title: A silicon-glass microwell platform for high-resolution imaging and high-content screening with single cell resolution.
abstract: We present a novel microwell array platform suited for various cell-imaging assays where single cell resolution is important. The platform consists of an exchangeable silicon-glass microchip for cell biological applications and a custom made holder that fits in conventional microscopes. The microchips presented here contain arrays of miniature wells, where the well sizes and layout have been designed for different applications, including single cell imaging, studies of cell-cell interactions or ultrasonic manipulation of cells. The device has been designed to be easy to use, to allow long-term assays (spanning several days) with read-outs based on high-resolution imaging or high-content screening. This study is focused on screening applications and an automatic cell counting protocol is described and evaluated. Finally, we have tested the device and automatic counting by studying the selective survival and clonal expansion of 721.221 B cells transfected to express HLA Cw6-GFP compared to untransfected 721.221 B cells when grown under antibiotic selection for 3 days. The device and automated analysis protocol make up the foundation for development of several novel cellular imaging assays.
authors.affiliation: Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Thomas Wilhelm
authors.initials: TW
authors.lastname: Frisk
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Khorshidi
authors.forename: Karolin
authors.initials: K
authors.lastname: Guldevall
authors.forename: Bruno
authors.initials: B
authors.lastname: Vanherberghen
authors.forename: Björn
authors.initials: B
authors.lastname: Önfelt
mesh_terms: D001402:B-Lymphocytes / Q000166:cytology; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D004867:Equipment Design; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D046228:Microarray Analysis / Q000295:instrumentation* / Q000379:methods*; D008854:Microscopy, Electron; D055616:Microtechnology; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods; D014162:Transfection