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检索条件"任意字段=Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXX 2023"

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Fast volumetric seizure model imaging in zebrafish brain with light sheet microscopy 31

Fast volumetric seizure model imaging in zebrafish brain wit...

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three-dimensional and multidimensional microscopy: image acquisition and processing xxxI 2024

作者： Liu, Bingxi Liu, Yang Duffy, Carly Lauderdale, James D. Kner, Peter School of Electrical and Computer Engineering University of Georgia AthensGA30602 United States Department of Cellular Biology University of Georgia AthensGA30602 United States

ISBN: (纸本)9781510669550

Light Sheet Fluorescence microscopy (LSFM) is an ideal tool for imaging model organisms that are hundreds of microns in size, providing high resolution and fast frame rates. In order to extend LSFM to rapid 3D volumetric imaging, a popular method is to add an electrically tunable lens (ETL) in the detection path of the LSFM. But for larger fields of view and higher NA objectives, the ETL introduces aberrations in the system. Here, we developed an LSFM with adaptive optics and an ETL for rapid focusing. We demonstrate that the system enables imaging over a volume of 499 × 499 × 140 μm3 with a volumetric speed of 4 Hz. We apply the system to image neural activity in the zebrafish larvae to capture rare seizure events. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

关键词： Fluorescence microscopy

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Compressive Time-resolved Multispectral Fluorescence microscopy with Single-Pixel Camera 30

Compressive Time-resolved Multispectral Fluorescence Microsc...

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three-dimensional and multidimensional microscopy: image acquisition and processing xxx 2023

作者： Ghezzi, Alberto Farina, Andrea Lenz, Armin J.M. Soldevila, Fernando Tajahuerce, Enrique Bassi, Andrea Valentini, Gianluca D’Andrea, Cosimo Politecnico di Milano Dipartimento di Fisica Piazza L. da Vinci 32 Milano20133 Italy Consiglio Nazionale delle Ricerche IFN Piazza L. da Vinci 32 Milan20133 Italy Universitat Jaume I Avda. Sos Baynat s/n Castelló12071 Spain UMR 8552 Collège de France 24 rue Lhomond Paris75005 France Center for NanoScience and Technology @PoliMi Istituto Italiano di Tecnologia via Pascoli 70/3 via Pascoli 70/3 Milan20133 Italy

ISBN: (纸本)9781510658752

Time-resolved multispectral fluorescence microscopy provides a 4D hypercube dataset with high specificity for cellular examination. However, this is generally obtained by significantly increasing the measurement time, which is quite limiting for in vivo measurement or with photosensitive sample. It is possible to reduce the measurement effort with a novel microscopy framework exploiting Compressive Sensing based on Single-Pixel Camera. In this work, we present a compressive sensing system and validate it with a cellular sample. Data fusion with a high-resolution camera image allows us to tackle the well-known problem of low-resolution in single-pixel imaging. © 2023 SPIE. All rights reserved.

关键词： Compressed sensing

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MAGNIFY: molecule anchorable gel-enabled nanoscale in-situ fluorescence microscopy for nanoscale imaging of biomolecules 30

MAGNIFY: molecule anchorable gel-enabled nanoscale in-situ f...

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three-dimensional and multidimensional microscopy: image acquisition and processing xxx 2023

作者： Klimas, Aleksandra Gallagher, Brendan R. DiBernardo, Emma Cheng, Zhangyu Zhao, Yongxin Department of Biological Sciences Carnegie Mellon University PittsburghPA United States

