Feb 22, 2024
Protocol for "non-human primate necropsy"
- 1Center for Applied Medical research (Cima) University of Navarra
Protocol Citation: jlanciego 2024. Protocol for "non-human primate necropsy". protocols.io https://dx.doi.org/10.17504/protocols.io.rm7vzxzw5gx1/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: February 22, 2024
Last Modified: February 22, 2024
Protocol Integer ID: 95602
Funders Acknowledgement:
Aligning Science Across Parkinson's through the Michael J. Fox Foundation for Parkinson's Research
Grant ID: ASAP020505
Disclaimer
Nothing to declare
Abstract
Here we describe the standard procedure conducted for performing a non-human primate necropsy, followed by removal of the brain and spinal cord from the skull and backbone, respectively.
Image Attribution
Jose L. Lanciego
Animal preparation
Animal preparation
Pre-surgical anesthesia: to be induced with ketamine (5 mg/Kg) and midazolam (0.5 mg/Kg), administered intramuscularly
Terminal anesthesia: to be induced with an overdose of sodium pentobarbital (200 mg/Kg).
Prefusion and pre-histological processing
Prefusion and pre-histological processing
Perform a T-shape incision with a scalpel through the thorax and abdomen.
Open and retract the chest cavity to visualize lungs and heart.
Open the left ventricle to allocate the infusion cannula at the level of the aortic
infundibulum and fixed it.
Open the right atrium, getting started with the perfusion protocol at a constant
rate of 3,500 ml/h) with an infusion pump.
First solution: 1,000 ml of saline Ringer solution.
Second solution: 3,000 ml of a buffered solution made of 4% paraformaldehyde and 0.1% glutaraldehyde in 0.125 M PB pH 7.4
Third solution: 1,000 ml of a buffered solution made of 1% dimethylsulphoxide (DMSO) and 10% glycerin in 0.125 M PB pH 7.4
Brain extraction: Once perfusion was completed, the brain and the spinal cord are removed from the skull and the backbone, respectively. 10 mm-thick brain blocks are made and the pia matter carefully removed under a dissection microscope. Brain blocks were stored for 48 h in a cryoprotectant solution containing 2% DMSO and 20% glycerin in 0.125 M PB 7.4