title: Fluorescent metal nanoshell and CK19 detection on single 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 10nm 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.
authors.affiliation: Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201, USA. jian@cfs.biomet.umaryland.edu
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
authors.forename: Yi
authors.initials: Y
authors.lastname: Fu
authors.forename: Ge
authors.initials: G
authors.lastname: Li
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Lakowicz
authors.forename: Richard Y
authors.initials: RY
authors.lastname: Zhao
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D002232:Carbocyanines / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D053539:Keratin-19 / Q000032:analysis*; D053768:Metal Nanoparticles / Q000737:chemistry*; D057145:Nanoshells / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry; D012834:Silver / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D002232:Carbocyanines; D005456:Fluorescent Dyes; D053539:Keratin-19; C085321:cyanine dye 5; D012834:Silver; D012822:Silicon Dioxide