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检索条件"任意字段=Conference on 3-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX"

共 559 条记录，以下是81-90 订阅

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White matter segmentation by estimating tissue optical attenuation from volumetric OCT massive histology of whole rodent brains 24

White matter segmentation by estimating tissue optical atten...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIV

作者： Lefebvre, Joel Castonguay, Alexandre Lesage, Frederic Ecole Polytech Montreal CP 6079Succ Ctr Ville Montreal PQ H3C 3A7 Canada Inst Cardiol Montreal 5000 Rue Belanger Est Montreal PQ H1T 1C8 Canada

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

A whole rodent brain was imaged using an automated massive histology setup and an Optical Coherence Tomography (OCT) microscope. Thousands of OCT volumetric tiles were acquired, each covering a size of about 2.5x2.5x0.8 mm3 with a sampling resolution of 4.9x4.9x6.5 microns. This paper shows the techniques for reconstruction, attenuation compensation and segmentation of the sliced brains. The tile positions within the mosaic were evaluated using a displacement model of the motorized stage and pairwise coregistration. Volume blending was then performed by solving the 3D Laplace equation, and consecutive slices were assembled using the cross-correlation of their 2D image gradient. This reconstruction algorithm resulted in a 3D map of optical reflectivity for the whole brain at micrometric resolution. OCT tissue slices were then used to estimate the local attenuation coefficient based on a single scattering photon model. The attenuation map obtained exhibits a high contrast for all white matter fibres, regardless of their orientation. The tissue optical attenuation from the intrinsic OCT reflectivity contributes to better white matter tissue segmentation. The combined 3D maps of reflectivity and attenuation is a step toward the study of white matter at a microscopic scale for the whole brain in small animals.

关键词： Massive histology Volume reconstruction image processing Optical Coherence Tomography Attenuation Coefficient White matter Segmentation

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Simple and fast spectral domain algorithm for quantitative phase imaging of living cells with digital holographic microscopy 24

Simple and fast spectral domain algorithm for quantitative p...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIV

作者： Min, Junwei Yao, Baoli Ketelhut, Steffi Kemper, Bjoern Univ Munster Biomed Technol Ctr Mendelstr 17 D-48149 Munster Germany Chinese Acad Sci Xian Inst Opt & Precis Mech State Key Lab Transient Opt & Photon Xian 710119 Shaanxi Peoples R China

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

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

关键词： Digital holographic microscopy aberration compensation frequency spectrum analysis quantitative phase imaging quantitative live cell analysis

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Probing neural tissue with Airy light-sheet microscopy: investigation of imaging performance at depth within turbid media 24

Probing neural tissue with Airy light-sheet microscopy: inve...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIV

作者： Nylk, Jonathan McCluskey, Kaley Aggarwal, Sanya Tello, Javier A. Dholakia, Kishan Univ St Andrews Sch Phys & Astron SUPA St Andrews KY16 9SS Fife Scotland Univ St Andrews Sch Med St Andrews KY16 9TF Fife Scotland

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Light-sheet microscopy (LSM) has received great interest for fluorescent imaging applications in biomedicine as it facilitates three-dimensional visualisation of large sample volumes with high spatiotemporal resolution whilst minimising irradiation of, and photo-damage to the specimen. Despite these advantages, LSM can only visualise superficial layers of turbid tissues, such as mammalian neural tissue. Propagation-invariant light modes have played a key role in the development of high-resolution LSM techniques as they overcome the natural divergence of a Gaussian beam, enabling uniform and thin light-sheets over large distances. Most notably, Bessel and Airy beam-based light-sheet imaging modalities have been demonstrated. In the single-photon excitation regime and in lightly scattering specimens, Airy-LSM has given competitive performance with advanced Bessel-LSM techniques. Airy and Bessel beams share the property of self-healing, the ability of the beam to regenerate its transverse beam profile after propagation around an obstacle. Bessel-LSM techniques have been shown to increase the penetration-depth of the illumination into turbid specimens but this effect has been understudied in biologically relevant tissues, particularly for Airy beams. It is expected that Airy-LSM will give a similar enhancement over Gaussian-LSM. In this paper, we report on the comparison of Airy-LSM and GaussianLSM imaging modalities within cleared and non-cleared mouse brain tissue. In particular, we examine image quality versus tissue depth by quantitative spatial Fourier analysis of neural structures in virally transduced fluorescent tissue sections, showing a three-fold enhancement at 50 p.m depth into non-cleared tissue with AiryLSM. Complimentary analysis is performed by resolution measurements in bead-injected tissue sections.

关键词： Light-sheet microscopy LSM Airy beam Tissue imaging turbid media neuroscience

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THREE-dimensional SEGMENTATION OF VESICULAR NETWORKS OF FUNGAL HYPHAE IN MACROSCOPIC microscopy image STACKS 24

THREE-DIMENSIONAL SEGMENTATION OF VESICULAR NETWORKS OF FUNG...

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24th IEEE International conference on image processing (ICIP)

作者： Saponaro, P. Treible, W. Kolagunda, A. Rhein, S. Caplan, J. Kambhamettu, C. Wisser, R. Univ Delaware Newark DE 19716 USA

ISBN: (纸本)9781509021758

Automating the extraction and quantification of features from three-dimensional (3-D) image stacks is a critical task for advancing many applications, such as the analysis of biological structures. The quantification of biological resistance of a plant tissue to fungal infection through the analysis of attributes such as fungal penetration depth, fungal mass, and branching of the fungal network at large scales requires the union of 3D image acquisition and analysis. From an image processing perspective, these tasks reduce to segmentation of vessel-like structures and the extraction of features from their skeletonization. In order to sample multiple infection events for analysis, we have developed an approach we refer to as macroscopic microscopy. However, macroscopic microscopy produces high-resolution image stacks that pose challenges to routine approaches and are difficult for a human to annotate to obtain ground truth data. We present a synthetic hyphal network generator, a comparison of several vessel segmentation methods, and a minimum spanning tree method for connecting small gaps resulting from imperfections in imaging or incomplete skeletonization of hyphal networks. Qualitative results are shown for real microscopic data. We believe the comparison of vessel detectors on macroscopic microscopy data, the synthetic vessel generator, and the gap closing technique are beneficial to the image processing community.

