Mar 25, 2024
Protocol Collection: Perfusing, Sectioning, IHC, Mounting and Coverslipping Mouse Brain Specimens
- 1Allen Institute for Neural Dynamics;
- 2Boston University
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
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
Mouse Cardiac Perfusion Fixation and Brain Collection V.5
Reference the Mouse Cardiac Perfusion Fixation and Brain Collection V.5 protocol.
Protocol
NAME
Mouse Cardiac Perfusion Fixation and Brain Collection
CREATED BY
Allen Institute
Sectioning Mouse Brain with Sliding Microtome
Sectioning Mouse Brain with Sliding Microtome
Reference the Sectioning Mouse Brain with Sliding Microtome protocol.
Protocol
NAME
Sectioning Mouse Brain with Sliding MicrotomeCREATED BY
Holly Myers
Immunohistochemistry (IHC) Staining Mouse Brain Sections
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.
Protocol
NAME
Immunohistochemistry (IHC) Staining Mouse Brain SectionsCREATED BY
Holly Myers
DAPI Staining Mouse Brain Sections
DAPI Staining Mouse Brain Sections
If DAPI staining only is needed, reference to DAPI Staining Mouse Brain Sections protocol.
Protocol
NAME
DAPI Staining Mouse Brain SectionsCREATED BY
daphne.toglia
Mounting and Coverslipping Mouse Brain Sections
Mounting and Coverslipping Mouse Brain Sections
Reference the Mounting and Coverslipping Mouse Brain Sections protocol.
Protocol
NAME
Mounting and Coverslipping Mouse Brain SectionsCREATED BY
Holly Myers