I'm working on organotypic cerebellar slice cultures from postnatal rats and I'm not sure how to tell if the slices are healthy at any given point. I know there should be a lot of microglial activation for the first couple of days after dissection, and I do see a lot of darker spots (Day 2) for a while, however, after that, the slices tend to become more transparent and lose some of their structural features (Day 4).
Could anyone with experience check the photos attached and give some comments?
Thanks in advance!
I mostly have experience with organotypic entorhino-hippocampal tissue cultures and usually it is hard to evaluate the quality of organotypic tissue cultures by eye. We evaluate the quality of the cultures based on propidium iodide stainings (but day 4 might be too early) and whole-cell patch clamp recordings. In general, only cultures that have a homogenous thickness proove suitable for later experiments.
We used trypan blue exclussion as an index of viability. In fact this essay allows to make a quantitative evaluation of viability.
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