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检索条件"任意字段=Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX"

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three-dimensional reconstruction and analysis of gastric malignancies by electronic slides of consecutive sections and virtual microscopy

Three-dimensional reconstruction and analysis of gastric mal...

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Proceedings of the 1999 three-dimensional and multidimensional microscopy: image acquisition and processing VI

作者： Molnar, Bela Papik, Kornel Tagscherer, Attila Csendes, Gabor Varga, Sebestyen V. Tulassay, Zsolt Schaefer, Rainer Mahoney, Walt Semmelweis Univ Budapest Hungary

ISBN: (纸本)0819430757

A study was carried out focusing on the application of an electronic slide and a software simulated virtual microscope to histological analysis. The electronic slide was developed using a fully computer controlled microscope, video camera and an Intel Pentium II based PC. A program called Pyramid was developed for the scanning of an entire slide. Autofocusing, image reduction and frame joining algorithms were implemented in the virtual microscope application. Results confirm that the electronic slide and the software contribute to a more efficient, convenient histological analysis.

关键词： Optical microscopy

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3-D image of protein visualization in a whole rice grain using an automatic precision microtome system

3-D image of protein visualization in a whole rice grain usi...

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three-dimensional and multidimensional microscopy: image acquisition processing VII

作者： Ogawa, Yukiharu Ohtani, Toshio Sugiyama, Junichi Hagiwara, Shoji Tanaka, Kunisuke Kudoh, Ken-ichi Higuchi, Toshiro Natl Food Research Inst Tsukuba Japan

The three-dimensional (3-D) image formation technique using confocal microscopy has allowed visualization of the 3-D chemical structure in small parts of the bio-body. However, the large-scale 3-D structure such as the distribution of chemical components throughout the whole body has not been shown. To allow such large scale visualization of the 3-D internal analysis technique for bio-body has been developed. The 3-D internal analysis method has three stages. In the first stage, sequential sections of a whole bio-body were made using the automatic precision microtome system. Although this system is that used for microscopy, the obtained sections were positioned on an adhesion tape for 3-D reconstruction. In the next stage, all sections were stained using a chemical or immunofluorescence technique for visualization of the chemical components. Each stained section was digitally imaged using a spectroscopic stereomicroscope with a CCD camera. The obtained 2-D digital images contained the spectroscopic information representing the chemical components. In the last stage, the 2-D digital images were reconstructed to produce a 3-D image. As a result, the distribution of chemical components in the whole bio-body could be visualized in 3-D. In this study, the distribution of protein in a whole rice grain was visualized using a 3-D image.

关键词： Medical imaging

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Hyperspectral imaging and adaptive thresholding to identify agonist-induced cAMP signals in pulmonary microvascular endothelial cells 29

Hyperspectral imaging and adaptive thresholding to identify ...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing Xxix

作者： Annamdevula, N. Johnson, S. Pleshinger, D. J. Castleberry, S. Russell, W. Britain, A. L. Francis, C. M. Rich, T. C. Leavesley, S. J. Univ S Alabama Pharmacol Mobile AL 36688 USA Univ S Alabama Physiol & Cell Biol Mobile AL 36688 USA Univ S Alabama Chem & Biomol Engn Mobile AL 36688 USA Univ West Alabama Livingston AL 35470 USA Auburn Univ Auburn AL 36849 USA

