流式细胞术演示稿

1、BD VantageSE DiVa option Beckman Coulter Epics Altra HypersortDakoCytomation MoFloBD FACSArian流式细胞仪就是进行流式细胞分析的仪器，它集电子技术、计算机技术、激光技术、流体理论于一体，是一种非常先进的检测仪器。细胞结构特异性抗原（细胞表面/胞浆/核）细胞内细胞因子细胞活性酶活性激素结合位点细胞受体细胞功能+ +Surface Phenotyping+ +IntracellularHOW?LaserLight488nmFL2 PESSCFSCFluidicsElectronicsComputerFL3

2、PerCP, Cy5 FL1 FITCHow does a flow cytometer work?Light detectors液流系统：流动室、液流驱动系统液流系统：流动室、液流驱动系统液流中心由单列匀速运动颗粒组成的液柱激光光源单波长、高强度、高稳定性多采用氩离子激光器或氦氖激光器一般选配2-4根激光，488nm 、633nm和355nm、407nmUV激光最多检测13个荧光参数 FALS SensorLaserFALS Sensor90LS SensorLaserLongpass Shortpass Bandpass460 500 540460 500 540460 500 540光收

3、集系统：滤光片 光收集系统：光电倍增管（光收集系统：光电倍增管（PMT）FACSCalibur 光路图Presenting and Interpreting DataDual parameter data can be displayed in two dimensions using dot, density or contour plots.Single parameters can be displayed as a histogram.Surface phenotypeCell cycleDNARNA Signal transductionIntracellular calcium p

4、HFactor productionGatingCompensationcompensationG0-G1SG2-MFluorescence Intensity# of EventsA typical DNA Histogram of cell proliferation488 nm laser+-Charged PlatesSingle cells sortedinto test tubesFALS SensorFluorescence detector+Antigen (cytokines)Fluorescence Detector antibodyCapture beads Captur

5、e antibodyBeadsShades of a colorAntibody coupled beads, emitting at distinct FL3 intensitiesVarious analytesAntibody coupled PE label, emitting at FL2 intensity proportional to analyte conc.Multiple sizesBeads provide an expandable assay platform for use with a flow cytometerDifferent intensitiesDifferent colors with different intensitiesIL-8IL-1bIL-6IL-10TNFIL-12Capture BeadsLysate or S