Hey, y'all! I am planning on staining mouse livers with picrosirius red, and I have found excellent protocols to do this and a sense that this absolutely can be done from frozen sections.
The way we do it here is we section from tissues fixed in OCT, and the slides are held in Normal Buffered Saline for ~10 mins, dried and held at -80C overnight before staining the next morning.
A lot of the protocols I see have the slides being dried out in toluene and ascending dilutions of EtOH. Is this necessary for frozen sections at all? Do I just bring 'em to RT, and start dipping? :-)
I'd appreciate any insight! Thanks, guys!
Best,
A.
Hello.
With frozen sections, you can leave them at RT for ~20min (immediately after cutting), wash the OCT with distilled water and start the staining (there's no need to pass through EtOH). If you are not able to start the staining within the same day, just store them at -20ºC and, when needed, bring them to RT and proceed.
The only thing you need to pay attention is if your sample is already fixed or not (with PFA or other). If it is, you can keep them at RT while you're cutting and proceed. If they're not, you need to fix your sections before starting the staining. So while cutting, keep them inside the cryostat chamber (so they don't rise to RT), afterwards fixed them as wished (metanol, acetone, ...) and proceed with the staining.
Good luck!
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining