Hi All, I am wondering what your thoughts are on this: Suppose you calculate a ratio between two separate FL distributions (the median, mode, or mean) on the same cell line (ratio 1). The acquisition for each cell line being set on the autofluorescent sample. Now suppose you want to compare ratio(1) with the ratio(2) of another cell line stained with the same set of mabs (or multiple cell lines for that matter). What are the considerations here? 1) How valid are the calculated ratios given the type of scale used to measure the range of FL? Linear conversions of log data? Channel number only? 2) There are obviously some limitations here with comparing very different cell lines? Say for example, kidney cell vs. liver cells, rather than liver cell samples from multiple animals? 3) Would a subtraction of the two distributions being compared, then a K-S test comparing the subtractions of two different cell lines be an alternative analysis? Somehow this doesn't seem right. 4) Is setting the instrument on the unstained sample for each line really useful? Why not just set on the highest signal of the whole bunch, and let the chips fall for all cell lines (if of course all cells being tested fall within the same analysis gate for scatter signals)? Different cell types would make this approach difficult. 5) And statistics? What is the comparison we are making? Ratio of two distribution's medians compared to the ratio of two distribution's medians--- ad nauseum for every cell line? What might be the best statistical test to approach the measure of significance? How would variation fit into the picture? Any ideas, any comments would be greatly appreciated. Thanks, Christopher Boyce Hybritech San Deigo, CA
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