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Quantum Dots — Characterization, Preparation and Usage in Biological Systems
Int. J. Mol. Sci. 2009, 10(4), 1930-1941; doi:10.3390/ijms10041930
Article

Fluorescence Lifetime Imaging of Quantum Dot Labeled DNA Microarrays

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1 School of Physics and Astronomy, The University of Edinburgh, The King’s Buildings, West Mains Road, Edinburgh EH9 3JZ, Scotland, UK 2 Division of Pathway Medicine, Medical School, The University of Edinburgh, Chancellor’s Building, Little France Crescent, Edinburgh EH16 4SB, Scotland, UK 3 School of Chemistry , The University of Edinburgh , Joseph Black Building, West Mains Road, Edinburgh, EH9 3JJ, Scotland, UK 4 Institute of Integrated Systems, School of Engineering, The University of Edinburgh, The King’s Buildings, West Mains Road, Edinburgh EH9 3JF, Scotland, UK 5 National Physical Laboratory, Hampton Road, Teddington, Middlesex TW11 0LW, UK
* Author to whom correspondence should be addressed.
Received: 4 March 2009 / Revised: 16 April 2009 / Accepted: 21 April 2009 / Published: 24 April 2009
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Abstract

Quantum dot (QD) labeling combined with fluorescence lifetime imaging microscopy is proposed as a powerful transduction technique for the detection of DNA hybridization events. Fluorescence lifetime analysis of DNA microarray spots of hybridized QD labeled target indicated a characteristic lifetime value of 18.8 ns, compared to 13.3 ns obtained for spots of free QD solution, revealing that QD labels are sensitive to the spot microenvironment. Additionally, time gated detection was shown to improve the microarray image contrast ratio by 1.8, achieving femtomolar target sensitivity. Finally, lifetime multiplexing based on Qdot525 and Alexa430 was demonstrated using a single excitation-detection readout channel.
Keywords: Quantum Dots (QD); Fluorescence Lifetime Imaging Microscopy (FLIM); DNA Microarray; Lifetime Multiplexing Quantum Dots (QD); Fluorescence Lifetime Imaging Microscopy (FLIM); DNA Microarray; Lifetime Multiplexing
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Giraud, G.; Schulze, H.; Bachmann, T.T.; Campbell, C.J.; Mount, A.R.; Ghazal, P.; Khondoker, M.R.; Ross, A.J.; Ember, S.W.J.; Ciani, I.; Tlili, C.; Walton, A.J.; Terry, J.G.; Crain, J. Fluorescence Lifetime Imaging of Quantum Dot Labeled DNA Microarrays. Int. J. Mol. Sci. 2009, 10, 1930-1941.

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