Digital microfluidics(DMF)shows great promise in addressing the need for miniaturization and automation in immunoassay detection.Despite recent advances,an automatically operated,multiplexed heterogeneous immunoassay ...Digital microfluidics(DMF)shows great promise in addressing the need for miniaturization and automation in immunoassay detection.Despite recent advances,an automatically operated,multiplexed heterogeneous immunoassay platform powered by DMF remains underdeveloped.Here we present a DMF platform for automated and multiplexed heterogeneous immunoassay detection by coupling spatial barcoding with automatic and uniform droplet dispensing.FluoroPel was selected as a robust hydrophobic reagent for coating the DMF top plate,and it also served as the substrate for the immuno-reaction.Its mechanical robustness was further enhanced with a Cytop CTL-809A adhesive layer under the top hydrophobic layer.Hourglass-shaped electrode patterns ensured consistent and uniform distribution of immunoassay reagents,with volume variation down to 1.0%.The analysis duration was significantly reduced from 75 min to 20 min after a heating module was integrated to elevate the immuno-reaction temperature to 37℃.Utilizing a compact instrument featuring a multi-droplet manipulation protocol,we successfully implemented fully operated,multi-sample,multiplexed immunoassays using recombinant proteins on cell culture supernatants on the DMF platform.This innovative platform significantly enhances the efficiency,reliability,and degree of automation of DMF-actuated multiplexed heterogeneous immunoassays,potentially providing a viable solution for field deployment and multi-sample parallel diagnosis.展开更多
基金supported by the National Key R&D Plan of China(No.2023YFB3210400)the National Natural Science Foundation of China(Nos.31927802,22574155)+2 种基金Liaoning Provincial Natural Science Foundation Young Scientists Fund Project(Category A,No.2025JH6/101100013)the fund from the Dalian Institute of Chemical Physics(Nos.DICP I202451,DMU-2&DICP UN202503)the Postgraduate Education Reform and Quality Improvement Project of Henan Province(No.YJS2023JD37)。
文摘Digital microfluidics(DMF)shows great promise in addressing the need for miniaturization and automation in immunoassay detection.Despite recent advances,an automatically operated,multiplexed heterogeneous immunoassay platform powered by DMF remains underdeveloped.Here we present a DMF platform for automated and multiplexed heterogeneous immunoassay detection by coupling spatial barcoding with automatic and uniform droplet dispensing.FluoroPel was selected as a robust hydrophobic reagent for coating the DMF top plate,and it also served as the substrate for the immuno-reaction.Its mechanical robustness was further enhanced with a Cytop CTL-809A adhesive layer under the top hydrophobic layer.Hourglass-shaped electrode patterns ensured consistent and uniform distribution of immunoassay reagents,with volume variation down to 1.0%.The analysis duration was significantly reduced from 75 min to 20 min after a heating module was integrated to elevate the immuno-reaction temperature to 37℃.Utilizing a compact instrument featuring a multi-droplet manipulation protocol,we successfully implemented fully operated,multi-sample,multiplexed immunoassays using recombinant proteins on cell culture supernatants on the DMF platform.This innovative platform significantly enhances the efficiency,reliability,and degree of automation of DMF-actuated multiplexed heterogeneous immunoassays,potentially providing a viable solution for field deployment and multi-sample parallel diagnosis.
