A novel approach for the detection of spectral data using a CytoFLEX mosaic 88-channel detection accessory for a CytoFLEX LX flow cytometer 4154

Abstract Description CytoFLEX LX is a conventional cytometer with up to 6 lasers (Ultraviolet (U): 355nm, Violet (V): 405nm, Blue (B): 488nm, Yellow (Y): 561nm, Red (R): 638nm, and Infrared (IR): 808nm) and 21 color parameters (U3, V5, B3, Y5, R3, IR2). Attaching an 88-channel spectral detection module (U20, V20, B16, Y12, R10, IR3) with 6 side scatter (USSC, VSSC, BSSC, YSSC, RSSC, and IRSSC) and 1 forward scatter detector allows for the expansion of the detection capabilities to at least 40 color parameters. Whole blood from normal donors was lysed using VersaLyse. The remaining white blood cells (WBC) were stained with ViaKrome 808 live/dead dye to take advantage of the CytoFLEX 808nm laser. The WBC were stained with a 40-color panel and single color controls. Target value settings for the 88 channels were established using CytoFLEX Ready to Use Daily QC and Daily IR QC Fluorospheres. Data was collected using CytoFLEX for Spectral software. At least 40 unique spectral signatures were detected using CytoFLEX Mosaic. Using the Poisson algorithm for unmixing resulted in reduced spreading compared to least squares method (LSM). Similar to previously published data, we were able to identify CD4+ and CD8+ T cells, Regulatory T cells, gamma delta T cells, NKT-Like cells, Innate Lymphocyte Cells, B cells, Basophils, Dendritic Cells, and Monocytes. The Beckman Coulter products mentioned herein are trademarks or registered trademarks of Beckman Coulter, Inc. in the United States, and other countries. Funding Sources Beckman Coulter Topic Categories Technological Innovations in Immunology (TECH)