I'm assuming you mean %[CD4/CD3 +/+] + %[CD8/CD3 +/+] doesn't add up to %CD3. %CD45 will equal (or be very close to) %B+%T+%NK determined by your choice of panel. There are several reasons why 3/4 and 3/8 won't add up to CD3, here are some examples: 1. NK-T cells may express CD3 and/or CD8. If they express CD3 but not CD8, you automatically are missing a couple % . Not fixable. 2. monocytes infiltrating your scatter gate. I'm assuming in your 4C panel you gate on SSC/45, but if you don't, you might be picking up extra monocytes thus elevating your CD4 count. Easily checked by presence of "dim" CD4 cells. Easily fixed by removing monos' from the gate. 3. NK cells expressing CD8 but not CD3. inverse of #2. 4. B/T cell aggregates. Every now and then we get specimens that are CD3/CD19 +/+ (which are the same size as the rest of the lymphs). Since I haven't seen any literature about CD3+ b Cells or CD19 positive T cells, I'm taking a wild guess that during vortexing in the staining preparation, B/T cell aggregates get torn apart and CD3 "sticks" and stays on the B cell. there are many other increasingly hypothetic scenarios I could come up with, but the point here is, my CD4's and CD8's don't always perfectly add up to my CD3's either. :) Maciej __________________________________________________ Do You Yahoo!? Kick off your party with Yahoo! Invites. http://invites.yahoo.com/
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