Hi all,

I am trying to take a tongue epithelium and generate single cells from it for flow. So far, I have had no luck. Getting the epithelium separated from the underlying tissue is easy enough with dispase treatment. After that, I just keep hitting walls. If I mechanically cut up the tissue (Via sterile scalpel/scissors) and place in a shaker w/trypsin and dispase, nothing really happens. If I try to treat with 1000-10000 units of Collagenases A and P in 10ml PBS with a gentleMACS C tube, nothing really happens either. No matter what I do, I wind up with 100-300000 cells that are usually mostly dead. Any help would be appreciated.