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Related Experiment Videos

Nanoparticle energy transfer on the cell surface.

László Bene1, Gergely Szentesi, László Mátyus

  • 1Department of Biophysics and Cell Biology, Medical and Health Science Center, Research Center for Molecular Medicine, University of Debrecen, Hungary. bene@jaguar.dote.hu

Journal of Molecular Recognition : JMR
|December 14, 2004
PubMed
Summary

This study demonstrates how immunogold nanoparticles enhance fluorescence, enabling proximity sensing of cell surface receptors. This technique offers a novel method for studying molecular interactions on living cells.

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Area of Science:

  • Biophysics
  • Cell Biology
  • Nanotechnology

Background:

  • Receptor topology is crucial for cell signaling.
  • Fluorescence resonance energy transfer (FRET) detects molecular proximities (2-10 nm).
  • Colloidal gold nanoparticles are used for cell labeling in electron microscopy.

Purpose of the Study:

  • To explore metal-fluorophore interactions for novel applications in fluorescence spectroscopy.
  • To investigate the distance-dependent enhancement of fluorophore emission by metal nanoparticles.
  • To demonstrate the feasibility of proximity sensors operating beyond conventional FRET ranges.

Main Methods:

  • Utilized immunogold nanoparticles and fluorescent dyes (rhodamine, cyanine) for labeling cell surface receptors (MHCI, MHCII, transferrin receptor, CD45).

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  • Employed flow cytometry and digital imaging microscopy to detect fluorescence enhancement and light scattering.
  • Performed mathematical simulations based on electrodynamic theory for fluorescence enhancement.

    • Main Results:

    • Observed enhanced fluorescence of dyes near immunogold nanoparticles on JY cells.
    • Demonstrated fluorescence enhancement sensitivity to excitation wavelength, gold bead size/amount, and fluorophore-nanoparticle distance.
    • Noted concentration- and size-dependent enhancement of 90-degree light scattering by immunogold.
    • Observed reduced photobleaching time constant of cyanine dye near gold particles.
    • Mathematical simulations showed consistency between measured values and theoretical predictions.

    Conclusions:

    • Metal nanoparticles can significantly enhance spontaneous emission of nearby fluorophores.
    • This phenomenon enables proximity sensing of cell surface receptors with extended detection ranges.
    • The developed method offers a versatile tool for studying molecular interactions on living cells.