文献类型：期刊文献

英文题名：Dielectrophoresis assisted high-throughput detection system for multiplexed immunoassays

作者：Yang, Shih-Mo[1];Lin, Qiang[1];Zhang, Hongbo[2];Yin, Ruixue[2];Zhang, Wenjun[3];Zhang, Minchao[4];Cui, Yubao[5]

机构：[1]Shanghai Univ, Biomed Sci & Technol Res Ctr, Sch Mechatron Engn & Automat, Shanghai, Peoples R China;[2]East China Univ Sci & Technol, Sch Mech & Power Engn, Shanghai, Peoples R China;[3]Univ Saskatchewan, Div Biomed Engn, Saskatoon, SK, Canada;[4]Colortech Suzhou Biotechnol, Suzhou, Peoples R China;[5]Nanjing Med Univ, Dept Clin Lab, Wuxi Peoples Hosp, Wuxi, Jiangsu, Peoples R China

年份：2021

卷号：180

外文期刊名：BIOSENSORS & BIOELECTRONICS

收录：;EI(收录号：20211110061692);WOS:【SCI-EXPANDED(收录号:WOS:000634109900005)】；

基金：This study was supported by the National Natural Science Foundation of China (Grant No. NSFC81971511), the 333 project of Jiangsu Province in 2017 (BRA2017216), the Primary Research and Development Plan of Jiangsu Province (Grant No. BE2018627), the Project of Wuxi Health Commission (Grant No. MS201949), and by 111 Project of China (D18003), the Shanghai Lianmeng Grant (LM201603), the Opening Project of the Shanghai Key Laboratory of Orthopedic Implant (KFKT2018001).

语种：英文

外文关键词：Multiplexed; Immunoassay; Dielelctrophoresis; High-throughput; Microfluidic

摘要：Digital ELISA is introduced as a novel platform with unique advantages for detecting multiple kinds of singlemolecule in the sample. How to improve the sensitivity of detection is the direction of current related research. Here, we report an immunoassay method that applied electrokinetic effects to isolate the individual encoded beads and confine in micro-wells to improve the efficiency of cytokines detection simultaneously. The microfluidic design provided a non-uniform electric field to induce dielectrophoresis (DEP) force and to manipulate the beads. Two wavelengths of excitation light excited the encoded beads for simultaneous detection of reporters. The light was confined to the bottom slide via the principle of total internal reflection. Finally, the concentration of captured cytokines was obtained by picking up each bead from the image and then integrating the intensity of fluorescent light emitted from the reporters. The results demonstrated that the fill percentage of encoded beads was raised from 10-20% to 60?80% via DEP effect. By comparing the fluorescence color of the particle, itself and its surface, the concentration of four target cytokines, IL-2, IL-6, IL-10 and TNF-?, were calculated to the pg/ml level. The spike and recovery experiments verified the efficiency, more than 70% of the target molecules were captured. The reliability of our method was verified by flow cytometry as well. In conclusion, we expect the application of DEP can increase the sensitivity of digital ELISA for multiple rapid detection.