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

共 561 条记录，以下是61-70 订阅

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Snapshot 3D tracking of insulin granules in live cells 25

Snapshot 3D tracking of insulin granules in live cells

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conference on Three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Wang, Xiaolei Huang, Xiang Gdor, Itay Daddysman, Matthew Yi, Hannah Selewa, Alan Haunold, Theresa Hereld, Mark Scherer, Norbert F. Univ Chicago James Franck Inst 929 E 57th St Chicago IL 60637 USA Argonne Natl Lab Math & Comp Sci Div 9700 S Cass Ave Lemont IL 60439 USA Univ Chicago Dept Chem 5801 S Ellis Ave Chicago IL 60637 USA Univ Chicago Biophys Sci 924 E 57th St Chicago IL 60637 USA

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Rapid and accurate volumetric imaging remains a challenge, yet has the potential to enhance understanding of cell function. We developed and used a multifocal microscope (MFM) for 3D snapshot imaging to allow 3D tracking of insulin granules labeled with mCherry in MIN6 cells. MFM employs a special diffractive optical element (DOE) to simultaneously image multiple focal planes. This simultaneous acquisition of information deteunines the 3D location of single objects at a speed only limited by the array detector's frame rate. We validated the accuracy of MFM imaging/tracking with fluorescence beads;the 3D positions and trajectories of single fluorescence beads can be detelinined accurately over a wide range of spatial and temporal scales. The 3D positions and trajectories of single insulin granules in a 3.2um deep volume were deteunined with imaging processing that combines 3D decovolution, shift correction, and finally tracking using the Imaris software package. We find that the motion of the granules is super diffusive, but less so in 3D than 2D for cells grown on coverslip surfaces, suggesting an anisotropy in the cytoskeleton (e.g. microtubules and action).

关键词： Multifocal microscopy diffraction optical element 3D tracking single particle tracking insulin granules

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Light-sheet enhanced resolution of light field microscopy for rapid imaging of large volumes 25

Light-sheet enhanced resolution of light field microscopy fo...

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conference on Three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Madrid-Wolff, Jorge Castro, Diego Arbelaez, Pablo Forero-Shelton, Manu Univ Los Andes Biophys Lab Bogota Colombia Univ Los Andes Biomed Comp Vis Grp Bogota Colombia

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Whole-brain imaging is challenging because it demands microscopes with high temporal and spatial resolution, which are often at odds, especially in the context of large fields of view. We have designed and built a light-sheet microscope with digital micromirror illumination and light-field detection. On the one hand, light sheets provide high resolution optical sectioning on live samples without compromising their viability. On the other hand, light field imaging makes it possible to reconstruct full volumes of relatively large fields of view from a single camera exposure;however, its enhanced temporal resolution comes at the expense of spatial resolution, limiting its applicability. We present an approach to increase the resolution of light field images using DMD-based light sheet illumination. To that end, we develop a method to produce synthetic resolution targets for light field microscopy and a procedure to correct the depth at which planes are refocused with rendering software. We measured the axial resolution as a function of depth and show a three-fold potential improvement with structured illumination, albeit by sacrificing some temporal resolution, also three-fold. This results in an imaging system that may be adjusted to specific needs without having to reassemble and realign it. This approach could be used to image relatively large samples at high rates.

关键词： Light field microscopy light sheet fluorescence microscopy resolution enhancement structured illumination Digital Micromirror Device

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Dual-view inverted selective plane illumination microscopy (diSPIM) with improved background rejection for accurate 3D digital pathology 25

Dual-view inverted selective plane illumination microscopy (...

引用

conference on Three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Hu, Bihe Bolus, Daniel Brown, J. Quincy Tulane Univ Dept Biomed Engn 500 Lindy Boggs Ctr New Orleans LA 70118 USA

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Current gold-standard histopathology for cancerous biopsies is destructive, time consuming, and limited to 2D slices, which do not faithfully represent true 3D tumor micro-morphology. Light sheet microscopy has emerged as a powerful tool for 3D imaging of cancer biospecimens. Here, we utilize the versatile dual-view inverted selective plane illumination microscopy (diSPIM) to render digital histological images of cancer biopsies. Dual-view architecture enabled more isotropic resolution in X, Y, and Z;and different imaging modes, such as adding electronic confocal slit detection (eCSD) or structured illumination (SI), can be used to improve degraded image quality caused by background signal of large, scattering samples. To obtain traditional H&E-like images, we used DRAQ5 and eosin (D&E) staining, with 488nm and 647nm laser illumination, and multi-band filter sets. Here, phantom beads and a D&E stained buccal cell sample have been used to verify our dual-view method. We also show that via dual view imaging and deconvolution, more isotropic resolution has been achieved for optical cleared human prostate sample, providing more accurate quantitation of 3D tumor architecture than was possible with single-view SPIM methods. We demonstrate that the optimized diSPIM delivers more precise analysis of 3D cancer microarchitecture in human prostate biopsy than simpler light sheet microscopy arrangements.