关键词： Segmentation Skeletonization Fungal Hyphae Macroscopic microscopy

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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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Three dimensional imaging using phase retrieval with two focus planes 13

Three dimensional imaging using phase retrieval with two foc...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Ilovitsh, Tali Ilovitsh, Asaf Weiss, Aryeh Meir, Rinat Zalevsky, Zeev Bar Ilan Univ Fac Engn IL-5290002 Ramat Gan Israel Bar Ilan Univ Bar Ilan Inst Nanotechnol & Adv Mat IL-5290002 Ramat Gan Israel

ISBN: (纸本)9781628419474

This work presents a technique for a full 3D imaging of biological samples tagged with gold-nanoparticles (GNPs) using only two images, rather than many images per volume as is currently needed for 3D optical sectioning microscopy. The proposed approach is based on the Gerchberg-Saxton (GS) phase retrieval algorithm. The reconstructed field is free space propagated to all other focus planes using post processing, and the 2D z-stack is merged to create a 3D image of the sample with high fidelity. Because we propose to apply the phase retrieving on nano particles, the regular ambiguities typical to the Gerchberg-Saxton algorithm, are eliminated. In addition, since the method requires the capturing of two images only, it can be suitable for 3D live cell imaging. The proposed concept is presented and validated both on simulated data as well as experimentally.

关键词： Three-dimensional microscopy Cellular imaging gold nanoparticle Phase retrieval

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Deformable mirror based remote focusing for fast three-dimensional microscopy 13

Deformable mirror based remote focusing for fast three-dimen...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Zurauskas, Mantas Frade, Maria Booth, Martin Univ Oxford Ctr Neural Circuits & Behav Mansfield Rd Oxford OX1 3SR England Univ Oxford Dept Engn Sci Parks Rd Oxford OX1 3PJ England

ISBN: (纸本)9781628419474

We present a deformable mirror based remote focusing method for three-dimensional imaging in high-resolution microscopy systems. The method relies on predefined mirror mode arrays that are obtained during initial mirror training step with a low complexity wavefront-sensing module. The imaging plane can be refocused over distances over a hundred times greater than the original depth of field of the objective lens along the optical axis at millisecond rates. We will demonstrate the combination of the remote focusing method with spatiotemporally focused two-photon excitation applied to three-dimensional imaging of biological samples.

关键词： Remote focusing adaptive optics three dimensional microscopy two photon microscopy

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3D imaging of the cleared intact murine colon with light sheet microscopy 13

3D imaging of the cleared intact murine colon with light she...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Zufiria, B. Bocancea, D. I. Gomez-Gaviro, M. V. Vaquero, J. J. Desco, M. Fresno, M. Ripoll, J. Arranz, A. Univ Autonoma Madrid CSIC Ctr Mol Biol Severo Ochoa CBMSO Madrid Spain Univ Carlos III Madrid Dept Bioengn & Aerosp Engn Madrid 28911 Spain Inst Invest Sanitaria Gregorio Maranon Madrid 28007 Spain

ISBN: (纸本)9781628419474

We here show 3D light sheet microscopy images of fixed and cleared murine colon tissue in-toto, which offer relevant cellular information without the need for physically sectioning the tissue. We have applied the recently developed CUBIC protocol (Susaki et al. Cell 157: 726, 2014) for colon tissues and have found that this clearing protocol enables imaging all the way to the central part of the lumen with cellular resolution, thus opening new ways for 3D imaging of colon samples

关键词： light sheet microscopy CUBIC protocol tissue clearing

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Fluorescence microscopy with isotropic resolution using three objectives 13

Fluorescence microscopy with isotropic resolution using thre...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Huelsnitz, Thomas Kner, Peter Univ Georgia Coll Engn Athens GA 30602 USA

ISBN: (纸本)9781628419474

Widefield and confocal fluorescence microscopy using a single objective suffer from poor resolution and a strong anisotropy between the lateral and axial resolution. Coherently combining the excitation and emission from two coaxial objectives improves the axial resolution up to sevenfold, but leaves the lateral resolution unchanged. Here we investigate the coherent combination of three objectives to create a point spread function (PSF) that is isotropic with higher resolution in the plane of the objectives. We develop a theoretical framework for simulating the performance of interferometric imaging with three objectives. Using three identical objectives with a large working distance and 0.9 numerical aperture (NA), the full-width half maximum of the confocal PSF is 135 nm compared to the lateral FWHM of 274 nm for imaging with a single objective at a wavelength of 515 nm.

关键词： microscopy fluorescence microscopy interferometric imaging PSF Engineering

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New Approaches for the analysis of confluent cell layers with quantitative phase digital holographic microscopy 13

New Approaches for the analysis of confluent cell layers wit...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Pohl, L. Kaiser, M. Ketelhut, S. Pereira, S. Goycoolea, F. Kemper, Bjoern Univ Munster Fac Med Biomed Technol Ctr Mendelstr 17 D-48149 Munster Germany Univ Munster Inst Plant Biol & Biotechnol Schlossgarten 3 D-48149 Munster Germany

ISBN: (纸本)9781628419474

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

关键词： digital holographic microscopy quantitative phase imaging live cell imaging minimally-invasive label-free stain-free confluent cell layers

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