It has been our experience that some of the transfection methode increase the autofluorescence of cells. We routinely analyze eGFP transduced EC cells which grwo as adherent cells and then are removed with trypsin. We recently compared the efficiency of transfection with different compounds compared to standard CaPO4 methods. We found that expression was enhanced significantly using Lipofectamine (PLUS) and Lipofectamine 2000. The increases were in both the percent of cells expressing the fluorescent protein and in the intensity of fluoresence per cell. We use a FACS Calibur and have not changed anything to detect this protein. Charles A. Kuszynski, Ph.D. Assistant Professor/Director Cell Analysis Facility University of Nebraska Medical Center 986495 Nebraska Medical Center Omaha, NE 68198-6495 402 559-6267 voice 402 559-4077 fax ckuszyns@unmc.edu
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