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检索条件"任意字段=Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIX 2022"

共 602 条记录，以下是101-110 订阅

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Correlative FRET: New method improves rigor and reproducibility in determining distances within synaptic nanoscale architecture 25

Correlative FRET: New method improves rigor and reproducibil...

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three-dimensional and multidimensional microscopy: image acquisition and processing XXV 2018

作者： Shinogle-Decker, Heather Martinez-Rivera, Noraida O'Brien, John Powell, Richard D. Joshi, Vishwas N. Connell, Samuel Rosa-Molinar, Eduardo Microscopy and Analytical Imaging Research Resource Core Laboratory University of Kansas LawrenceKS66045 United States Biological Imaging Group University of Kansas LawrenceKS66045 United States Department of Ophthalmology and Visual Science University of Texas Health Science Center at Houston TX77030 United States Nanoprobes Inc. YaphankNY11980 United States Anvesha Laboratories SetauketNY11733 United States Intelligent Imaging Innovations DenverCO80216 United States

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614833

A new correlative Förster Resonance Energy Transfer (FRET) microscopy method using FluoroNanogold™, a fluorescent immunoprobe with a covalently attached Nanogold® particle (1.4nm Au), overcomes resolution limitations in determining distances within synaptic nanoscale architecture. FRET by acceptor photobleaching has long been used as a method to increase fluorescence resolution. The transfer of energy from a donor to an acceptor generally occurs between 10-100Å, which is the relative distance between the donor molecule and the acceptor molecule. For the correlative FRET microscopy method using FluoroNanogold™, we immuno-labeled GFP-tagged-HeLa-expressing Connexin 35 (Cx35) with anti-GFP and with anti-Cx35/36 antibodies, and then photo-bleached the Cx before processing the sample for electron microscopic imaging. Preliminary studies reveal the use of Alexa Fluor® 594 FluoroNanogold™ slightly increases FRET distance to 70Å, in contrast to the 62.5Å using AlexaFluor 594®. Preliminary studies also show that using a FluoroNanogold™ probe inhibits photobleaching. After one photobleaching session, Alexa Fluor 594® fluorescence dropped to 19% of its original fluorescence;in contrast, after one photobleaching session, Alexa Fluor 594® FluoroNanogold™ fluorescence dropped to 53% of its original intensity. This result confirms that Alexa Fluor 594® FluoroNanogold™ is a much better donor probe than is Alexa Fluor 594®. The new method (a) creates a double confirmation method in determining structure and orientation of synaptic architecture, (b) allows development of a two-dimensional in vitro model to be used for precise testing of multiple parameters, and (c) increases throughput. Future work will include development of FluoroNanogold™ probes with different sizes of gold for additional correlative microscopy studies. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

关键词： Forster resonance energy transfer

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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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Spatially-controlled illumination with rescan confocal microscopy enhances image quality, resolution and reduces photodamage 24

Spatially-controlled illumination with rescan confocal micro...

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

作者： Krishnaswami, Venkataraman De Luca, Giulia M. R. Breedijk, Ronald M. P. Van Noorden, Cornelis J. F. Manders, Erik M. M. Hoebe, Ron A. Univ Amsterdam Acad Med Ctr van Leeuwenhoek Ctr Adv Microscopy Cellular ImagingDept Cell Biol & Histol Amsterdam Netherlands Univ Amsterdam Swammerdam Inst Life Sci van Leeuwenhoek Ctr Adv Microscopy Amsterdam Netherlands

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Fluorescence microscopy is an important tool in biomedical imaging. An inherent trade-off lies between image quality and photodamage. Recently, we have introduced rescan confocal microscopy (RCM) that improves the lateral resolution of a confocal microscope down to 170 nm. Previously, we have demonstrated that with controlled-light exposure microscopy, spatial control of illumination reduces photodamage without compromising image quality. Here, we show that the combination of these two techniques leads to high resolution imaging with reduced photodamage without compromising image quality. Implementation of spatially-controlled illumination was carried out in RCM using a line scanning-based approach. Illumination is spatially-controlled for every line during imaging with the help of a prediction algorithm that estimates the spatial profile of the fluorescent specimen. The estimation is based on the information available from previously acquired line images. As a proof-of-principle, we show images of N1E-115 neuroblastoma cells, obtained by this new setup with reduced illumination dose, improved resolution and without compromising image quality.

