Picoliter droplet array based on bioinspired microholes for in situ single-cell analysis

The division of aqueous samples into microdroplet arrays has many applications in biochemical and medical analysis. Inspired by biological features, we propose a method to produce picoliter droplet arrays for single-cell analysis based on physical structure and interface. A 0.9 pL droplet array with an RSD (relative standard deviation) less than 6.3% and a density of 49,000 droplets/cm² was successfully generated on a PDMS chip (polydimethylsiloxane) from a micromachined glass mold. The droplet generation principle of the wetting behavior in the microholes with splayed sidewalls on the PDMS chip by liquid smearing was exploited. The feasibility of the picoliter droplets for bacterial single-cell analysis was verified by the separation of mixed bacteria into single droplets and isolated in situ bacteria propagation.