High-throughput Secondary Screening at the Single-cell Level

Overview

Journal J Lab Autom

Date 2012 Sep 13

PMID 22968419

Citations 6

Authors

J Paul Robinson

Valery Patsekin

Cheryl Holdman

Kathy Ragheb

Jennifer Sturgis

Ray Fatig

Larisa V Avramova

Bartek Rajwa

V Jo Davisson

Nicole Lewis

Padma Narayanan

Nianyu Li

C W Qualls Jr

Affiliations

Soon will be listed here.

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.

Citing Articles

Comparative analysis between zebrafish and an automated live-cell assay to classify developmental neurotoxicant chemicals.

St Mary L, Truong L, Bieberich A, Fatig 3rd R, Rajwa B, Tanguay R Toxicol Appl Pharmacol. 2023; 476:116659.

PMID: 37604412 PMC: 10529185. DOI: 10.1016/j.taap.2023.116659.

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition).

Cossarizza A, Chang H, Radbruch A, Acs A, Adam D, Adam-Klages S Eur J Immunol. 2019; 49(10):1457-1973.

PMID: 31633216 PMC: 7350392. DOI: 10.1002/eji.201970107.

Ultra-high throughput functional enrichment of large monoamine oxidase (MAO-N) libraries by fluorescence activated cell sorting.

Sadler J, Currin A, Kell D Analyst. 2018; 143(19):4747-4755.

PMID: 30199078 PMC: 6156879. DOI: 10.1039/c8an00851e.

Guidelines for the use of flow cytometry and cell sorting in immunological studies.

Cossarizza A, Chang H, Radbruch A, Akdis M, Andra I, Annunziato F Eur J Immunol. 2017; 47(10):1584-1797.

PMID: 29023707 PMC: 9165548. DOI: 10.1002/eji.201646632.

Flow Cytometry: Impact on Early Drug Discovery.

Edwards B, Sklar L J Biomol Screen. 2015; 20(6):689-707.

PMID: 25805180 PMC: 4606936. DOI: 10.1177/1087057115578273.

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