Hi, we are generating primary neurons - I was curious if the remainder of the brain could be fixed in PFA and used for IHC analysis without prior perfusion fixation?
unless you slice fresh and dump in pfa perfusion definetely make a difference.
We do both pre and post fixation of our brain the tissues. Overall, they all work to some degree. Post fixation in PFA always gives us a weaker signal after staining but works, nonetheless. (The signal is mostly weaker for larger proteins). One important note is that post fixation in PFA is a bit tricky for larger brains. Depending on the size of your brain the time in PFA is crucial. The usual rule is that PFA penetrates up to 2-3 mm per 24 hours so calculate the time you need according the size of your brain. Excessive post fixation really messes up with IHC.
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