T细胞亚群检测解析.pptVIP

* * 流式细胞仪通过前向散射与侧向散射、荧光信号这两种属性将细胞进行分群、分析的。 * Dichroic,二色的 fliters Bandpass filters * Their fluorescence emission wavelengths are: FITC = 530 nm, PE = 575 nm, PE–CY5 = 670 nm PE–TR = 613 nm, phycoerythrin–cyanine 5 tandem complex:PE-CY5 fluorescein isothiocyanate (FITC), phycoerythrin (PE), * Their fluorescence emission wavelengths are: FITC = 530 nm, PE = 575 nm, PE–CY5 = 670 nm PE–TR = 613 nm, phycoerythrin–cyanine(花青)5 tandem complex:PE-CY5 fluorescein isothiocyanate (FITC),异硫氰酸荧光素 phycoerythrin (PE),藻红蛋白 Detection of T cell subtypes by Flow Cytometry * BD公司流式细胞仪 FACSCalibur LSRII FACSCanto FACSCount FACSVantageDiVa FACSAria * What is Flow Cytometry? Principals and Applications FLOW CYTOMETRY FLOW =FLUID CYTO =CELLS METRY =MEASUREMENT Definition Flow Cytometry * Measuring properties of cells in flow Flow sorting * Sorting (separating) cells based on properties measuring in flow * Also called Fluorescence-Activated Cell Sorting (FACS) * FSC and SSC Forward Scatter (FSC) ---Approximate cell size Side Orthogonal Scatter (SSC)--- Cell complexity or granularity Scatter are used for preliminary identification of cells. In a peripheral blood sample, lymphocyte, monocyte and granulocyte populations can be defined on the basis of forward and side scatter. Forward and side scatter are used to exclude debris and dead cells. * FSC与SSC检测器 * FSC and SSC * FSC and SSC are used for preliminary identification of cells. ???????????????????????????????????????????????????????????????????????????????????????????????? ?????? ?????? ?????? ?????? ???? ?????? ?????? ???????????????????????????????????????????????????????????????????????????????????????????????? ?????? ?????? ?????? ?????? ???? ?????? ?????? * ???????????????????????????????????????????????????????????????????????????????????????????????? ?????? ?????? ?????? ?????? ??? ?????? ?????? ???????????????????????????????????