Good morning!
Just a quick question out of paranoia: for *fixed* tissue (specifically brains already having been fixed in 4% paraformaldehyde) that are cryosectioned what are the storage conditions (long/short) do you keep them at?
RNA stability is *not* a concern for these samples (Regular protein-IHC will be done, not FISH or any nucleus acid detection that would warrant more care involved in sample prep)
I left some of the cut cryosections in the hood for a few days- do you suppose these samples have severely degraded? Or should it not be a concern? I was always taught that as long as a tissue was fixed it is stable following cryosection but I’m paranoid and would like to make sure.
Thanks for your input! Much appreciated
Hi Anthony Undisclosed
Dry the slides for 30-45 mins post sectioning by cryotome. For long term storage it is best to keep the slides in a packed slide box in -80 freezer.
Thanks,
It depends on what kind of technique you will use for your IHC. If you are working with more or less 30µm thick sections I would recommand to use the free floating techninique. In that case you should store the sections not longer than 4 weeks in PBS at 4°C. If you like to keep the sections longer stored then deep freez them in a protecting solution of 30% sucrose in PBS and Glycerol equal parts at -20°C. You can store this material under this conditions for years.
If you you are working with sections thicknesses of around 10µm then the slide mthode is highly recommanded. Mount the sections on superfrost plus slides . They have to be dried out and completely transparent othewise they coud swim away while you are doing your immuno. I would recommand to staore them at -20°C.
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