Cell-Based Screening Using High-Throughput Flow Cytometry

Abstract

This review describes the use of high-throughput flow cytometry for performing multiplexed cell-based and bead-based screens. With the many advances in cell-based analysis and screening, flow cytometry has historically been underutilized as a screening tool largely due to the limitations in handling large numbers of samples. However, there has been a resurgence in the use of flow cytometry due to a combination of innovations around instrumentation and a growing need for cell-based and bead-based applications. The HTFC™ Screening System (IntelliCyt Corporation, Albuquerque, NM) is a novel flow cytometry-based screening platform that incorporates a fast sample-loading technology, HyperCyt®, with a two-laser, six-parameter flow cytometer and powerful data analysis capabilities. The system is capable of running multiplexed screening assays at speeds of up to 40 wells per minute, enabling the processing of a 96- and 384-well plates in as little as 3 and 12 min, respectively. Embedded in the system is HyperView®, a data analysis software package that allows rapid identification of hits from multiplexed high-throughput flow cytometry screening campaigns. In addition, the software is incorporated into a server-based data management platform that enables seamless data accessibility and collaboration across multiple sites. High-throughput flow cytometry using the HyperCyt technology has been applied to numerous assay areas and screening campaigns, including efflux transporters, whole cell and receptor binding assays, functional G-protein-coupled receptor screening, in vitro toxicology, and antibody screening.

Identification of neutrophil surface marker changes in health and inflammation using high-throughput screening flow cytometry

“…In summary, the use of HTS-FC gave us the opportunity to rapidly evaluate all known CD markers simultaneously [43]. The primary aim of this study was to identify a unique neutrophil cellsurface signature, independent of location and activation state that would permit isolation of mature neutrophils.…”

Assessment of Cell Cycle Inhibitors by Flow Cytometry

“…Hand-free automations and the introduction of 96 well plate autosamplers have increased throughput capabilities in mammalian or plant cell (Cappella et al, 2010) (Cousin et al, 2009). Advancements in high throughput FCM have been implemented following the introduction of BD "Multiwell Autosampler" MAS™ and HTS™ , or efficient micro fluidic devices such as "plug-FCM" systems HyperCyt™ have been introduced and whereas traditional autosamplers load samples as single entities, automated sampling systems for FCM allowed individual samples to be assayed sequentially (Black et al, 2010).…”

Comparison of Flow Cytometry and Image-Based Screening for Cell Cycle Analysis

“…IBHCS utilizes the biological sample more efficiently. However, new FCs that require a lower cell number are becoming available [7]. In the near future we will observe more systems that merge the strengths of microscopy with those of FC.…”