Integration and Quantitative Visualization of 3,3′,5,5′-Tetramethylbenzidine-Probed Enzyme-Linked Immunosorbent Assay-like Signals in a Photothermal Bar-Chart Microfluidic Chip for Multiplexed Immunosensing
Guanglei Fu, Ruixia Hou, Xianbo Mou, Xiujun Li
Abstract
The photothermal effect shows significant promise for various biomedical applications but is rarely exploited for microfluidic lab-on-a-chip bioassays. Herein, a photothermal bar-chart microfluidic immunosensing chip, with the integration of the conventional 3,3′,5,5′-tetramethylbenzidine (TMB)-probed enzyme-linked immunosorbent assay (ELISA)-like system, was developed based on exploiting the photothermal pumping technique for visual bar-chart microfluidic immunosensing. Both the sandwich ELISA-like system and the photothermal pumping protocol were integrated into a single photothermal bar-chart chip. On-chip immunocaptured iron oxide nanoparticles catalyzed the oxidation of the chromogenic substrate, TMB, to produce a sensitive photothermal and chromogenic dual-functional probe, oxidized TMB. As the result of heat generation and the subsequent production of elevating vapor pressure in the sealed microfluidic environment, the on-chip near-infrared laser-driven photothermal effect of the probe served as a dose-dependent pumping force to drive the multiplexed quantitative display of the immunosensing signals as visual dye bar charts. Prostate-specific antigen as a model analyte was tested at a limit of detection of 1.9 ng·mL–1, lower than the clinical diagnostic threshold of prostate cancer. This work presents a new perspective for microfluidic integration and multiplexed quantitative bar-chart visualization of the conventional TMB-probed ELISA signals possibly by means of an affordable handheld laser pointer in a lab-on-a-chip format.