CD (compact disc) - Based DNA hybridization and detection

Guangyao Jia, Kuo Sheng Ma, Jitae Kim, Jim V. Zoval, Marc J. Madou, Sapna K. Deo, Sylvia Daunert, Régis Peytavi, Michel G. Bergeron

Research output: Contribution to journalConference articlepeer-review

3 Scopus citations

Abstract

A DNA hybridization and detection unit was developed for a compact disc (CD) platform. The compact disc was used as the fluidic platform for sample and reagent manipulation using centrifugal force. Chambers for reagent storage and conduits for fluidic functions were replicated from polydimethylsiloxane (PDMS) using an SU-8 master mold fabricated with a 2-level lithography process we developed specially for the microfluidic structures used in this work. For capture probes, we used self-assembled DNA oligonucleotide monolayers (SAMs) on gold pads patterned on glass slides. The PDMS flow cells were aligned with and sealed against glass slides to form the DNA hybridization detection units. Both an enzymatic-labeled fluorescence technique and a bioluminescent approach were used for hybridization detection. An analytical model was introduced to quantitatively predict the accumulation of hybridized targets. The flow-through hybridization units were tested using DNA samples (25-mers) of different concentrations down to 1 pM and passive assays (no flow), using samples of the same concentrations, were performed as controls. At low concentrations, with the same hybridization time, a significantly higher relative fluorescence intensity was observed in both enzymatic and bioluminescent flow-through assays compared to the corresponding passive hybridization assays. Besides the fast hybridization rate, the CD-based method has the potential for enabling highly automated, multiple and self-contained assays for DNA detection.

Original languageEnglish (US)
Pages (from-to)341-352
Number of pages12
JournalProceedings of SPIE - The International Society for Optical Engineering
Volume5455
DOIs
StatePublished - Dec 2 2004
Externally publishedYes
EventMEM, MOEMS, and Micromachining - Strasbourg, France
Duration: Apr 29 2004Apr 30 2004

Keywords

  • Bioluminescence
  • Cd platform
  • Centrifugal
  • ELF 97
  • Flow cells
  • Flow-through dna hybridization
  • Microfluidics
  • Self-assembled monolayers (sams)
  • Su-8 photolithography

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

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