A Covalent Fluorescent–Gold Immunoprobe: Simultaneous Detection of a Pre-mRNA Splicing Factor by Light and Electron Microscopy

Powell, Richard D.; Halsey, Carol; Spector, David L.; Kaurin, Shelley L.; McCann, John D.; Hainfeld, James F.

doi:10.1177/002215549704500704

Cited by 73 publications

(54 citation statements)

References 26 publications

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“…This methodology has not been widely used because the colloidal gold tends to quench the fluorescence signal (e.g., Goodman et al 1991). FNG was developed as an alternative immunoprobe (Powell et al 1997). This fluorescent immunoprobe is distinct from the colloidal gold-fluorescent probes.…”

Section: Discussionmentioning

confidence: 99%

“…FNG has several advantageous properties as an immunoprobe for correlative microscopy: (a) the fluorescence signal does not appear to be reduced because of proximity to the gold cluster; (b) the fluorochrome and antibody do not readily dissociate from a 1.4-nm gold particles because they are covalently linked [dissociation of antibodies from colloidal gold immunoprobes has been reported (Kramarcy and Sealock 1991)]; and (c) FNG penetrates into cells as readily as conventional immunofluorescence probes (Robinson and Vandré 1997). Previous studies have shown FNG to be a valuable immunoprobe for correlative microscopy (Powell et al 1997;Robinson and Vandré 1997). Although these approaches for correlative microscopy have been helpful for morphological analysis, they did not fully explore the capabilities of the methodology.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, a unique ultrasmall immunogold probe, FluoroNanogold (FNG), has been developed for use as a secondary antibody in immunocytochemical applications (Powell et al 1997). It consists of a 1.4-nm gold cluster compound to which antibodies and fluorochromes are covalently conjugated.…”

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confidence: 99%

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Correlative Microscopy Using FluoroNanogold on Ultrathin Cryosections: Proof of Principle

Tajima

Suzuki

Robinson

1998

J Histochem Cytochem.

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SUMMARYWe demonstrate a fluorescent ultrasmall immunogold probe, FluoroNanogold (FNG), to be a versatile reporter system for immunocytochemical labeling of ultrathin cryosections. FNG-labeled molecules in the same ultrathin cryosections can be resolved by two imaging techniques (i.e., fluorescence and electron microscopy). Lactoferrin, a marker protein for the specific granules in human neutrophils, was employed as the target for FNG immunolabeling. The spatial resolution of the fluorescence signal from FNG-labeled specific granules was compatible with that of silver-enhanced gold signal from the same granules in electron microscopy. Our results confirm that FNG can be used as a probe for highresolution correlation between immunofluorescence and electron microscopy.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Correlative Microscopy Using FluoroNanogold on Ultrathin Cryosections: Proof of Principle

Tajima

Suzuki

Robinson

1998

J Histochem Cytochem.

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“…Additional tools for immunolabeling are fluorescent derivatives of NG known as FluoroNanogold (FNG) (Powell et al 1997). These bifunctional probes consist of a 1.4-nm gold cluster compound to which an antibody [IgG, F(ab Ј ) 2 , or Fab] is conjugated; fluorochromes are conjugated to the antibody.…”

Section: Applications Using Fluoronanogoldmentioning

confidence: 99%

Enhanced Labeling Efficiency Using Ultrasmall Immunogold Probes: Immunocytochemistry

Robinson

Tajima

Vandré

2000

J Histochem Cytochem.

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“…Resonant energy transfer (RET) systems consisting of organic dye molecules and noble metal nanoparticles have recently gained considerable interest in biophotonics [1][2][3][4] as well as in materials science [5,6]. Closely related are donor-acceptor pairs of organic dye molecules forming Förster resonant energy transfer (FRET) systems.…”

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confidence: 99%

Fluorescence Quenching of Dye Molecules near Gold Nanoparticles: Radiative and Nonradiative Effects

Dulkeith¹,

Morteani²,

et al. 2002

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The radiative and nonradiative decay rates of lissamine dye molecules, chemically attached to differently sized gold nanoparticles, are investigated by means of time-resolved fluorescence experiments. A pronounced fluorescence quenching is observed already for the smallest nanoparticles of 1 nm radius. The quenching is caused not only by an increased nonradiative rate but, equally important, by a drastic decrease in the dye's radiative rate. Assuming resonant energy transfer to be responsible for the nonradiative decay channel, we compare our experimental findings with theoretical results derived from the Gersten-Nitzan model. DOI: 10.1103/PhysRevLett.89.203002 PACS numbers: 33.50.-j, 81.07.Pr Resonant energy transfer (RET) systems consisting of organic dye molecules and noble metal nanoparticles have recently gained considerable interest in biophotonics [1][2][3][4] as well as in materials science [5,6]. Closely related are donor-acceptor pairs of organic dye molecules forming Förster resonant energy transfer (FRET) systems. They have been theoretically modeled [7] and applied in biophysics extensively during the past decade (see, e.g., [8]). Yet these classical purely dye-based systems show disadvantages regarding quenching efficiency [4] and photostability [9].If the donor molecule is placed in the vicinity of a metal surface instead of an organic acceptor, not only resonant energy transfer takes place but also the radiative lifetime of the donor molecule changes. For metal films this has been investigated extensively [10 -13]. Much less is known about donor molecules in the vicinity of metal nanoparticles. Theoretical treatments of the moleculenanoparticle problem [14 -17] predict energy transfer rates and radiative decay rates that deviate substantially from what has been found for dye molecules in front of a metal film. Both radiative and nonradiative rates are expected to depend critically on size and shape of the nanoparticle, the distance between the dye molecule and the nanoparticle, the orientation of the molecular dipole with respect to the dye-nanoparticle axis, and the overlap of the molecule's emission with the nanoparticle's absorption spectrum. Recent experimental investigations deal with metal island films or rough surfaces only (see [18,19] and references in [20]), where the above mentioned parameters are undefined.Here we report results of time-resolved fluorescence experiments on a donor-acceptor system composed of lissamine molecules (donor) chemically attached to a gold nanoparticle (acceptor). The distance between the lissamine molecule and the surface of the nanoparticle is kept constant at 1 nm, whereas the nanoparticle radius is varied between 1 and 30 nm. We find time constants for the energy transfer on a picosecond time scale which turn out to decrease with increasing nanoparticle size. In addition, the dye's radiative rate is reduced by more than an order of magnitude. Both effects are responsible for the drastic quenching of the fluorescence yield as predicted by the so-called...

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A Covalent Fluorescent–Gold Immunoprobe: Simultaneous Detection of a Pre-mRNA Splicing Factor by Light and Electron Microscopy

Cited by 73 publications

References 26 publications

Correlative Microscopy Using FluoroNanogold on Ultrathin Cryosections: Proof of Principle

Correlative Microscopy Using FluoroNanogold on Ultrathin Cryosections: Proof of Principle

Enhanced Labeling Efficiency Using Ultrasmall Immunogold Probes: Immunocytochemistry

Fluorescence Quenching of Dye Molecules near Gold Nanoparticles: Radiative and Nonradiative Effects

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