Jul 12, 2024
Tissue Processing
- daniel.dautan daniel1,2,
- Per Svenningsson1,2
- 1Department of Clinical Neuroscience, Karolinska Institutet, 171 76 Stockholm, Sweden;
- 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA
Protocol Citation: daniel.dautan daniel, Per Svenningsson 2024. Tissue Processing. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l622zzgqe/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: June 04, 2024
Last Modified: July 12, 2024
Protocol Integer ID: 101200
Keywords: ASAPCRN, mouse, brain, tissue
Funders Acknowledgement:
Aligning Science Across Parkinson's
Grant ID: 020608
Abstract
Protocol for processing mouse tissue for immunohistochemistry.
After completion of live experiments, anesthetize the mouse with institution-approved substance at correct dosage.
Ensure the mouse is deeply anesthetized via the toe-pinch test and that there is no reaction. Place the mouse with its chest facing up above a basin to capture blood, and secure the paws so that the body doesn't move.
While there is still a heart beat, open chest cavity using small scissors and forceps, making sure not to puncture organs.
With a small needle/syringe attached to a pump, pierce the left (on your right side) ventricle. Pierce the left atrium to allow blood to flow through.
Perfuse the mouse with 1x PBS to clear the blood.
Once blood has been cleared, perfuse the mouse with 4% PFA.
Once the whole body is perfused, dissect all needed tissues and organs.
Post-fix the dissected tissues in 4% PFA for 4-6 hours.
After post-fixation, gently dry the tissue.
Embed each tissue to OCT block.
Section tissue at 40um thickness using cryostat.