16 August 2004 CD (compact disc)-based DNA hybridization and detection
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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.
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Guangyao Jia, Guangyao Jia, Kuo-Sheng Ma, Kuo-Sheng Ma, Jitae Kim, Jitae Kim, Jim V. Zoval, Jim V. Zoval, Marc J. Madou, Marc J. Madou, Sapna K. Deo, Sapna K. Deo, Sylvia Daunert, Sylvia Daunert, Regis Peytavi, Regis Peytavi, Michel G. Bergeron, Michel G. Bergeron, } "CD (compact disc)-based DNA hybridization and detection", Proc. SPIE 5455, MEMS, MOEMS, and Micromachining, (16 August 2004); doi: 10.1117/12.548754; https://doi.org/10.1117/12.548754
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