Mixing during Trapping Enabled a Continuous-Flow Microfluidic Smartphone Immunoassay Using Acoustic Streaming.

Smartphone-enabled microfluidic chemiluminescence immunoassay is a promising portable system for point-of-care (POC) biosensing applications. However, due to the rather faint emitted light in such a limited sample volume, it is still difficult to reach the clinically accepted range when the smartphone serves as a standalone detector. Besides, the multiple separation and washing steps during sample preparation hinder the immunoassay's applications for POC usage. Herein, we proposed a novel acoustic streaming tweezers-enabled microfluidic immunoassay, where the probe particles' purification, reaction, and sensing were simply achieved on the same chip at continuous-flow conditions. The dedicatedly designed high-speed microscale vortexes not only enable dynamic trapping and washing of the probe particles on-demand but also enhance the capture efficiency of the heterogeneous particle-based immunoassay through active mixing during trapping. The enriched probe particles and enhanced biomarker capture capability increase the local chemiluminescent light intensity and enable direct capture of the immunobinding signal by a regular smartphone camera. The system was tested for prostate-specific antigen (PSA) sensing both in buffer and serum, where a limit of detection of 0.2 ng/mL and a large dynamic response range from 0.3 to 10 ng/mL using only 10 μL of sample were achieved in a total assay time of less than 15 min. With the advantages of on-chip integration of sample preparation and detection and high sensing performance, the developed POC platform could be applied for many on-site diagnosis applications.