BD流式细胞仪细胞周期分析艺术.pdfVIP

CELL CYCLE ANALYSIS Analysis of a cell population’s state of replication can be achieved by fluorescently labeling the cell nuclei then analyzing the fluorescence properties of each cell in the population by flow cytometry. Quiescent and G1 cells will have one copy of DNA and will therefore have 1X fluorescence intensity. Cells in G2/M phase of the cell cycle will have two copies of DNA and accordingly will have 2X intensity. Since the cells in S phase are synthesizing DNA they will have fluorescence values between the 1X and 2X populations. 1X 2X S G0/1 = 1X S Phase G2/M = 2X DYE FLUORESCENCE The resulting histrogram consists of three populations: two Gaussian curves (1X and 2X peaks) and the S- phase population. Adjacent populations overlap each other. Because of this, a modeling program is required to de-convolute the populations and assign percentage values to each population. Expert and subjective review of the modeling software’s cell cycle phase percentage assignment is the final stage of cell cycle analysis prior to reporting the results. G0G1: 33.94 % Mean: 49.19 G0G1 CV: 2.97 % G2M G2M: S-Phase 15.28 % Mean: 96.41 G2/G1: 1.96 S-Phase: 50.78 % Mean: 69.20 Density Plot and Histogram generated with ModFit LT 3.0 TM