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Pathology Flow Cytometry Facility

Flow Cytometry data is technically a discontinuous variable function as the values recorded in your FCS data files are the results from analogue to digital converters (ADCs)The number of bins (or channels as flow jocks call them) varies greatly for today's cytometers..E,g, Facscan 1024

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Pathology Flow Cytometry Facility

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    6. When are cells positive?

    7. Basic KS comparisons are too sensitive to distinguish real from instrument variation

    9. The Mann-Whitney U test: a simple test for reproducibility Mann-Whitney U test will be generally applicable wherever you want to compare univariate distributions for a test sample and a control sample If the ranges do not overlap, you only need 3 samples of each to get p<0.05 Overlapping ranges require more samples but for example median FI Controls test

    10. Where inter- experiment variability is too high, the Wilcoxon’s signed rank test will still deliver significance at 5% for 5 pairs of samples Provided that which the control value is always lower than the test sample within each experimental pair

    11. A comparison of multiple treatments

    12. An example of Friedmann’s test

    13. Median GFP fluorescence

    14. Convert these to Ranks within each experiment

    16. Dunn’s Post test for pairwise comparison

    19. Use common sense Biological significance not statistical significance Compare like with like Reduce heterogeneity as far as possible Use non-parametric tests Mann-Whitney Wilcoxon’s Friedmann + Dunn’s Clear, reproducible cytometry data does not need stats; but if pushed you can risk using parametric stats with care

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