The project I'm working on involves dissecting out a small piece of epithelial tissue that we're going to run through FACS and single cell RNA seq. My usual way of dissociating tissue is in 50ul of 100ug/ml Liberase:PBS for ~1 minute at 37*C and then adding 50ul of PBS and placing the tube on ice to kill the reaction. However, the cells were clumped together when my collaborators tried doing FACS and they were only able to get 5 cells. My lab doesn't have a protocol for this small of a tissue dissociation and there is no way for me to up the number of cells. We have a protocol for adult mouse organs but that has not been optimized and has not worked on previous experiments.

I was going to try again with an EDTA:PBS solution at 2g/L and couple that with Liberase, but if anyone has a better suggestion I would love to hear it!