I am separating neuron populations from mouse brains using FACS. I am trying to find a fixation technique for intracellular antigen staining and FACS that will allow long term storage afterwards for RNAseq. I have looked at ZBF which allows long term storage of cells with little change in RNA integrity, but they have to be taken out of the fixation buffer for flow, so RNA integrity will likely be affected during flow and I am not sure if they can be 'refixed' after. For any other fixation methods I have read that mention the shelf life of cells, they refer to how long they can be stored BEFORE staining and FACS, and do not mention any storage AFTER facs.
Hi,
This thread could probably help!
https://www.researchgate.net/post/RNA-seq_after_FACS_sorting
Best of luck
See doi: 10.1186/1756-0500-3-328 for spring the cells using RNAlater solution.
After the FACS, you can centrifuge the cells down to remove the solution from the FACS running, and then add the RNA later onto cell pellet and then store your cells at negative 80 for later RNA isolation. For RNA isolation, just do the same thing centrifuge the cells down after thawing, then remove the RNA later and go on with RNA isolation kit instruction on the cell pellet.
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