Hi Rochelle. In the past I have sorted yeast without much problem through a 76 micron flow cell at rates of up to 10,000/sec without much clogging, so (in my experience, at least) clumping shouldn't be a problem. However, I have been told that certain species of yeast can form long tubules that would make sorting problematic, but I didn't have that problem. As far as decontamination goes, they are really hard to kill. One client was sorting into 15 ml conicals and added a little water from a wash bottle to the bottom of the tube. Except it wasn't water but 10% clorox. But, he washed them and spun them down and they seemed to be none the worse for wear. As a rule we kept special tubing for the yeast applications and never sorted anything else with "yeast tubing". We worked with cryptosporidia oocysts once upon a time and the investigator told us that even straight clorox wouldn't kill them. However, straight lysol (in the brown bottle) would. Maybe that would work on the yeast as well. I don't recall anything of interest about their autofluorescence. Happy flowing! David McFarland Howard Hughes Medical Institute Flow Cytometry Facility Vanderbilt University Medical Center _______________________________________________________________________________ Subject: sorting live yeast From: "Rochelle A. Diamond" <diamond@cco.caltech.edu> at +inet Date: 01/07/1998 3:23 PM Hi Flow Community- I would like to get some information about sorting live yeast cells. In particular I would like to know the following: How badly do they reaggregate after filtering thru nylon mesh ( do I need to worry about the size of my orifice?) How do you decontaminate the sorter after the run (preferred cleansing agents)? What kind of autofluorescence do yeast cells (S. Cere.) exhibit? Any experiences good or bad would be useful. The experiment is to sort out high and low expressing GFP (EGFP-like) cells. Thanks ever so much Rochelle Diamond Caltech Biology Flow Cytometry/Cell Sorting Facility diamond@cco.caltech.edu
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