Single-molecule tracing on a fluidic microchip for quantitative detection of low-abundance nucleic acids

Tza Huei Wang, Yahui Peng, Chunyang Zhang, Pak Kin Wong, Chih Ming Ho

Research output: Contribution to journalArticle

109 Citations (Scopus)

Abstract

Here, we report a method capable of quantitative detection of low-abundance DNA/RNA molecules by incorporating confocal fluorescence spectroscopy, molecular beacons, and a molecular-confinement microfluidic reactor. By using a combination of ac and dc fields via a trio of 3-D electrodes in the microreactor, we are able to precisely direct the transport of individual molecules to a minuscule laser-focused detection volume (∼1 fL). A burst of fluorescence photons is detected whenever a molecular beacon-target hybrid flows through the detection region, and the amount of targets can be directly quantified according to the number of recorded single-molecule flow-through events. This assay consumes only attomoles of molecular probes and is able to quantitatively detect subpicomolar DNA targets. A measurement time of less than 2 min is sufficient to complete the detection.

Original languageEnglish (US)
Pages (from-to)5354-5359
Number of pages6
JournalJournal of the American Chemical Society
Volume127
Issue number15
DOIs
StatePublished - Apr 20 2005
Externally publishedYes

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Nucleic acids
Fluidics
Nucleic Acids
Molecular Probes
Molecules
Microfluidics
DNA
Fluorescence Spectrometry
Photons
Electrodes
Lasers
Fluorescence
Fluorescence spectroscopy
RNA
Time measurement
Assays

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Single-molecule tracing on a fluidic microchip for quantitative detection of low-abundance nucleic acids. / Wang, Tza Huei; Peng, Yahui; Zhang, Chunyang; Wong, Pak Kin; Ho, Chih Ming.

In: Journal of the American Chemical Society, Vol. 127, No. 15, 20.04.2005, p. 5354-5359.

Research output: Contribution to journalArticle

Wang, Tza Huei ; Peng, Yahui ; Zhang, Chunyang ; Wong, Pak Kin ; Ho, Chih Ming. / Single-molecule tracing on a fluidic microchip for quantitative detection of low-abundance nucleic acids. In: Journal of the American Chemical Society. 2005 ; Vol. 127, No. 15. pp. 5354-5359.
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