title: Electroactive microwell arrays for highly efficient single-cell trapping and analysis.
abstract: We present a novel method, implemented in the form of a microfluidic device, for arraying and analyzing large populations of single cells. The device contains a large array of electroactive microwells where manipulation and analysis of large population of cells are carried out. On the device, single cells can be actively trapped in the microwells by dielectrophoresis (DEP) and then lysed by electroporation (EP) for subsequent analysis of the confined cell lysates. The DEP force in the selected dimensions of the microwells could achieve efficient trapping in nearly all the microwells (95%) in less than three minutes. Moreover, the positions of the cells in the microwells are maintained even when unstable flow of liquid is applied. This makes it possible to exchange the DEP buffer to a solution that will be subsequently used for stimulating or analyzing the trapped cells. After closing the microwells, EP is conducted to lyse the trapped cells by applying short electric pulses. Tight enclosure is critical to prevent dilution, diffusion and cross contamination of the cell lysates. We demonstrated the feasibility of our approach with an enzymatic assay measuring the intracellular-galactosidase activity. The use of this method should greatly help analysis of large populations of cells at the single-cell level. Furthermore, the method offers rapidity in the trapping and analysis of multiple cell types in physiological conditions that will be important to ensure the relevance of single cell analyses.
authors.affiliation: Institute of Industrial Science, The University of Tokyo, 4-6-1, Komaba, Meguro-ku, Tokyo 153-8505, Japan.
authors.forename: Soo Hyeon
authors.initials: SH
authors.lastname: Kim
authors.forename: Takatoki
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Dominique
authors.initials: D
authors.lastname: Fourmy
authors.forename: Teruo
authors.initials: T
authors.lastname: Fujii
mesh_terms: D002458:Cell Fractionation; D057075:Enzyme Assays; D057809:HEK293 Cells; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D008856:Microscopy, Fluorescence; D059010:Single-Cell Analysis / Q000295:instrumentation* / Q000379:methods*; D059008:Time-Lapse Imaging; D020298:U937 Cells