (c) a software package with onthe- fly processing capabilities. With several tens of patients examined during routine GI endoscopy procedures, the most relevant clinical parameters could be assessed in a doubled-blinded fashion between the endoscopist and a pathologist and results showing very high accuracy in the differentiation of neoplasia from normal and hyperplastic tissue were obtained. In the field of pulmonology, the micro-autofluorescence properties of tissues could be assessed and structures never before accessed in vivo were observed. Cellvizio® may be useful to study bronchial remodeling in asthma and chronic obstructive pulmonary diseases. Using appropriate topical fluorescent dye, the Confocal Miniprobes may also make it possible to perform optical biopsy of precancerous and superficial bronchial cancers. Cellvizio® is as a new tool towards "targeted biopsies", leading to earlier, more reliable and cost effective diagnostic procedures. Other applications, specifically in molecular imaging are also made possible by the miniaturization of the probe (combination with biopsy needle for solid organs use or lymph node detection) and by the compatibility of the system with other imaging modalities (auto-fluorescence and narrow-band imaging endoscopy, MRI, PET, etc).

  20. Digital adaptive optics line-scanning confocal imaging system

    NASA Astrophysics Data System (ADS)

    Liu, Changgeng; Kim, Myung K.

    2015-11-01

    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.

  1. A dual modality fluorescence confocal and optical coherence tomography microendoscope

    NASA Astrophysics Data System (ADS)

    Makhlouf, Houssine; Rouse, Andrew R.; Gmitro, Arthur F.

    2010-02-01

    We demonstrate the implementation of a Fourier domain optical coherence tomography (OCT) imaging system incorporated into the optical train of a fluorescence confocal microendoscope. The slit-scanning confocal system has been presented previously and achieves 3?m lateral resolution and 25?m axial resolution over a field of view of 430?m. Its multi-spectral mode of operation captures images with 6nm average spectral resolution. To incorporate OCT imaging, a common-path interferometer is made with a super luminescent diode and a reference coverslip located at the distal end of the fiber bundle catheter. The infrared diode spectral width allows a theoretical OCT axial resolution of 12.9?m. Light from the reference and sample combine, and a diffraction grating produces a spectral interferogram on the same 2D CCD camera used for confocal microendoscopic imaging. OCT depth information is recovered by a Fourier transform along the spectral dispersion direction. Proper operation of the system scan mirrors allows rapid switching between confocal and OCT imaging modes. The OCT extension takes advantage of the slit geometry, so that a 2D image is acquired without scanning. Combining confocal and OCT imaging modalities provides a more comprehensive view of tissue and the potential to improve disease diagnosis. A preliminary bench-top system design and imaging results are presented.

  2. Fused oblique incidence reflectometry and confocal fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Risi, Matthew D.; Rouse, Andrew R.; Gmitro, Arthur F.

    2011-03-01

    Confocal microendoscopy provides real-time high resolution cellular level images via a minimally invasive procedure, but relies on exogenous fluorophores, has a relatively limited penetration depth (100 ?m) and field of view (700 ?m), and produces a high rate of detailed information to the user. A new catheter based multi-modal system has been designed that combines confocal imaging and oblique incidence reflectometry (OIR), which is a non-invasive method capable of rapidly extracting tissue absorption, ?a, and reduced scattering, ?'s, spectra from tissue. The system builds on previous developments of a custom slit-scan multi-spectral confocal microendoscope and is designed to rapidly switch between diffuse spectroscopy and confocal fluorescence imaging modes of operation. An experimental proof-of-principle catheter has been developed that consists of a fiber bundle for traditional confocal fluorescence imaging and a single OIR source fiber which is manually redirected at +/- 26 degrees. Diffusely scattered light from each orientation of the source fiber is collected via the fiber bundle, with a frame of data representing spectra collected at a range of distances from the OIR source point. Initial results with intralipid phantoms show good agreement to published data over the 550-650 nm spectral range. We successfully imaged and measured the optical properties of rodent cardiac muscle.

  3. Total Internal Reflection Fluorescence (TIRF) Microscopy of Chlamydomonas Flagella

    PubMed Central

    Engel, Benjamin D.; Lechtreck, Karl-Ferdinand; Sakai, Tsuyoshi; Ikebe, Mitsuo; Witman, George B.; Marshall, Wallace F.

    2013-01-01

    The eukaryotic flagellum is host to a variety of dynamic behaviors, including flagellar beating, the motility of glycoproteins in the flagellar membrane, and intraflagellar transport (IFT), the bidirectional traffic of protein particles between the flagellar base and tip. IFT is of particular interest, as it plays integral roles in flagellar length control, cell signaling, development, and human disease. However, our ability to understand dynamic flagellar processes such as IFT is limited in large part by the fidelity with which we can image these behaviors in living cells. This chapter introduces the application of total internal reflection fluorescence (TIRF) microscopy to visualizing the flagella of Chlamydomonas reinhardtii. The advantages and challenges of TIRF are discussed in comparison to confocal and differential interference contrast (DIC) techniques. This chapter also reviews current IFT insights gleaned from TIRF microscopy of Chlamydomonas and provides an outlook on the future of the technique, with particular emphasis on combining TIRF with other emerging imaging technologies. PMID:20409817

  4. Identifying joints from measured reflection coefficients in beam-like structures with application to a pipe support

    NASA Astrophysics Data System (ADS)

    Zhang, Bing; Waters, Timothy P.; Mace, Brian R.

    2010-04-01

    The properties of joints in mechanical systems are notoriously uncertain causing corresponding uncertainty in the systems' dynamic responses. A piping system is one such example where an accurate knowledge of joint properties is useful for the purposes of structure-borne sound transmission, fatigue considerations and structural health monitoring. This paper presents an inverse technique that is applicable to joint estimation in one-dimensional structures such as a pipe. Measured wave reflection coefficients are used which have several advantages over modal information. First, they characterise just the joint and adjacent pipes and are independent of the rest of the built-up system. Second, they are potentially more sensitive to the joint parameters in question than are modal parameters. The method is illustrated by means of an experimental case study featuring a straight pipe suspended by a cantilevered hanger. The stiffness and inertia of the hanger are accurately identified from measured data at frequencies significantly higher than the fundamental modes of the structure.

  5. Application of Fresnel diffraction from a 2D array of reflective disks in optical profilometry of a flat surface

    NASA Astrophysics Data System (ADS)

    Darudi, Ahmad; Asgari, Pegah; Pourvais, Yousef

    2015-05-01

    Optical methods of three-dimensional profilometry have been of growing interest in both industrial and scientific applications. These techniques provide absolutely non-destructive measurement due to their non-contact nature and maintain their high precision in a large field of view. Most of these techniques however, are based on interferometry which happens to be considerably sensitive to environmental noises such as turbulence and vibration. We have used the phenomena of Fresnel diffraction from phase-steps instead of interferometry to maintain a higher precision and reduce sensitivity to environmental noises. This phenomena has been recently introduced as a method for precise measurement of wavelength, thickness and refractive index. A 2D array of reflective disks are placed above the test surface to provide the required phase-steps. In this paper, theoretical principles of Fresnel diffraction from phase-steps are discussed and the experimental results of testing an optical flat surface are presented. A flat mirror surface has been tested as an optical test surface and is been profiled. The results show that the method is precise and is not sensitive to environmental noises such as vibration and turbulence. Furthermore, the method seems to be a powerful means for testing of curved surfaces, too.

