Fabrication of microwell arrays based on two-dimensional ordered polystyrene microspheres for high-throughput single-cell analysis.
This paper describes a method of fabricating rounded bottom microwell arrays (MA) in poly(dimethylsiloxane) (PDMS) by molding a monolayer of ordered polystyrene (PS) microspheres. PS microspheres were self-assembled on a glass slide and partially melted mainly from the bottom at 240 °C to increase adhesive force with the substrate. The partially melted PS arrays were used as master to generate MA. Microwell sizes are tunable in the 10-20 μm range with rounded bottoms; such a 3D structure is not readily obtainable through conventional soft lithography. Both adherent and nonadherent cell types can be retained in the microwells with high efficiency. As a demonstration of the advantage of real-time cell screening with this MA, single cell enzyme kinetic analysis was also carried out on trapped single cells. The PDMS MA may find applications in high-throughput drug screening, guided formation of cell clusters, and multicellular communication.
The Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Department of Chemistry, Tsinghua University, Beijing 100084, China.
Chuansen
C
Liu
Jiangjiang
J
Liu
Dan
D
Gao
Mingyu
M
Ding
Jin-Ming
JM
Lin
D043182:Carboxylesterase / Q000378:metabolism; D057075:Enzyme Assays; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D046228:Microarray Analysis / Q000191:economics / Q000379:methods*; D008863:Microspheres; D055616:Microtechnology / Q000379:methods*; D011137:Polystyrenes / Q000737:chemistry*; D059010:Single-Cell Analysis / Q000191:economics / Q000295:instrumentation*; D013997:Time Factors
D011137:Polystyrenes; D043182:Carboxylesterase