Hello! I am trying to take punches from mouse CEA and BLA. Our lab has typically taken whole amygdala but we would like to do qPCR on the subregions as opposed to whole amygdala. Anybody have a good method to use, or tips/tricks? Thanks!
Hello Sadie,
I've never done punches, but you could try slicing the brain with a vibratome and then cutting the specific part you need with a scalpel under a magnfiying lens. It will not be as precise as laser micro-dissection but it can work and is quite easy. I have successfully done it to get tissue samples from the CeA for Western Blots. I attached two pictures of a rough dissection as reference (slices were cut in horizontal plane).
Good luck with your experiments,
Ivan
Hi Sadie,
We have had success taking punches from rat CeA and MeA by punching the subregions while the brains were on a cryostat. If I remember correctly, we used 0.5mm diameter punches, but that is something you can determine from the mouse atlas. The risk with doing it on the cryostat is with variation in the depth of the punches. If you have concerns about this, slicing, then punching might be best. A challenge with mice is going to be the size of the areas, but I do see that Thermo sells a 0.3mm punch.
- Badges
- Science topic
- Similar topics
- Psychology