A Software Framework Enabling Analysis of Plate-Based Flow Cytometry Data for High-Throughput Screening

Abstract

Flow cytometry (FCM) is an important technology with a broad spectrum of applications ranging from basic research to clinical diagnostics. In a typical FCM experiment, thousands of cells are queried with respect to size, shape, and abundance of multiple cell surface antigens. Recent advances in FCM techniques and instrumentation have enabled researchers to raise the throughput of experimentation dramatically. However, data analysis has remained a time-consuming activity requiring significant manual intervention for gating as well as for overall data reduction and interpretation. Presented in this article is a novel, algorithmically flexible, internally developed, software framework for the analysis of plate-based FCM data for high-throughput screening (HTS). Utilizing a post-treatment pooling strategy, >87,000 individual wells representing over 240,000 compounds were automatically gated, percent of control (POC) calculated, results assembled, deconvolved, and sorted, allowing researchers to visually assess wells of interest in minutes.