关键词： dual-view inverted selective plane illumination microscopy electronic confocal slit detection structured illumination improved background rejection isotropic resolution 3D digital pathology pseudo-H&E cleared prostate biopsy

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Imaging a seizure model in zebrafish with structured illumination light sheet microscopy 25

Imaging a seizure model in zebrafish with structured illumin...

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conference on Three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Liu, Yang Dale, Savannah Ball, Rebecca VanLeuven, Ariel J. Baraban, Scott Sornborger, Andrew Lauderdale, James D. Kner, Peter Univ Georgia Coll Engn Athens GA 30602 USA Clemson Univ Dept Bioengn Clemson SC 29634 USA Univ Georgia Dept Cellular Biol Athens GA 30602 USA Univ Calif San Francisco Dept Neurol Surg San Francisco CA 94143 USA Univ Calif Davis Dept Math Davis CA 95616 USA

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Zebrafish are a promising vertebrate model for elucidating how neural circuits generate behavior under normal and pathological conditions. The Baraban group first demonstrated that zebrafish larvae are valuable for investigating seizure events and can be used as a model for epilepsy in humans. Because of their small size and transparency, zebrafish embryos are ideal for imaging seizure activity using calcium indicators. Light-sheet microscopy is well suited to capturing neural activity in zebrafish because it is capable of optical sectioning, high frame rates, and low excitation intensities. We describe work in our lab to use light-sheet microscopy for high-speed long-time imaging of neural activity in wildtype and mutant zebrafish to better understand the connectivity and activity of inhibitory neural networks when GABAergic signaling is altered in vivo. We show that, with light-sheet microscopy, neural activity can be recorded at 23 frames per second in two-colors for over 10 minutes allowing us to capture rare seizure events in mutants. We have further implemented structured illumination to increase resolution and contrast in the vertical and axial directions during high-speed imaging at an effective frame rate of over 7 frames per second.

关键词： fluorescence microscopy light sheet microscopy structured illumination microscopy neurophotonics

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Investigation of second harmonic generation and multispectral imaging as new contrast mechanisms in scanning laser optical tomography 25

Investigation of second harmonic generation and multispectra...

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conference on Three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Nolte, Lena Antonopoulos, Georgios C. Heisterkamp, Alexander Ripken, Tammo Meyer, Heiko Laser Zentrum Hannover eV Ind & Biomed Opt Dept Hollerithallee 8 D-30419 Hannover Germany Leibniz Univ Hannover Inst Quantenopt Hannover Germany

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Scanning laser optical tomography (SLOT) is a 3D imaging technique, based on the principle of computed tomography to visualize samples up to magnitude of several centimeters. Intrinsic contrast mechanisms as absorption, scattering and autofluorescence provide information about the 3D architecture and composition of the sample. Another valuable intrinsic contrast mechanism is second harmonic generation (SHG), which is generated in non-centrosymmetric materials and commonly used to image collagen in biological samples. The angular dependence of the SHG signal, however, produces artifacts in reconstructed optical tomography datasets (OPT, SLOT). Thus, successful use of this intrinsic contrast mechanism is impaired. We investigate these artifacts by simulation and experiment and propose an elimination procedure that enables successful reconstruction of SHG-SLOT data. Nevertheless, in many cases specific labeling of certain structures is necessary to make them visible. Using multiple dyes in one sample can lead to crosstalk between the different channels and reduce contrast of the images. Also autofluorescence of the sample itself can account for that. By using multispectral imaging in combination with spectral unmixing techniques, this loss can be compensated. Therefore either a spectrally resolved detection path, or spectrally resolved excitation is required. Therefore we integrated a white supercontinuum light source in our SLOT-setup that enables a spectral selection of the excitation beam and extended the detection path to a four channel setup. This enables the detection of three fluorescence channels and one absorption channel in parallel, and increases the contrast in the reconstructed 3D images significantly.

关键词： Scanning Laser Optical Tomography 3D microscopy Tomographic imaging Nonlinear Optics Second Harmonic Generation Multispectral imaging Spectral unmixing

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Preprocessing method to correct illumination pattern in sinusoidal-based structured illumination microscopy 25

Preprocessing method to correct illumination pattern in sinu...

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conference on Three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Shabani, H. Doblas, A. Saavedra, G. Preza, C. Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA Univ Valencia Dept Opt E-46100 Burjassot Spain

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

The restored images in structured illumination microscopy (SIM) can be affected by residual fringes due to a mismatch between the illumination pattern and the sinusoidal model assumed by the restoration method. When a Fresnel biprism is used to generate a structured pattern, this pattern cannot be described by a pure sinusoidal function since it is distorted by an envelope due to the biprism's edge. In this contribution, we have investigated the effect of the envelope on the restored SIM images and propose a computational method in order to address it. The proposed approach to reduce the effect of the envelope consists of two parts. First, the envelope of the structured pattern, determined through calibration data, is removed from the raw SIM data via a preprocessing step. In the second step, a notch filter is applied to the images, which are restored using the well-known generalized Wiener filter, to filter any residual undesired fringes. The performance of our approach has been evaluated numerically by simulating the effect of the envelope on synthetic forward images of a 6-ium spherical bead generated using the real pattern and then restored using the SIM approach that is based on an ideal pure sinusoidal function before and after our proposed correction method. The simulation result shows 74% reduction in the contrast of the residual pattern when the proposed method is applied. Experimental results from a pollen grain sample also validate the proposed approach.

