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Use of Dual Optical Tweezers and Microfluidics for Single-Molecule Studies
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Improving Flow Bead Assay: Combination of Near-Infrared Optical Tweezers Stabilizing and Upconversion Luminescence

Bei Zheng1, Ya-Feng Kang1, Ting Zhang1

  • 1Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, People's Republic of China.

Analytical Chemistry
|March 12, 2020
PubMed
Summary

This study introduces a novel microfluidic platform using optical tweezers and upconversion luminescence for stable, sensitive detection of microRNAs (miRNAs). The method enables simultaneous quantification of specific miRNA sequences in complex biological samples.

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Area of Science:

  • Biomedical Engineering
  • Nanotechnology
  • Analytical Chemistry

Background:

  • Conventional flow bead-based fluorescence detection suffers from signal instability and background interference.
  • Existing methods often involve complex apparatus and signal fluctuations.
  • There is a need for enhanced stability and reduced background in fluorescence detection.

Purpose of the Study:

  • To develop a microfluidic chip-assisted platform for enhanced signal acquisition stability.
  • To diminish background interference in flow bead-based fluorescence detection.
  • To enable simultaneous quantitative determination of specific miRNA sequences.

Main Methods:

  • Integration of near-infrared optical tweezers with upconversion luminescence encoding.
  • Utilizing a single 980 nm laser for optical trapping and excitation of upconversion nanoparticles (UCNPs).
  • Employing two-color UCNPs (Er-UCNPs and Tm-UCNPs) with minimal spectral overlap for multiplexed detection.
  • Fabrication of UCNPs-miRNAs-microbead sandwich constructs via one-step nucleic acid hybridization.

Main Results:

  • Achieved stable signal acquisition and reduced background interference.
  • Demonstrated automatic and simultaneous quantitative determination of miRNA-205 and miRNA-21 sequences.
  • Obtained uniform terrace peaks with a picomolar (pM) level detection limit.
  • Successfully applied the technique to analyze cell lysates and human tissue lysates.

Conclusions:

  • The developed platform offers enhanced stability and sensitivity for miRNA detection.
  • The technique shows potential for disease diagnosis through analysis of biological samples.
  • The platform can be extended for investigating other biomolecules and analyzing particle heterogeneity in biological fluids.