CytoFLEX Instrument Evaluation Using Biological Specimens
Evaluation of the CytoFLEX flow cytometer, by independent research flow cytometry laboratories and Beckman Coulter senior research and development scientists, was performed using flow cytometers equipped with 12 parameters and 9 fluorescent detectors. Instruments were equipped with the enhanced Side Scatter channel (VSSC) option. Evaluation of biological specimens included fresh human whole blood, human leukocyte control cells, and smaller cells and particles, such as RBCs, platelets, microbes, liposomes, and vesicles. A 9-color data set was collected, using a prototype human leukocyte control cells, developed by BCI. Analysis of the human control leukocyte cells clearly demonstrates expected populations of (T cells, NKT cells, NK cells, and B cells). To explore the instrument capabilities to resolve small particles, analysis of red blood cell micro particles was performed. A three month old human blood sample, stored under standard blood banking conditions, was stained with Annexin V. Forward scatter extinction and side scatter revealed red blood cell micro particles, transition events and intact red blood cells. A homogeneous population of Listeria innocua, a rod shaped bacteria with 0.5 μm in width and 1–1.5 μm in length was stained with Propidium Iodide and Syto9. The specimen was analyzed for live cell versus dead cell populations. In another experiment, micro particles were examined using vesicles isolated from rat plasma. Vesicles were stained with AnnV Dy488 and CD42d. Resolution of vesicles from instrument noise was achieved through the VSSC (Violet laser side scatter signal) and fluorescence. An increase in scatter signal sensitivity was achieved using the VSSC collection channel, as well as an increase in background noise. Data and figures presented in this poster are a collaborative effort, through the efforts of scientists representing the following laboratories: Purdue University Cytometry Laboratory (PUCL), La Jolla Bioengineering Institute, University of Pittsburgh, and Beckman Coulter, Miami Site. The flow cytometry platforms used for the evaluations are for research use only.