Mar 26, 2024
Protocol Collection: Perfusing, Sectioning, IHC, Mounting and Coverslipping Mouse Brain Specimens
- Naveen Ouellette1,
- daphne.toglia 1,2,
- Holly Myers1
- 1Allen Institute for Neural Dynamics;
- 2Boston University
- Allen Institute for Neural Dynamics
Protocol Citation: Naveen Ouellette, daphne.toglia , Holly Myers 2024. Protocol Collection: Perfusing, Sectioning, IHC, Mounting and Coverslipping Mouse Brain Specimens. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygx3yxkg8j/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: September 05, 2023
Last Modified: June 18, 2024
Protocol Integer ID: 87408
Keywords: Immunohistochemistry, IHC, Histology, Immunolabeling, Microtome, Coverslip, Mounting Tissue, Antibody, whole mouse brain specimen, whole brain into section, coverslipping mouse brain, anatomical structures in brain region, microscope, brain region, brain removal, anatomical structure, sliding microtome, intracardiac perfusion fixation, perfusing, whole brain, brain, section, sectioning, slide, laser channel
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.
Mouse Cardiac Perfusion Fixation and Brain Collection V.5
Reference the Mouse Cardiac Perfusion Fixation and Brain Collection V.5 protocol.
Sectioning Mouse Brain with Sliding Microtome
Reference the Sectioning Mouse Brain with Sliding Microtome protocol.
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.
DAPI Staining Mouse Brain Sections
If DAPI staining only is needed, reference to DAPI Staining Mouse Brain Sections protocol.
Mounting and Coverslipping Mouse Brain Sections
Reference the Mounting and Coverslipping Mouse Brain Sections protocol.