ISBN: (纸本)9781510658752

Expansion microscopy (ExM) is a powerful imaging strategy that offers a low-cost solution for interrogating biological systems at the nanoscale using conventional optical microscopes. It achieves this by physically and isotropically magnifying preserved biological specimens embedded in a cross-linked water-swellable hydrogel. However, most reported techniques are unable to preserve endogenous epitopes due to strong protease digestion used to expand samples. In addition, these protocols rely on mechanically fragile hydrogels that only expand by at most 4.5× linearly. We present a new ExM framework, Molecule Anchorable Gel-enabled Nanoscale In-situ Fluorescence microscopy (MAGNIFY), that exhibits a broad retention of nucleic acids, proteins, and lipids without the need for a separate anchoring step. By using a mechanically sturdy hydrogel, MAGNIFY is capable of expanding biological specimens up to 11×. This facilitates nanoscale imaging (~25-nm effective resolution) using an ∼280-nm diffraction-limited objective lens on a conventional optical microscope and can be furthered to ~15 nm effective resolution if combined with computational methods such as Super-resolution Optical Fluctuation Imaging (SOFI). Here, we demonstrate that MAGNIFY provides a generalized solution for imaging nanoscale subcellular features of a broad range of biological specimens. We also show that MAGNIFY provides a novel, accessible tool for improving the precision, utility, and generality of nanoscopy. © 2023 SPIE. All rights reserved.

关键词： Fluorescence microscopy

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three-dimensional multi-directional light-sheet microscopy with structured illumination 31

Three-dimensional multi-directional light-sheet microscopy w...

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three-dimensional and multidimensional microscopy: image acquisition and processing xxxI 2024

作者： Song, Ming Liu, Yang Wu, Hsueh-Fu Zeltner, Nadja Kner, Peter A. School of Electrical and Computer Engineering University of Georgia AthensGA United States Chan Zuckerberg Biohub San FranciscoCA United States Center for Molecular Medicine University of Georgia AthensGA United States Department of Biochemistry and Molecular Biology University of Georgia AthensGA United States Department of Cellular Biology University of Georgia AthensGA United States

ISBN: (纸本)9781510669550

Light-Sheet Fluorescence microscopy (LSFM) has demonstrated its effectiveness in imaging many model organisms. However, traditional LSFMs often exhibit low resolution and encounter striping artifacts. Here, we introduce a novel LSFM design to address these challenges of imaging large biological samples. Our system employs a single-objective light-sheet geometry, incorporating Structured Illumination microscopy (SIM) to allow multi-direction light-sheet illumination. This combination aims to rectify striping artifacts and achieve improved lateral resolutions of better than 200 nm over a 277 × 277 × 500 μm3 volume. Our system is demonstrated through imaging beads, C. elegans and cerebellum organoids. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

关键词： Fluorescence microscopy

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Imaging the brain in vivo with reflectance phase-contrast confocal microscopy at 1650nm 31

Imaging the brain in vivo with reflectance phase-contrast co...

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three-dimensional and multidimensional microscopy: image acquisition and processing xxxI 2024

作者： Delafontaine Martel, P. Zhang, Cong Linninger, Andreas A. Lesage, Frédéric Biomedical Engineering Institute Polytechnique Montreal Montreal Canada Montreal Heart Institute Montreal Canada Departement of Biomedical Engineering and Neurosurgery University of Illinois ChicagoIL United States

ISBN: (纸本)9781510669550

Recent advancements in sources and detectors operating in the NIR-II wavelengths have driven the emergence of NIR-II intrinsic microscopy. These significant technological strides were necessary because longer wavelengths are known to experience reduced scattering and absorption in biological tissue. Leveraging this optical advantage, the application of the NIR-II spectral domain in microscopy holds the potential to improve the depth of imaging and preserve coherence depth. In this study, we showcase the integration of phase-contrast imaging into a NIR-II reflectance confocal microscope for cortical imaging. By capturing images of cortical cell bodies at depths of up to 800 μm, we demonstrate that the implementation of phase contrast provides clear delineation of cortical cell edges, including myelinated axons, blood vessels, and cortical cell bodies. Additionally, we devised a computational method to enhance dynamic components and generate a digitized vascular network from the acquired images. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

关键词： Reflection

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image projection through a multimode fiber with reference-lessly retrieved transmission matrix 31

Image projection through a multimode fiber with reference-le...

