Mouse perfusions and brain tissue processing
Ayse Ulusoy, Rita Pinto-Costa, Angela Rollar, Donato Di Monte, Michael Klinkenberg, Michael Helwig, Shirley Lee, Sinead O'Sullivan
Abstract
The protocol describes tissue processing for histological analyses and includes perfusions, brain fixation, and tissue harvesting from perfused rodents.
Steps
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) using a freezing microtome.
