Automated study on kinetics and biosensing of glow-type luminescence reaction via digital microfluidics–chemiluminescence

Automated manipulation of discrete droplets by digital microfluidics (DMF) combined with chemiluminescence (CL) is promising to achieve automated and sensitive biosensing and bioanalysis. Herein, a DMF-CL device was built to automatically study CL kinetics and biosensing of a glow-type CL reaction. Copper-cysteine nanoparticles (Cu/CysNP) were synthesized as a new CL catalyst to extend the CL reaction of luminol-H2O2 to more than 10 min. The automated manipulation of droplets reduced reagent costs and manual errors, leading to real-time, automated, and reliable study of CL kinetics. The CL kinetics curve collected by the DMF-CL integration device is in accordance with that of a commercial CL analyser. The long-lasting luminescence ensured automated, sensitive, and reliable detection of H2O2 as a direct or indirect analyte of the cascade catalytic reaction. Moreover, an innovative asymmetrical splitting method is proposed to quickly and precisely generate daughter droplets to ensure uniformity of the droplets and good repeatability of the DMF-CL measurements. Therefore, the DMF-CL analysis holds great potential for achieving online and automatic analysis and biosensing.