Near-isotropic, reconstructed volume electron microscopy (FIB-SEM) of Drosophila ventral nerve cord (jrc_fly-vnc-1)

<strong>Sample</strong>: VNC of adult male Drosophila (Genome type: iso Canton S G1 x w1118)
<strong>Sample Description</strong>: 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.
<strong>Protocol</strong>: Chemical Fixation, ORTO-Lead-EPTA staining via progressive lowering of temperature and low temperature staining (PLT-LTS) heavy metal enhancement protocol.
<strong>Contributions</strong>: Sample provided by David Stern (HHMI/Janelia), prepared by Zhiyuan Lu (HHMI/Janelia), imaged by Wei Qiu (HHMI/Janelia), with post-processing by Gleb Shtengel and Harald Hess(HHMI/Janelia).
<strong>Acquisition ID</strong>: jrc_fly-vnc-1
<strong>Voxel size (nm)</strong>: 4 x 4 x 4.7 (X, Y, Z)
<strong>Data dimensions (µm)</strong>:<strong> </strong>40.2 x 20.5 x 16.2 (X, Y, Z)
<strong>Imaging start date</strong>: 2018-12-05
<strong>Imaging duration (days)</strong>: 12
<strong>Landing energy (eV)</strong>: 900
<strong>Imaging current (nA)</strong>: 0.25
<strong>Scanning speed (MHz)</strong>: 0.200
<strong>Dataset URL (Redirect)</strong>: https://data.janelia.org/Acbea
<strong>Visualization Website</strong>: https://openorganelle.janelia.org/datasets/jrc_fly-vnc-1
<strong>Publication</strong>: Scheffer, et al., 2020; Xu et al., 2017; Xu et al., 2021