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Luminescence Resonance Energy Transfer to Study Conformational Changes in Membrane Proteins Expressed in Mammalian Cells
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Strategy for photostable proximity bioassays using lanthanides.

Jeanne P Haushalter1, Gregory W Faris

  • 1Molecular Physics Laboratory, SRI International, CA 94025, USA.

Applied Optics
|March 16, 2007
PubMed
Summary

Researchers developed photostable lanthanide chelate reporters for proximity assays. These novel reporters utilize nanoparticle interactions to achieve high fluorescence enhancement without photobleaching, improving assay performance.

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Preparation, Purification, and Characterization of Lanthanide Complexes for Use as Contrast Agents for Magnetic Resonance Imaging
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Published on: July 21, 2011

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Preparation, Purification, and Characterization of Lanthanide Complexes for Use as Contrast Agents for Magnetic Resonance Imaging
13:21

Preparation, Purification, and Characterization of Lanthanide Complexes for Use as Contrast Agents for Magnetic Resonance Imaging

Published on: July 21, 2011

Area of Science:

  • Biochemistry
  • Materials Science
  • Analytical Chemistry

Background:

  • Proximity assays require reporters with high sensitivity and stability.
  • Photobleaching and limited enhancement ratios hinder current assay technologies.
  • Lanthanide chelates offer unique photophysical properties for reporter development.

Purpose of the Study:

  • To create photostable lanthanide chelate reporters for proximity assays.
  • To leverage fluorescence enhancement near noble metal nanoparticles.
  • To achieve high on/off enhancement ratios and overcome photobleaching.

Main Methods:

  • Synthesized lanthanide chelates as fluorescent reporters.
  • Investigated reporter performance on silver island films.
  • Evaluated reporters in colloidal suspensions near nanoparticles.
  • Measured fluorescence enhancement factors and photostability.

Main Results:

  • Achieved fluorescence enhancement factors of 1-2 orders of magnitude.
  • Demonstrated significant enhancement for strongly quenched lanthanides.
  • Observed reporter stability, indicating photostability.

Conclusions:

  • Photostable lanthanide chelate reporters show promise for advanced proximity assays.
  • Nanoparticle-enhanced fluorescence offers a viable strategy for high-performance assays.
  • This approach overcomes limitations of photobleaching and low enhancement ratios.