Confinement-enhanced microalgal individuals biosensing for digital atrazine assay

Microalgal sensors are widely recognized for their high sensitivity, accessibility, and low cost. However, the current dilemma of motion-induced spatial phase changes and concentration-related multiple scattering interferes with induced test instability and limited sensitivity, which has hindered their practical applications. Here, a differentiated strategy, named confinement-enhanced microalgal biosensing (C-EMB), is developed and proposed to pave the way. The in-situ printed microgel trap is designed to confine Chlamydomonas reinhardtii individuals, stabilizing their spatial phase. The microgel trap arrays are introduced to eliminate the multiple scattering of microalgae, breaking the existing effective concentration in traditional microalgal sensing and enabling sensitive assays. The integration with lab-on-a-chip technology and a developed digital imaging algorithm empower portable and automated detection. With this system, a microalgae analyzer is developed for atrazine detection, featuring a linear range of 0.04-100 μg/L. We assess the system's performance through practical atrazine assays on commercial food, using a double-blind test against a standard instrument. Our results demonstrate the good accuracy and test stability of this system with the mean bias atrazine detection in corn and sugarcane juice samples (SD) were 1.661 μg/L (3.122 μg/L) and 3.144 μg/L (4.125 μg/L), respectively. This method provides a new paradigm of microalgal sensors and should advance the further applications of microalgal sensors in commercial and practical settings.