关键词： Spatially-controlled illumination Rescan-confocal fluorescence biomedical imaging image quality photodamage

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Remote focusing multifocal plane microscopy for the imaging of 3D single molecule dynamics with cellular context 24

Remote focusing multifocal plane microscopy for the imaging ...

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

作者： Chao, Jerry Velmurugan, Ramraj You, Sungyong Kim, Dongyoung Ward, E. Sally Ober, Raimund J. Texas A&M Univ Dept Biomed Engn College Stn TX 77843 USA Texas A&M Univ Hlth Sci Ctr Dept Mol & Cellular Med College Stn TX 77843 USA Texas A&M Univ Hlth Sci Ctr Dept Microbial Pathogenesis & Immunol Bryan TX 77807 USA Univ Texas Southwestern Med Ctr Dallas Biomed Engn Grad Program Dallas TX 75390 USA

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

three-dimensional (3D) single molecule fluorescence microscopy affords the ability to investigate subcellular trafficking at the level of individual molecules. An imaged single molecule trajectory, however, often reveals only limited information about the underlying biological process when insufficient information is available about the organelles and other cellular structures with which the molecule interacts. A new 3D fluorescence microscopy imaging modality is described here that enables the simultaneous imaging of the trajectories of fast-moving molecules and the associated cellular context. The new modality is called remote focusing multifocal plane microscopy (rM UM), as it extends multi focal plane microscopy (MUNI) with a remote focusing module. MUM is a modality1 uses multiple detectors to image distinct focal planes within the specimen at the same time, and it has been demonstrated to allow the determination of 31) single molecule trajectories with high accuracy. Remote focusing is a method that makes use of two additional objective lenses to enable the acquisition of a z -stack of the specimen without having to move the microscope's objective lens or sample stage, components which are required by MUM to be fixed in place. rMUM's remote focusing module thus allows the cellular context to be imaged in the form of z -stacks as the trajectories of molecules or other objects of interest are imaged by MUM. In addition to a description of the modality, a discussion of rMUM data analysis and an example of data acquired using an rMUM setup are provided in this paper.

关键词： Cellular context fluorescence microscopy multifocal plane microscopy remote focusing single molecule localization single molecule microscopy single molecule tracking three-dimensional microscopy

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Toward multi-focal spot remote focusing two-photon microscopy for high speed imaging 24

Toward multi-focal spot remote focusing two-photon microscop...

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

作者： Li, Bei Corbett, Alexander D. Chong, Ee Mann, Edward Wilson, Tony Booth, Martin J. Bub, Gil Univ Oxford Dept Engn Sci Oxford England Univ Exeter Dept Phys & Astron Exeter Devon England Univ Oxford Dept Physiol Anat & Genet Oxford England McGill Univ Dept Physiol Montreal PQ Canada

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Optical sectioning techniques using two-photon excitation of fluorescent indicators are central to diverse imaging applications. The limitations of the technique are low speed and undesirable specimen agitation. In our design, highspeed axial scanning is carried out by moving a reference objective to axially displace the focal spot without introducing significant spherical aberration and any agitation of the specimen. Further, the system is configured to allow switching between single spot and multiple focal spot remote focusing to allow either high dynamic range or high speed imaging.

关键词： two-photon excitation remote focusing temporal pixel multiplexing

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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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Fast axial scanning for 2-photon microscopy using liquid lens technology 24

Fast axial scanning for 2-photon microscopy using liquid len...

