Heterogeneous immunoassays in microfluidic format using fluorescence detection with integrated amorphous silicon photodiodes |
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Authors: | Pereira A T Novo P Prazeres D M F Chu V Conde J P |
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Institution: | 1INESC Microsistemas e Nanotecnologias and IN-Institute of Nanoscience and Nanotechnology, R. Alves Redol, 9, 1000-029 Lisbon, Portugal;2IBB-Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Av. Rovisco Pais, 1, 1049-001 Lisbon, Portugal;3Department of Chemical and Biological Engineering, Instituto Superior Técnico, Av. Rovisco Pais, 1, 1049-001 Lisbon, Portugal |
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Abstract: | Miniaturization of immunoassays through microfluidic technology has the potential to decrease the time and the quantity of reactants required for analysis, together with the potential of achieving multiplexing and portability. A lab-on-chip system incorporating a thin-film amorphous silicon (a-Si:H) photodiode microfabricated on a glass substrate with a thin-film amorphous silicon-carbon alloy directly deposited above the photodiode and acting as a fluorescence filter is integrated with a polydimethylsiloxane-based microfluidic network for the direct detection of antibody-antigen molecular recognition reactions using fluorescence. The model immunoassay used consists of primary antibody adsorption to the microchannel walls followed by its recognition by a secondary antibody labeled with a fluorescent quantum-dot tag. The conditions for the flow-through analysis in the microfluidic format were defined and the total assay time was 30 min. Specific molecular recognition was quantitatively detected. The measurements made with the a-Si:H photodiode are consistent with that obtained with a fluorescence microscope and both show a linear dependence on the antibody concentration in the nanomolar-micromolar range. |
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