Near-isotropic, aligned volume electron microscopy (eFIB-SEM) of Mouse hippocampus (jrc_mus-hippocampus-3)

<p dir="ltr"><b>Sample</b>: 2–3 month old male mouse, strain: C57BL/6J from Jackson Lab.<br></p><p><br></p><p dir="ltr"><b>Sample Description</b>: Tissue samples from the CA1 region of the mouse hippocampus were collected from 2–3-month-old male C57BL/6J mice. Samples were perfused with 3% paraformaldehyde, post-fixed, vibratome sectioned, and further fixed overnight in 1% PFA and 2% glutaraldehyde. 2 mm tissue punches were high-pressure frozen in 1-hexadecene, freeze-substituted with osmium tetroxide, uranyl acetate, and either 3-amino-1,2,4-triazole or imidazole, and embedded in Durcupan resin. The sample preparation was performed by Shu-Hsien Sheu. Imaging and post-processing were carried out by C. Shan Xu (Yale), Gleb Shtengel, and Harald Hess (HHMI/Janelia).</p><p dir="ltr"><b>Protocol</b>: Two datasets were acquired from the CA1 region of the mouse hippocampus using a hybrid fixation and staining protocol optimized for primary cilia visualization. 2–3-month-old male C57BL/6 mice were perfused with 3% paraformaldehyde and post-fixed in modified PFA buffer for 2 hours. Tissue was rinsed in 400 mOsM buffer and sectioned (100 µm) on a vibratome. Sections were fixed overnight in 1% PFA and 2% glutaraldehyde. 2 mm punches were high-pressure frozen in 1-hexadecene, freeze-substituted in osmium tetroxide, uranyl acetate, and either 3-amino-1,2,4-triazole or imidazole. Samples were embedded in Durcupan resin.<br></p><p><br></p><p dir="ltr"><b>Contributions</b>: Sample prepared by Shu-Hsien Sheu. Imaging and post-processing by C. Shan Xu (Yale), Gleb Shtengel, and Harald Hess(HHMI/Janelia).<br></p><p><br></p><p dir="ltr"><b>Acquisition ID</b>: jrc_mus-hippocampus-3</p><p dir="ltr"><b>Voxel size (nm)</b>: 8 x 8 x 8 (x, y, z)</p><p dir="ltr"><b>Data dimensions (µm)</b>:<b> </b>60.9 x 81.28 x 45.07 (x, y, z)</p><p dir="ltr"><b>Imaging start date</b>: 2018-22-02</p><p dir="ltr"><b>Imaging duration (days)</b>: 7</p><p dir="ltr"><b>Landing energy (eV)</b>: 1200</p><p dir="ltr"><b>Imaging current (nA)</b>: 2</p><p dir="ltr"><b>Scanning speed (MHz)</b>: 2.083</p><p dir="ltr"><b>Dataset URL</b>: <a href="https://data.janelia.org/LmJNw5" rel="noreferrer" target="_blank">s3://janelia-cosem-datasets/jrc_mus-hippocampus-3/jrc_mus-hippocampus-3.zarr/recon-1/em/</a></p><p dir="ltr"><b>Visualization Website</b>: <a href="https://openorganelle.janelia.org/datasets/jrc_mus-hippocampus-3" rel="noreferrer noopener" target="_blank">https://openorganelle.janelia.org/datasets/jrc_mus-hippocampus-3</a></p><p dir="ltr"><b>Publication</b>:<b> </b>Sheu et al., 2022;<b> </b>Xu et al., 2017</p><p><br></p>