Jul 23, 2020
Snap freezing brains
This protocol is a draft, published without a DOI.
- 1University of California, San Diego
- George Lab @ UCSDTech. support email: olgeorge@ucsd.edu
Protocol Citation: Molly Brennan, Olivier George 2020. Snap freezing brains. protocols.io https://protocols.io/view/snap-freezing-brains-biv4ke8w
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: July 22, 2020
Last Modified: July 23, 2020
Protocol Integer ID: 39580
Snap freezing brains
euthanize animal in either a bell jar of Isoflorane or in C02 chamber.
once animal is dead you then can use scissors and or a gullitoine to remove head at nape of neck
Brain Removal
-Use a pair of scissors to cut connective tissue under the skin, reaching in between the eyes
-Cut skin open, exposing muscles on top of skull to the eyes
-Use scalpel, cut muscle connecting to the skull bone on both sides
-Carefully place bone cutter against the inside of the skull, and make cuts following the scalpel incision
-Lift the occipital, parietal, and frontal bones using forceps, fully exposing the brain
-Cut membrane on top and scoop brain out of the skull; cut optical nerves and connective tissue
Once brain is removed place it in -38-40 degree tempeture of Ispentane and or 2 methylbutane
Once brain is submerged into the 2 methylbutane it needs to stay there for 30-45 seconds or until brain is frozen
do not have temp lower than -40 degrees or the brain will crack and or split in two. VERY IMPORTANT
take brain out and place in foil with a label or vial. your choice. store in -80 freezer.