This is a very interesting question, thanks for posting it. Isotype must be applied to and analysed on each cell type separately. When I stain for markers Y and Z (we don't do much work on X, sorry) when looking at lymphocytes and monocytes, I do 2 separate acquisitions from the same tube, from the same patient, one optimized for monocytic isotype and one for lymps (monos isotype with lymp settings would be in 2nd decade). As far as statistics go we usually look for absolute numbers, such as setting M1 in a isotype histogram to 98% cells and pasting that same M1 in the fluorochrome. Also you can do relative shift analysis, such as how far has your fluorochrome expression changed before and after stimulation etc. This also would involve setting your M1 in one and pasting it in the other. In cellquest, I look for % gated and GM. I would be interested if you could post replies to your query on the newsgroup as a summary Regards, Maciej __________________________________________________ Do You Yahoo!? Yahoo! Messenger - Talk while you surf! It's FREE. http://im.yahoo.com/
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