1 intro.ppt

1、23-13536-00 Rev. 01,Introduction to Multicolor Research Applications,23-13536-00 Rev. 01,2,Common Multicolor Research Applications,Intracellular cytokine staining Regulatory T cells (Tregs) Protein phosphorylation (BD Phosflow) Leukemia and lymphoma phenotyping Stem cell research,23-13536-00 Rev. 01

2、,3,Benefits of Multicolor Applications,Fewer tubes means less sample needed. Each additional color can significantly increase the number of populations that can be detected and quantified.,23-13536-00 Rev. 01,4,Challenges of Multicolor Applications,Loss of sensitivity due to spillover Limited availa

3、bility of marker-fluorochrome combinations Cytometer optimization and experimental design are critical Complexity of analysis,23-13536-00 Rev. 01,5,Meeting the Challenge,Know your cytometer and verify its performance. Design your panels to make best use of available reagents. Optimize titers and use

4、 effective assay controls. Use best practices to optimize cytometer settings and to acquire and analyze your data.,23-13536-00 Rev. 01,6,Digital Cytometry Advances,More colors, efficient light collection, and flexible configuration of detectors.,Improved detection of dim populations and more accurat

5、e compensation.,Improved fluidics and optics support high flow rates without compromising the data.,Advanced electronics and new software algorithms improve data accuracy and precision.,23-13536-00 Rev. 01,7,Key Cytometer Performance Factors,Sensitivityability to resolve dim populations. Dynamic ran

6、geability to accurately measure very bright populations. Linearityproportionality of fluorescence measurements. Trackingcytometer QC and reproducibility of results. Standardizationreproducibility of results across time and among laboratories.,23-13536-00 Rev. 01,8,Experimental Design Considerations,

7、Know your cytometer Reagent selection Optimal titer Appropriate controls Events to acquire Analysis and gating strategy,23-13536-00 Rev. 01,9,Visualizing Multicolor Data,Log Scaling,Biexponential Scaling,23-13536-00 Rev. 01,10,Visualizing Multicolor Data,Log Scaling,Biexponential Scaling,23-13536-00 Rev. 01,11,Biexponential Scaling,Approximates log scaling at the upper end. Approximates linear scaling at the lower end. Can display events at or below zero. Improves visual separation of populations