A flexible multiwell format for immunofluorescence screening microscopy of small-molecule inhibitors. Assay Drug Dev Technol 2010 Oct;8(5):571-80
Date
07/30/2010Pubmed ID
20666660DOI
10.1089/adt.2009.0260Scopus ID
2-s2.0-77958561850 (requires institutional sign-in at Scopus site) 3 CitationsAbstract
Large-scale screens in mammalian cells demand for flexible high-throughput screening platforms that allow to analyze cellular traits on a genome-wide level or to identify small-molecule inhibitors (SMIs) from complex compound libraries. In this study we developed and tested high-density cell arrays made out of polydimethylsiloxane (PDMS) that support cell growth directly on standard glass microscope objective slides. We analyzed the effect of 3 reference inhibitors (MLN-8054, VX-680, and flavopiridol) and 4 exploratory, cell permeable small-molecule kinase inhibitors (two benzothiophene-based and two 4-amino-6-arylpyrimidine-based compounds) on different cell lines, using prototype 5 × 5 and 9 × 9 array carpets. We found that high-density PDMS cell arrays support growth of a broad range of cell types, are well suited for compound screens, and are compatible with high-content screening platforms. This novel array format is of particular advantage for compound screening to identify SMIs, when a combination of flexibility with respect to culture volume, well density, and high-resolution imaging is required. In addition, we demonstrated the suitability of this format for reverse transfection and siRNA experiments.
Author List
Scholz AK, Klebl BM, Morkel M, Lehrach H, Dahl A, Lange BMAuthor
Alexandra K. Scholz PsyD Assistant Professor in the Psychiatry and Behavioral Medicine department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsAntineoplastic Agents
COS Cells
Caco-2 Cells
Cell Line, Tumor
Cell Proliferation
Cell Survival
Centrosome
Dimethylpolysiloxanes
Drug Screening Assays, Antitumor
High-Throughput Screening Assays
Humans
Interphase
Microscopy, Fluorescence
Mitotic Index
Protein Kinase Inhibitors
RNA, Small Interfering
Small Molecule Libraries
Tissue Array Analysis
Transfection
