I have collected brains for IHC in PBS instead of PFA, and after 24h I changed media to 30%sucrose. Can I fix that with putting them in PFA?
Were the brains perfused with PFA before collection (e.g., by animal perfusion)? This would ensure some internal fixation in the tissue, but if they were in just PBS for hours, they are probably damaged, as fixation should be performed as soon as the tissue is collected.
This link might help to clarify: https://www.ndbbio.com/post/tissue_fixation
Thank you Gabriella, It is perfused with PFA but get collected in PBS and left in there for 24 h followed by sucrose over the weekend!
Hello Salma, since the tissues were perfused with PFA you may try to go ahead with your protocol, and hopefully there is no relevant damage. Fingers crossed!
Thank you Gabriella. Fortunately it did work fine and with no damage.
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