Protocols

Here you will find information regarding the different protocols used at the Primate Brain Bank to conserve and store brain material. The links below will take you to the different protocols.

If you have any questions or comments regarding these protocols or if your study requires a different conservation and/or storage method please DO NOT HESITATE to contact us, either via email or regular mail.

Perfusion through the carotid or heart

Immersion

Storage

These are our most commonly used methods to conserve the brain of the deceased animal. We strive to do this as soon as possible after the animal has passed away.

For these protocols the fixation fluids are pumped through the brain either via both common carotid arteries or via the heart

The following solutions are pumped through the brain

A) Preperfusion: 500 ml saline (0.9% NaCl) containing 2500 I.U. heparin, room temperature

B) Fixation: 500 ml 4% formaldehyde in a 0.1 M phosphate buffer, pH 7.4, 4ºC

C) Rinsing: 500 ml 15% sucrose in a 0.1 M phosphate buffer, pH 7.4, 4ºC

We multiply the volume of these solutions in relation to the bodyweight of the animal, using the tables below

After the fixation the brain is removed from the skull and checked to see if fixation was sufficient, if not, an overnight post-fixation in applied, in an ample volume of the following fluid:

D) Post-Fixation: sufficient volume of 4% formaldehyde in a 0.1 M phosphate buffer, pH 7.4, 4ºC

Finaly the brain is stored at 4ºC in a closed canister in a sufficient amount of the following fluid:

E) Storage: 15% sucrose in a 0.1 M phosphate buffer, pH 7.4, 4ºC

Animals that have passed away should be perfused as soon as possible, this protocol is used only when fixation through the heart or carotids is NOT possible.

The brain is first removee from the skull

3) The brain is submerged in a sufficient volume of the following solutions

A) Saline (0.9% NaCl), room temperature

B) Fixation: 4% formaldehyde in a 0.1 M phosphate buffer, pH 7.4, 4ºC. Leave the brain in this solution for: 2 to 4 days (depending on size of brain)

C) Rinsing: 15% sucrose in a 0.1 M phosphate buffer, pH 7.4, 4ºC. Leave the brain in this solution for: 3 hours, then: replace the solution with fresh solution and leave the brain in it for another 3 hours

Finaly the brain is stored at 4ºC in a closed canister in a sufficient amount of the following fluid:

D) Storage: 15% sucrose in a 0.1 M phosphate buffer, pH 7.4, 4ºC

1) Brains should be stored at 4ºC in a closed canister in a sufficient amount of the following fluid:

1A) Storage: 15% sucrose in a 0.1 M phosphate buffer, pH 7.4, 4ºC

2) All brains should be checked every 2 months for signs of fungal growth in the sucrose solution (a layer forms on the surface).

3) The sucrose solution should be replaced every 5 months, or earlier in case or doubt.

4) The replacement of sucrose solution should be reported to the Primate Brain Bank administrator, using the sucrose Refreshment Report.

Back to the Primate Brain Bank start page