Analysis of Flow Cytometry Data

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Analysis of Flow Cytometry Data

Analysis of Flow Cytometry Data

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Susan O. Sharrow1

1 National Cancer Institute, Bethesda, Maryland

John E. Coligan, Ada M. Kruisbeek, David H. Margulies, Ethan M. Shevach, and Warren Strober (eds.)
Current Protocols in Immunology

This unit provides several approaches to flow cytometry data analysis. Frequency determinations based on analysis of single-parameter fluorescence histograms and dual-parameter contour plots are presented. Next, steps are described for calculating values for signal-to-noise ratios when logarithmic amplification is used for data collection. This calculation can be used to compare the amounts of antigenic determinants per cell between different cells stained with the same reagent. Finally, a procedure is described for constructing a calibration curve for logarithmic amplifiers. This calibration curve is required for calculation of signal-to-noise values and can also be used to determine if the amplifiers are working properly.