Multi-color Flow Cytometry
Flow Cytometry technology can give you accurate and quantitative data, but only if the proper controls are done prior to sample analysis. This is especially important when multi-color analysis is undertaken (i.e. more than one color).
Spectral overlap between fluorochromes in multi-color experiments requires the use of compensation where mathematics is used to calculate and remove spectral overlap.
An important note: Remember to optimize your sample preparation by:
- titrating out the antibodies
- factoring spectral overlap and spillover spread when designing your multi-color panel
- factoring in autofluorescence of your samples
- use transformed data on a biexponential scale to see all your compensated data
A few good reads:
- Quantifying spillover spreading for comparing instrument performance and aiding in multicolor panel design
- Quality assurance for polychromatic flow cytometry
Please Note: The Facility’s mission is to serve investigators in their quest to obtain accurate data. To do so the Core staff will not assist with running samples when the necessary compensation controls are not provided by the investigator. If you are unsure about what kinds of controls you need, come speak with the Core staff.