  6. Spinning Disk Confocal Imaging of Neutrophil Migration in Zebrafish

    PubMed Central

    Lam, Pui-ying; Fischer, Robert S; Shin, William D.; Waterman, Clare M; Huttenlocher, Anna

    2014-01-01

    Live-cell imaging techniques have been substantially improved due to advances in confocal microscopy instrumentation coupled with ultrasensitive detectors. The spinning disk confocal system is capable of generating images of fluorescent live samples with broad dynamic range and high temporal and spatial resolution. The ability to acquire fluorescent images of living cells in vivo on a millisecond timescale allows the dissection of biological processes that have not previously been visualized in a physiologically relevant context. In vivo imaging of rapidly moving cells such as neutrophils can be technically challenging. In this chapter, we describe the practical aspects of imaging neutrophils in zebrafish embryos using spinning disk confocal microscopy. Similar setups can also be applied to image other motile cell types and signaling processes in translucent animals or tissues. PMID:24504955

  7. Frequency Division Multiplexed Multichannel High-Speed Fluorescence Confocal Microscope

    PubMed Central

    Wu, Fei; Zhang, Xueqian; Cheung, Joseph Y.; Shi, Kebin; Liu, Zhiwen; Luo, Claire; Yin, Stuart; Ruffin, Paul

    2006-01-01

    In this article, we report a new type of fluorescence confocal microscope: frequency division multiplexed multichannel fluorescence confocal microscope, in which we encode the spatial location information into the frequency domain. In this microscope, the exciting laser beam is first split into multiple beams and each beam is modulated at a different frequency. These multiple beams are focused at different locations of the target to form multiple focal points, which further generate multiple fluorescent emission spots. The fluorescent emissions from different focal points are also modulated at different frequencies, because the exciting beams are modulated at different frequencies (or difference carrier frequency). Then, all the fluorescent emissions (modulated at different frequencies) are collected together and detected by a highly sensitive, large-dynamic-range photomultiplier tube. By demodulating the detected signal (i.e., via the Fourier transform), we can distinguish the fluorescent light emitted from the different locations by the corresponding carrier frequencies. The major advantage of this unique fluorescence confocal microscope is that it not only has a high sensitivity because of the use of photomultiplier tube but also can get multiple-point data simultaneously, which is crucial to study the dynamic behavior of many biological process. As an initial step, to verify the feasibility of the proposed multichannel confocal microscope, we have developed a two-channel confocal fluorescence microscope and applied it to study the dynamic behavior of the changes of the calcium ion concentration during the single cardiac myocyte contraction. Our preliminary experimental results demonstrated that we could indeed realize multichannel confocal fluorescence microscopy by utilizing the frequency division multiplexed microscope, which could become an effective tool to study the dynamic behavior of many biological processes. PMID:16815894

  8. Nonlinear Confocal Microscopy for High-Resolution Measurement

    NASA Astrophysics Data System (ADS)

    Egami, Chikara; Ito, Atsuo; Liu, Yingzhi

    2008-08-01

    We demonstrate cell imaging with a new confocal nonlinear optical microscope using a low-power cw laser. The confocal nonlinear optical microscope, employing degenerate four-wave mixing geometry, can detect the fine structure of submicron objects with nanoscale contents such as biological cells. The optical signal, which is given by the third-order susceptibility tensor, is confined to the focal region of the focusing incident beam, because the absorption of the object is dependent on the third power of the excitation laser intensity. We have observed a thylakoid membrane in a chloroplast by scanning the tensor element inside.

  9. Confocal laser endomicroscopy: technical status and current indications.

    PubMed

    Hoffman, A; Goetz, M; Vieth, M; Galle, P R; Neurath, M F; Kiesslich, R

    2006-12-01

    Confocal laser endomicroscopy is a newly introduced endoscopic tool that makes it possible to carry out confocal microscopic examination of the mucosal layer during ongoing endoscopy. Different types of tissue and diseases can be diagnosed immediately, facilitating early diagnosis of gastrointestinal cancer. Analysis of the in vivo microarchitecture is helpful in targeting biopsies to relevant areas. In addition, subsurface imaging can unmask microscopic diseases - (microscopic colitis) or bacterial infection ( HELICOBACTER PYLORI), for example. Molecular imaging is becoming feasible, and this will shortly open the door to new indications in gastrointestinal endoscopy (e.g., in vivo receptor analysis). PMID:17163333

  10. Confocal Raman microscopy for identification of bacterial species in biofilms

    NASA Astrophysics Data System (ADS)

    Beier, Brooke D.; Quivey, Robert G.; Berger, Andrew J.

    2011-03-01

    Implemented through a confocal microscope, Raman spectroscopy has been used to distinguish between biofilm samples of two common oral bacteria species, Streptococcus sanguinis and mutans, which are associated with healthy and cariogenic plaque, respectively. Biofilms of these species are studied as a model of dental plaque. A prediction model has been calibrated and validated using pure biofilms. This model has been used to identify the species of transferred and dehydrated samples (much like a plaque scraping) as well as hydrated biofilms in situ. Preliminary results of confocal Raman mapping of species in an intact two-species biofilm will be shown.

  11. Optimized extended depth of focus simulated analysis for confocal microscopy.

    PubMed

    Duadi, Hamootal; Margalit, Ofer; Zalevsky, Zeev; Sarafis, Vassilios

    2010-06-01

    In imaging systems and especially in confocal microscopy systems there is a trade-off between the lateral resolution and the obtained depth of focus. The use of complex pupils to improve the lateral resolution by engineering the point spread function is a common approach; however the lateral improvement reduces the effective depth of focus and therefore the fluorescence efficiency. In this work we analytically develop an optimized approach for obtaining a complex pupil with an extended depth of focus. The proposed solution is numerically applied and tested in designing an improved focal depth in confocal microscope configuration. PMID:20508707

  12. Confocal Raman imaging of crystalline an glassy materials

    SciTech Connect

    Bradley, N.L.; Morris, M.D.

    1995-12-31

    Spatial distribution of materials components can be measured by confocal Raman imaging. We describe a confocal line-imaging system in which the spectrograph entrance slit functions as a spatial filter. The instrument uses a scanning galvanometer mirror to generate uniform intensity line illumination. A flexure mount with better than 0.1 micrometer positioning accuracy moves the sample under the fixed optical system. The Raman scatter is collected and projected along the entrance slit of an axial transmissive spectrograph. A CCD collects spatially resolved spectra.

  13. Optimization of confocal laser induced fluorescence in a plasma.

    PubMed

    VanDervort, R; Elliott, D; McCarren, D; McKee, J; Soderholm, M; Sears, S; Scime, E

    2014-11-01

    Laser Induced Fluorescence (LIF) provides measurements of flow speed, temperature, and density of ions or neutrals in a plasma. Traditionally, a LIF measurement requires two ports on a plasma device; one for laser injection and one for emission collection. Proper alignment of LIF optics is time consuming and sensitive to mechanical vibration. We describe a confocal configuration for LIF that requires a single port and requires no alignment. The measurement location is scanned radially by physically moving the entire optical structure. Confocal LIF measurements are compared to traditional LIF measurements over the same radial range. PMID:25430315

  14. Optimization of confocal laser induced fluorescence in a plasma

    NASA Astrophysics Data System (ADS)

    VanDervort, R.; Elliott, D.; McCarren, D.; McKee, J.; Soderholm, M.; Sears, S.; Scime, E.

    2014-11-01

    Laser Induced Fluorescence (LIF) provides measurements of flow speed, temperature, and density of ions or neutrals in a plasma. Traditionally, a LIF measurement requires two ports on a plasma device; one for laser injection and one for emission collection. Proper alignment of LIF optics is time consuming and sensitive to mechanical vibration. We describe a confocal configuration for LIF that requires a single port and requires no alignment. The measurement location is scanned radially by physically moving the entire optical structure. Confocal LIF measurements are compared to traditional LIF measurements over the same radial range.

  15. Confocal shift interferometry of coherent emission from trapped dipolar excitons

    SciTech Connect

    Repp, J.; Schinner, G. J.; Schubert, E.; Rai, A. K.; Wieck, A. D.; Reuter, D.; Wurstbauer, U.; Holleitner, A. W.; and others

    2014-12-15

    We introduce a confocal shift-interferometer based on optical fibers. The presented spectroscopy allows measuring coherence maps of luminescent samples with a high spatial resolution even at cryogenic temperatures. We apply the spectroscopy onto electrostatically trapped, dipolar excitons in a semiconductor double quantum well. We find that the measured spatial coherence length of the excitonic emission coincides with the point spread function of the confocal setup. The results are consistent with a temporal coherence of the excitonic emission down to temperatures of 250 mK.

  16. An invertebrate embryologist's guide to routine processing of confocal images.

    PubMed

    von Dassow, George

    2014-01-01

    It is almost impossible to use a confocal microscope without encountering the need to transform the raw data through image processing. Adherence to a set of straightforward guidelines will help ensure that image manipulations are both credible and repeatable. Meanwhile, attention to optimal data collection parameters will greatly simplify image processing, not only for convenience but for quality and credibility as well. Here I describe how to conduct routine confocal image processing tasks, including creating 3D animations or stereo images, false coloring or merging channels, background suppression, and compressing movie files for display. PMID:24567209

  17. Full-field interferometric confocal microscopy using a VCSEL array

    PubMed Central

    Redding, Brandon; Bromberg, Yaron; Choma, Michael A.; Cao, Hui

    2014-01-01

    We present an interferometric confocal microscope using an array of 1200 VCSELs coupled to a multimode fiber. Spatial coherence gating provides ~18,000 continuous virtual pinholes allowing an entire en face plane to be imaged in a snapshot. This approach maintains the same optical sectioning as a scanning confocal microscope without moving parts, while the high power of the VCSEL array (~5 mW per laser) enables high-speed image acquisition with integration times as short as 100 µs. Interferometric detection also recovers the phase of the image, enabling quantitative phase measurements and improving the contrast when imaging phase objects. PMID:25078199

  18. Reflection-contrast limit of fiber-optic image guides

    PubMed Central

    Lane, Pierre M.; MacAulay, Calum E.

