The latest development of commercial routine flow cytometers (FCM) is that they are equipped with three (blue, red,
violet) or more lasers and many PMT detectors. Nowadays routine clinical instruments are capable of detecting 10 or
more fluorescence colors simultaneously. Thereby, presenting opportunities for getting detailed information on the single
cell level for cytomics and systems biology for improve diagnostics and monitoring of patients. The University Leipzig
(Germany) recently started a cluster of excellence to study the molecular background of life style and environment
associated diseases, enrolling 25000 individuals (LIFE). To this end the most comprehensive FCM protocol has to be
developed for this study. We aimed to optimize fluorochrome and antibody combinations to the characteristics of the
instrument for successful 10-color FCM. Systematic review of issues related to sampling, preparation, instrument settings,
spillover and compensation matrix, reagent performance, and general principles of panel construction was performed.
10-color FCM enables for increased accuracy in cell subpopulation identification, the ability to obtain detailed
information from blood specimens, improved laboratory efficiency, and the means to consistently detect major and rare
cell populations. Careful attention to details of instrument and reagent performance allows for the development of panels
suitable for screening of samples from healthy and diseased donors. The characteristics of this technique are particularly
well suited for the analysis of broad human population cohorts and have the potential to reach the everyday practice in a
standardized way for the clinical laboratory.