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Nat Protoc
. 2006;1(3):1160-4. doi: 10.1038/nprot.2006.162.
Creating Self-Illuminating Quantum Dot Conjugates
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PMID: 17406398
Abstract
Semiconductor quantum dots are inorganic fluorescent nanocrystals that, because of their unique optical properties compared with those of organic fluorophores, have become popular as fluorescent imaging probes. Although external light excitation is typically required for imaging with quantum dots, a new type of quantum dot conjugate has been reported that can luminesce with no need for external excitation. These self-illuminating quantum dot conjugates can be prepared by coupling of commercially available carboxylate-presenting quantum dots to the light-emitting protein Renilla luciferase. When the conjugates are exposed to the luciferase's substrate coelenterazine, the energy released by substrate catabolism is transferred to the quantum dots through bioluminescence resonance energy transfer, leading to quantum dot light emission. This protocol describes step-by-step procedures for the preparation and characterization of these self-illuminating quantum dot conjugates. The preparation process is relatively simple and can be done in less than 2 hours. The availability of self-illuminating quantum dot conjugates will provide many new possibilities for in vivo imaging and detection, such as monitoring of in vivo cell trafficking, multiplex bioluminescence imaging and new quantum dot-based biosensors.
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Publication types
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
MeSH terms
Energy Transfer
Imidazoles
Luciferases, Renilla
Luminescence*
Nanotechnology / methods*
Pyrazines
Quantum Dots*
Substances
Imidazoles
Pyrazines
coelenterazine
Luciferases, Renilla
Grant support
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Acknowledgements
This work was supported by the Burroughs Wellcome Fund and the National Cancer Institute Centers of Cancer Nanotechnology Excellence (1U54CA119367-01).
Author information
Affiliations
Department of Radiology and Bio-X Program, Molecular Imaging Program at Stanford, Stanford University School of Medicine, 1201 Welch Road, Stanford, 94305-5484, California, USA
Min-Kyung So
, Andreas M Loening
, Sanjiv S Gambhir
& Jianghong Rao
Department of Bioengineering, Stanford University School of Medicine, 1201 Welch Road, Stanford, 94305-5484, California, USA
Andreas M Loening
& Sanjiv S Gambhir
Departments of Biophysics and Cancer Biology Program, Stanford University School of Medicine, 1201 Welch Road, Stanford, 94305-5484, California, USA
Jianghong Rao
Contributions
M.-K.S. did the conjugation and imaging experiments and prepared figures; A.M.L. prepared Luc8 and wrote the manuscript; S.S.G. provided discussions; and J.R. designed and coordinated research, analyzed data and wrote the manuscript.
Corresponding author
Correspondence to Jianghong Rao.
Ethics declarations
Competing interests
Stanford University is seeking to file a patent application covering part of the information contained in the paper.
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Cite this article
So, M., Loening, A., Gambhir, S. et al. Creating self-illuminating quantum dot conjugates. Nat Protoc 1, 1160–1164 (2006). https://doi.org/10.1038/nprot.2006.162
Published07 September 2006
Issue DateAugust 2006
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