Hi, I am working with a FACS Vantage with TS option in a hematology/oncology lab. We are sorting stem cell subpopulations in human leukapharesis products. Some of the cells are frozen in plasma and 10 % DMSO like normal blood cells for stem cell transplantation. After thawing in warm water I quickly dilute the cells in PBS with 5mM EDTA and 10 %FCS and wash them. But within a short time (about half an hour) cells will agregate so that staining and sorting is impossible. Has anyone suggestions how to avoid this clumping? I have tried trypsin and nylon mesh to disconnect the cell masses, but that was not sucessful. Any proposal is highly welcome since we have deepfreezed many interesting samples from leukemia patients. Thanks in advance, Conny Brendel
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