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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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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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Super-resolution in confocal microscopy using generative adversarial networks with paired and unpaired data 30

Super-resolution in confocal microscopy using generative adv...

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

作者： Patra, S. Trujillo, C. Doblas, A. Department of Electrical and Computer Engineering The University of Memphis TN United States School of Applied Sciences and Engineering Universidad EAFIT Medellin Colombia

ISBN: (纸本)9781510658752

The advantages of confocal microscopy over widefield microscopy are their ability to produce optically sectioned images and their ability to produce multi-color imaging in which different organelles within the biological specimen are stained using multiple dyes, enabling colocalization studies. These features make confocal microscopy a widely used tool to provide valuable morphological and functional information within cells and tissues. One of the major drawbacks of confocal microscopy is its limited spatial resolution. Here, we train two generative adversarial networks using paired and unpaired data of low- and high-resolution images to improve the spatial resolution in confocal microscopy. © 2023 SPIE. All rights reserved.

关键词： Confocal microscopy

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Utilizing fluorescent beads to mimic single fluorophores 30

Utilizing fluorescent beads to mimic single fluorophores

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

作者： Feng, Sherry Yi-Ting Alemán-Castañeda, Luis A. Gutiérrez-Cuevas, Rodrigo Herrera, Isael Brown, Thomas G. Brasselet, Sophie Alonso, Miguel A. The Institute of Optics University of Rochester RochesterNY14627 United States Aix Marseille Univ CNRS Centrale Marseille Institut Fresnel MarseilleF-13013 France Institut Langevin ESPCI Paris Université PSL CNRS Paris75005 France

In this paper, numerical and semi-analytical methods are used for studying how well a fluorescent bead with a given radius in a polarizer-supplemented microscopy setup, can emulate a single fluorescent molecule. ©... 详细信息

ISBN: (纸本)9781510658752

关键词： Fluorescence microscopy

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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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High-speed light-sheet microscopy imaging and data post-processing using custom hardware and software solutions 30

High-speed light-sheet microscopy imaging and data post-proc...

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

作者： Mazzamuto, Giacomo Silvestri, Ludovico Sancataldo, Giuseppe Gavryusev, Vladislav Scardigli, Marina Costantini, Irene Pavone, Francesco S. Sesto Fiorentino Italy Sesto Fiorentino Italy Department of Physics and Astronomy University of Florence Italy Department of Physics and Chemistry University of Palermo Italy Department of Biology University of Florence Italy

ISBN: (纸本)9781510658752

High-speed imaging with Light-Sheet Fluorescence microscopy poses several challenges throughout the whole pipeline, from data acquisition in the lab to image stitching and post-processing. Here we present our custom hardware and software solutions that allow us to map large biological samples at the cellular level, e.g. large portions of human brain cortex. Our custom optical setup — a dual-view, inverted, light-sheet microscope — is capable of simultaneous two-color acquisition at a data rate of 1 GB/s. Our open source tools include the instrument’s data acquisition and control software and also cover volumetric image stitching and post-processing. © 2023 SPIE. All rights reserved.

关键词： Brain

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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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Light-field microscopy to study cardiac arrhythmias in zebrafish larvae 31

Light-field microscopy to study cardiac arrhythmias in zebra...

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

作者： Saberigarakani, Alireza Zhang, Xinyuan Almasian, Milad Ding, Yichen Department of Bioengineering The University of Texas at Dallas RichardsonTX75080 United States Center for Imaging and Surgical Innovation The University of Texas at Dallas RichardsonTX75080 United States Hamon Center for Regenerative Science and Medicine UT Southwestern Medical Center DallasTX75390 United States

ISBN: (纸本)9781510669550

Paroxysmal arrhythmias caused by medications are challenging to be prospectively identified. Zebrafish have emerged as an ideal model organism for screening small molecule compounds to study cardiac abnormalities, due to their rapid development, optical transparency during early stages, and similarities to the human heart. To overcome the challenges associated with observing cardiac abnormalities in zebrafish, we sought to develop a light-field microscope, a rapid imaging method with high photon efficiency, for the volumetric acquisition in a single snapshot. This method, along with its variations, utilizes a multi-lens array (MLA) to capture angular information. We have customized a light-field system and developed a pipeline to incorporate the MLA into the detection path. A program based on wave optics has also been developed to calculate the point spread function at different depths. The program involves two main steps: calculating the wide-field PSF and applying the MLA effect as a mask. This enables us to simulate the impact of the MLA on the imaging system. The comparison between the simulated and experimental data allows for the determination of MLA position. We aim to capture drug-induced arrhythmias in zebrafish larvae using this method, exploring the contractile dysfunction across the atrium and ventricle during multiple cardiac cycles. This research aims to deepen our understanding of the mechanisms underlying these arrhythmias and their connection to drug effects. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

关键词： Optical transfer function

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Single-shot capture bright-field and digital holographic microscopy for the analysis of living cells 30

Single-shot capture bright-field and digital holographic mic...

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

作者： Picazo-Bueno, José Ángel Barroso, Álvaro Ketelhut, Steffi Schnekenburger, Jürgen Micó, Vicente Kemper, Björn Biomedical Technology Center University of Muenster Mendelstraβe 17 MuensterD-48149 Germany Department of Optics and Optometry and Vision Sciences University of Valencia C/Doctor Moliner 50 Burjassot46100 Spain

ISBN: (纸本)9781510658752

Digital holographic microscopy (DHM) is an emerging label-free modality for quantitative imaging of semi-transparent biological specimens. DHM provides quantitative phase imaging (QPI) of the light transmitted or scattered by an object, which allows to extract information about morphology and dynamics of investigated specimens. DHM can be implemented with other optical microscopy techniques like bright field (BF) or fluorescence, for example, to provide complementary information about cellular processes. Multimodal microscopy is often achieved by sequential acquisition of information from different imaging modalities or simultaneously by capturing images with different acquisition devices. However, such arrangements can be cost-intensive and/or limited in speed for imaging of dynamic processes. Here, we investigate a single-snapshot multimodal approach that enables simultaneous acquisition of DHM-based QPI and BF images and is compatible with common optical microscopes for the analysis of living cells. The technique is based on spatially multiplexed interferometric microscopy (SMIM), where off-axis holograms are generated by spatial multiplexing of the illuminated sample plane utilizing a diffraction grating, which is extended by additional superposition of complementary white-light image information. image separation is achieved by Fourier-transformation-based numerical demultiplexing procedures. The technique is firstly characterized by investigations on microspheres. Then, the application on living adherent and suspended cells is illustrated. © 2023 SPIE. All rights reserved.

关键词： Holograms

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three-dimensional and multidimensional microscopy: image acquisition and processing XXVII

Three-Dimensional and Multidimensional Microscopy: Image Acq...

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three-dimensional and multidimensional microscopy: image acquisition and processing XXVII 2020

ISBN: (纸本)9781510632530

The proceedings contain 14 papers. The topics discussed include: multi-immersion open-top light-sheet microscopy;compact spectrograph for hyperspectral imaging with overlapping fluorescent reporters;computational super-resolution microscopy: leveraging noise model, regularization and sparsity to achieve highest resolution;wavelet-based characterization of the spatial relationship of nerve and collagen in neuropathic adipose tissue;multifocus imaging via polarization wavefront shaping;a study for accelerating the speed of all-in-focus image processing;excitation-scan mirror array system advancements to hyperspectral imaging applications;and 3D refractive index tomography with quantitative oblique back-illumination microscopy.

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