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蔡泊諺 (Po-Yen Tsai)
蔡泊諺 (Po-Yen Tsai)
Digital micro-fluidic chip for bead-based immunoassay with low bead number
Digital micro-fluidic chip for bead-based immunoassay with low bead number
低磁珠顆數之數位微流體免疫檢測晶片的研發
Bead-based immunoassay on digital micro-fluidic (DMF) device leads to higher sensitivity than conventional immunoassay in 96 well plate due to higher ratio of surface/volume. Most literatures of bead-based DMF immunoassay chip were focused on the pre-process of immunoassay, improvement of post-assay detection was rarely investigated. In addition, bead number applied in each immunoassay were mostly higher than 106 beads. Applying low bead number in DMF immunoassay chip is investigated in this research. Lower bead number leads to higher signal intensity on each bead, which lowers detection limit of immunoassay according to literature. But the least bead number reported was 250, the tendency is first evaluated in conditions of bead number under 100 in this research.
Dealing with bead washing on-chip with low bead number, techniques of droplet operation with electro-wetting on dielectric (EWOD) are investigated. “Double-sided EWOD bead washing” is developed in this research for low bead number cases by reducing bead loss and improving washing efficiency. Signal enhancement by bead aggregating, which was first proposed by our group, is integrated in the process. In order to compare on-chip results to off-chip ones in prior research, same detection target should be applied. Therefore human soluble tumor necrosis factor receptor-1 (sTNF-R1) is adopted as detection target.
Prior researches applied single-sided EWOD bead washing, which could be achieved only with high bead number of magnetic beads. Being tested in this research, if apply 6 μm diameter beads, 2,550 is the least number to achieve single-sided EWOD. Washing with lower bead number could only be achieved by double-sided EWOD. Washing efficiency of the improved double-sided EWOD washing could exceed 98% in only 1 cycle of wash and cause no bead loss. The whole assay process is accomplished on DMF chip, and proved the bead number effect still works as bead number lower than 100. Compared to prior results of our lab, the assay time is reduced into 1 hour, sample consumption is reduced into 200 nL and detection limit is reduced from 15 pg/mL to 3.14 pg/mL.
Journal papers
Journal papers
1. C.-Y. Huang, P.-Y. Tsai, I-C. Lee, H.-Y. Hsu, H.-Y. Huang, S.-K. Fan, D.-J. Yao, C.-H. Liu, W. Hsu, "A highly efficient bead extraction technique with low bead number for digital microfluidic immunoassay," Biomicrofluidics 2016, 10, 011901. (IF=3.357, N/M=3/31)
1. C.-Y. Huang, P.-Y. Tsai, I-C. Lee, H.-Y. Hsu, H.-Y. Huang, S.-K. Fan, D.-J. Yao, C.-H. Liu, W. Hsu, "A highly efficient bead extraction technique with low bead number for digital microfluidic immunoassay," Biomicrofluidics 2016, 10, 011901. (IF=3.357, N/M=3/31)
2. 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)
2. 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. Y.-H. Chung, C.-Y. Huang, P.-Y. Tsai, H.-Y. Hsu, and W. Hsu, "Investigation of three-dimension bead aggregation effect for optical sensing in digital microfluidics platform,” in The 5th International Conference on Optofluidics (Optofluidics 2015), Taipei, Taiwan, July 2015. (Oral)
1. Y.-H. Chung, C.-Y. Huang, P.-Y. Tsai, H.-Y. Hsu, and W. Hsu, "Investigation of three-dimension bead aggregation effect for optical sensing in digital microfluidics platform,” in The 5th International Conference on Optofluidics (Optofluidics 2015), Taipei, Taiwan, July 2015. (Oral)
2. Y.-H. Chung, P.-Y. Tsai, C.-Y. Huang, H.-Y. Hsu, and W. Hsu, "Effects of gap size and bead aggregation on fluorescence detection intensity in digital microfluidic immunoassay," in The 40th International Conference on Micro and Nano Engineering (MNE 2014), Lausanne, Switzerland, September 2014.
2. Y.-H. Chung, P.-Y. Tsai, C.-Y. Huang, H.-Y. Hsu, and W. Hsu, "Effects of gap size and bead aggregation on fluorescence detection intensity in digital microfluidic immunoassay," in The 40th International Conference on Micro and Nano Engineering (MNE 2014), Lausanne, Switzerland, September 2014.
3. C.-Y. Huang, P.-H. Shih, P.-Y. Tsai, I-C. Lee, H.-Y. Hsu, S.-K. Fan, and W. Hsu, "Enhanced fluorescence detection through post-immunoassay beads aggregation on bead-based immunoassay using digital microfluidics," in The 9th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE-NEMS 2014), Hawaii, USA, April 2014. (Oral)
3. C.-Y. Huang, P.-H. Shih, P.-Y. Tsai, I-C. Lee, H.-Y. Hsu, S.-K. Fan, and W. Hsu, "Enhanced fluorescence detection through post-immunoassay beads aggregation on bead-based immunoassay using digital microfluidics," in The 9th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE-NEMS 2014), Hawaii, USA, April 2014. (Oral)
4. P.-Y. Tsai, C.-Y. Huang, H.-Y. Hsu, and W. Hsu, "Low bead number washing for bead-based immunoassay using double sided EWOD and magnetic force," in The 9th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE-NEMS 2014), Hawaii, USA, April 2014. (Oral)
4. P.-Y. Tsai, C.-Y. Huang, H.-Y. Hsu, and W. Hsu, "Low bead number washing for bead-based immunoassay using double sided EWOD and magnetic force," in The 9th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE-NEMS 2014), Hawaii, USA, April 2014. (Oral)
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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