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Temporal-spatial-color Multi-resolved 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, on-site 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 multi-resolved CL imaging strategy for multiplex label-free immunoassays using a smartphone coupled with microfluidic chip. Temporal-spatial-color multi-resolved CL imaging signals were produced by sequentially transporting co-reactant 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 devel-oped for multiplex analysis of three model cancer biomarkers with 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 multi-resolved imaging for multiplex analysis.