Quantitative characterization of quantum dot-labeled lambda phage for Escherichia coli detection §

Authors

  • Peter B. Yim,

    1. National Institute of Standards and Technology, 100 Bureau Drive, Mailstop 8443, Gaithersburg, Maryland 20899; telephone: 301-975-4580; fax: 301-975-6991
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  • Matthew L. Clarke,

    1. National Institute of Standards and Technology, 100 Bureau Drive, Mailstop 8443, Gaithersburg, Maryland 20899; telephone: 301-975-4580; fax: 301-975-6991
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  • Michael McKinstry,

    1. Biological Defense Research Directorate, Naval Medical Research Center, Silver Spring, Maryland
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  • Silvia H. De Paoli Lacerda,

    1. National Institute of Standards and Technology, 100 Bureau Drive, Mailstop 8443, Gaithersburg, Maryland 20899; telephone: 301-975-4580; fax: 301-975-6991
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  • Leonard F. Pease III,

    1. National Institute of Standards and Technology, 100 Bureau Drive, Mailstop 8443, Gaithersburg, Maryland 20899; telephone: 301-975-4580; fax: 301-975-6991
    2. Chemical Engineering Department, University of Utah, Salt Lake City, Utah
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  • Marina A. Dobrovolskaia,

    1. Nanotechnology Characterization Laboratory, SAIC-Frederick/NCI-Frederick, Frederick, Maryland
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  • HyeongGon Kang,

    1. National Institute of Standards and Technology, 100 Bureau Drive, Mailstop 8443, Gaithersburg, Maryland 20899; telephone: 301-975-4580; fax: 301-975-6991
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  • Timothy D. Read,

    1. Department of Human Genetics, Division of Infectious Diseases, Emory University School of Medicine, Atlanta, Georgia
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  • Shanmuga Sozhamannan,

    1. Biological Defense Research Directorate, Naval Medical Research Center, Silver Spring, Maryland
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  • Jeeseong Hwang

    Corresponding author
    1. National Institute of Standards and Technology, 100 Bureau Drive, Mailstop 8443, Gaithersburg, Maryland 20899; telephone: 301-975-4580; fax: 301-975-6991
    • National Institute of Standards and Technology, 100 Bureau Drive, Mailstop 8443, Gaithersburg, Maryland 20899; telephone: 301-975-4580; fax: 301-975-6991.
    Search for more papers by this author

  • Certain commercial equipment, instruments, or materials are identified in this paper to foster understanding and does not imply recommendation or endorsement by NIST, nor does it imply that the materials or equipment identified are necessarily the best available for the purpose. The views expressed in this article are those of the authors and do not necessarily reflect the official policy or position of the Department of the Navy, Department of Defense, nor the U.S. Government. The BDRD authors are employees of the U. S. Government. This work was prepared as part of their official duties. Title 17 U.S.C. §105 provides that “Copyright protection under this title is not available for any work of the United States Government.” Title 17 U.S.C. §101 defines a U.S. Government work as a work prepared by a military service member or employee of the U.S. Government as part of that person's official duties.

  • Peter B. Yim and Matthew L. Clarke contributed equally to this work.

  • §

    This article is a U.S. Goverment work and is in the public domain in the U.S.A.

Abstract

We characterize CdSe/ZnS quantum dot (QD) binding to genetically modified bacteriophage as a model for bacterial detection. Interactions among QDs, lambda (λ) phage, and Escherichia coli are examined by several cross-validated methods. Flow and image-based cytometry clarify fluorescent labeling of bacteria, with image-based cytometry additionally reporting the number of decorated phage bound to cells. Transmission electron microscopy, image-based cytometry, and electrospray differential mobility analysis allow quantization of QDs attached to each phage (4–17 QDs) and show that λ phage used in this study exhibits enhanced QD binding to the capsid by nearly a factor of four compared to bacteriophage T7. Additionally, the characterization methodology presented can be applied to the quantitative characterization of other fluorescent nanocrystal-biological conjugates. Biotechnol. Bioeng. 2009;104: 1059–1067. Published 2009 Wiley Periodicals, Inc.

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