I am somewhat new to flow cytometry and I just ran my first set of cells that I stained myself. I have confusing results though. I am using an indirect staining method to look at a cell surface receptor on a mouse endothelial cell line. When I ran my controls everything fell into the negative area on the histogram (the first decade) for the PE signal. When I ran my positives, it was as if the negative control peak shifted to the right by about 3/4 of a decade. My positives therefore only have one peak (instead of two). I'm not sure what went wrong. Something I have thought of: I did not complete a fc receptor blocking step. Could this be the reason for not getting two distinct peaks? Your insight is much appreciated. Jess Jankowsky
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