I want to fix acute brain slice after cutting and want to do immune and histology. My question how will I fix slices for these experiments? Can anyone here for giving advice?
I would just drop them straight into 4% PFA. I do brain slices and I only fix them for 15-20 minutes in 4%PFA at RT. Basically I cut vibratome sections and do elecrtrophysiology on them for many hours and then fix them, and it works everytime. Make sure your 4%PFA is fresh or frozen vials. 1 ml vial per slice is plentiful in little petri-dishes. For technical details see my papaers listed below.
best wishes, Refik
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Thanks Refik, I was also thinking like this but from literature I found some confusing thoughts. Some has said 4%PFA in PBS for one hour and on the other hand some 4% PFA, PBS also other solution as well for 2 hours thats why I posed that question. Now, I should be clear I think. Thanks for your help. Be fine and will ask more in the future thats for sure. Bye
in my studies I did the fixation of brain slices with ice cold 4%PFA on ice for 15 minutes. You may check which option gives the best result for your staining.