Bio-assay based on single molecule fluorescence detection in microfluidic channels

Christopher W. Hollars, Jana Puls, Olgica Bakajin, Brad Olsan, Chad E. Talley, Stephen M. Lane, Thomas R Huser

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

A rapid bioassay is described based on the detection of colocalized fluorescent DNA probes bound to DNA targets in a pressure-driven solution flowing through a planar microfluidic channel. By employing total internal reflection excitation of the fluorescent probes and illumination of almost the entire flow channel, single fluorescent molecules can be efficiently detected leading to the rapid analysis of nearly the entire solution flowed through the device. Cross-correlation between images obtained from two spectrally distinct probes is used to determine the target concentration and efficiently reduces the number of false positives. The rapid analysis of DNA targets in the low pM range in less than a minute is demonstrated.

Original languageEnglish (US)
Pages (from-to)1384-1388
Number of pages5
JournalAnalytical and Bioanalytical Chemistry
Volume385
Issue number8
DOIs
StatePublished - Aug 2006

Keywords

  • Biomedical assay
  • Colocalization
  • Image cross-correlation
  • Microfluidics
  • Single molecule fluorescence
  • TIRF

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry

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