Near-isotropic, reconstructed volume electron microscopy (FIB-SEM) of Nocodazole treated HeLa cell (jrc_hela-nz-1)

<b>Sample</b>: Nocodazole treated interphase HeLa cell (ATCC CCL-2) overexpressing mEmerald-ManII<b>Sample Description</b>: Near isotropic FIB-SEM imaging of cultured HeLa cells transiently transfected with mEmerald-ManII. Cells incubated with 10 µM Nocodazole on ice then incubated for 4 hours before high pressure freezing. Widefield fluorescence microscopy used to confirm Golgi fragmentation after Nocodazole treatment.<b>Protocol</b>: HeLa cells transiently transfected with mEmerald-ManII. Cells incubated with 10 µM Nocodazole on ice then incubated for 4 hours before high pressure freezing. Widefield fluorescence microscopy used to confirm Golgi fragmentation after Nocodazole treatment. High pressure freezing, freeze-substitution resin embedding with 2% OsO₄ 0.1% UA 3% H₂O in acetone; resin embedding in Eponate 12.<b>Contributions</b>: Sample provided by Aubrey Weigel (HHMI/Janelia), prepared for imaging H. Amalia Pasolli (Rockefeller), imaging and post-processing by C. Shan Xu (Yale), and manual segmentation by Aubrey Weigel.<b>Acquisition ID</b>: jrc_hela-nz-1<b>Voxel size (nm)</b>: 8 x 8 x 7.9 (x, y, z)<b>Data dimensions (µm)</b>:<b> </b>99.2 x 4.0 x 73.1 (x, y, z)<b>Imaging start date</b>: 2018-10-04<b>Imaging duration (days)</b>: 8<b>Landing energy (eV)</b>: 1200<b>Imaging current (nA)</b>: 2.00<b>Scanning speed (MHz)</b>: 0.500<b>Dataset URL</b>: s3://janelia-cosem-datasets/jrc_hela-nz-1/jrc_hela-nz-1.zarr/recon-1/em/<b>Visualization Website</b>: https://openorganelle.janelia.org/datasets/jrc_hela-nz-1<b>Publication</b>: Xu et al., 2017; Xu et al., 2021