The BD Biosciences Webinar Series is an ongoing, informative program in which senior scientists present online seminars covering a broad range of topics.
Optimization of Multicolor Panel Design Incorporating Low Receptor Density Antigens
Date: Wednesday, May 29, 2013
Time: 9 AM PDT/12 PM EDT
Speaker: Robert Balderas, Vice President, Biological Sciences, BD Biosciences
Today scientists choose dyes based on the capabilities of the instrumentation available, the number of surface receptors on the cells they are studying, and their brightness and spillover value.
This webinar will present a methodology for designing multicolor experiments, including demonstrations of how effective use of fluorochromes allows for the identification of cell populations with lower receptor density than was previously possible. A wide range of data using this methodology will be presented.
Topics will include:
- The impact of optical design of the cytometer on signal detection
- Methodology for choosing fluorochromes for optimum multicolor panels—with data from 6 to 18 colors
- Importance of standardization of instruments and optimization of instrument setup to data quality
- Importance of the use of appropriate controls
How to Analyze More Markers Without Adding Detectors
Date: Wednesday, June 5, 2013
Time: 8 AM PDT/11 AM EDT
Speaker: Koen Verbrugghe, PhD, Senior Scientist, BD Biosciences
In multicolor flow cytometry, researchers conventionally assign no more than one marker to each detector. However, by taking advantage of biology and spillover, you can increase the number of markers detected, identify more populations, and collect more data.
This webinar will begin by showing how to double-up markers in one detector when populations can be separated by other parameters such as light scatter. It will go on to demonstrate that fluorescence spillover—far from being a nuisance—can be exploited to analyze more fluorochromes with a given number of detectors. Using staining protocols based on this strategy, three human lymphocyte populations (B cells, NK cells, and T cells) can successfully be identified using two detectors and this can be extended to identify 7 lymphocyte markers on the 4 detectors of the BD Accuri™ C6 personal flow cytometer. This strategy is especially suited to an instrument like the BD Accuri C6, in which detector voltage, optical filters, and laser power are fixed and spillover is predictable.
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