Hi, Lee Anne Talbot We have worke with dog lekocytes for flow cytometric analysis of CD markers using antibodies from SEROTEC. We have developed a protocol that we can lyse the whole blood using comercial FLS from BD and then kept the washed cells "fixed by Lysing solution" sterile in PBS 10% of FBS for over a week at 4 degrees. We did not find any increase on the autofluorescence for lymphocytes. We found out that CD14 for monocytes are lost after the fixation process and unfortunately if we need to quantify them we need to label cells right the way. However, for CD5, Thy, CD4, CD8, CD45RA, CD45RB, HLA-DR and CD21 no problems were detect comparing results before and after fixation. If you need any aditional information I will be glad to help. Unfortunately I do not have experience with rat cells. Sincerely yours, Olindo Assis Centro de Pesquisas Rene Rachou - FIOCRUZ/BH Brazil
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