    2009-01-01

    Fiber-optic image guides in confocal reflectance endomicroscopes introduce background backscatter that limits the achievable contrast in these devices. We show the dominant source of backscatter from the image guide is due to Rayleigh scattering at short wavelengths and terminal reflections of the fibers at long wavelengths. The effective Rayleigh scattering coefficient and the wavelength-independent reflectivity due terminal reflections are measured experimentally in a commercial image guide. The Rayleigh scattering component of backscatter can be accurately predicted using the fractional refractive-index difference and length of the fibers in the image guide. We also presented a simple model that can be used to predict signal-to-background ratio in a fiber-optic confocal reflectance endomicroscope for biologically relevant tissues and contrast agents that cover a wide range of reflectivity. PMID:20059266

  19. Confocal mosaicing microscopy of human skin ex vivo: spectral analysis for digital staining to simulate histology-like appearance

    NASA Astrophysics Data System (ADS)

    Bini, Jason; Spain, James; Nehal, Kishwer; Hazelwood, Vikki; Dimarzio, Charles; Rajadhyaksha, Milind

    2011-07-01

    Confocal mosaicing microscopy enables rapid imaging of large areas of fresh tissue, without the processing that is necessary for conventional histology. Mosaicing may offer a means to perform rapid histology at the bedside. A possible barrier toward clinical acceptance is that the mosaics are based on a single mode of grayscale contrast and appear black and white, whereas histology is based on two stains (hematoxylin for nuclei, eosin for cellular cytoplasm and dermis) and appears purple and pink. Toward addressing this barrier, we report advances in digital staining: fluorescence mosaics that show only nuclei, are digitally stained purple and overlaid on reflectance mosaics, which show only cellular cytoplasm and dermis, and are digitally stained pink. With digital staining, the appearance of confocal mosaics mimics the appearance of histology. Using multispectral analysis and color matching functions, red, green, and blue (RGB) components of hematoxylin and eosin stains in tissue were determined. The resulting RGB components were then applied in a linear algorithm to transform fluorescence and reflectance contrast in confocal mosaics to the absorbance contrast seen in pathology. Optimization of staining with acridine orange showed improved quality of digitally stained mosaics, with good correlation to the corresponding histology.

  20. Confocal mosaicing microscopy of human skin ex vivo: spectral analysis for digital staining to simulate histology-like appearance.

    PubMed

    Bini, Jason; Spain, James; Nehal, Kishwer; Hazelwood, Vikki; DiMarzio, Charles; Rajadhyaksha, Milind

    2011-07-01

    Confocal mosaicing microscopy enables rapid imaging of large areas of fresh tissue, without the processing that is necessary for conventional histology. Mosaicing may offer a means to perform rapid histology at the bedside. A possible barrier toward clinical acceptance is that the mosaics are based on a single mode of grayscale contrast and appear black and white, whereas histology is based on two stains (hematoxylin for nuclei, eosin for cellular cytoplasm and dermis) and appears purple and pink. Toward addressing this barrier, we report advances in digital staining: fluorescence mosaics that show only nuclei, are digitally stained purple and overlaid on reflectance mosaics, which show only cellular cytoplasm and dermis, and are digitally stained pink. With digital staining, the appearance of confocal mosaics mimics the appearance of histology. Using multispectral analysis and color matching functions, red, green, and blue (RGB) components of hematoxylin and eosin stains in tissue were determined. The resulting RGB components were then applied in a linear algorithm to transform fluorescence and reflectance contrast in confocal mosaics to the absorbance contrast seen in pathology. Optimization of staining with acridine orange showed improved quality of digitally stained mosaics, with good correlation to the corresponding histology. PMID:21806269

  1. Polymer Cholesteric Liquid Crystal (PCLC) Flake/Fluid Host Suspensions: A Novel Electro-Optical Medium for Reflective Color Display Applications

    SciTech Connect

    Marshall, K.L.; Trajkovska-Petkoska, A.; Kosc, T.Z.; Jacobs, S.D.

    2006-04-17

    Polymer cholesteric liquid crystal (PCLC) flake/fluid host suspensions are a new and promising particle display technology for both full-color flexible display applications and electronic paper. Devices containing these "polarizing pigments" switch rapidly at very low voltages and produce highly saturated, circularly polarized reflectance colors without requiring polarizers or color filters.

  2. Impact of immersion oils and mounting media on the confocal imaging of dendritic spines

    PubMed Central

    Peterson, Brittni M.; Mermelstein, Paul G.; Meisel, Robert L.

    2015-01-01

    Background Structural plasticity, such as changes in dendritic spine morphology and density, reflect changes in synaptic connectivity and circuitry. Procedural variables used in different methods for labeling dendritic spines have been quantitatively evaluated for their impact on the ability to resolve individual spines in confocal microscopic analyses. In contrast, there have been discussions, though no quantitative analyses, of the potential effects of choosing specific mounting media and immersion oils on dendritic spine resolution. New Method Here we provide quantitative data measuring the impact of these variables on resolving dendritic spines in 3D confocal analyses. Medium spiny neurons from the rat striatum and nucleus accumbens are used as examples. Results Both choice of mounting media and immersion oil affected the visualization of dendritic spines, with choosing the appropriate immersion oil as being more imperative. These biologic data are supported by quantitative measures of the 3D diffraction pattern (i.e. point spread function) of a point source of light under the same mounting medium and immersion oil combinations. Comparison with Existing Method Although not a new method, this manuscript provides quantitative data demonstrating that different mounting media and immersion oils can impact the ability to resolve dendritic spines. These findings highlight the importance of reporting which mounting medium and immersion oil are used in preparations for confocal analyses, especially when comparing published results from different laboratories. Conclusion Collectively, these data suggest that choosing the appropriate immersion oil and mounting media is critical for obtaining the best resolution, and consequently more accurate measures of dendritic spine densities. PMID:25601477

  3. Sensitivity and Specificity of Cardiac Tissue Discrimination Using Fiber-Optics Confocal Microscopy

    PubMed Central

    Huang, Chao; Sachse, Frank B.; Hitchcock, Robert W.; Kaza, Aditya K.

    2016-01-01

    Disturbances of the cardiac conduction system constitute a major risk after surgical repair of complex cases of congenital heart disease. Intraoperative identification of the conduction system may reduce the incidence of these disturbances. We previously developed an approach to identify cardiac tissue types using fiber-optics confocal microscopy and extracellular fluorophores. Here, we applied this approach to investigate sensitivity and specificity of human and automated classification in discriminating images of atrial working myocardium and specialized tissue of the conduction system. Two-dimensional image sequences from atrial working myocardium and nodal tissue of isolated perfused rodent hearts were acquired using a fiber-optics confocal microscope (Leica FCM1000). We compared two methods for local application of extracellular fluorophores: topical via pipette and with a dye carrier. Eight blinded examiners evaluated 162 randomly selected images of atrial working myocardium (n = 81) and nodal tissue (n = 81). In addition, we evaluated the images using automated classification. Blinded examiners achieved a sensitivity and specificity of 99.2±0.3% and 98.0±0.7%, respectively, with the dye carrier method of dye application. Sensitivity and specificity was similar for dye application via a pipette (99.2±0.3% and 94.0±2.4%, respectively). Sensitivity and specificity for automated methods of tissue discrimination were similarly high. Human and automated classification achieved high sensitivity and specificity in discriminating atrial working myocardium and nodal tissue. We suggest that our findings facilitate clinical translation of fiber-optics confocal microscopy as an intraoperative imaging modality to reduce the incidence of conduction disturbances during surgical correction of congenital heart disease. PMID:26808149