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

作者： Tehrani, Kayvan Forouhesh Sun, Min Kyoung Karumbaiah, Lohitash Mortensen, Luke J. Univ Georgia Rhodes Ctr ADS Regenerat Biosci Ctr Athens GA 30602 USA Univ Georgia Coll Engn Athens GA 30602 USA

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Scanning microscopy methods require movement of the focus in Z coordinates to produce an image of a 3-dimensional volume. In a typical imaging system, the optical setup is kept fixed and either the sample or the objective is translated with a mechanical stage driven by a stepper motor or a piezoelectric element. Mechanical Z scanning is precise, but its slow response and vulnerability to mechanical vibrations and stress make it disadvantageous to image dynamic, time-varying samples such as live cell structures. An alternative method less susceptible to these problems is to change the focal plane using conjugate optics. Deformable mirrors, acousto-optics, and electrically tunable lenses have been experimented with to achieve this goal and have attained very fast and precise Z-scanning without physically moving the sample. Here, we present the use of a liquid lens for fast axial scanning. Liquid lenses have a long functional life, high degree of phase shift, and low sensitivity to mechanical stress. They work on the principle of refraction at a liquid-liquid interface. At the boundary of a polar and an apolar liquid a spherical surface is formed whose curvature can be controlled by adjusting its relative wettability using electro-wetting. We characterize the effects of the lens on attainable Z displacement, beam spectral characteristics, and pulse duration as compared with mechanical scanning.

关键词： Remote focusing Liquid lens Active optics Multi-photon microscopy

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Time-resolved wavelet-based acquisitions using a single pixel camera 24

Time-resolved wavelet-based acquisitions using a single pixe...

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

作者： Rousset, Florian Ducros, Nicolas Farina, Andrea Valentini, Gianluca D'Andrea, Cosimo Peyrin, Francoise Univ Lyon INSA Lyon INSERM UCBLCNRS 5220U1206CREATIS 7 Ave Jean Capelle F-69621 Villeurbanne France Politecn Milan Dipartimento Fis Piazza Leonardo da Vinci 32 I-20133 Milan Italy CNR IFN Piazza Leonardo da Vinci 32 I-20133 Milan Italy Ist Italiano Tecnol Ctr Nano Sci & Technol Polimi Via Pascoli 70-3 I-20133 Milan Italy

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

We propose to couple a single-pixel camera with a photon counting board in order to obtain an inexpensive time-resolved imaging system having a high spatial and temporal resolution. As an alternative to compressive sensing, a wavelet-based adaptive acquisition strategy is employed which allows high compression rates with little degradation of the image quality. The applicability of our approach is demonstrated for fluorescence lifetime imaging. Experimental results obtained by imaging samples embedding several fluorophores are provided. The proposed imaging system with the wavelet-based strategy can be suitable for a microscope in order to perform fluorescence lifetime imaging microscopy measurements.

关键词： single pixel camera computational imaging fluorescence lifetime imaging time-resolved measurements wavelets adaptive acquisition

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Comparison of 3D structured patterns with tunable frequency for use in Structured Illumination microscopy 24

Comparison of 3D structured patterns with tunable frequency ...

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

作者： Doblas, Ana Shabani, Hasti Saavedra, Genaro Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA Univ Valencia Dept Opt E-46100 Burjassot Spain

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

Structured illumination microscopy (SIM) doubles the lateral resolution and produces optically-sectioned images. In SIM, the illumination system is modified in order to illuminate the sample by a structured pattern. Previously, axially-localized high-contrast sinusoidal patterns generated using a slit-prism illumination system based on a Fresnel biprism were investigated. In this contribution, we propose a Wollaston prism to replace the Fresnel biprism and produce the corresponding 3D structured illumination pattern. In this study, both optical elements are illuminated by the light emerging from an axial point source. Our results show that the benefits of using a Wollaston prism instead of a Fresnel biprism are twofold: (1) there is no envelope modulation perturbing the sinusoidal patterns and thereby reducing their visibility, and (2) the region of interference fringes is significantly larger than the one created by the Fresnel biprism.

关键词： structured illumination interference diffraction theory

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