Near-isotropic, reconstructed volume electron microscopy (FIB-SEM) of Drosophila brain: ellipsoid body (jrc_fly-ellipsoid-body)

<b>Sample:</b> Ellipsoid body of a 5 day-old adult male Drosophila (Genome type: iso Canton S G1 x w1118)<b>Sample Description:</b> 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.<b>Protocol:</b> Chemical Fixation, ORTO-Lead-EPTA staining via progressive lowering of temperature and low temperature staining (PLT-LTS) heavy metal enhancement protocol.<b>Contributions:</b> Sample provided by Zhiyuan Lu (HHMI/Janelia), prepared, imaged and post-processed by Song Pang (Yale), with post-processing by C. Shan Xu (Yale).<b>Acquisition ID:</b> jrc_fly-ellipsoid-body<b>Voxel size (nm):</b> 4 x 4 x 4 (X, Y, Z)<b>Data dimensions (µm): </b>7.6 x 7.3 x 9.4 (X, Y, Z)<b>Imaging start date:</b> 2016-06-08<b>Imaging duration (days):</b> 6<b>Landing energy (eV): </b>200<b>Imaging current (nA): </b>0.22<b>Scanning speed (MHz): </b>0.200<b>Dataset URL: </b>s3://janelia-cosem-datasets/jrc_fly-ellipsoid-body/jrc_fly-ellipsoid-body.zarr/recon-2/em/<b>Visualization Website</b>: https://openorganelle.janelia.org/datasets/jrc_fly-ellipsoid-body<b>Publication: </b>Scheffer, et al., 2020; Xu et al., 2017; Xu et al., 2021<br>