A 13-color parametric analysis for TBNK subset for the CytoFLEX instrument (TECH3P.935)

Abstract

Abstract CytoFLEX is the newest cytometer offered by Beckman Coulter, Inc. that provides up to 3 lasers, 13 colors for detection in a compact system. This instrument incorporates the latest technology, Fiber array photodiode, FADP, to provide high resolution, dynamic range and sensitivity in signal detection. To demonstrate the ability of its multicolor detection, the standard multicolor panel design principles were followed to develop a 14 marker, 13 color experiment. The cocktail design process also took into account the desired immunophenotypic markers, the available fluorochromes, and the use of well-defined control populations to identify the classic TBNK populations. Data was collected from normal, whole blood stained with single color reagents and lysed with VersaLyseTM Lysing Solution. Data was analyzed using the CytExpert software and is presented here with special emphasis on the optimization of flow analysis and compensation visualization. Results show that all TBNK populations were well resolved, notably the CD25+ subset in CD4+ T-cells, even though this maker was placed in the PC7 channel, which normally has poor resolution. The data presented demonstrates the ability of the CytoFLEX to provide sound detection and resolution for even the most dim versus negative population.