Hello everyone,
Just a brief question: what would you suggest if I have a bunch of tissue samples floating in formalin for ~ a year. Mainly, I have two options:
1. go with freezing (wash from formalin, then run through saccharose gradients, etc.)
2. go with paraffin embedding.
Of course, both variants go with antigen unmasking.
What would you suggest?
Thank you!
Hello!
In our laboratory we fix brain (previously perfused with pbs and then with formalin) in the formalin+sucrose(30%) for a month and then freeze it using O.C.T. compound by Sakura Finetek. After that we slice it and go IHC on freezed section (unmasking in IHC protocol). That's all. If tou interested I may show you our protocol. But I not sure that your formalin fixed tissue will be stained follow it.
The other way we are fixed brains is in formalin and then in sucrose (not remember how long, sorry, but if you need it I may try to find it). But this methods we used for free floating slices.
I think it depends on the kind of investigation you like to do. It depends on antibodies you will use when you are doing immunohistochemistry or immunfluorescence or insitu hybridization. I would recommand to check with 2 brains the quality of frozen and paraffinsections. Compare them and use those technique which gives the best result for your further investigations.
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