Transmission electron microscopy quality checks of P7 mouse thymus for jrc_mus-thymus-1 (tem/4)

<p dir="ltr"><b>Description</b>: Transmission electron microscopy quality checks of P7 mouse thymus for jrc_mus-thymus-1.</p><p dir="ltr"><b>TEM ID</b>: tem/4</p><p dir="ltr"><b>Protocol</b>: All procedures were approved by the Institutional Animal Care and Use Committee at Janelia Research Campus (protocol 21-0206). Postnatal day 7 (P7) wild-type C57BL/6J male mice were anesthetized, perfused with a fixative solution, and tissues were dissected and fixed for 24 hours at 4°C. The tissues were sectioned into 300-μm slices, postfixed with osmium tetroxide, stained with thiocarbohydrazide and osmium tetroxide, and then dehydrated and embedded in resin for polymerization.</p><p dir="ltr">After fixation, samples were dehydrated in ethanol, infiltrated with resin, and polymerized. Ultrathin sections (70-90 nm) were cut and mounted on grids. TEM imaging was performed using an FEI Tecnai Spirit BioTWIN at 80 kV to assess en bloc staining quality for future FIB-SEM imaging. Initial images were captured at 440× magnification, with final images taken at 2,900× magnification.</p><p dir="ltr"><b>Contributions</b>: Sample collection was performed by Wei-Ping Li (HHMI/Janelia/EMSR). Wei-Ping Li and Zhiyuan Lu (HHMI/Janelia/EMSR) prepared the samples for imaging and performed TEM imaging for quality check. Rebeccca Vorimo (HHMI/Janelia/CellMap Project Team) and Yurii Zubov (HHMI/Janelia/CellMap Project Team) curated the data. The rest of the CellMap Project Team contributed to general development and review of the project. Aubrey Weigel (HHMI/Janelia/CellMap Project Team) oversaw general project development.</p><p dir="ltr"><b>TEM voxel size (nm)</b>: 6 x 6 (x, y)</p><p dir="ltr"><b>TEM data dimensions (µm)</b>: 24.58 x 24.58 (x, y)</p><p dir="ltr"><b>Resulting FIB-SEM volume</b>: <a href="https://doi.org/10.25378/janelia.24085287" rel="noreferrer noopener" target="_blank">https://doi.org/10.25378/janelia.24085287</a><br></p><p dir="ltr"><b>Visualization website</b>: <a href=" https://openorganelle.janelia.org/datasets/jrc_mus-thymus-1" rel="noreferrer" target="_blank">https://openorganelle.janelia.org/datasets/jrc_mus-thymus-1</a></p>