Fluorescence lifetime imaging of quantum dot labeled DNA microarrays.
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.
Giraud
Gerard
G
School of Physics and Astronomy, The University of Edinburgh, The King's Buildings, West Mains Road, Edinburgh EH9 3JZ, Scotland, UK.
Schulze
Holger
H
Division of Pathway Medicine, Medical School, The University of Edinburgh, Chancellor's Building, Little France Crescent, Edinburgh EH16 4SB, Scotland, UK.
Bachmann
Till T
TT
Division of Pathway Medicine, Medical School, The University of Edinburgh, Chancellor's Building, Little France Crescent, Edinburgh EH16 4SB, Scotland, UK.
Campbell
Colin J
CJ
Division of Pathway Medicine, Medical School, The University of Edinburgh, Chancellor's Building, Little France Crescent, Edinburgh EH16 4SB, Scotland, UK.
Mount
Andrew R
AR
School of Chemistry , The University of Edinburgh , Joseph Black Building, West Mains Road, Edinburgh, EH9 3JJ, Scotland, UK.
Ghazal
Peter
P
Division of Pathway Medicine, Medical School, The University of Edinburgh, Chancellor's Building, Little France Crescent, Edinburgh EH16 4SB, Scotland, UK.
Khondoker
Mizanur R
MR
Division of Pathway Medicine, Medical School, The University of Edinburgh, Chancellor's Building, Little France Crescent, Edinburgh EH16 4SB, Scotland, UK.
Ross
Alan J
AJ
Division of Pathway Medicine, Medical School, The University of Edinburgh, Chancellor's Building, Little France Crescent, Edinburgh EH16 4SB, Scotland, UK.
Ember
Stuart W J
SWJ
Division of Pathway Medicine, Medical School, The University of Edinburgh, Chancellor's Building, Little France Crescent, Edinburgh EH16 4SB, Scotland, UK.
Ciani
Ilenia
I
School of Chemistry , The University of Edinburgh , Joseph Black Building, West Mains Road, Edinburgh, EH9 3JJ, Scotland, UK.
Tlili
Chaker
C
School of Chemistry , The University of Edinburgh , Joseph Black Building, West Mains Road, Edinburgh, EH9 3JJ, Scotland, UK.
Walton
Anthony J
AJ
Institute of Integrated Systems, School of Engineering, The University of Edinburgh, The King's Buildings, West Mains Road, Edinburgh EH9 3JF, Scotland, UK.
Terry
Jonathan G
JG
Institute of Integrated Systems, School of Engineering, The University of Edinburgh, The King's Buildings, West Mains Road, Edinburgh EH9 3JF, Scotland, UK.
Crain
Jason
J
School of Physics and Astronomy, The University of Edinburgh, The King's Buildings, West Mains Road, Edinburgh EH9 3JZ, Scotland, UK.|National Physical Laboratory, Hampton Road, Teddington, Middlesex TW11 0LW, UK.
D003587:Cytomegalovirus / Q000235:genetics; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism; D016174:Hepacivirus / Q000235:genetics; D006801:Humans; D008856:Microscopy, Fluorescence; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D045663:Quantum Dots; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism
D004279:DNA, Viral; D012367:RNA, Viral; D004247:DNA