关键词： structured illumination microscopy sinusoidal pattern residual fringe Fresnel biprism

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Hybrid of two-photon microscopy and optical multimodality imaging for multi-scale imaging of small animals 25

Hybrid of two-photon microscopy and optical multimodality im...

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conference on Three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Li, Tianmeng Hui, Hui Ma, He Yang, Xin Tian, Jie Northeastern Univ Sinodutch Biomed & Informat Engn Sch Shenyang Liaoning Peoples R China Chinese Acad Sci Inst Automat Key Lab Mol Imaging Beijing 100190 Peoples R China Beijing Key Lab Mol Imaging Inst Automat Beijing 100190 Peoples R China

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Non-invasive imaging technologies, such as magnetic resonance imaging (MRI) and optical multimodality imaging methods, are commonly used for diagnosing and supervising the development of inflammatory bowel disease (IBD). These in vivo imaging methods can provide morphology changes information of IBD in macro-scale. However, it is difficult to investigate the intestinal wall in molecular and cellular level. State-of-art light-sheet and two-photon microscopy have the ability to acquire the changes for IBD in micro-scale. The aim of this work is to evaluate the size of the enterocoel and the thickness of colon wall using both MRI for in vivo imaging, and light-sheet and two-photon microscope for in vitro imaging. C57BL/6 mice were received 3.5% Dextran sodium sulfate (DSS) in the drinking water for 5 days to build IBD model. Mice were imaged with MRI on days 0, 6 to observe colitis progression. After MRI imaging, the mice were sacrificed to take colons for tissue clearing. Then, light-sheet and two-photon microscopies are used for in vitro imaging of the cleared samples. The experimental group showed symptoms of bloody stools, sluggishness and weight loss. It showed that the colon wall was thicker while the enterocoel was narrower compare to control group. The more details are observed using light-sheet and two-photon microscope. It is demonstrated that hybrid of MRI in macro-scale and light-sheet and two-photon microscopy in micro-scale imaging is feasible for colon inflammation diagnosing and supervising.

关键词： Multi-scale molecular imaging Inflammatory bowel disease Magnetic resonance imaging Two-photon microscopy Light-sheet microscopy

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Coherent light 3D materials microscopy without phase retrieval 24

Coherent light 3D materials microscopy without phase retriev...

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

作者： Xing, Jian Yu, Jiun-Yann Chen, Simeng Cogswell, Carol Univ Colorado Dept Elect Comp & Energy Engn 425 UCB Boulder CO 80309 USA

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

We report a novel microscopy technique, utilizing our previously reported expanded point information content (EPIC) concept [1], to extend the technique into the coherent regime. Preliminary data shows coherent EPIC (CoEPIC) can image reflective samples successfully, and can recover the 3D structure without the need to acquire an image stack at multiple depths. Numerical simulation demonstrates the potential super-resolution capability of CoEPIC.

关键词： 3D materials inspection PSF engineering phase retrieval super-resolution coherent illumination

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Single image sectioning using spatial frequency domain imaging 24

Single image sectioning using spatial frequency domain imagi...

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

作者： Hoffman, Zachary R. DiMarzio, Charles A. Northeastern Univ 360 Huntington Ave Boston MA 02115 USA Draper Lab 555 Technol Sq Cambridge MA 02139 USA

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Structured illumination microscopy (SIM) works well to produce depth information by removing undesired light from out of focus planes within a specimen. However, it generally requires multiple images in order to produce sectioning. We show that, under the right conditions, we can leverage Spatial Frequency Domain Imaging (SFDI) to produce both sectioning and relative depth information about a specimen using only a single image. After processing, we find that the axial resolution is comparable to that of traditional 3 phase SIM. With this technique, we are able to produce real-time videos in-vivo at 21 frames per second. Additionally, no moving parts are needed to produce sectioning at depth.

关键词： Structured Illumination optical sectioning super-resolution incoherent SFDI

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Focus-tunable lens in limited-angle holographic tomography 24

Focus-tunable lens in limited-angle holographic tomography

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

作者： Kus, A. Krauze, W. Kujawinska, M. Warsaw Univ Technol Inst Micromech & Photon Sw Boboli 8 St PL-02525 Warsaw Poland

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

In this paper a new, hardware-based solution for extending the depth of field in holographic tomography is presented. The solution is based on a 4f system and an electric, focus-tunable lens, which provides fast, motion-free defocusing of the plane conjugate with the camera, which acquires holograms. The optimum parameters for the required axial scanning are provided for a specific model of a commercially available tunable lens. Then, the quality of the system equipped with the designed module is analyzed and the reconstruction of a standard object (microsphere) scanned by the 4f-based defocusing system is presented. Finally, the result of the increased depth of field in the measurement domain is demonstrated with a reconstruction of a mouse fibroblast cell.

关键词： Tomographic phase microscopy digital holography diffraction tomography quantitative phase imaging 3D imaging refractive index

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