Making Panel Design Easier in the Era of High Dimensional Flow Cytometry

With the advent of the series of BD Horizon Brilliant™ Ultraviolet dyes, long gone are the teenage days of flow cytometry. Once limited by the availability of dyes to less than 18 colors, simultaneous detection of more than 25 markers is now possible, allowing immunophenotyping of multiple cell populations in a single sample or delving deeply into a particular cell type of interest. With any multicolor flow cytometry experiment, spectral overlap of fluorophores is a potential issue that has to be accounted for. As more fluorophores are included in the panel, broadening of the negative population due to spillover spreading error becomes a concern which may adversely affect the resolution of dim marker sunless this is factored into the panel design process. This can make high dimensional panel design seem challenging and intimidating. In this webinar, we will cover the main concepts to integrate when designing high dimensional flow cytometry panels. We will also propose a trio of “core panels” for the analysis of 1) T cells, 2) B cells / NK cells and 3) Myeloid cells. We have designed these core panels to include common markers of interest paired to specific fluorophores, based on antigen expression levels, fluorophore brightness and marker co-expression. These core panels manage potentially problematic fluorophore interactions while leaving more common, non-problematic detector channels open. This way, the core panels provide an approachable and standardized entry into the high parameter realm whilst allowing flexibility for additions to the core panels for researchers to address their own markers of interest. We will also cover other advanced aspects of panel design such as the sentinel approach. Key topics discussed in the webinar will be: Design of a 28 color panel The need for flexibility in panel design Immunophenotyping with T, B / NK and Myeloid core panels Use of high dimensional tools to analyze the many clusters derived from the panels. Speaker: Anis Larbi, PhD Head of A*STAR Flow Cytometry Facility, Immuno-monitoring Platform and Principal Investigator, Biology of Aging Program SIgN, A*STAR, Singapore Original Broadcast: October 8th, 2019