Near-isotropic, reconstructed volume electron microscopy (FIB-SEM) of Drosophila mushroom body (jrc_fly-mb-1a)

This acquisition is part of the CellMap 2024 Segmentation Challenge
Challenge DOI: https://doi.org/10.25378/janelia.c.7456966Challenge Website: https://cellmapchallenge.janelia.org/Sample: Alpha lobe of CantonS G1x w1118 female fly, 6 days old
Sample Description: The sample consists of the alpha lobe from a 6-day-old CantonS G1x w1118 female fly. The tissue was prepared using high-pressure freezing and freeze substitution with 1% OsO₄, 0.2% uranyl acetate, and 3% water in acetone, followed by resin embedding in Durcupan. A 235 μm frontal slice was obtained using a vibrating microtome (Leica VT1000) and prefixed in 2.5% glutaraldehyde and 2.5% paraformaldehyde in 0.1M cacodylate buffer for 15 minutes. After embedding, the sample was polymerized, mounted on a copper stub, and trimmed to the alpha lobe region with an ultramicrotome (Leica UC7) for FIB-SEM imaging. The sample was provided by Yoshinori Aso (HHMI/Janelia), prepared for imaging by Zhiyuan Lu (HHMI/Janelia), imaged by Song Pang and C. Shan Xu (HHMI/Janelia), and post-processed and stitched by Eric Trautman and Stephan Preibisch (HHMI/Janelia).
Protocol: High-pressure freezing, freeze substitution with 1% OsO₄, 0.2% UA, and 3% H₂O in acetone; resin embedding in Durcupan. Fly tissue was sectioned using a vibrating microtome (Leica VT1000) to obtain a 235 μm frontal slice, prefixed in 2.5% glutaraldehyde and 2.5% paraformaldehyde in 0.1M cacodylate buffer for 15 minutes. After embedding, the sample was polymerized, mounted on a copper stub, and trimmed to the a-lobe region with an ultramicrotome (Leica UC7) for FIB-SEM imaging.

Contributions: Sample provided by Yoshinori Aso (HHMI/Janelia), prepared for imaging by Zhiyuan Lu (HHMI/Janelia), imaged by Song Pang and C. Shan Xu (HHMI/Janelia), and post-processed and stitched by Eric Trautman and Stephan Preibisch (HHMI/Janelia).
Acquisition ID: jrc_fly-mb-1a
Voxel size (nm): 4 x 4 x 4 (x, y, z)

Data dimensions (µm): 44.02 x 37.14 x 38.02 (x, y, z)
Imaging start date: 2020-12-20
Imaging duration (days): 38
Landing energy (eV): 700
Imaging current (nA): .25
Scanning speed (MHz): .5
Dataset URL: s3://janelia-cosem-datasets/jrc_fly-mb-1a/jrc_fly-mb-1a.zarr/recon-1/em/
EM DOI: https://doi.org/10.25378/janelia.27075448
Visualization Website: https://openorganelle.janelia.org/datasets/jrc_fly-mb-1a
Publication: Xu et al., 2021