Hello dear Flowers, I work at Pasteur Institute of Cambodia. I try to transfecte human PBMC by siRNA-Cy5 by using oligofectamine. This tranfected cells have been verify by Flow cytometry ( type FACScalibur). But by this technique, I'm not sure whether my siRNA-Cy5 enter into the cell or they adhere on the external surface of PBMC. Have any one have expriences with this. I would like to know that by which mean I could be sure that my siRNA-Cy5 are really go into the cell in using Flow cytometry technique. Another question is I try to label siRNA-Cy5 transfected PBMC with CD4 APC. I've encountered a problem about emission spectrum overlapping of Cy5 and APC. So I try to pass only siRNA-Cy5 transfected PBMC without CD4 APC labeling. I see that the emission spectrum of Cy5 can detected in both detector is FL3 and FL4 but different gate percentage ( % gate in FL4 is higher than in FL3). I don't know which one I should use it to detected Cy5 . All idea and suggestion have been appreciated. best regard POLIDY PEAN Institute Pasteur of Cambodia 5, boulevard Monivong Tel : 855 12 552 182 Fax : 855 23 725 606Received on Thu Mar 25 11:58:00 2004
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