Hello Mathi, A very interesting question that presents other questions! When we gate on Lymphocytes do we include Blast cells in this region? Are Monoblasts or Myeloblasts included? Is the gate large enough to include all the immature cells? It all depends on what you are screening for! Another interesting question is : When you calculate the absolute numbers, do you do the calculation based on total leukocytes or the differential count? If you base it on specific cells that are not in the gated region, you may get a count of zero because your gate does not include them! Any comments or corrections are most gratefully received Warmest Regards From Fareed Al-Gurg Dubai - UAE. Mathi wrote: > Dear Flow Ers, > > I am working with punch of smart people who has no flow backgrounds. > They are experts in their fields. > > we manufacture activated lymphocytes to implant into various tumors. > FDA needs validation for all my test methods. > > 1. CD 69 to measure early activation T3+ cells > 2. RNA and DNA stainig by AO ( acridine orange) > > They want me to show a)precision b)repeatability c) robustness d) > accuracy. > When I talk about gating: i say gate on lymphocyte. They ask me to > prove that the cells in the gate are lymphocyte. > > Please any body save me. I appreciate all your help and any help at all. > I promise i wiil post the out come. > > Thanks for saving my soul in advance. > > Mathi > > Cytometry > Meyer Pharmaceuticals > mathi@meyerpharm.com
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