ISBN: (纸本)9781510648043;9781510648036

Cyclic AMP (cAMP) is a second messenger that regulates a wide variety of cellular functions. There is increasing evidence suggesting that signaling specificity is due in part to cAMP compartmentalization. In the last 15 years, development of cAMP-specific Forster resonance energy transfer (FRET) probes have allowed us to visualize spatial distributions of intracellular cAMP signals. The use of FRET-based sensors is not without its limitations, as FRET probes display low signal to noise ratio (SNR). Hyperspectral imaging and analysis approaches have, in part, allowed us to overcome these limitations by improving the SNR of FRET measurements. Here we demonstrate that the combination of hyperspectral imaging approaches, linear unmixing, and adaptive thresholding allow us to visualize regions of elevated cAMP (regions of interest - ROIs) in an unbiased manner. We transfected cDNA encoding the H188 FRET-based cAMP probe into pulmonary microvascular endothelial cells. Application of isoproterenol and prostaglandin E1 (PGE(1)) triggered complex cAMP responses. Spatial and temporal aspects of cAMP responses were quantified using an adaptive thresholding approach and compared between agonist treatment groups. Our data indicate that both the origination sites and spatial/temporal distributions of cAMP signals are agonist dependent in PMVECs. We are currently analyzing the data in order to better quantify the distribution of cAMP signals triggered by different agonists.

关键词： FRET fluorescence microscopy spectral imaging adaptive thresholding second messengers cAMP signals localized signals

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Method for measurement of collagen monomer orientation in fluorescence microscopy 28

Method for measurement of collagen monomer orientation in fl...

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three-dimensional and multidimensional microscopy: image acquisition and processing XXVIII 2021

作者： Alzola, Rodrigo P. Delpiano, Jose Gajjar, Anuj Stureborg, Rickard Siadat, Seyed Mohammad Ruberti, Jeffrey W. Dimarzio, Charles A. Department of Electrical and Computer Engineering Northeastern University BostonMA United States Optical Communications Lab School of Engineering and Applied Sciences Universidad de los Andes Santiago Chile Department of Bioengineering Northeastern University BostonMA United States Department of Mechanical and Industrial Engineering Northeastern University BostonMA United States

ISBN: (数字)9781510641341

ISBN: (纸本)9781510641334

Collagen is one of the most important proteins in mammals, conforming most animal tissues. This work explores how a basic collagen monomer unit is visualized using fluorescence microscopy and how its spatial orientation is determined. Defining the orientation of collagen monomers is not a trivial problem, as the particle has a weak contrast and is relatively small. Possible attach fluorescence tags for contrast, but the size is still a problem for detecting orientation using fluorescence microscopy. This document presents a simulation of the visualization of collagen monomers and two methods for detecting monomer and classifying its orientation. A modify Gabor filter set, and an automatic classifier, trained by convolutional neuronal network (CNN), were used. By evaluating the performance of these two approaches compare to human observation, our results show that it is possible to determine the location and orientation of a single monomer with fluorescence microscopy. These findings can contribute to understanding collagen elements as collagen fibril. © 2021 Copyright SPIE.

关键词： Collagen

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The Gray Institute Open Microscopes applied to radiobiology and protein interaction studiesZ

The Gray Institute Open Microscopes applied to radiobiology ...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Barber, P. R. Tullis, I. D. C. Rowley, M. I. Martins, C. D. Weitsman, G. Lawler, K. Coffey, M. Woodman, N. Gillett, C. E. Ng, T. Vojnovic, B. Univ Oxford Gray Inst Radiat Oncol & Biol Dept Oncol Oxford OX1 2JD England Inst Math & Molecular Biomed Kings Coll London London England Kings Coll London Div Cancer Studies London England Kings Coll London Randall Div London England Kings Coll London Breakthrough Breast Cancer Res Unit London England Kings Coll London Guys & St Thomas Breast Tissue & Data Bank London England