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three-dimensional and multidimensional microscopy: image acquisition and processing xxxI 2024

作者： Zhong, Jingshan Wen, Zhong Li, Quanzhi Deng, Qilin Yang, Qing Research Center for Humanoid Sensing Zhejiang Lab Hangzhou311100 China State Key Laboratory of Extreme Photonics and Instrumentation College of Optical Science and Engineering International Research Center for Advanced Photonics Zhejiang University Hangzhou310027 China

ISBN: (纸本)9781510669550

image projection through a multimode fiber (MMF) or scattering media has applications ranging from optogenetics to near eye-displays. It requires developing computer-generated holography algorithm to obtain phase pattern of spatial light modulator. In order to accurately project light, conventional methods measured the transmission matrix (TM) of the imaging system by interference. However, it is sensitive to phase instability, easily caused by thermal drift and mechanical vibration. In this work, we proposed to use the TM retrieved from intensity-only measurements and develop a nonlinear optimization algorithm to obtain the displayed phase patterns. Our method formulates the forward model with the retrieved TM, derives the analytical derivative and adopts a second-order optimization method. We validate the improved quality of the projected intensity image by an experiment setup with a MMF. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

关键词： Multimode fibers

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Multispectral digital holographic microscopy with tunable supercontinuum lasers 31

Multispectral digital holographic microscopy with tunable su...

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three-dimensional and multidimensional microscopy: image acquisition and processing xxxI 2024

作者： Kurtz, Tobias Barroso, Álvaro Picazo-Bueno, José Ángel Ketelhut, Steffi Schnekenburger, Jürgen Kemper, Björn Biomedical Technology Center of the Medical Faculty University of Muenster MuensterD-48149 Germany Department Department of Optics Optometry and Vision Science Faculty of Physics University of Valencia Burjassot46100 Spain

ISBN: (纸本)9781510669550

Supercontinuum (SC) lasers in combination with tunable acousto-optic filters facilitate multiwavelength digital holographic microscopy (DHM) with a broad spectral range. However, to achieve optimized quantitative phase imaging (QPI) in off-axis DHM the coherence properties of the tunable light source and the holographic carrier interference fringe pattern formation have to be considered as they are wavelength dependent. We thus determined experimentally the wavelength dependency of coherence length and off-axis carrier fringe frequency in a Michelson interferometer-based multi-spectral DHM setup with a continuously tunable SC laser and moreover analyzed the impact of both parameters on hologram formation and QPI image quality. The results are found in good agreement with theoretical predictions and provide essential information for a reliable adjustment of SC laser based multispectral off-axis DHM systems. Finally, the application of multispectral-DHM for reduction of coherence induced disturbances in quantitative phase images of living pancreatic tumor cells in illustrated. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

关键词： Michelson interferometers

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Hyperspectral microscopy as a particle exposure assessment tool 31

Hyperspectral microscopy as a particle exposure assessment t...

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three-dimensional and multidimensional microscopy: image acquisition and processing xxxI 2024

作者： Blanch-Perez Del Notario, Carolina Bertier, Pieter Jayapala, Murali Lambrechts, Andy Imec Kapeldreef 75 Leuven3001 Belgium Belgian Center for Occupational Hygiene BeCOH Brusselsesteenweg 46 Leuven3000 Belgium

