標題: AMPFLUID: Aggregation Magnified Post-Assay Fluorescence for Ultrasensitive Immunodetection on Digital Microfluidics
作者: Huang, Cheng-Yeh
Shih, Po-Huai
Tsai, Po-Yen
Lee, I-Chin
Hsu, Hsin-Yun
Huang, Hong-Yuan
Fan, Shih-Kang
Hsu, Wensyang
機械工程學系
應用化學系
Department of Mechanical Engineering
Department of Applied Chemistry
關鍵字: Aggregation;bead-based immunoassay;concentration;digital microfluidic (DMF);electrowetting on dielectric (EWOD);electrowetting;immunoassay;immunochip;lab-on-a-chip;magnetic beads;micro total analysis system (mu TAS);microfluidics
公開日期: 1-Feb-2015
摘要: Several strategies are currently employed to enhance the detection limit of bead-based assays, but all these approaches improve the sensitivity by varying the assay procedures chemically or biologically. In previous digital microfluidic setups for bead-based immunoassay, the magnetic beads were suspended for detection. We investigated the effect of bead aggregation in such an immunoassay system and propose the aggregation magnified post-assay fluorescence for ultrasensitive immunodetection on digital microfluidics (AMPFLUID). The detection signal and sensitivity are further enhanced even at the post-assay stage without altering the original assay protocol on employing magnetically triggered post-assay aggregation of beads in a digital microfluidic setup followed by processing of the fluorescent signal. This method is shown to enhance the fluorescent signal with increased consistency and sensitivity after appropriate charge-coupled device (CCD) calibration. This method of on-chip detection allows the fulfilment of consumption of a volume at the nanoliter level and a limit of detection in the range picogram/mL. In our sTNF-RI model immunoassay, only 2.5 nL of sample is required; a detection limit 15 pg/mL is achieved. The decreased uncertainty of the measure is indicated by the error bars and coefficient of variation.
URI: http://dx.doi.org/10.1109/JPROC.2014.2376967
http://hdl.handle.net/11536/124572
ISSN: 0018-9219
DOI: 10.1109/JPROC.2014.2376967
期刊: PROCEEDINGS OF THE IEEE
Volume: 103
起始頁: 225
結束頁: 235
Appears in Collections:Articles