Aug 02, 2023
Mouse perfusions and brain tissue processing
- Ayse Ulusoy1,
- Shirley Lee1,
- Angela Rollar1,
- Michael Helwig1,
- Michael Klinkenberg1,
- Sinead O'Sullivan1,
- Rita Pinto-Costa1,
- Donato Di Monte1
- 1DZNE
Protocol Citation: Ayse Ulusoy, Shirley Lee, Angela Rollar, Michael Helwig, Michael Klinkenberg, Sinead O'Sullivan, Rita Pinto-Costa, Donato Di Monte 2023. Mouse perfusions and brain tissue processing. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l69kn1lqe/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: August 25, 2022
Last Modified: May 31, 2024
Protocol Integer ID: 69172
Keywords: ASAPCRN
Abstract
The protocol describes tissue processing for histological analyses and includes perfusions, brain fixation, and tissue harvesting from perfused rodents.
Inject sodium pentobarbital (600 mg/kg, i.p.) to sacrifice the mouse
Once the respiration is stopped, open the chest and place the cannula connected to a peristaltic pump into the heart. Make sure that the cannula has access to the ascending aorta.
Perfuse, first with 20 ml saline solution, kept at room temperature, and then with ice-cold 60 ml 4% (w/v) paraformaldehyde.
Quickly remove the brain and immerse in 4% paraformaldehyde for 24 hours.
For cryopreservation, place the brain in a 30% (w/v) sucrose solution.
Once sunk, section the brain on the coronal plane (35 undetermined ) using a freezing microtome.