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

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 b...

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Bibliographic Details
Published in:Biotechnology and bioengineering 2009-12, Vol.104 (6), p.1059-1067
Main Authors: Yim, Peter B, Clarke, Matthew L, McKinstry, Michael, De Paoli Lacerda, Silvia H, Pease, Leonard F. III, Dobrovolskaia, Marina A, Kang, HyeongGon, Read, Timothy D, Sozhamannan, Shanmuga, Hwang, Jeeseong
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacteria
bacteria detection
Bacteriophage lambda - metabolism
Bacteriophage lambda - physiology
Bacteriophages
Base Sequence
Binding
Binding sites
Biological and medical sciences
Biotechnology
Capsid Proteins - metabolism
Cells
Cytometry
E coli
electron-spray differential mobility assay
Escherichia coli
Escherichia coli - isolation & purification
Escherichia coli - virology
Flow Cytometry
Fluorescence
Fundamental and applied biological sciences. Psychology
Genetics
Mathematical models
microscopy
Microscopy, Electron, Transmission
Molecular Sequence Data
nanocrystal
Nanocrystals
Phage
Quantum Dots
Staining and Labeling - methods
Virus Attachment
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Summary: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.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.22488