Hi all,

I am trying to investigate an organoid system cultured in a pure collagen I hydrogel using mass cytometry techniques.

Our mass cytometry collaborators usually use a solution of 0.5 mg/mL dispase, 0.2 mg/mL collagenase IV, and 0.2 mg/mL DNase I on an OctoDissociator using a well-tested 1 hour protocol to digest organoids cultured in Matrigel with great success.

I need to digest the organoid aggregate into a single cell suspension AFTER fixing the intact structure with 4% PFA to preserve the intracellular signalling state that we are investigating.

We do get many single cells using the above protocol but are still left with large clumps of material at the end, even even when we increase the collagenase and collagenase to 1 mg/mL each.

Any suggestions would be much appreciated.

Thanks,

Brendan