A Single-Droplet Multiplex Bioassay on a Robust and Stretchable Extreme Wetting Substrate through Vacuum-Based Droplet Manipulation.

Herein, a droplet manipulation system with a superamphiphobic (SPO)-superamphiphilic (SPI) patterned polydimethylsiloxane (PDMS) substrate is developed for a multiplex bioassay from single-droplet samples. The SPO substrate is fabricated by sequential spraying of adhesive and fluorinated silica nanoparticles onto a PDMS substrate. It is subsequently subjected to oxygen plasma with a patterned mask to form SPI patterns. The SPO layer exhibits extreme liquid repellency with a high contact angle (>150°) towards low-surface-tension and viscous biofluidic droplets (e.g., ethylene glycol, blood, dimethyl sulfoxide, and alginate hydrogel). In contrast, the SPI exhibits liquid adhesion with a near zero contact angle. Using the droplet manipulation system, various liquid droplets can be precisely manipulated and dispensed onto the predefined SPI patterns on the SPO PDMS substrate. This system enables a multiplex colorimetric bioassay, capable of detecting multiple analytes, including glucose, uric acid, and lactate, from a single sample droplet. In addition, the detection of glucose concentrations in a plasma droplet of diabetic and healthy mice are performed to demonstrate the feasibility of the proposed system for efficient clinical diagnostic applications.