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施博懷 (Po-Huai Shih)
Ultrasensitive Fluorescence Detection of Bead-Based Immunoassay on Digital Microfluidics by Aggregating Magnetic Beads
Ultrasensitive Fluorescence Detection of Bead-Based Immunoassay on Digital Microfluidics by Aggregating Magnetic Beads
以磁珠聚集法提升數位微流體式免疫晶片之螢光偵測能力
以磁珠聚集法提升數位微流體式免疫晶片之螢光偵測能力
Most of the references proposed that manipulating micro-beads to get involved in mixing and washing of immunoassay pre-process, but few about detecting sensitivity enhancement. In the research, a novel digital micro-fluidic chip (DMF chip) for immunoassay had been developed, which can create sample droplet in nano-scale by electro-wetting on dielectric (EWOD) force and aggregating bio-conjugated beads by magnetic force. The features of DMF chip are high sensitivity and low limit of detection (LOD). Take fluorescent labeled tumor-necrosis factor receptor-1 (TNFR1) as a detecting target antigen. Firstly, calibrate CCD black balance to the proper range of fluorescent exposure time from 60 to 80 ms to reduce the interference of unwanted background noise. Then the result indicated that the longer fluorescent exposure time in the proper range the more fluorescent sensitivity and the lower LOD. In addition, the fluorescent detecting sensitivity can also be enhanced by aggregating bio-conjugated magnetic beads. Finally, take flow cytometry as a TNFR1 detecting control group to prove the functions of high fluorescent sensitivity and low LOD on DMF chip. The results of flow cytometry shows that LOD is 146 pg/ml and detecting sample consumption is 50 μl. In contrast with the flow cytometry, LOD and detecting sample consumption of DMF chip platform are 31.47 pg/ml and 2.5 nl. In summary, the advantages of DMF chip in the research are high fluorescent detecting sensitivity, low limit of detection and sample consumption.
Most of the references proposed that manipulating micro-beads to get involved in mixing and washing of immunoassay pre-process, but few about detecting sensitivity enhancement. In the research, a novel digital micro-fluidic chip (DMF chip) for immunoassay had been developed, which can create sample droplet in nano-scale by electro-wetting on dielectric (EWOD) force and aggregating bio-conjugated beads by magnetic force. The features of DMF chip are high sensitivity and low limit of detection (LOD). Take fluorescent labeled tumor-necrosis factor receptor-1 (TNFR1) as a detecting target antigen. Firstly, calibrate CCD black balance to the proper range of fluorescent exposure time from 60 to 80 ms to reduce the interference of unwanted background noise. Then the result indicated that the longer fluorescent exposure time in the proper range the more fluorescent sensitivity and the lower LOD. In addition, the fluorescent detecting sensitivity can also be enhanced by aggregating bio-conjugated magnetic beads. Finally, take flow cytometry as a TNFR1 detecting control group to prove the functions of high fluorescent sensitivity and low LOD on DMF chip. The results of flow cytometry shows that LOD is 146 pg/ml and detecting sample consumption is 50 μl. In contrast with the flow cytometry, LOD and detecting sample consumption of DMF chip platform are 31.47 pg/ml and 2.5 nl. In summary, the advantages of DMF chip in the research are high fluorescent detecting sensitivity, low limit of detection and sample consumption.
Journal papers
Journal papers
1. C.-Y. Huang, P.-H. Shih, P.-Y. Tsai, I-C. Lee, H.-Y. Hsu, H.-Y. Huang, S.-K. Fan, and W. Hsu, "AMPFLUID: Aggregation magnified post-assay fluorescence for ultrasensitive immunodetection on digital microfluidics." Proceedings of the IEEE 2015, 103, 225-235. (IF=5.466, N/M=6/248)
1. C.-Y. Huang, P.-H. Shih, P.-Y. Tsai, I-C. Lee, H.-Y. Hsu, H.-Y. Huang, S.-K. Fan, and W. Hsu, "AMPFLUID: Aggregation magnified post-assay fluorescence for ultrasensitive immunodetection on digital microfluidics." Proceedings of the IEEE 2015, 103, 225-235. (IF=5.466, N/M=6/248)
International conference papers
1. T.-M. Chuo, P.-H. Shih, W. Hsu, C.-Y. Huang, Y.-S. Huang, Y.-T. Chen, and S.-K. Fan (November 2012). Signal Enhancement by Particle Concentration Using Dielectrophoresis in an Electrowetting-driven Droplet. The 6th IEEE International Conference on Nano/Molecular Medicine and Engineering, Bangkok, Thailand. (Best Student Paper Finalist)
1. T.-M. Chuo, P.-H. Shih, W. Hsu, C.-Y. Huang, Y.-S. Huang, Y.-T. Chen, and S.-K. Fan (November 2012). Signal Enhancement by Particle Concentration Using Dielectrophoresis in an Electrowetting-driven Droplet. The 6th IEEE International Conference on Nano/Molecular Medicine and Engineering, Bangkok, Thailand. (Best Student Paper Finalist)
2. C.-Y. Huang, P.-H. Shih, P.-Y. Tsai, I-C. Lee, H.-Y. Hsu, S.-K. Fan, and W. Hsu (April 2014). Enhanced fluorescence detection through post-immunoassay beads aggregation on bead-based immunoassay using digital microfluidics. The 9th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems, Hawaii, U.S.A..
2. C.-Y. Huang, P.-H. Shih, P.-Y. Tsai, I-C. Lee, H.-Y. Hsu, S.-K. Fan, and W. Hsu (April 2014). Enhanced fluorescence detection through post-immunoassay beads aggregation on bead-based immunoassay using digital microfluidics. The 9th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems, Hawaii, U.S.A..
Patents
Patents
1. 徐文祥,黄正鄴,蔡泊諺,施博懷,范士岡,饒達仁,劉承賢,黃泓淵. 磁珠式數位微流體免疫分析裝置與其方法. Taiwan Patent. (Pending)
1. 徐文祥,黄正鄴,蔡泊諺,施博懷,范士岡,饒達仁,劉承賢,黃泓淵. 磁珠式數位微流體免疫分析裝置與其方法. Taiwan Patent. (Pending)
2. W. Hsu, C.-Y. Huang, P.-Y. Tsai, P.-H. Shih, S.-K. Fan, D.-J. Yao, C.-H. Liu, and H.-Y. Huang. Magnetic bead–based digital microfluidic immunoanalysis device and method thereof. US Patent. (Pending)
2. W. Hsu, C.-Y. Huang, P.-Y. Tsai, P.-H. Shih, S.-K. Fan, D.-J. Yao, C.-H. Liu, and H.-Y. Huang. Magnetic bead–based digital microfluidic immunoanalysis device and method thereof. US Patent. (Pending)
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