Alternate header for print version

Public Domain: This image is in the public domain and thus free of any copyright restrictions. However, as is the norm in scientific publishing and as a matter of courtesy, any user should credit the content provider for any public or private use of this image whenever possible. Learn more
*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:50453*  Cite 

In this study, we developed an electron microscopy method, called CryoChem, to optimally preserve the morphology of genetically labeled tissues. This method also allows us to perform 3D correlated light and electron microscopy (CLEM) in the well-preserved tissue. This dataset contains the tdTomato confocal volume of the CLEM experiment in a mouse brain expressing tdTomato in corticotropin releasing factor-expressing neurons.

Technical Details

Brief description of sample preparation
After cryofixation by high-pressure freezing and freeze-substitution, cryofixedmouse brain slices were rehydrated gradually. Rehydrated samples were then imaged with confocal microscopy to capture the DRAQ5 and tdTomato signals. Next, the samples were stained using a high-contrast en bloc staining protocol. Then the samples were dehydrated for resin infiltration and embedding, followed by imaging with X-ray microscopy and then SBEM.

Confocal Microscopy
After freeze-substitution and rehydration, the mouse brain slice was placed in ice-cold 0.15 M sodium cacodylate for imaging. Confocal volumes of DRAQ5 and tdTomato signals were collected on an Olympus FluoView 1000 confocal microscope with a 20X air and 60X water objectives using 561 nm and 633 nm excitation.

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
Mouse brain tissue
Cellular Component
Mouse brain tissue expressing tdTomato in corticotropin releasing factor-expressing neurons
Tin Ki Tsang
Eric Bushong
Daniela Boassa
Junru Hu
Benedetto Romoli
Sebastien Phan
Davide Dulcis
Chih-Ying Su
Mark Ellisman
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Image Type
recorded image
Image Mode
confocal microscopy
Spatial Axis Image Size Pixel Size
X 1024px ——
Y 1024px ——
Z 73px ——