At 03:30 PM 11/26/01 +1030, you wrote: Hi there, I've got another problem for you all... I'm trying to isolate nuclei from CFSE stained cells so that I can then fix and permeabilize the nuclei, perform intranuclear staining, and analyse using flow cytometry. <SNIP> The DNA versus incorporated halogenated pyrimidine (BrdUrd etc) techniques routinely utilize nuclei. Try pepsin to digest your cells. The tricks to avoid clumping include low speed spins and no ++ ions . See our suggested recipes in Methods in Cell Biology - Cytometry 3'rd edition or call me for info. Nick Nicholas Terry, M.A., Ph.D., Associate Professor, Experimental Radiation Oncology - 066, The University of Texas M. D. Anderson Cancer Center, 1515 Holcombe Blvd., Houston, Texas 77030. 'Phone: 713-792-3424, 'Fax: 713-794-5369. http://drad52.mdacc.tmc.edu. ntflow@odin.mdacc.tmc.edu.
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