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Proceedings Paper

Superiorities of time-correlated single-photon counting against standard fluorimetry in exploiting the potential of fluorochromized oligonucleotide probes for biomedical investigation
Author(s): Marco Lamperti; Luca Nardo; Maria Bondani
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Paper Abstract

Site-specific fluorescence-resonance-energy-transfer donor-acceptor dual-labelled oligonucleotide probes are widely used in state-of-art biotechnological applications. Such applications include their usage as primers in polymerase chain reaction. However, the steady-state fluorescence intensity signal emitted by these molecular tools strongly depends from the specificities of the probe conformation. For this reason, the information which can be reliably inferred by steady-state fluorimetry performed on such samples is forcedly confined to a semi-qualitative level. Namely, fluorescent emission is frequently used as ON/OFF indicator of the probe hybridization state, i.e. detection of fluorescence signals indicates either hybridization to or detachment from the template DNA of the probe. Nonetheless, a fully quantitative analysis of their fluorescence emission properties would disclose other exciting applications of dual-labelled probes in biosensing. Here we show how time-correlated single-photon counting can be applied to get rid of the technical limitations and interpretational ambiguities plaguing the intensity analysis, and to derive information on the template DNA reaching single-base.

Paper Details

Date Published: 6 May 2015
PDF: 8 pages
Proc. SPIE 9504, Photon Counting Applications 2015, 95040L (6 May 2015); doi: 10.1117/12.2179002
Show Author Affiliations
Marco Lamperti, Univ. of Insubria (Italy)
Luca Nardo, Univ. of Milano Bicocca (Italy)
Maria Bondani, Institute for Photonics and Nanotechnologies, CNR (Italy)
CNISM (Italy)

Published in SPIE Proceedings Vol. 9504:
Photon Counting Applications 2015
Ivan Prochazka; Roman Sobolewski; Ralph B. James, Editor(s)

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