Temporal-Spatial-Color Multiresolved Chemiluminescence Imaging for Multiplex Immunoassays Using a Smartphone Coupled with Microfluidic Chip

Chemiluminescence (CL) bioassays have become a main technology in clinical diagnosis. However, multiplex analysis for sensitive and simultaneous determination of multiple biomarkers related to one disease in one detection run on a single chip in order to achieve fast and accurate diagnosis of diseases remains a great challenge. Herein, we report temporal-spatial-color multiresolved CL imaging strategy for multiplex label-free immuno-assays using a smartphone coupled with a microfluidic chip. Temporal-spatial-color multiresolved CL imaging signals were produced by sequentially transporting coreactant H2O2 to the detection zones to initiate cobalt-based zeolitic imidazolate frameworks ZIF-67 catalyzed luminol-H2O2 CL and CL resonance energy transfer reactions. As proof-of-concept, a ZIF-67 regulated sensing strategy was developed for multiplex analysis of three model cancer biomarkers with a low detection limit of pg/mL to fg/mL, good selectivity, and low-cost, avoiding complicated labeling procedures and interferences from adjacent detection zones. This work opens a door for temporal-spatial-color multiresolved imaging for multiplex analysis.