Isotropic 3D electron microscopy reference data of wild-type HeLa cell undergoing mitosis (jrc_hela-4)

Understanding cellular architecture is essential for understanding biology. Electron microscopy (EM) uniquely visualizes cellular structure with nanometer resolution. However, traditional methods, such as thin-section EM or EM tomography, have limitations inasmuch as they only visualize a single slice or a relatively small volume of the cell, respectively. Here, we overcome these limitations by long-term imaging whole cells and tissues via the enhanced Focus Ion Beam Scanning Electron Microscopy (FIB-SEM) platform in high resolution mode with month-long acquisition duration. We use this approach to generate reference 3D image data sets at 4-nm isotropic voxels. Together with subsequent segmentation, we hope to create a reference library to explore comprehensive quantification of whole cells and all their constituents, thus addressing questions related to cell identities, cell morphologies, cell-cell interactions, as well as intracellular organelle organization and structure. Sample: Wild-type HeLa cell undergoing mitosis Protocol: 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 by Gleb Shtengel (HHMI/Janelia), with imaging and post-processing by C. Shan Xu (HHMI/Janelia). Dataset ID: jrc_hela-4 Final voxel size (nm): 4.00 x 4.00 x 4.28 (X, Y, Z) Dimensions (µm): 30 x 14 x 35 (X, Y, Z) Acquisition date: 2019-05-30 Dataset URL: s3://janelia-cosem-datasets/jrc_hela-4/jrc_hela-4.zarr/recon-1/em/ Visualization Website: https://openorganelle.janelia.org/datasets/jrc_hela-4 Publication: “Isotropic 3D electron microscopy reference library of whole cells and tissues” by C. Shan Xu, et al. (in preparation)