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

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.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-7A9F-B Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-7AA1-3
Genre: Journal Article
Alternative Title : Assay Drug Dev. Technol.

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 Creators:
Scholz, Anne-Kathrin1, Author              
Klebl, Bert M., Author
Morkel, Markus2, Author              
Lehrach, Hans3, Author              
Dahl, Andreas3, Author              
Lange, Bodo M.H.4, Author
Affiliations:
1• Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              
2Dept. of Developmental Genetics (Head: Bernhard G. Herrmann), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433548              
3Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              
4Max Planck Society, ou_persistent13              

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 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.

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Language(s): eng - English
 Dates: 2010-07-28
 Publication Status: Published in print
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 Identifiers: eDoc: 536202
DOI: 10.1089/adt.2009.0260
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Title: Assay and Drug Development Technologies.
  Alternative Title : Assay Drug Dev. Technol.
Source Genre: Journal
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Pages: - Volume / Issue: 8 (5) Sequence Number: - Start / End Page: 571 - 580 Identifier: ISSN: 1540-658X