Hi, I have been using WINMDI 2.8 for analysis of intracellular/extracellular cytokine detection. Usually I display my data in histograms overlaying specific staining on to isotype controls. Although this works for most of my samples, the differences in some samples is quite discrete. I have decided to try using MFI or gMFI instead of histograms, but I have run into some difficulties. When using WINMDI 2.8 my histograms will only show geometric means. When I change to a linear scale, my program crashes. Question 1. How do I properly determine the linear mean fluorescent intensity of my samples using WINMDI 2.8? Question 2. How do I compare geometric mean fluorescent intensities between sample and control? Can I subtract background gMFI from specific? I am afraid that log transformed data can not be linearly compared, this is true? Question 3. If I want to pool all of my gMFI data from different experiments to determine the "average" value, how do I do this, again my fear is that gMFI can not be compared arithmetically. Thanks in advance, Greg Neely Graduate student University of Calgary Canada
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