  4. Sensitivity and Specificity of Cardiac Tissue Discrimination Using Fiber-Optics Confocal Microscopy.

    PubMed

    Huang, Chao; Sachse, Frank B; Hitchcock, Robert W; Kaza, Aditya K

    2016-01-01

    Disturbances of the cardiac conduction system constitute a major risk after surgical repair of complex cases of congenital heart disease. Intraoperative identification of the conduction system may reduce the incidence of these disturbances. We previously developed an approach to identify cardiac tissue types using fiber-optics confocal microscopy and extracellular fluorophores. Here, we applied this approach to investigate sensitivity and specificity of human and automated classification in discriminating images of atrial working myocardium and specialized tissue of the conduction system. Two-dimensional image sequences from atrial working myocardium and nodal tissue of isolated perfused rodent hearts were acquired using a fiber-optics confocal microscope (Leica FCM1000). We compared two methods for local application of extracellular fluorophores: topical via pipette and with a dye carrier. Eight blinded examiners evaluated 162 randomly selected images of atrial working myocardium (n = 81) and nodal tissue (n = 81). In addition, we evaluated the images using automated classification. Blinded examiners achieved a sensitivity and specificity of 99.2±0.3% and 98.0±0.7%, respectively, with the dye carrier method of dye application. Sensitivity and specificity was similar for dye application via a pipette (99.2±0.3% and 94.0±2.4%, respectively). Sensitivity and specificity for automated methods of tissue discrimination were similarly high. Human and automated classification achieved high sensitivity and specificity in discriminating atrial working myocardium and nodal tissue. We suggest that our findings facilitate clinical translation of fiber-optics confocal microscopy as an intraoperative imaging modality to reduce the incidence of conduction disturbances during surgical correction of congenital heart disease. PMID:26808149

  5. A confocal microscope position sensor for micron-scale target alignment in ultra-intense laser-matter experiments

    NASA Astrophysics Data System (ADS)

    Willis, Christopher; Poole, Patrick L.; Akli, Kramer U.; Schumacher, Douglass W.; Freeman, Richard R.

    2015-05-01

    A diagnostic tool for precise alignment of targets in laser-matter interactions based on confocal microscopy is presented. This device permits precision alignment of targets within the Rayleigh range of tight focusing geometries for a wide variety of target surface morphologies. This confocal high-intensity positioner achieves micron-scale target alignment by selectively accepting light reflected from a narrow range of target focal planes. Additionally, the design of the device is such that its footprint and sensitivity can be tuned for the desired chamber and experiment. The device has been demonstrated to position targets repeatably within the Rayleigh range of the Scarlet laser system at The Ohio State University, where use of the device has provided a marked increase in ion yield and maximum energy.

  6. The Reflective Learning Continuum: Reflecting on Reflection

    ERIC Educational Resources Information Center

    Peltier, James W.; Hay, Amanda; Drago, William

    2005-01-01

    The importance of reflection to marketing educators is increasingly recognized. However, there is a lack of empirical research that considers reflection within the context of both the marketing and general business education literature. This article describes the use of an instrument that can be used to measure four identified levels of a

  7. Application of CO2 Snow Jet Cleaning in Conjunction with Laboratory Based Total Reflection X-Ray Fluorescence

    NASA Technical Reports Server (NTRS)

    Schmeling, M.; Burnett, D. S.; Allton, J. H.; Rodriquez, M.; Tripa, C. E.; Veryovkin, I. V.

    2013-01-01

    The Genesis mission was the first mission returning solar material to Earth since the Apollo program [1,2]. Unfortunately the return of the space craft on September 8, 2004 resulted in a crash landing, which shattered the samples into small fragments and exposed them to desert soil and other debris. Thus only small fragments of the original collectors are available, each having different degrees of surface contamination. Thorough surface cleaning is required to allow for subsequent analysis of solar wind material embedded within. An initial cleaning procedure was developed in coordination with Johnson Space Center which focused on removing larger sized particulates and a thin film organic contamination acquired during collection in space [3]. However, many of the samples have additional residues and more rigorous and/or innovative cleaning steps might be necessary. These cleaning steps must affect only the surface to avoid leaching and re-distribution of solar wind material from the bulk of the collectors. To aid in development and identification of the most appropriate cleaning procedures each sample has to be thoroughly inspected before and after each cleaning step. Laboratory based total reflection X-ray fluorescence (TXRF) spectrometry lends itself to this task as it is a non-destructive and surface sensitive analytical method permitting analysis of elements from aluminum onward present at and near the surface of a flat substrate [4]. The suitability of TXRF has been demonstrated for several Genesis solar wind samples before and after various cleaning methods including acid treatment, gas cluster ion beam, and CO2 snow jet [5 - 7]. The latter one is non-invasive and did show some promise on one sample [5]. To investigate the feasibility of CO2 snow jet cleaning further, several flown Genesis samples were selected to be characterized before and after CO2 snow application with sample 61052 being discussed below.

  8. Two-photon fluorescence properties of curcumin as a biocompatible marker for confocal imaging

    NASA Astrophysics Data System (ADS)

    Kumar, Abhishek; Li, Lian; Chaturvedi, Akanksha; Brzostowski, Joseph; Chittigori, Joshna; Pierce, Susan; Samuelson, Lynne A.; Sandman, Daniel; Kumar, Jayant

    2012-05-01

    Two-photon (TP) fluorescence properties of an antioxidant and anti-tumor molecule, curcumin, were investigated. The two-photon absorption (TPA) action cross-section was measured in organic solvents and found to be 6 GM in tetrahydrofuran and 2 GM in dimethyl sulfoxide. The measured TPA cross-section is comparable to that of rhodamine 6G. One-photon and TP confocal microscopy has demonstrated that curcumin is internalized in cells and can be used for imaging applications. Our investigation indicates that curcumin is a viable biocompatible TP fluorescent marker.

  9. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: FOUNDATIONS FOR CALIBRATION, QUANTITATION AND SPECTROSCOPY

    EPA Science Inventory

    The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. The goal of a CLSM is to acquire and quantify fluorescence and in some instruments acquire spectral characterization of emitted signals. The accuracy of these measurements demands that...

  10. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: QA TESTS, QUANTITATION AND SPECTROSCOPY

    EPA Science Inventory

    Confocal Microscopy System Performance: QA tests, Quantitation and Spectroscopy.

    Robert M. Zucker 1 and Jeremy M. Lerner 2,
    1Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research Development, U.S. Environmen...

  11. Laser excited confocal microscope fluorescence scanner and method

    DOEpatents

    Mathies, R.A.; Peck, K.

    1992-02-25

    A fluorescent scanner is designed for scanning the fluorescence from a fluorescence labeled separated sample on a sample carrier. The scanner includes a confocal microscope for illuminating a predetermined volume of the sample carrier and/or receiving and processing fluorescence emissions from the volume to provide a display of the separated sample. 8 figs.

  12. Automated stromal nerve rejection in corneal confocal images in vivo

    NASA Astrophysics Data System (ADS)

    Brown, Eric N.; Camp, Jon J.; Patel, Sanjay V.; McLaren, Jay W.; Bourne, William M.; Robb, Richard A.

    2000-06-01

    With the advent of corneal confocal microscopy, investigators can determine keratocyte density in the corneal stroma in vivo. We and others have written automated algorithms to measure keratocyte density from human corneal confocal images. Such algorithms are only accurate if they exclude images of stromal nerve bundles (elongated objects) that would otherwise be counted as keratocytes. In this study we devised an algorithm to identify stromal nerve bundles and exclude them from measurements of keratocyte density. Nerve bundles were detected based on their size and aspect ratio, and were then subtracted from images by using a combination of morphology operations and direction calculations. The validity of nerve removal on measurements of keratocyte density was assessed. Keratocyte density was measured from confocal images of three normal human corneas in vivo by using our algorithm with nerve removal. After the same eyes underwent enucleation, density was measured manually from histologic sections. Keratocyte density was also measured from confocal images of 57 normal corneas in vivo (57 subjects) with and without nerve removal. In the three enucleated eyes, there was no significant difference between keratocyte density measured by automated counting with nerve removal and by histologic methods (P equals 0.75). However, in the 57 normal corneas, use of the nerve-removal algorithm reduced estimates of density by 57.0 +/- 164.6 cells/mm3 (mean +/- SD, p < 0.038) in the anterior two-thirds of the stroma.

  13. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: FOUNDATIONS FOR MEASUREMENTS, QUANTITATION AND SPECTROSCOPY

    EPA Science Inventory

    The confocal laser-scanning microscopy (CLSM) has enormous potential in many biological fields. The goal of a CLSM is to acquire and quantify fluorescence and in some instruments acquire spectral characterization of the emitted signal. The accuracy of these measurements demands t...

  14. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: SPECTROSCOPY AND FOUNDATIONS FOR QUANTITATION

    EPA Science Inventory

    The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. The reliability of the CLSM to obtain specific measurements and quantify fluorescence data is dependent on using a correctly aligned machine that contains a stable laser power. For man...

