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Title: Fluorescent metal nanoshell and CK19 detection on single cell image

Abstract

Highlights: {yields} Novel metal nanoshell as fluorescence imaging agent. {yields} Fluorescent mAb-metal complex with enhanced intensity and shortened lifetime. {yields} Immuno-interactions of mAb-metal complexes with CK19 molecules on CNCAP and HeLa cell surfaces. {yields} Isolation of conjugated mAb-metal complexes from cellular autofluorescence on cell image. -- Abstract: In this article, we report the synthesis strategy and optical properties of a novel type of fluorescence metal nanoshell when it was used as imaging agent for fluorescence cell imaging. The metal nanoshells were made with 40 nm silica cores and 10 nm silver shells. Unlike typical fluorescence metal nanoshells which contain the organic dyes in the cores, novel metal nanoshells were composed of Cy5-labelled monoclonal anti-CK19 antibodies (mAbs) on the external surfaces of shells. Optical measurements to the single nanoparticles showed that in comparison with the metal free labelled mAbs, the mAb-Ag complexes displayed significantly enhanced emission intensity and dramatically shortened lifetime due to near-field interactions of fluorophores with metal. These metal nanoshells were found to be able to immunoreact with target cytokeratin 19 (CK19) molecules on the surfaces of LNCAP and HeLa cells. Fluorescence cell images were recorded on a time-resolved confocal microscope. The emissions from the metal nanoprobes could bemore » clearly isolated from the cellular autofluorescence backgrounds on the cell images as either individuals or small clusters due to their stronger emission intensities and shorter lifetimes. These emission signals could also be precisely counted on single cell images. The count number may provide an approach for quantifying the target molecules in the cells.« less

Authors:
 [1];  [1];  [2];  [1];  [2];  [3];  [3]
  1. Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201 (United States)
  2. Division of Molecular Pathology, Department of Pathology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201 (United States)
  3. (United States)
Publication Date:
OSTI Identifier:
22207493
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 413; Journal Issue: 1; Other Information: Copyright (c) 2011 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANTIBODIES; DYES; FLUORESCENCE; HELA CELLS; IMAGES; NANOSTRUCTURES; OPTICAL PROPERTIES; SILICA; SILVER; SILVER COMPLEXES; TIME RESOLUTION

Citation Formats

Zhang, Jian, E-mail: jian@cfs.biomet.umaryland.edu, Fu, Yi, Li, Ge, Lakowicz, Joseph R., Zhao, Richard Y., E-mail: rzhao@som.umaryland.edu, Department of Microbiology-Immunology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201, and Institute of Human Virology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201. Fluorescent metal nanoshell and CK19 detection on single cell image. United States: N. p., 2011. Web. doi:10.1016/J.BBRC.2011.08.042.
Zhang, Jian, E-mail: jian@cfs.biomet.umaryland.edu, Fu, Yi, Li, Ge, Lakowicz, Joseph R., Zhao, Richard Y., E-mail: rzhao@som.umaryland.edu, Department of Microbiology-Immunology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201, & Institute of Human Virology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201. Fluorescent metal nanoshell and CK19 detection on single cell image. United States. doi:10.1016/J.BBRC.2011.08.042.
Zhang, Jian, E-mail: jian@cfs.biomet.umaryland.edu, Fu, Yi, Li, Ge, Lakowicz, Joseph R., Zhao, Richard Y., E-mail: rzhao@som.umaryland.edu, Department of Microbiology-Immunology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201, and Institute of Human Virology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201. Fri . "Fluorescent metal nanoshell and CK19 detection on single cell image". United States. doi:10.1016/J.BBRC.2011.08.042.
@article{osti_22207493,
title = {Fluorescent metal nanoshell and CK19 detection on single cell image},
author = {Zhang, Jian, E-mail: jian@cfs.biomet.umaryland.edu and Fu, Yi and Li, Ge and Lakowicz, Joseph R. and Zhao, Richard Y., E-mail: rzhao@som.umaryland.edu and Department of Microbiology-Immunology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201 and Institute of Human Virology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201},
abstractNote = {Highlights: {yields} Novel metal nanoshell as fluorescence imaging agent. {yields} Fluorescent mAb-metal complex with enhanced intensity and shortened lifetime. {yields} Immuno-interactions of mAb-metal complexes with CK19 molecules on CNCAP and HeLa cell surfaces. {yields} Isolation of conjugated mAb-metal complexes from cellular autofluorescence on cell image. -- Abstract: In this article, we report the synthesis strategy and optical properties of a novel type of fluorescence metal nanoshell when it was used as imaging agent for fluorescence cell imaging. The metal nanoshells were made with 40 nm silica cores and 10 nm silver shells. Unlike typical fluorescence metal nanoshells which contain the organic dyes in the cores, novel metal nanoshells were composed of Cy5-labelled monoclonal anti-CK19 antibodies (mAbs) on the external surfaces of shells. Optical measurements to the single nanoparticles showed that in comparison with the metal free labelled mAbs, the mAb-Ag complexes displayed significantly enhanced emission intensity and dramatically shortened lifetime due to near-field interactions of fluorophores with metal. These metal nanoshells were found to be able to immunoreact with target cytokeratin 19 (CK19) molecules on the surfaces of LNCAP and HeLa cells. Fluorescence cell images were recorded on a time-resolved confocal microscope. The emissions from the metal nanoprobes could be clearly isolated from the cellular autofluorescence backgrounds on the cell images as either individuals or small clusters due to their stronger emission intensities and shorter lifetimes. These emission signals could also be precisely counted on single cell images. The count number may provide an approach for quantifying the target molecules in the cells.},
doi = {10.1016/J.BBRC.2011.08.042},
journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 1,
volume = 413,
place = {United States},
year = {2011},
month = {9}
}