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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:54612*  Cite 
Description

The biophysical properties of sensory neurons are influenced by their morphometric and morphological features, whose precise measurements require high-quality volume electron microscopy (EM). However, systematic surveys of these nanoscale characteristics for identified neurons are scarce. Taking advantage of the CryoChem method, which permits high-quality ultrastructural preservation of cryofixed and genetically labeled samples for volume EM, we acquired serial block-face scanning electron microscopy (SBEM) images of antennal tissues in which select ORNs expressed the APEX2 EM marker. These SEBEM datasets allow a systematic morphometric and morphological analysis of identified olfactory receptor neurons in Drosophila melanogaster.

Technical Details

Drosophila antennae were prepared as described in Tsang et al. After cryofixation by high-pressure freezing and freeze-substitution in a cocktail containing uranyl acetate and glutaraldehyde, cryofixed antennae were rehydrated gradually at ice temperature. Rehydrated samples were then subjected to DAB labeling reaction and stained using a high-contrast en bloc staining protocol. Next, the samples were dehydrated for resin infiltration and embedded in Durcupan epoxy, followed by imaging with X-ray microscopy and then serial block-face scanning electron microscopy.
The antenna was imaged using a Zeiss Merlin scanning electron microscope equipped with a Gatan 3View2XP and OnPoint backscatter detector. Images were acquired at 2.0 kV accelerating voltage with a 30 μm condenser aperture and 1.0 μsec dwell time; Z step size was 40 nm; pixel size was 4.38 nm; raster size was 14k x 16k; Z dimension was 999 sections, and local gas injection was set to 85%. Volume was aligned using cross correlation, segmented, and visualized using IMOD.

Biological Sources
NCBI Organism Classification
Drosophila
Cell Type
olfactory receptor neuron
Cellular Component
vacuole
sensillum lumen
dendrite
mitochondria
Attribution
Names
Eric A Bushong
Mark H Ellisman
Cesar Nava Gonzales
Matthew Madany
Quintyn McKaughan
Chih-Ying Su
Tin Ki Tsang
Link
Article
OTHER
Or47a-GAL4>10XUAS-myc-APEX2-Orco
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL54612
Archival Resource Key (ARK)
ark:/b7295/w9cil54612
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
serial block face SEM (SBFSEM)