title: High-throughput secondary screening at the single-cell level.
abstract: We have developed an automated system for drug screening using a single-cell-multiple functional response technology. The approach uses a semiautomated preparatory system, high-speed sample collection, and a unique analytical tool that provides instantaneous results for compound dilutions using 384-well plates. The combination of automation and rapid robotic sampling increases quality control and robustness. High-speed flow cytometry is used to collect single-cell results together with a newly defined analytical tool for extraction of IC(50) curves for multiple assays per cell. The principal advantage is the extreme speed of sample collection, with results from a 384-well plate being completed for both collection and data processing in less than 10 min. Using this approach, it is possible to extract detailed drug response information in a highly controlled fashion. The data are based on single-cell results, not populations. With simultaneous assays for different functions, it is possible to gain a more detailed understanding of each drug/compound interaction. Combined with integrated advanced data processing directly from raw data files, the process from sampling to analytical results is highly intuitive. Direct PubMed links allow review of drug structure and comparisons with similar compounds.
authors.affiliation: Purdue University Cytometry Laboratories, Department of Basic Medical Sciences, Purdue University, West Lafayette, IN 47907, USA. jpr@flowcyt.cyto.purdue.edu
authors.forename: J Paul
authors.initials: JP
authors.lastname: Robinson
authors.forename: Valery
authors.initials: V
authors.lastname: Patsekin
authors.forename: Cheryl
authors.initials: C
authors.lastname: Holdman
authors.forename: Kathy
authors.initials: K
authors.lastname: Ragheb
authors.forename: Jennifer
authors.initials: J
authors.lastname: Sturgis
authors.forename: Ray
authors.initials: R
authors.lastname: Fatig
authors.forename: Larisa V
authors.initials: LV
authors.lastname: Avramova
authors.forename: Bartek
authors.initials: B
authors.lastname: Rajwa
authors.forename: V Jo
authors.initials: VJ
authors.lastname: Davisson
authors.forename: Nicole
authors.initials: N
authors.lastname: Lewis
authors.forename: Padma
authors.initials: P
authors.lastname: Narayanan
authors.forename: Nianyu
authors.initials: N
authors.lastname: Li
authors.forename: C W
authors.initials: CW
authors.lastname: Qualls
mesh_terms: D001331:Automation; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005434:Flow Cytometry; D018922:HL-60 Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D020128:Inhibitory Concentration 50; D008928:Mitochondria / Q000378:metabolism; D059010:Single-Cell Analysis / Q000379:methods*; D013997:Time Factors