Fluorescent avidin-bound silver particle: a strategy for single target molecule detection on a cell membrane.
Cy5-avidin conjugate-bound silver nanoparticles were prepared as a fluorescence molecular reagent for the cell imaging. Compared with the metal-free avidin conjugate, the avidin-metal complex was observed to display a stronger emission intensity, shorter lifetime, and better photostability. The avidin-metal complexes were conjugated with the biotin-sites on the surfaces of PM1 cell lines, and the cell images were recorded using scanning confocal microscopy. It was noticed that the avidin-metal complexes bound on the cell surfaces could be identified as the isolated emission spots distinct from the cellular autofluorescence. The emission intensity over the cell image was increased with an increase of the number of avidin-metal complexes on the cell surface but the lifetime was decreased. A quantitative regression curve was achieved between the amount of avidin-metal complex on the cell surface and the emission intensity or lifetime over the entire cell image. On the basis of this curve, we expect to develop an approach that can be used to quantify the amount of target molecules on the cell surfaces using the cell intensity and lifetime images at the single cell level.
Zhang
Jian
J
Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, Maryland 21201, USA.
Fu
Yi
Y
Liang
Dong
D
Zhao
Richard Y
RY
Lakowicz
Joseph R
JR
D001360:Avidin / Q000737:chemistry*; D001710:Biotin / Q000378:metabolism; D002232:Carbocyanines / Q000737:chemistry; D002462:Cell Membrane / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D018613:Microscopy, Confocal; D012834:Silver / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013601:T-Lymphocytes / Q000166:cytology
D002232:Carbocyanines; D005456:Fluorescent Dyes; C085321:cyanine dye 5; D001360:Avidin; D012834:Silver; D001710:Biotin