Methods to Simplify the Design of Multi-color Panels.

Flow cytometry has long been a critical tool for immunologists. As the number of parameters used in flow cytometry continues to increase, one of the more difficult aspects of this practice is panel design. Fully resolving dim populations of interest can be challenging. Low antigen density, fluorescence spillover, available fluorochromes and limited catalog offerings are just a few of the factors that make panel design difficult and lead to sub-optimal resolution. Here, we present a basic set of principles that can help simplify panel design. Following these principles helps reduce the number of iterations needed to maximize the resolution for populations of interest. Additionally, we present a basic methodology that can be used to characterize your flow cytometer and generate a matrix showing the relative impact of fluorochrome combinations on population resolution. Using this Resolution Impact Matrix [RIM], low complexity, no/low compensation panels can be more easily created. Similarly, when performing higher order multicolor experiments, this characterization is critical to help design the optimum panel for that instrument. When used together, these two methods can help simplify the panel design process and lead to higher quality data.