Cell-based screening using high-throughput flow cytometry.

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.

[1]  L A Sklar,et al.  Plug flow cytometry extends analytical capabilities in cell adhesion and receptor pharmacology. , 2001, Cytometry.

[2]  John A. Tallarico,et al.  Multi-parameter phenotypic profiling: using cellular effects to characterize small-molecule compounds , 2009, Nature Reviews Drug Discovery.

[3]  Terry D. Foutz,et al.  Ligand-receptor-G-protein molecular assemblies on beads for mechanistic studies and screening by flow cytometry. , 2003, Molecular pharmacology.

[4]  Tudor I. Oprea,et al.  High-throughput flow cytometry to detect selective inhibitors of ABCB1, ABCC1, and ABCG2 transporters. , 2008, Assay and drug development technologies.

[5]  Tudor I. Oprea,et al.  High-throughput flow cytometry for drug discovery , 2007, Expert opinion on drug discovery.

[6]  Howard M. Shapiro,et al.  Practical Flow Cytometry , 1985 .

[7]  S. Burchiel,et al.  A multifunctional androgen receptor screening assay using the high‐throughput Hypercyt® flow cytometry system , 2008, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[8]  R. Neubig,et al.  Identification of Small-Molecule Inhibitors of RGS4 Using a High-Throughput Flow Cytometry Protein Interaction Assay , 2007, Molecular Pharmacology.

[9]  Garry P Nolan,et al.  Fluorescent cell barcoding in flow cytometry allows high-throughput drug screening and signaling profiling , 2006, Nature Methods.

[10]  L A Sklar,et al.  HTPS Flow Cytometry: A Novel Platform for Automated High Throughput Drug Discovery and Characterization , 2001, Journal of biomolecular screening.

[11]  P. Chattopadhyay,et al.  Seventeen-colour flow cytometry: unravelling the immune system , 2004, Nature Reviews Immunology.

[12]  Tudor I. Oprea,et al.  Integration of Virtual Screening with High-Throughput Flow Cytometry to Identify Novel Small Molecule Formylpeptide Receptor Antagonistss⃞ , 2005, Molecular Pharmacology.

[13]  H. Shapiro Practical Flow Cytometry: Shapiro/Flow Cytometry 4e , 2005 .

[14]  John P Nolan,et al.  The flow of cytometry into systems biology. , 2007, Briefings in functional genomics & proteomics.

[15]  Christian D Muller,et al.  Can flow cytometry play a part in cell based high‐content screening? , 2007, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[16]  Tudor I. Oprea,et al.  Flow cytometry for high-throughput, high-content screening. , 2004, Current opinion in chemical biology.

[17]  Tudor I. Oprea,et al.  Detection of Intracellular Granularity Induction in Prostate Cancer Cell Lines by Small Molecules Using the HyperCyt® High-Throughput Flow Cytometry System , 2009, Journal of biomolecular screening.

[18]  P. Luciw,et al.  Multiplexed microbead immunoassays by flow cytometry for molecular profiling: Basic concepts and proteomics applications , 2009, Critical reviews in biotechnology.

[19]  Bruce S Edwards,et al.  High‐throughput flow cytometry: Validation in microvolume bioassays , 2003, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[20]  Johan Lindberg,et al.  Determination of Binding Specificities in Highly Multiplexed Bead-based Assays for Antibody Proteomics *S , 2007, Molecular & Cellular Proteomics.