Hello, this is Denise Zinniel a research technician from Vet College at the University of Nebraska-Lincoln. My boss, Dr. Raul Barletta, would like to sort unfixed cells of Mycobacterium paratuberculosis. We have two unique cultures of K-10 cells with the Green Fluorescent Protein and Fire-fly Luciferase reporter genes. We are attempting to mix these two cultures and sort them using a fluorescent-activated cell sorter. I have talked to Dr. Aru here at the Beadle Center and Dr. Kuszynski at the University of Nebraska-Omaha Medical Center. Both facilities at the present time are unable to sort biosafety level 2 pathogens. The problem is that we want to keep the cells alive not fix them. Dr. Aru gave me a paper entitled "Biosafety Guidelines for Sorting of Unfixed Cell" from Cytometry 28:99-117(1997) which provides recommendations for handling and sorting of potentially biohazardous specimens. Both of these gentlemen said we could attempt the protocol given, but we are hoping to find a facility already capable of attempting this project. Please let me know if you have any ideas for facilities or comments. Thanks. Denise
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