Dear Flowers, I have a basic question on mean fluorescence channel analysis: I always used this approach to estimate on a semi quantitative way the binding of fluorescent reagents on my supports (cells, beads, etc). I usually apply this tool to analysis populations the present a unimodal distribution on single color histograms. In order to do that I only convert my acquired data, at the time I analyze then to linear scale (or channel values) in order to get the mean fluoresce channel on a scale 256 or 1024 on a BD FACScalibur. However, last week I was told that if I really want to analyze data based on mean fluorescence channel I am supposed to acquire data using linear gains to fluorescence. I have noticed that on a BD instrument always we need to use log gains to fluorescence. Otherwise we do not get any signal. Does anyone know a situation when linear gains for fluorescence are indicated to acquire data. We have started a discussion here in our institution and we are not going anywhere. Could anyone give some support to answer this question. My major point is that MFC is only a tool that we can use to analyze the intensity of fluorescence presented by a population (most useful to analyze unimodal distribution) and that can be applied after acquisition. Moreover, I think that log gains are essential to acquisition on flow cytometry. We would appreciate any comments, Thanks in advance, Olindo
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