Dear all, The protocol I use for cryo-sectioning of rat spinal cord after injury includes: keeping the tissues in 4% paraformaldehyde made in PBS O/N then in 30% sucrose. I don't know if I can keep them in sucrose for a long time or not? any help? Thanks
Hi Mahdieh,
In my experience, you can keep tissue in sucrose for a few weeks before freezing it and it has a negligible effect in immunohistochemistry experiments, as far as I can tell. From others, I've heard that you can keep tissue in sucrose for a few months. However, I would advise caution. I have never kept tissue in sucrose more than 3 days if that tissue is part of an important experiment because over time, tissue in sucrose will break down.
Cheers
Brett
I've previously used tissue that was in sucrose for months and it was ok. I wouldn't advise it though. Just freeze it down as soon as it's ready, keep it in the -80 and have peace of mind.
Hello Mahdieh.
I agree with Niels. You should not keep the tissue too long in sucrose. I have made some bad experiences with tissue kept too long in sucrose: without Na-azide, the solution was contaminated with mold...
Greetings
Thanks all,
Niels, you mean I should keep the intact paraformaldehyde and sucrose treated tissues at -80 c or it's better to cut them and keep the mounted slides at -80.
Thanks
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