ISBN: (纸本)9780819498625

We describe an 'open' design methodology for wide-field fluorescence, confocal and fluorescence lifetime imaging microscopy (FLIM), and how the resulting microscopes are being applied to radiation biology and protein activity studies in cells and human tissue biopsies. The design approach allows easy expansion as it moves away from the use of a monolithic microscope body to small, commercial off-the-shelf and custom made modular components. Details have been made available under an open license for non-commercial use at http://***/similar to atdgroup. Two radiobiology 'end-stations' have been constructed which enable fast radiation targeting and imaging of biological material opening up completely novel studies, where the consequences of ionising radiation (signaling and protein recruitment) can be studied in situ, at short times following irradiation. One is located at Surrey University, UK, where radiation is a highly focused in beam (e.g. protons, helium or higher mass ions). The second is installed at the Gray Institute linear accelerator facility, Oxford University, which uses sub-microsecond pulses of 6 MeV electrons. FLIM capabilities have enhanced the study of protein-protein interactions in cells and tissues via Frster Resonance Energy Transfer (FRET). Extracting FRET signals from breast cancer tissue is challenging because of endogenous and fixation fluorescence;we are investigating novel techniques to measure this robustly. Information on specific protein interactions from large numbers of patient tumors will reveal prognostic and diagnostic information.

关键词： Tissues Fluorescence resonance energy transfer Microscopes image filtering Atrial fibrillation Fluorescence lifetime imaging Optical filters Proteins Luminescence Linear filtering

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3D image restoration for confocal microscopy: Towards a wavelet deconvolution for the study of complex biological structures

3D image restoration for confocal microscopy: Towards a wave...

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three-dimensional and multidimensional microscopy: image acquisition processing VII

作者： Boutet de Monvel, J. Le Calvez, S. Ulfendahl, M. Karolinska Hospital Stockholm Sweden

image restoration algorithms provide efficient tools for recovering part of the information lost in the imaging process of a microscope. We describe recent progress in the application of deconvolution to confocal microscopy. The point spread function of a Biorad-MRC1024 confocal microscope was measured under various imaging conditions, and used to process 3D-confocal images acquired in an intact preparation of the inner ear developed at Karolinska Institutet. Using these experiments we investigate the application of deionizing methods based on wavelet analysis as a natural regularization of the deconvolution process. Within the Bayesian approach to image restoration, we compare wavelet denoising with the use of a maximum entropy constraint as another natural regularization method. Numerical experiments performed with test images show a clear advantage of the wavelet deionizing approach, allowing to `cool down' the image with respect to the signal, while suppressing much of the fine-scale artefacts appearing during deconvolution due to the presence of noise, incomplete knowledge of the point spread function, or undersampling problems. We further describe a natural development of this approach, which consists of performing the Bayesian inference directly in the wavelet domain.

关键词： Medical imaging

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Fast IR imaging with sub-wavelength resolution using a transient near-field probe (tipless near-field microscopy)

Fast IR imaging with sub-wavelength resolution using a trans...

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Proceedings of the 1999 three-dimensional and multidimensional microscopy: image acquisition and processing VI

作者： Palanker, Daniel V. Smith, Todd I. Schwettman, H.Alan Stanford Univ Stanford CA United States

ISBN: (纸本)0819430757

A technique that enables infrared (IR) imaging with the resolution of a visible microscope is introduced. The technique uses conventional IR and no near-field tips. The resolution of the IR microscope is determined by the dimensions of the transient mirror. To prevent resolution degradation due to diffusion of the photo-excited carriers in the substrate, the probe (IR) pulse duration should not exceed a few tens of picoseconds.

关键词： Optical microscopy

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Method for intracellular imaging of ion concentrations, using confocal microscopy and fluorophore lifetimes

Method for intracellular imaging of ion concentrations, usin...

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three-dimensional and multidimensional microscopy: image acquisition processing VII

作者： Carlsson, Kjell Liljeborg, Anders Andersson, Ronnie M. Brismar, Hjalmar Royal Inst of Technology Stockholm Sweden

There exist a number of fluorescent probes whose lifetimes change in response to ion concentrations (for example H+ and Ca2+) in the surrounding medium. We describe a technique for utilizing this property in a confocal scanning laser microscope. The technique is based on intensity-modulated laser illumination of the specimen, and phase-sensitive lock-in detection of the fluorescent light. In this way we get a lifetime-dependent output signal which, after calibration, can provide information concerning ion concentrations. In the current study we have used a pH sensitive fluorophore, SNAFL-2, to study the performance of this technique. We find that the sensitivity is such that a pH difference of 0.1 units can easily be detected in an 8-bit digital image. Noise measurements show that under realistic conditions we can expect a pixel-to-pixel standard deviation of approximately one to two pH units.