ISBN: (纸本)9781510669550

Hyperspectral imaging combines the characteristics of computer vision and point spectroscopy by obtaining an image with both spatial and spectral information. Therefore, in combination with microscopy, it can increase material discrimination possibilities with respect to regular microscopy imaging. We explore this increased discrimination potential to assess exposure to particle contamination, since workplace exposure to specific particle materials poses well-known health hazards. In this respect, we are focusing on discriminating more health relevant particles such as silica in the respirable size fraction. For this purpose, a particle sampling protocol has been proposed and hyperspectral imaging in combination with transmission microscopy is used for particle material identification. We use a Snapscan visual near-infrared (VNIR) camera providing high spectral and spatial resolution in the 460-900 nm range, 150 spectral bands and up to 7 Mpixels of spatial resolution and high acquisition speed. The hyperspectral microscopy system has been tested for discrimination of fifteen different particle materials, such as silica, coal, dolomite, barite, or rutile, among others. The combined analysis of spatial and spectral information shows potential to accurately discriminate the 15 tested particle materials so far by means of a random forest classifier. In addition, a band relevance analysis is performed showing that only a few specific bands are needed to provide accurate discrimination of the tested materials. The hyperspectral hardware and method presented could lead to a faster exposure assessment than traditional techniques used for occupational exposure estimation. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

关键词： Health hazards

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Normalized level set model for segmentation of low-contrast objects in 2- and 3-dimensional images 31

Normalized level set model for segmentation of low-contrast ...

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three-dimensional and multidimensional microscopy: image acquisition and processing xxxI 2024

作者： Baig, Mirza M. Junaid Wang, Yao L. Chung, Samuel H. Stepanyants, Armen Department of Physics Northeastern University 360 Huntington Ave BostonMA02115 United States Department of Bioengineering Northeastern University 360 Huntington Ave BostonMA02115 United States

ISBN: (纸本)9781510669550

Analyses of biomedical images often rely on accurate segmentation of structures of interest. Traditional segmentation methods based on thresholding, watershed, fast marching, and level set perform well in high-contrast images containing structures of similar intensities. However, such methods can under-segment or miss entirely low-intensity objects on noisy backgrounds. Machine learning segmentation methods promise superior performance but require large training datasets of labeled images which are difficult to create, particularly in 3D. Here, we propose an algorithm based on the Local Binary Fitting level set method and its application specifically designed to improve the segmentation accuracy for low-contrast structures even with significant noise levels present. The proposed algorithm, the Normalized Local Binary Fitting level set method, shows promise in enhancing the segmentation of low-contrast structures in biomedical images, addressing the limitations of traditional segmentation methods, and offering an alternative to machine learning approaches that require extensive training datasets. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

关键词： Machine learning

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On-the-fly deconvolution of diagonally-scanned Airy lightsheet microscopy 31

On-the-fly deconvolution of diagonally-scanned Airy lightshe...

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three-dimensional and multidimensional microscopy: image acquisition and processing xxxI 2024

作者： Vettenburg, Tom Valantinas, Laurynas Center for Medical Engineering and Technology School of Science and Engineering University of Dundee Nethergate DD14HN United Kingdom

ISBN: (纸本)9781510669550

Its subcellular resolution and minimal sample exposure make light-sheet microscopy the ideal tool to study biological specimen during their early development. A light-sheet microscope scans the sample with a plane of light and collects fluorescence with an objective orthogonal to the illumination. However, tightly focused Gaussian light-sheets suffer from a shallow depth of focus and are susceptible to scattering-induced aberrations. Light-sheets created by non-diffractive Airy beams can overcome this to yield isotropic sub-cellular resolution over a ten-fold larger field-of-view. Airy beam light-sheets have a characteristically curved structure and a broad transverse structure with side-lobes. Digital deconvolution of the raw data is thus essential to obtain high-fidelity images. Provided that the scan is along the direction of the detection axis, and all recorded data fits within working memory, a simple and efficient Wiener filter can recover accurate 3D images. However, multi-millimeter sized samples must be scanned with a light-sheet that is diagonal to the sample surface. The diagonal movement prevents the use of standard Wiener filtering. Moreover, the associated data sets can become too large to fit within the working memory of a consumer-grade GPU. This demands slow off-line processing, thus breaking a rapid experimental feedback-loop. Here, we investigate the potential of on-the-fly deconvolution of diagonally-scanned Airy light-sheet microscopy. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

关键词： Fluorescence

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