  15. In vivo confocal microscopy of the human cornea

    PubMed Central

    Jalbert, I; Stapleton, F; Papas, E; Sweeney, D F; Coroneo, M

    2003-01-01

    Aims: To describe the optics of in vivo confocal microscopy, its advantages over previous methods, and to summarise the literature that arose from its use for the observation of the human cornea. A critical review of the clinical usefulness of this new technology for the corneal examination is undertaken. Methods: Confocal microscopes obtain increased resolution by limiting the illumination and observation systems to a single point. Rapid scanning is used to reconstruct a full field of view and allows for real time viewing. Results: Coronal sections of the in situ epithelium, Bowmans membrane, stroma, and endothelium can be visualised at a resolution of 12 ?m. A backscattered light intensity curve allows objective measurements of sublayer thickness and corneal haze to be taken. In vivo confocal microscopy is therefore particularly useful in the areas of infective keratitis, corneal dystrophies, refractive surgery, and contact lens wear, where it aids in differential diagnosis and detection of subtle short and long term changes. Real time endothelial cell assessment can also be performed. Conclusion: Because of their ability to visualise living tissue at cellular levels, confocal microscopes have proved useful additions to the current clinical tools. PMID:12543757

  16. Laser excited confocal microscope fluorescence scanner and method

    DOEpatents

    Mathies, Richard A. (Contra Costa, CA); Peck, Konan (Contra Costa, CA)

    1992-01-01

    A fluorescent scanner for scanning the fluorescence from a fluorescence labeled separated sample on a sample carrier including a confocal microscope for illuminating a predetermined volume of the sample carrier and/or receiving and processing fluorescence emissions from said volume to provide a display of the separated sample.

  17. In vivo subsurface morphological and functional cellular and subcellular imaging of the gastrointestinal tract with confocal mini-microscopy

    PubMed Central

    Goetz, Martin; Memadathil, Beena; Biesterfeld, Stefan; Schneider, Constantin; Gregor, Sebastian; Galle, Peter R; Neurath, Markus F; Kiesslich, Ralf

    2007-01-01

    AIM: To evaluate a newly developed hand-held confocal probe for in vivo microscopic imaging of the complete gastrointestinal tract in rodents. METHODS: A novel rigid confocal probe (diameter 7 mm) was designed with optical features similar to the flexible endomicroscopy system for use in humans using a 488 nm single line laser for fluorophore excitation. Light emission was detected at 505 to 750 nm. The field of view was 475 ?m 475 ?m. Optical slice thickness was 7 ?m with a lateral resolution of 0.7 ?m. Subsurface serial images at different depths (surface to 250 ?m) were generated in real time at 1024 1024 pixels (0.8 frames/s) by placing the probe onto the tissue in gentle, stable contact. Tissue specimens were sampled for histopathological correlation. RESULTS: The esophagus, stomach, small and large intestine and meso, liver, pancreas and gall bladder were visualised in vivo at high resolution in n = 48 mice. Real time microscopic imaging with the confocal mini-microscopy probe was easy to achieve. The different staining protocols (fluorescein, acriflavine, FITC-labelled dextran and L. esculentum lectin) each highlighted specific aspects of the tissue, and in vivo imaging correlated excellently with conventional histology. In vivo blood flow monitoring added a functional quality to morphologic imaging. CONCLUSION: Confocal microscopy is feasible in vivo allowing the visualisation of the complete GI tract at high resolution even of subsurface tissue structures. The new confocal probe design evaluated in this study is compatible with laparoscopy and significantly expands the field of possible applications to intra-abdominal organs. It allows immediate testing of new in vivo staining and application options and therefore permits rapid transfer from animal studies to clinical use in patients. PMID:17465494

  18. Scanning microphotolysis: a new photobleaching technique based on fast intensity modulation of a scanned laser beam and confocal imaging.

    PubMed

    Wedekind, P; Kubitscheck, U; Peters, R

    1994-10-01

    The fluorescence photobleaching method has been widely used to study molecular transport in single living cells and other microsystems while confocal microscopy has opened new avenues to high-resolution, three-dimensional imaging. A new technique, scanning microphotolysis (Scamp), combines the potential of photobleaching, beam scanning and confocal imaging. A confocal scanning laser microscope was equipped with a sufficiently powerful laser and a novel device, the 'Scamper'. This consisted essentially of a filter changer, an acousto-optical modulator (AOM) and a computer. The computer was programmed to activate the AOM during scanning according to a freely defined image mask. As a result, almost any desired pattern could be bleached ('written') into fluorescent samples at high definition and then imaged ('read') at non-bleaching conditions, employing full confocal resolution. Furthermore, molecular transport could be followed by imaging the dissipation of bleach patterns. Experiments with living cells concerning dynamic processes in cytoskeletal filaments and the lateral mobility of membrane lipids suggest a wide range of potential biological applications. Thus, Scamp offers new possibilities for the optical manipulation and analysis of both technical and biological microsystems. PMID:7799426

  19. Development and Beam-Shape Analysis of an Integrated Fiber-Optic Confocal Probe for High-Precision Central Thickness Measurement of Small-Radius Lenses

    PubMed Central

    Sutapun, Boonsong; Somboonkaew, Armote; Amarit, Ratthasart; Chanhorm, Sataporn

    2015-01-01

    This work describes a new design of a fiber-optic confocal probe suitable for measuring the central thicknesses of small-radius optical lenses or similar objects. The proposed confocal probe utilizes an integrated camera that functions as a shape-encoded position-sensing device. The confocal signal for thickness measurement and beam-shape data for off-axis measurement can be simultaneously acquired using the proposed probe. Placing the probe’s focal point off-center relative to a sample’s vertex produces a non-circular image at the camera’s image plane that closely resembles an ellipse for small displacements. We were able to precisely position the confocal probe’s focal point relative to the vertex point of a ball lens with a radius of 2.5 mm, with a lateral resolution of 1.2 µm. The reflected beam shape based on partial blocking by an aperture was analyzed and verified experimentally. The proposed confocal probe offers a low-cost, high-precision technique, an alternative to a high-cost three-dimensional surface profiler, for tight quality control of small optical lenses during the manufacturing process. PMID:25871720

  20. Confocal Laser Microscope Scanning Applied To Three-Dimensional Studies Of Biological Specimens.

    NASA Astrophysics Data System (ADS)

    Franksson, Olof; Liljeborg, Anders; Carlsson, Kjell; Forsgren, Per-Ola

    1987-08-01

    The depth-discriminating property of confocal laser microscope scanners can be used to record the three-dimensional structure of specimens. A number of thin sections (approx. 1 ?m thick) can be recorded by a repeated process of image scanning and refocusing of the microscope. We have used a confocal microscope scanner in a number of feasibility studies to investigate its possibilities and limitations. It has proved to be well suited for examining fluorescent specimens with a complicated three-dimensional structure, such as nerve cells. It has also been used to study orchid seeds, as well as cell colonies, greatly facilitating evaluation of such specimens. Scanning of the specimens is performed by a focused laser beam that is deflected by rotating mirrors, and the reflected or fluorescent light from the specimen is detected. The specimen thus remains stationary during image scanning, and is only moved stepwise in the vertical direction for refocusing between successive sections. The scanned images consist of 256*256 or 512*512 pixels, each pixel containing 8 bits of data. After a scanning session a large number of digital images, representing consecutive sections of the specimen, are stored on a disk memory. In a typical case 200 such 256*256 images are stored. To display and process this information in a meaningful way requires both appropriate software and a powerful computer. The computer used is a 32-bits minicomputer equipped with an array processor (FPS 100). The necessary software was developed at our department.

  1. In vivo fluorescence confocal microscopy: indocyanine green enhances the contrast of epidermal and dermal structures

    NASA Astrophysics Data System (ADS)

    Skvara, Hans; Kittler, Harald; Schmid, Johannes A.; Plut, Ulrike; Jonak, Constanze

    2011-09-01

    In recent years, in vivo skin imaging devices have been successfully implemented in skin research as well as in clinical routine. Of particular importance is the use of reflectance confocal microscopy (RCM) and fluorescence confocal microscopy (FCM) that enable visualization of the tissue with a resolution comparable to histology. A newly developed commercially available multi-laser device in which both technologies are integrated now offers the possibility to directly compare RCM with FCM. The fluorophore indocyanine green (ICG) was intradermally injected into healthy forearm skin of 10 volunteers followed by in vivo imaging at various time points. In the epidermis, accurate assessment of cell morphology with FCM was supplemented by identification of pigmented cells and structures with RCM. In dermal layers, only with FCM connective tissue fibers were clearly contoured down to a depth of more than 100 μm. The fluorescent signal still provided a favorable image contrast 24 and 48 hours after injection. Subsequently, ICG was applied to different types of skin diseases (basal cell carcinoma, actinic keratosis, seborrhoeic keratosis, and psoriasis) in order to demonstrate the diagnostic benefit of FCM when directly compared with RCM. Our data suggest a great impact of FCM in combination with ICG on clinical and experimental dermatology in the future.

