A flexible multiwell format for immunofluorescence screening microscopy of small-molecule inhibitors.
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.
Scholz
Anne-Kathrin
AK
Department of Vertebrate Genomics, Max-Planck Institute for Molecular Genetics, Berlin, Germany.
Klebl
Bert M
BM
Morkel
Markus
M
Lehrach
Hans
H
Dahl
Andreas
A
Lange
Bodo M H
BM
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D019556:COS Cells; D018938:Caco-2 Cells; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D018385:Centrosome / Q000187:drug effects; D002522:Chlorocebus aethiops; D004129:Dimethylpolysiloxanes; D004354:Drug Screening Assays, Antitumor; D057166:High-Throughput Screening Assays; D006801:Humans; D007399:Interphase / Q000187:drug effects; D008856:Microscopy, Fluorescence; D008940:Mitotic Index; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000235:genetics; D054852:Small Molecule Libraries; D046888:Tissue Array Analysis; D014162:Transfection
D000970:Antineoplastic Agents; D004129:Dimethylpolysiloxanes; D047428:Protein Kinase Inhibitors; D034741:RNA, Small Interfering; D054852:Small Molecule Libraries; C013830:baysilon