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3 results for “TIRF microscopy”

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zenodo52/100

Example Microscopy Metadata JSON files produced using Micro-Meta App to document the acquisition of example images using a custom-built TIRF Epifluorescence Structured Illumination Microscope

<p><strong>Example Microscopy Metadata JSON files produced using the <a href="https://wu-bimac.github.io/MicroMetaApp.github.io/">Micro-Meta App</a> documenting an example raw-image file acquired using the custom-built TIRF Epifluorescence Structured Illumination Microscope.</strong></p> <p>For this use case, which&nbsp;is presented in Figure 5 of <a href="http://doi: https://doi.org/10.1101/2021.05.31.446382">Rigano et al., 2021</a>,&nbsp;Micro-Meta App was utilized to document:</p> <p>1)&nbsp;The <strong>Hardware Specifications</strong>&nbsp;of the&nbsp;custom build&nbsp;TIRF Epifluorescence Structured light Microscope (TESM; <a href="https://www.pnas.org/content/109/8/E471.long">Navaroli et al., 2010</a>)&nbsp;developed,&nbsp;built on the basis of the based on Olympus IX71 microscope stand, and owned by the&nbsp;Biomedical Imaging&nbsp;Group (http://big.umassmed.edu/)&nbsp;at the Program in Molecular Medicine&nbsp;of the&nbsp;University of Massachusetts Medical School. Because TESM was custom-built the most appropriate documentation level is&nbsp;<strong>Tier 3</strong>&nbsp;(<em>Manufacturing/Technical Development/Full Documentation</em>) as specified by the&nbsp;<a href="https://doi.org/10.5281/zenodo.4710731">4DN-BINA-OME</a>&nbsp;Microscopy Metadata model&nbsp;(<a href="https://doi.org/10.1101/2021.04.25.441198">Hammer et al., 2021</a>).</p> <p>The TESM Hardware Specifications are stored in:&nbsp;<strong>Rigano et al._Figure 5_UseCase_Biomedical Imaging Group_TESM.JSON</strong></p> <p>2) The <strong>Image Acquisition Settings</strong> that were applied to the TESM microscope for the acquisition of an example image (FSWT-6hVirus-10minFIX-stk_4-EPI.tif.ome.tif)&nbsp;obtained by Nicholas Vecchietti and Caterina Strambio-De-Castillia. For this image,&nbsp;TZM-bl human cells were infected with HIV-1 retroviral three-part vector (FSWT+PAX2+pMD2.G). Six hours post-infection cells were fixed for 10 min with 1% formaldehyde in PBS, and permeabilized. Cells were stained with mouse anti-p24 primary antibody followed by DyLight488-anti-Mouse secondary antibody, to detect HIV-1 viral Capsid. In addition, cells were counterstained using rabbit anti-Lamin B1 primary antibody followed by DyLight649-anti-Rabbit secondary antibody, to visualize the nuclear envelope and with DAPI to visualize the nuclear chromosomal DNA.</p> <p>The Image Acquisition Settings used to acquire the&nbsp;FSWT-6hVirus-10minFIX-stk_4-EPI.tif.ome.tif image&nbsp;are stored in:&nbsp;<strong>Rigano et al._Figure 5_UseCase_AS_fswt-6hvirus-10minfix-stk_4-epi.tif.JSON</strong></p> <p><em><strong>Instructional video tutorials on how to use these example data files:</strong></em><br> Use these videos to get started with using Micro-Meta App after downloading the example data files available here.</p> <ul> <li><a href="https://vimeo.com/562022222">Part 1/2</a></li> <li><a href="https://vimeo.com/562022281">Part 2/2</a></li> </ul>

opencc-by-4.0May 2021View details →
zenodo36/100

TIRF Microscopy Data Files. "Multiple RNA- and DNA-binding proteins exhibit direct transfer of polynucleotides: Implications for target site search"

<p>TIRF-microscopy images and analysis data from single-molecule experiments assessing the direct transfer phenomenon in the TREX1 exonuclease.&nbsp;</p>

opencc-by-4.0Dec 2021View details →
zenodo36/100

High-speed TIRF and 2D super-resolution structured illumination microscopy with large field of view based on fiber optic components

<p>Super-resolved structured illumination microscopy (SR-SIM) is among the most flexible, fast, and least perturbing fluorescence microscopy techniques capable of surpassing the optical diffraction limit. Current custom-built instruments are easily able to deliver two-fold resolution enhancement at video-rate frame rates, but the cost of the instruments is still relatively high, and the physical size of the instruments based on the implementation of their optics is still rather large. Here, we present our latest results towards realizing a new generation of compact, cost-efficient, and high-speed SR-SIM instruments. Tight integration of the fiber-based structured illumination microscope capable of multi-color 2D- and TIRF-SIM imaging, allows us to demonstrate SR-SIM with a field of view of up to 150 &times; 150 &mu;m<sup>2</sup>&nbsp;and imaging rates of up to 44 Hz while maintaining highest spatiotemporal resolution of less than 100 nm. We discuss the overall integration of optics, electronics, and software that allowed us to achieve this, and then present the fiberSIM imaging capabilities by visualizing the intracellular structure of rat liver sinusoidal endothelial cells, in particular by resolving the structure of their trans-cellular nanopores called fenestrations.</p>

opencc-by-4.0May 2023View details →

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