Protocol Collection: Perfusing, Sectioning, IHC, Mounting and Coverslipping Mouse Brain Specimens
Naveen Ouellette, Holly Myers, daphne.toglia
Abstract
This protocol collection details the steps for a whole mouse brain specimen to be perfused, sliced, stained with antibodies, DAPI, or both, and made into slides. The intracardiac perfusion fixation includes anesthesia, exsanguination, fixation, brain removal, and post-fixation storage. A sliding microtome is used to slice the whole brain into sections, and the sections are stained, mounted on glass slides, and coverslipped. The result is that anatomical structures in brain regions of interest may be labeled in a way that can be viewed by laser channels on a microscope and imaged for data collection.
Before start
Steps
Mouse Cardiac Perfusion Fixation and Brain Collection V.5
Reference the Mouse Cardiac Perfusion Fixation and Brain Collection V.5 protocol.
Mouse Cardiac Perfusion Fixation and Brain Collection
Sectioning Mouse Brain with Sliding Microtome
Reference the Sectioning Mouse Brain with Sliding Microtome protocol.
Sectioning Mouse Brain with Sliding Microtome
Immunohistochemistry (IHC) Staining Mouse Brain Sections
If antibody staining or both antibody and DAPI staining is needed, reference the Immunohistochemistry (IHC) Staining Mouse Brain Sections protocol.
Immunohistochemistry (IHC) Staining Mouse Brain Sections
DAPI Staining Mouse Brain Sections
If DAPI staining only is needed, reference to DAPI Staining Mouse Brain Sections protocol.
DAPI Staining Mouse Brain Sections
Mounting and Coverslipping Mouse Brain Sections
Reference the Mounting and Coverslipping Mouse Brain Sections protocol.
Mounting and Coverslipping Mouse Brain Sections