  2. Spectral imaging technique for retinal perfusion detection using confocal scanning laser ophthalmoscopy

    NASA Astrophysics Data System (ADS)

    Rasta, Seyed Hossein; Manivannan, Ayyakkannu; Sharp, Peter F.

    2012-11-01

    To evaluate retinal perfusion in the human eye, a dual-wavelength confocal scanning laser ophthalmoscope (cSLO) was developed that provides spectral imaging of the fundus using a combination of red (670 nm) and near-infrared (810 nm) wavelengths. The image of the ocular fundus was analyzed to find out if quantitative measurements of the reflectivity of tissue permit assessment of the oxygen perfusion of tissue. We explored problems that affect the reproducibility of patient measurements such as non-uniformity errors on the image. For the first time, an image processing technique was designed and used to minimize the errors of oxygen saturation measurements by illumination correction in retina wide field by increasing SNR. Retinal images were taken from healthy and diabetic retinopathy eyes using the cSLO with a confocal aperture of 100 ?m. The ratio image (RI) of red/IR, as oxygen saturation (SO2) index, was calculated for normal eyes. The image correction technique improved the reproducibility of the measurements. Average RI intensity variation of healthy retina tissue was determined within a range of about 5.5%. The capability of the new technique to discriminate oxygenation levels of retinal artery and vein was successfully demonstrated and showed good promise in the diagnosis of the perfused retina.

  3. Automated detection of malignant features in confocal microscopy on superficial spreading melanoma versus nevi

    PubMed Central

    Gareau, Dan; Hennessy, Ricky; Wan, Eric; Pellacani, Giovanni; Jacques, Steven L.

    2010-01-01

    In-vivo reflectance confocal microscopy (RCM) shows promise for the early detection of superficial spreading melanoma (SSM). RCM of SSM shows pagetoid melanocytes (PMs) in the epidermis and disarray at the dermal-epidermal junction (DEJ), which are automatically quantified with a computer algorithm that locates depth of the most superficial pigmented surface [DSPS(x,y)] containing PMs in the epidermis and pigmented basal cells near the DEJ. The algorithm uses 200 noninvasive confocal optical sections that image the superficial 200 μm of ten skin sites: five unequivocal SSMs and five nevi. The pattern recognition algorithm automatically identifies PMs in all five SSMs and finds none in the nevi. A large mean gradient ψ (roughness) between laterally adjacent points on DSPS(x,y) identifies DEJ disruption in SSM ψ = 11.7 ± 3.7 [−] for n = 5 SSMs versus a small ψ = 5.5 ± 1.0 [−] for n = 5 nevi (significance, p = 0.0035). Quantitative endpoint metrics for malignant characteristics make digital RCM data an attractive diagnostic asset for pathologists, augmenting studies thus far, which have relied largely on visual assessment. PMID:21198161

  4. Automated detection of malignant features in confocal microscopy on superficial spreading melanoma versus nevi

    NASA Astrophysics Data System (ADS)

    Gareau, Dan; Hennessy, Ricky; Wan, Eric; Pellacani, Giovanni; Jacques, Steven L.

    2010-11-01

    In-vivo reflectance confocal microscopy (RCM) shows promise for the early detection of superficial spreading melanoma (SSM). RCM of SSM shows pagetoid melanocytes (PMs) in the epidermis and disarray at the dermal-epidermal junction (DEJ), which are automatically quantified with a computer algorithm that locates depth of the most superficial pigmented surface [DSPS(x,y)] containing PMs in the epidermis and pigmented basal cells near the DEJ. The algorithm uses 200 noninvasive confocal optical sections that image the superficial 200 ?m of ten skin sites: five unequivocal SSMs and five nevi. The pattern recognition algorithm automatically identifies PMs in all five SSMs and finds none in the nevi. A large mean gradient ? (roughness) between laterally adjacent points on DSPS(x,y) identifies DEJ disruption in SSM ? = 11.7 +/- 3.7 [-] for n = 5 SSMs versus a small ? = 5.5 +/- 1.0 [-] for n = 5 nevi (significance, p = 0.0035). Quantitative endpoint metrics for malignant characteristics make digital RCM data an attractive diagnostic asset for pathologists, augmenting studies thus far, which have relied largely on visual assessment.

  5. Technique of laser confocal and Raman spectroscopy for living cell analysis

    NASA Astrophysics Data System (ADS)

    Meng, Xiaochen; Zhu, Lianqing

    2013-10-01

    Because of the shortcomings of the main methods used to analysis single cell, the need of single living cell analysis with no damage, unmarked and in situ dynamic multi-parameter measurement is urgent in the life sciences and biomedical advanced research field. And the method of for living cells analysis is proposed. The spectral pretreatment technology of living cell is the key work of laser confocal Raman spectroscopy. To study the spectrum processing methods for Raman spectrum on single living cell and develop the pre-process techniques to enhance the signal-to-noise ratio, sensitivity, and decrease the influence of fluorescence, elimination the cosmic rays was used to improve the spectrum. The classification, average and filtration of spectrum were applied to enhance signal-to-noise ratio. The fluorescence was depressed for quantity analysis or utilized for analysis by comparing the background and the spectrum. The results show that the proposed technique for laser confocal Raman spectrum of single cell can perform the sensitive and weak intensity peaks and reflect the information of molecules structures very well.

  6. Plasmon resonance and the imaging of metal-impregnated neurons with the laser scanning confocal microscope.

    PubMed

    Thompson, Karen J; Harley, Cynthia M; Barthel, Grant M; Sanders, Mark A; Mesce, Karen A

    2015-01-01

    The staining of neurons with silver began in the 1800s, but until now the great resolving power of the laser scanning confocal microscope has not been utilized to capture the in-focus and three-dimensional cytoarchitecture of metal-impregnated cells. Here, we demonstrate how spectral confocal microscopy, typically reserved for fluorescent imaging, can be used to visualize metal-labeled tissues. This imaging does not involve the reflectance of metal particles, but rather the excitation of silver (or gold) nanoparticles and their putative surface plasmon resonance. To induce such resonance, silver or gold particles were excited with visible-wavelength laser lines (561 or 640 nm), and the maximal emission signal was collected at a shorter wavelength (i.e., higher energy state). Because the surface plasmon resonances of noble metal nanoparticles offer a superior optical signal and do not photobleach, our novel protocol holds enormous promise of a rebirth and further development of silver- and gold-based cell labeling protocols. PMID:26670545

  7. High throughput, detailed, cell-specific neuroanatomy of dendritic spines using microinjection and confocal microscopy

    PubMed Central

    Dumitriu, Dani; Rodriguez, Alfredo; Morrison, John H.

    2012-01-01

    Morphological features such as size, shape and density of dendritic spines have been shown to reflect important synaptic functional attributes and potential for plasticity. Here we describe in detail a protocol for obtaining detailed morphometric analysis of spines using microinjection of fluorescent dyes, high resolution confocal microscopy, deconvolution and image analysis using NeuronStudio. Recent technical advancements include better preservation of tissue resulting in prolonged ability to microinject, and algorithmic improvements that compensate for the residual Z-smear inherent in all optical imaging. Confocal imaging parameters were probed systematically for the identification of both optimal resolution as well as highest efficiency. When combined, our methods yield size and density measurements comparable to serial section transmission electron microscopy in a fraction of the time. An experiment containing 3 experimental groups with 8 subjects in each can take as little as one month if optimized for speed, or approximately 4 to 5 months if the highest resolution and morphometric detail is sought. PMID:21886104

  8. Plasmon resonance and the imaging of metal-impregnated neurons with the laser scanning confocal microscope

    PubMed Central

    Thompson, Karen J; Harley, Cynthia M; Barthel, Grant M; Sanders, Mark A; Mesce, Karen A

    2015-01-01

    The staining of neurons with silver began in the 1800s, but until now the great resolving power of the laser scanning confocal microscope has not been utilized to capture the in-focus and three-dimensional cytoarchitecture of metal-impregnated cells. Here, we demonstrate how spectral confocal microscopy, typically reserved for fluorescent imaging, can be used to visualize metal-labeled tissues. This imaging does not involve the reflectance of metal particles, but rather the excitation of silver (or gold) nanoparticles and their putative surface plasmon resonance. To induce such resonance, silver or gold particles were excited with visible-wavelength laser lines (561 or 640 nm), and the maximal emission signal was collected at a shorter wavelength (i.e., higher energy state). Because the surface plasmon resonances of noble metal nanoparticles offer a superior optical signal and do not photobleach, our novel protocol holds enormous promise of a rebirth and further development of silver- and gold-based cell labeling protocols. DOI: http://dx.doi.org/10.7554/eLife.09388.001 PMID:26670545

