Transmission electron microscopy quality checks of P7 mouse skin for jrc_mus-skin-1 (tem/25)

Description: Transmission electron microscopy quality checks of P7 mouse skin for jrc_mus-skin-1.

TEM ID: tem/25

Protocol: 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.

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.

Contributions: 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.

TEM voxel size (nm):

TEM data dimensions (µm):

Resulting FIB-SEM volume: https://doi.org/10.25378/janelia.24085269

Visualization website: https://openorganelle.janelia.org/datasets/jrc_mus-skin-1