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

Sample: Nocodazole treated interphase HeLa cell (ATCC CCL-2) overexpressing mEmerald-ManII

Sample Description: 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.

Protocol: 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.

Contributions: 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.

Acquisition ID: jrc_hela-nz-1

Voxel size (nm): 8 x 8 x 7.9 (x, y, z)

Data dimensions (µm): 99.2 x 4.0 x 73.1 (x, y, z)

Imaging start date: 2018-10-04

Imaging duration (days): 8

Landing energy (eV): 1200

Imaging current (nA): 2.00

Scanning speed (MHz): 0.500

Dataset URL (Redirect): https://data.janelia.org/bco9J

Visualization Website: https://openorganelle.janelia.org/datasets/jrc_hela-nz-1

Publication: Xu et al., 2017; Xu et al., 2021