My question has two parts re the organotypic slice culture technique:
How long can tissue (in this case brain) remain embedded in agarose prior to slicing on vibratome (4% low-melt agarose in ACSF).
If this isn't viable, then after slicing can the slices be stored in ACSF for a period of time prior to being placed in culture?
I have a number of animals I need for my pilot experiment and I can't hold them in quarantine so I need to cull all of them at the same time, hence my dilemma! Thanks
Hi, there.
It depends on the molecule what you are interedted in. For immunohistochemistry, we use the slice in a week after cutting or we will lose the activiry. If your material keeps in the pareffin block, they are safe longer (even 1 year) but after cutting and being exposing the air, it causes damage to proteins. If you are interested in DNA, that much easier to keep in for longer time. If you are interested in mRNA expression, you need to keep them in a frozen block. For DNA/RNA extraction, they are safe in -80C for longer time and qRT-PCR works well.
I hope this comment could be usuful for you.
MASA
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