For mouse brain, can I do post-fixation and 30% sucrose incubation simultaneously and get as good a result as the standard post-fixation followed by sucrose?
That may not a good idea. You should not over fix the brain tissue for immunostaining. Otherwise, you cannot get good staining. Sucrose treatment for a mouse brain needs a to 2 night in 15% sucrose and then 2 to 3 days in 30% sucrose.
Why don't you do fixation and cryoprotection separately?
It should be done separately because the length of time for cryoprotection with sucrose is different than post-fixing with paraformaldehyde.
@ Guoxiang, I'm not doing IHC on these brains, so overfixation shouldn't be a problem. I typically do overnight post-fixation anyway, and same with sucrose, so I figured by combining them I could save a lot of time. I tried it yesterday and the brain sections looked great, so I may stick to this for non-IHC brains in the future.
- Badges
- Science topic