  9. Fault localization and analysis in semiconductor devices with optical-feedback infrared confocal microscopy

    SciTech Connect

    Sarmiento, Raymund; Cemine, Vernon Julius; Tagaca, Imee Rose; Salvador, Arnel; Mar Blanca, Carlo; Saloma, Caesar

    2007-11-01

    We report on a cost-effective optical setup for characterizing light-emitting semiconductor devices with optical-feedback confocal infrared microscopy and optical beam-induced resistance change.We utilize the focused beam from an infrared laser diode to induce local thermal resistance changes across the surface of a biased integrated circuit (IC) sample. Variations in the multiple current paths are mapped by scanning the IC across the focused beam. The high-contrast current maps allow accurate differentiation of the functional and defective sites, or the isolation of the surface-emittingp-i-n devices in the IC. Optical beam-induced current (OBIC) is not generated since the incident beam energy is lower than the bandgap energy of the p-i-n device. Inhomogeneous current distributions in the IC become apparent without the strong OBIC background. They are located at a diffraction-limited resolution by referencing the current maps against the confocal reflectance image that is simultaneously acquired via optical-feedback detection. Our technique permits the accurate identification of metal and semiconductor sites as well as the classification of different metallic structures according to thickness, composition, or spatial inhomogeneity.

  10. A UV laser-scanning confocal microscope for the measurement of intracellular Ca2+.

    PubMed

    Kuba, K; Hua, S Y; Hayashi, T

    1994-09-01

    Modifications to the optics of a conventional confocal laser-scanning microscope were made to allow imaging intracellular Ca(2+)-dependent fluorescence with a UV laser (351 or 364 nm). Modifications included: (1) a chromatic compensation lens in the laser path; (2) the design of a practically achromatic relay lens; (3) a longer tube length for the objective; and (4) highly reflective mirrors maximizing fluorescence measurement. This UV laser-scanning confocal microscope (UV-CLSM) yielded a lateral resolution of < 0.3 micron and an axial resolution of < 1.5 microns and a relevant field size of 100 microns in diameter for a 40X objective). The effects of varying the focal length of a compensation lens, the degree of the correction for the coverglass thickness of objective and the detector aperture size on the quality of image formation were examined. Finally, UV-CLSM revealed optical sections of fine and complex structures of bullfrog sympathetic neurones loaded with a Ca(2+)-sensitive fluorescent probe. Changes in intracellular free Ca2+ distribution in response to high [K+] or caffeine were demonstrated. In addition, an increase in the intracellular concentration of caffeine applied externally was clearly imaged in space and time and distinguished from a resultant rise in [Ca2+]i. Thus, the UV-CLSM developed is suitable for ratiometric intracellular Ca2+ measurements and other biological studies. PMID:7828174

  11. Quantitative visualization of colloidal and intracellular gold nanoparticles by confocal microscopy

    NASA Astrophysics Data System (ADS)

    Klein, Sabine; Petersen, Svea; Taylor, Ulrike; Rath, Detlef; Barcikowski, Stephan

    2010-05-01

    Gold nanoparticles (AuNPs) have the potential to become a versatile biomarker. For further use of AuNPs labeled with functionalized molecules, their visualization in biological systems by routine laboratory tools such as light microscopy is crucial. However, the size far below the diffraction limit affords specialized parameters for microscopical detection, which stimulated the current study, aimed to determine from which size onward AuNPs, either in dispersion or cell-associated, can be reliably detected by standard confocal microscopy. First, gold colloids of size-restricted fractions are examined in dispersion. At a minimum particle size of 60 nm, detection appears to be reliable. Particle counts in dilution series confirm these results by revealing single particle detection of 60-nm colloids. Second, AuNPs are visualized and quantified in cells, which interestingly cause a phase shift in the reflection of AuNPs. Gold mass spectroscopy confirms the number of AuNPs counted microscopically inside cells. Furthermore, it demonstrates for the first time a very high diffusion rate of 15-nm particles into the cells. In conclusion, the results back the suitability of confocal microscopy for the quantitative tracking of colloidal and intracellular gold nanoparticles sized 60 nm.

  12. Solar Confocal interferometers for Sub-Picometer-Resolution Spectral Filters

    NASA Technical Reports Server (NTRS)

    Gary, G. Allen; Pietraszewski, Chris; West, Edward A.; Dines. Terence C.

    2007-01-01

    The confocal Fabry-Perot interferometer allows sub-picometer spectral resolution of Fraunhofer line profiles. Such high spectral resolution is needed to keep pace with the higher spatial resolution of the new set of large-aperture solar telescopes. The line-of-sight spatial resolution derived for line profile inversions would then track the improvements of the transverse spatial scale provided by the larger apertures. In particular, profile inversion allows improved velocity and magnetic field gradients to be determined independent of multiple line analysis using different energy levels and ions. The confocal interferometer's unique properties allow a simultaneous increase in both etendue and spectral power. The higher throughput for the interferometer provides significant decrease in the aperture, which is important in spaceflight considerations. We have constructed and tested two confocal interferometers. A slow-response thermal-controlled interferometer provides a stable system for laboratory investigation, while a piezoelectric interferometer provides a rapid response for solar observations. In this paper we provide design parameters, show construction details, and report on the laboratory test for these interferometers. The field of view versus aperture for confocal interferometers is compared with other types of spectral imaging filters. We propose a multiple etalon system for observing with these units using existing planar interferometers as pre-filters. The radiometry for these tests established that high spectral resolution profiles can be obtained with imaging confocal interferometers. These sub-picometer spectral data of the photosphere in both the visible and near-infrared can provide important height variation information. However, at the diffraction-limited spatial resolution of the telescope, the spectral data is photon starved due to the decreased spectral passband.

  13. Characterization of reflectance variability in the industrial paint application of automotive metallic coatings by using principal component analysis

    NASA Astrophysics Data System (ADS)

    Medina, Jos M.; Daz, Jos A.

    2013-05-01

    We have applied principal component analysis to examine trial-to-trial variability of reflectances of automotive coatings that contain effect pigments. Reflectance databases were measured from different color batch productions using a multi-angle spectrophotometer. A method to classify the principal components was used based on the eigenvalue spectra. It was found that the eigenvalue spectra follow distinct power laws and depend on the detection angle. The scaling exponent provided an estimation of the correlation between reflectances and it was higher near specular reflection, suggesting a contribution from the deposition of effect pigments. Our findings indicate that principal component analysis can be a useful tool to classify different sources of spectral variability in color engineering.

  14. Photometric phase functions of common geologic minerals and applications to quantitative analysis of mineral mixture reflectance spectra

    NASA Technical Reports Server (NTRS)

    Mustard, John F.; Pieters, Carle M.

    1989-01-01

    The mass fractional abundance of components in intimately mixed, particulate surfaces is calculated from laboratory reflectance spectra using Hapke's (1981) model for bidirectional reflectance. It is found that a simplified version of the model is accurate to within 7 percent for mixtures not containing low albedo components. The model is not appropriate for mixtures with very low and high albedo components. Consideration is given to the possible improvement of the method's accuracy by using an empirical single-particle phase function to describe the scattering characteristics of all minerals, or by solving for the single-particle phase function of the minerals involved exactly using extensive bidirectional reflectance measurements and Hapke's equations for bidirectional reflectance. Also, the results are used to examine the general scattering behavior of particulate mineral surfaces.

  15. Advances in the reduction and compensation of film stress in high-reflectance multilayer coatings for extreme ultraviolet lithography applications

    SciTech Connect

    Mirkarimi, P.B., LLNL

    1998-02-20

    Due to the stringent surface figure requirements for the multilayer-coated optics in an extreme ultraviolet (EUV) projection lithography system, it is desirable to minimize deformation due to the multilayer film stress. However, the stress must be reduced or compensated without reducing EUV reflectivity, since the reflectivity has a strong impact on the throughput of a EUV lithography tool. In this work we identify and evaluate several leading techniques for stress reduction and compensation as applied to Mo/Si and Mo/Be multilayer films. The measured film stress for Mo/Si films with EUV reflectances near 67.4% at 13.4 nm is approximately - 420 MPa (compressive), while it is approximately +330 MPa (tensile) for Mo/Be films with EUV reflectances near 69.4% at 11.4 nm. Varying the Mo-to-Si ratio can be used to reduce the stress to near zero levels, but at a large loss in EUV reflectance (> 20%). The technique of varying the base pressure (impurity level) yielded a 10% decrease in stress with a 2% decrease in reflectance for our multilayers. Post-deposition annealing was performed and it was observed that while the cost in reflectance is relatively high (3.5%) to bring the stress to near zero levels (i.e., reduce by 1 00%), the stress can be reduced by 75% with only a 1.3% drop in reflectivity at annealing temperatures near 200{degrees}C. A study of annealing during Mo/Si deposition was also performed; however, no practical advantage was observed by heating during deposition. A new non-thermal (athermal) buffer-layer technique was developed to compensate for the effects of stress. Using this technique with amorphous silicon and Mo/Be buffer-layers it was possible to obtain Mo/Be and Mo/Si multilayer films with a near zero net film stress and less than a 1% loss in reflectivity. For example a Mo/Be film with 68.7% reflectivity at 11.4 nm and a Mo/Si film with 66.5% reflectivity at 13.3 nm were produced with net stress values less than 30 MPa.

