Adult mouse lung digestion test. Adult mouse lung digestion test

This experiment was performed to help optimize a digestion protocol for FACS of adult mouse lung cells. The sample JH2 was used to help generate lung epithelial cell signatures. Overall design: The goal of this experiment was to compare the various lung cell types yielded from two enzymatic digestion protocols: Morrisey Lab protocol (collagenase and dispase) and Jessie Huang's optimized protocol (a combination of Nan Tang and Jay Mizgerd's protocols: collagenase, dispase, elastase, with agarose instillation in the lungs) The four samples and their target cell recovery are the following: JH1 (non-EpCAM), 4000 cells, JH2 (EpCAM), 1000 cells, EM1 (non-EpCAM), 4000 cells, EM2 (EpCAM), 1000 cells I'll be adding 2 extra lanes this time to include cells digested through Ed Morrisey's lung digestion protocol. The other change is that I'm sorting the immune/endothelial cells separately from the mesenchymal population, and recombining them 1:1 in my non-EpCAM lanes (so that the former population doesn't overwhelm the latter in terms of cell numbers).