关键词： Medical imaging

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5D imaging approaches reveal the formation of distinct intracellular cAMP spatial gradients 24

5D imaging approaches reveal the formation of distinct intra...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXIV

作者： Rich, Thomas C. Annamdevula, Naga Trinh, Kenny Britain, Andrea L. Mayes, Samuel A. Griswold, John R. Deal, Joshua Hoffman, Chase West, Savannah Leavesley, Silas J. Univ S Alabama Pharmacol Mobile AL 36688 USA Univ S Alabama Ctr Lung Biol Mobile AL 36688 USA Univ S Alabama Basic Med Sci Grad Program Mobile AL 36688 USA Univ S Alabama Chem & Biomol Engn Mobile AL 36688 USA Univ S Alabama Biomed Sci Mobile AL 36688 USA

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Cyclic AMP (cAMP) is a ubiquitous second messenger known to differentially regulate many cellular functions. Several lines of evidence suggest that the distribution of cAMP within cells is not uniform. However, to date, no studies have measured the kinetics of 3D cAMP distributions within cells. This is largely due to the low signal-tonoise ratio of FRET-based probes. We previously reported that hyperspectral imaging improves the signal-to-noise ratio of FRET measurements. Here we utilized hyperspectral imaging approaches to measure FRET signals in five dimensions (5D) -three spatial (x, y, z), wavelength (lambda), and time (t) -allowing us to visualize cAMP gradients in pulmonary endothelial cells. cAMP levels were measured using a FRET-based sensor (H188) comprised of a cAMP binding domain sandwiched between FRET donor and acceptor - Turquoise and Venus fluorescent proteins. We observed cAMP gradients in response to 0.1 or 1 mu M isoproterenol, 0.1 or 1 mu M PGE1, or 50 mu M forskolin. Forskolin-and isoproterenol-induced cAMP gradients formed from the apical (high cAMP) to basolateral (low cAMP) face of cells. In contrast, PGE1-induced cAMP gradients originated from both the basolateral and apical faces of cells. Data suggest that 2D (x, y) studies of cAMP compartmentalization may lead to erroneous conclusions about the existence of cAMP gradients, and that 3D (x, y, z) studies are required to assess mechanisms of signaling specificity. Results demonstrate that 5D imaging technologies are powerful tools for measuring biochemical processes in discrete subcellular domains. This work was supported by NIH P01HL066299, R01HL058506, S10RR027535, AHA 16PRE27130004 and the Abraham Mitchell Cancer Research Fund.

关键词： Hyperspectral imaging spectral imaging microscopy FRET Forster resonance energy transfer cAMP second messenger signaling pulmonary endothelium

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Influence of the background estimation on the superresolution properties of non-linear image restoration algorithms

Influence of the background estimation on the superresolutio...

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Proceedings of the 1999 three-dimensional and multidimensional microscopy: image acquisition and processing VI

作者： van Kempen, G.M.P. van Vliet, L.J. Unilever Research Vlaardingen Netherlands

ISBN: (纸本)0819430757

A study was carried out focusing on the effect of the background estimation on the performance of the Tikhonov-Miller algorithm (ICTM), Carrington, and Richardson-Lucy algorithms. A comparison was also made with the linear Tikhonov-Miller restoration filter. It was found that an underestimation of the background will make the non-negativity constraint ineffective which results in a performance that does not differ much from the performance obtained by the linear restoration filter. A novel method was devised to estimate the background based on the dependency of non-linear restoration algorithms on the background.

关键词： image reconstruction

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