  16. Optimal detection pinhole for lowering speckle noise while maintaining adequate optical sectioning in confocal reflectance microscopes

    PubMed Central

    Glazowski, Christopher; Rajadhyaksha, Milind

    2012-01-01

    Abstract. Coherent speckle influences the resulting image when narrow spectral line-width and single spatial mode illumination are used, though these are the same light-source properties that provide the best radiance-to-cost ratio. However, a suitable size of the detection pinhole can be chosen to maintain adequate optical sectioning while making the probability density of the speckle noise more normal and reducing its effect. The result is a qualitatively better image with improved contrast, which is easier to read. With theoretical statistics and experimental results, we show that the detection pinhole size is a fundamental parameter for designing imaging systems for use in turbid media. PMID:23224184

  17. Identification of Nodal Tissue in the Living Heart Using Rapid Scanning Fiber-optics Confocal Microscopy and Extracellular Fluorophores

    PubMed Central

    Huang, Chao; Kaza, Aditya K.; Hitchcock, Robert W.; Sachse, Frank B.

    2014-01-01

    Background Risks associated with pediatric reconstructive heart surgery include injury of the sinoatrial node (SAN) and atrioventricular node (AVN), requiring cardiac rhythm management using implantable pacemakers. These injuries are result of difficulties in identifying nodal tissues intraoperatively. Here, we describe an approach based on confocal microscopy and extracellular fluorophores to quantify tissue microstructure and identify nodal tissue. Methods and Results Using conventional three-dimensional confocal microscopy we investigated the microstructural arrangement of SAN, AVN, and atrial working myocardium (AWM) in fixed rat heart. AWM exhibited a regular striated arrangement of the extracellular space. In contrast, SAN and AVN had an irregular, reticulated arrangement. AWM, SAN and AVN tissue were beneath a thin surface layer of tissue that did not obstruct confocal microscopic imaging. Subsequently, we imaged tissues in living rat hearts with real-time fiber-optics confocal microscopy (FCM). FCM images resembled images acquired with conventional confocal microscopy. We investigated spatial regularity of tissue microstructure from Fourier analysis and 2nd order image moments. Fourier analysis of FCM images showed that the spatial regularity of AWM was greater than that of nodal tissues (37.55.0% versus 24.33.9% for SAN and 23.83.7% for AVN, P<0.05). Similar differences of spatial regularities were revealed from 2nd order image moments (50.07.3% for AWM versus 29.36.7% for SAN and 27.35.5% for AVN; P<0.05). Conclusions The study demonstrates feasibility of identifying nodal tissue in living heart using extracellular fluorophores and FCM. Application of the approach in pediatric reconstructive heart surgery may reduce risks of injuring nodal tissues. PMID:23811748

  18. Three-Dimensional Visualization of Interfacial Phenomena Using Confocal Microscopy

    NASA Astrophysics Data System (ADS)

    Shieh, Ian C.

    Surfactants play an integral role in numerous functions ranging from stabilizing the emulsion in a favorite salad dressing to organizing the cellular components that make life possible. We are interested in lung surfactant, which is a mixture of lipids and proteins essential for normal respiration because it modulates the surface tension of the air-liquid interface of the thin fluid lining in the lungs. Through this surface tension modulation, lung surfactant ensures effortless lung expansion and prevents lung collapse during exhalation, thereby effecting proper oxygenation of the bloodstream. The function of lung surfactant, as well as numerous interfacial lipid systems, is not solely dictated by the behavior of materials confined to the two-dimensional interface. Rather, the distributions of materials in the liquid subphase also greatly influence the performance of interfacial films of lung surfactant. Therefore, to better understand the behavior of lung surfactant and other interfacial lipid systems, we require a three-dimensional characterization technique. In this dissertation, we have developed a novel confocal microscopy methodology for investigating the interfacial phenomena of surfactants at the air-liquid interface of a Langmuir trough. Confocal microscopy provides the excellent combination of in situ, fast, three-dimensional visualization of multiple components of the lung surfactant system that other characterization techniques lack. We detail the solutions to the numerous challenges encountered when imaging a dynamic air-liquid interface with a high-resolution technique like confocal microscopy. We then use confocal microscopy to elucidate the distinct mechanisms by which a polyelectrolyte (chitosan) and nonadsorbing polymer (polyethylene glycol) restore the function of lung surfactant under inhibitory conditions mimicking the effects of lung trauma. Beyond this physiological model, we also investigate several one- and two-component interfacial films of the various lipid constituents of lung surfactant. Confocal microscopy allows us to use a water-soluble, cationic fluorophore that partitions into the disordered phases of lipid monolayers. By exploiting the properties of this water-soluble fluorophore, we investigate both the phase behavior and electrostatics of the interfacial lipid systems. Overall, we believe the work presented in this dissertation provides the building blocks for establishing confocal microscopy as a ubiquitous characterization technique in the interfacial and surface sciences.

  19. Application of the specular and diffuse reflection analysis for in vitro diagnostics of dental erosion: correlation with enamel softening, roughness, and calcium release

    PubMed Central

    Rakhmatullina, Ekaterina; Bossen, Anke; Hschele, Christoph; Wang, Xiaojie; Beyeler, Barbara; Meier, Christoph; Lussi, Adrian

    2011-01-01

    We present assembly and application of an optical reflectometer for the analysis of dental erosion. The erosive procedure involved acid-induced softening and initial substance loss phases, which are considered to be difficult for visual diagnosis in a clinic. Change of the specular reflection signal showed the highest sensitivity for the detection of the early softening phase of erosion among tested methods. The exponential decrease of the specular reflection intensity with erosive duration was compared to the increase of enamel roughness. Surface roughness was measured by optical analysis, and the observed tendency was correlated with scanning electron microscopy images of eroded enamel. A high correlation between specular reflection intensity and measurement of enamel softening (r2 ? ?0.86) as well as calcium release (r2 ? ?0.86) was found during erosion progression. Measurement of diffuse reflection revealed higher tooth-to-tooth deviation in contrast to the analysis of specular reflection intensity and lower correlation with other applied methods (r2 = 0.420.48). The proposed optical method allows simple and fast surface analysis and could be used for further optimization and construction of the first noncontact and cost-effective diagnostic tool for early erosion assessment in vivo. PMID:22029364

  20. Application of the specular and diffuse reflection analysis for in vitro diagnostics of dental erosion: correlation with enamel softening, roughness, and calcium release

    NASA Astrophysics Data System (ADS)

    Rakhmatullina, Ekaterina; Bossen, Anke; Höschele, Christoph; Wang, Xiaojie; Beyeler, Barbara; Meier, Christoph; Lussi, Adrian

    2011-10-01

    We present assembly and application of an optical reflectometer for the analysis of dental erosion. The erosive procedure involved acid-induced softening and initial substance loss phases, which are considered to be difficult for visual diagnosis in a clinic. Change of the specular reflection signal showed the highest sensitivity for the detection of the early softening phase of erosion among tested methods. The exponential decrease of the specular reflection intensity with erosive duration was compared to the increase of enamel roughness. Surface roughness was measured by optical analysis, and the observed tendency was correlated with scanning electron microscopy images of eroded enamel. A high correlation between specular reflection intensity and measurement of enamel softening (r2 >= -0.86) as well as calcium release (r2 >= -0.86) was found during erosion progression. Measurement of diffuse reflection revealed higher tooth-to-tooth deviation in contrast to the analysis of specular reflection intensity and lower correlation with other applied methods (r2 = 0.42-0.48). The proposed optical method allows simple and fast surface analysis and could be used for further optimization and construction of the first noncontact and cost-effective diagnostic tool for early erosion assessment in vivo.