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A Flexible Multiwell Format for Immunofluorescence Screening Microscopy of Small-Molecule Inhibitors.

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http://pubman.mpdl.mpg.de/cone/persons/resource/persons50528

Scholz,  Anne-Kathrin
• Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons50435

Morkel,  Markus
Dept. of Developmental Genetics (Head: Bernhard G. Herrmann), Max Planck Institute for Molecular Genetics, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons50409

Lehrach,  Hans
Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons50130

Dahl,  Andreas
Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

Lange,  Bodo M.H.
Max Planck Society;

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Citation

Scholz, A.-K., Klebl, B. M., Morkel, M., Lehrach, H., Dahl, A., & Lange, B. M. (2010). A Flexible Multiwell Format for Immunofluorescence Screening Microscopy of Small-Molecule Inhibitors. Assay and Drug Development Technologies., 8(5), 571-580. doi:10.1089/adt.2009.0260.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0010-7A9F-B
Abstract
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.