From: Albert Tai (acktai@exelixis.com)
Date: Fri Feb 14 2003 - 20:51:38 EST
Hi Cytometrists, As a relatively new comer to 2-color flow cytometry, I have experienced some observation that I cannot explain and hope to get some insights from the experts in this forum. The experiment involves PI staining fixed cells that are transfected with ZsGreen vector expressing a transgene of interest. I am intersted in looking at the cell cycle profile of green cells (which expressed the transgene) vs non-green cells (control). As an indication of the transfection efficiency, I also trasfected the same cells with the same vector but did not stain the cells with PI (they were also fixed with 0.5% paraformaldehyde). The transfection efficiency of the fixed cells without PI was consistently higher than the ones that stained with PI, roughly 2X. For staining with PI, I also added detergent @ 0.05% as final conc. I would think if it is a compensation issue, it would be the other way. Any thought or comment is much appreciated. Hope you all have a nice weekend. Regards, Albert Tai Exelixis, Inc. South San Francisco, CA
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