20 February 2014 Amplification and modulation of fluorescent signals by using hybridization chain reactions for multiplexed sensing of biomolecules in a one-pot
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Fluorescence readout of molecular information is a promising approach for biomolecular sensing. For detection of enormous biomolecules via uorescence, biomolecular information should be converted to codes that can be readout easily and simultaneously. For the purpose, we study a biomolecule uorescence color (B/F) encoders that modulate uorescence signals by control of uorescence resonance energy transfer (FRET). The B/F encoder converts biomolecular signals into uorescent color codes represented with uorescent wavelengths and intensity levels. The combination offers a great number of codes for representing the biomolecular information. In this study, we discuss multiplexed detection of target biomolecules using B/F encoders. Use of the B/F encoders would offer a multiplexed biomolecular sensing in a one-pot without micro-fabrication like DNA microarray. In the experiments, we prepared B/F encoders based on two kinds of hybridization chain reactions (HCR) that make long double-stranded DNA polymers to control positions of uorescence and quencher molecules. In the B/F encoders, target molecules trigger to start assembling the polymer structures. The uorescent molecules in the absence of the targets are near the quenchers and the output uorescence is suppressed by FRET. The polymerization process separates the uorescent and quencher dyes and the uorescent signal increase. The experimental results show that the B/F encoders based on HCRs have linear and independent response to each target, and temporal signals during the encoding reactions are usable for multiplexed readout. This result leads to the multiplexed sensing in a one-pot by uorescent ampli cation and multiple uorescent color-coding.
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Takahiro Nishimura, Takahiro Nishimura, Yusuke Ogura, Yusuke Ogura, Kenji Yamada, Kenji Yamada, Yuko Ohno, Yuko Ohno, Jun Tanida, Jun Tanida, "Amplification and modulation of fluorescent signals by using hybridization chain reactions for multiplexed sensing of biomolecules in a one-pot", Proc. SPIE 8954, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications XI, 89540P (20 February 2014); doi: 10.1117/12.2041436; https://doi.org/10.1117/12.2041436

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