Mapping leucocyte populations in mouse lymphoid tissues and blood using a 28-color panel on the BD FACSymphonyTM System

Abstract

Abstract Multiparameter flow cytometric analysis of single cells allows the enumeration and characterization of heterogeneous cell populations in a variety of infections, blood cancers, immunodeficiencies and other disorders. Supported by advances in reagent development and instrumentation, this approach has increased dimensionality in single-cell data collection, and accelerated the discovery and testing of new therapies. Here we show a broad immunophenotyping panel that identifies the major leucocyte populations and various sub-populations from different mouse tissues. The panel can also be divided into focused modules for specific analysis of T cell, B cell or myeloid/natural killer cell lineages. BD FACSymphony combined with BD Horizon Brilliant Dyes allowed for high resolution identification of known leucocyte populations. Dimensionality reduction with t-SNE and automated clustering algorithms simplified cell analysis, enabling a faster and more efficient comparison of cell phenotypes across the tissues. Importantly, automated cell clustering analyses, while recognizing the predicted cell populations, also identified cell subsets missed during supervised analysis of the dataset. Thus, we describe a comprehensive single-cell profiling application capable of identifying well-characterized leucocyte populations and that has the potential to discover unique cell phenotypes.