I have successfully been doing ratiometric Ca++ measurements by flow for about two years for our own purposes. However, data analysis has always been a problem, since I have not been able to find a clear description of what people are really dividing to get their ratio numbers, and if I do the math myself (divide means from X & Y), I get very different numbers compared to software which automatically calculates a ratio. When a paper says "linear", I can't tell if they mean a true linear scale, or what BD calls a linear scale (in its "preferences" box on Lysys) which is actually a "log" scale. On a true linear scale, the fluorescent events are too spread out. However, I have often seen people convert the "log" (linear) scale to a "channel" scale, which superimposes a false linear axis over the true "log" values during data analysis. How can that be a true representation of the fluorescence values, especially when calculating ratios? Please respond to the list, or preferably, directly to me. Any help with your experiences will be very, very much appreciated. Please include references to literature which deals with this, if available. Also, if this or a similar topic has been discussed on this list in the past, please give approximate dates & message titles & I'll look it up on the net. Many thanks. Darren Hickerson, ECU School of Medicine
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