The emerging field of high-throughput (HT) flow cytometry is extending the capabilities of cell-based screening technologies into the profiling of compound libraries. The screening of molecules for effects in living cell populations has become an integral step in virtually every drug discovery program. High-throughput flow cytometry is a particularly useful approach for screening compounds in cell models where multiple readouts are desired. This can be performed using the Invitrogen™ Attune™ NxT Flow Cytometer with Attune™ NxT Autosampler, where 96- and 384-well plates can be acquired and analyzed. In addition, up to 16 different parameters per cell can be measured on the Attune NxT Flow Cytometer, and thousands to millions of cells can be analyzed in minutes at up to 35,000 cells/sec and 1 mL/min sample input.
The Attune NxT Flow Cytometer uses a unique volumetric sample and sheath fluid delivery system. Samples are introduced to the Attune NxT Flow Cytometer with syringes, producing accurate measurements of the volumes of acquired samples and thus accurate calculation of cell concentrations. Given time savings, high content, and assay robustness, high-throughput flow cytometry provides the capacity needed as part of an effective and practical systems biology approach. These recent developments expand the role of flow cytometry in drug discovery and life science research and are a key factor in understanding cellular and molecular networks.
In this application note, consistency across 96-well plates was examined. In brief, lysed human whole blood was labeled with fluorophore-conjugated monoclonal antibodies for CD45, CD3, CD4, and CD8 targets. The labeled cells were fixed, then added in equal amounts to each well of two 96-well plates, and acquired on a 4-laser Attune NxT Flow Cytometer with Autosampler. The concentration statistic representative of events/μL of any given population and percent positive are examined in depth.