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Proceedings Paper

Toward the development of optical nucleic acid biosensors based on TIRF and TCSPC for high sensitivity determinations
Author(s): Paul A.E. Piunno; Virginijus Barzda; Sarah C. Jantzi; Christopher C. Kotoris; Arkady Major; Sergei Musikhin; Sandeep Raha; Ulrich J. Krull
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Paper Abstract

The results of preliminary investigations toward the design of an optical biosensor instrument for the selective and direct analysis of low copy numbers of target nucleic acids in native form are reported. A concept development prototype was constructed based on a total internal reflection fluorescence (TIRF) configuration and the use of time correlated single photon counting (TCSPC). Selective detection of interfacial hybrid formation was done by identification of luminescence of characteristic (20ns) lifetime from the intercalant fluorophore ethidium bromide associated with nucleic acid hybrids formed at the interaction surface of optical sensor elements. Results of these investigations suggest that detection limits on the order of 107 dye:dsDNA complexes can be achieved when an effective sensor interaction surface of 150 µm diameter is used. The presence of interfacial nucleic acid duplexes at a sensor surface was further verified by thermal denaturation studies. The sensitivity of this concept design prototype was found to be most limited by long lifetime fluorescence intrinsic to the detection optics in conjunction with large amounts of scatter dispersed from the sensor cartridge. Future directions for continued device development are discussed.

Paper Details

Date Published: 13 October 2005
PDF: 9 pages
Proc. SPIE 5969, Photonic Applications in Biosensing and Imaging, 59690R (13 October 2005); doi: 10.1117/12.628696
Show Author Affiliations
Paul A.E. Piunno, Univ. of Toronto at Mississauga (Canada)
Virginijus Barzda, Univ. of Toronto at Mississauga (Canada)
Univ. of Toronto (Canada)
Sarah C. Jantzi, Univ. of Toronto at Mississauga (Canada)
Christopher C. Kotoris, Univ. of Toronto at Mississauga (Canada)
Arkady Major, Univ. of Toronto at Mississauga (Canada)
Univ. of Toronto (Canada)
Sergei Musikhin, Univ. of Toronto at Mississauga (Canada)
Sandeep Raha, Univ. of Toronto at Mississauga (Canada)
Ulrich J. Krull, Univ. of Toronto at Mississauga (Canada)

Published in SPIE Proceedings Vol. 5969:
Photonic Applications in Biosensing and Imaging
Brian C. Wilson; Richard I. Hornsey; Warren C. W. Chan; Ulrich J. Krull; Robert A. Weersink; Kui Yu, Editor(s)

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