Dielectrophoresis assisted high-throughput detection system for multiplexed immunoassays.
Dielectrophoresis assisted high-throughput detection system for multiplexed immunoassays.
复制标题
DOI：
10.1016/j.bios.2021.113148
复制
发表时间：
2021-03
影响因子：
12.6
通讯作者：
Shih-Mo Yang;Q. Lin;Hongbo Zhang;R. Yin;Wenjun Zhang;Minchao Zhang;Yubao Cui
中科院分区：
文献类型：
--
作者：
Shih-Mo Yang;Q. Lin;Hongbo Zhang;R. Yin;Wenjun Zhang;Minchao Zhang;Yubao Cui
数字化酶联免疫吸附试验是一种新型的检测平台，具有独特的优势，可以检测样品中的多种单分子。如何提高检测的灵敏度是当前相关研究的方向。在这里，我们报告了一种免疫分析方法，应用电动效应分离单个编码珠并限制在微孔中，以同时提高细胞因子检测的效率。微流控设计提供了一个非均匀的电场诱导介电泳（DEP）力和操纵的珠子。两种波长的激发光激发编码的珠粒，用于同时检测报告基因。光通过全内反射原理被限制在底部载玻片上。最后，通过从图像中拾取每个珠，然后对从报告子发射的荧光强度进行积分，获得捕获的细胞因子的浓度。结果表明，通过DEP效应，编码珠的填充率从10-20%提高到60-80%。通过比较颗粒、颗粒本身及其表面的荧光颜色，计算出4种靶细胞因子IL-2、IL-6、IL-10和TNF-α的浓度，达到pg/ml水平。通过加标和回收实验验证了该方法的有效性，超过70%的目标分子被捕获。流式细胞术也验证了该方法的可靠性。总之，我们期望DEP的应用可以提高数字化ELISA的灵敏度，用于多重快速检测。
Digital ELISA is introduced as a novel platform with unique advantages for detecting multiple kinds of single-molecule 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-α, 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.