Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Fluorescence energy transfer in the rapid-diffusion limit.

D D Thomas1, W F Carlsen, L Stryer

  • 1Department of Structural Biology, Sherman Fairchild Center, Stanford University School of Medicine, Stanford, California 94305.

Proceedings of the National Academy of Sciences of the United States of America
|December 1, 1978
PubMed
Summary

Researchers achieved rapid-diffusion energy transfer using long-lived terbium donors. This method is highly sensitive to molecular distances, enabling precise probing of membrane dynamics and chromophore localization.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

HTS driven by fluorescence lifetime detection of FRET identifies activators and inhibitors of cardiac myosin.

SLAS discovery : advancing life sciences R & D·2023
Same author

Allomonal functions of steroid hormone, antheridiol, in water moldAchlya.

Journal of chemical ecology·2013
Same author

Electrophotographic mirror contact printing.

Applied optics·2010
Same author

Protein-lipid interactions within purified and reconstituted cytochrome C reductase and oxidase.

Biophysical journal·2009
Same author

High-Resolution Detection of muscle Crossbridge Orientation by Electron Paramagnetic Resonance.

Biophysical journal·2009
Same author

Harden M. McConnell: The Science Speaks for Itself.

Biophysical journal·2009

Area of Science:

  • Photochemistry
  • Biophysics
  • Materials Science

Background:

  • Energy transfer efficiency is influenced by translational diffusion of donor and acceptor molecules.
  • Previous studies focused on the static limit (Dtau(0)/s(2) << 1), where diffusion has minimal impact.
  • Achieving the rapid-diffusion limit (Dtau(0)/s(2) >> 1) requires long donor lifetimes and high diffusion rates.

Purpose of the Study:

  • To experimentally demonstrate energy transfer in the rapid-diffusion limit.
  • To utilize long-lived energy donors for probing translational motion in complex systems like membranes.
  • To establish a technique for determining the transverse location of chromophores in membranes.

Main Methods:

  • Steady-state and kinetic fluorescence experiments.

Related Experiment Videos

  • Utilizing terbium (Tb(3+)) chelated to dipicolinate as a long-lived energy donor (tau(0) = 2.2 msec).
  • Employing rhodamine B as the energy acceptor in a membrane vesicle model system.

    • Main Results:

    • Achieved the rapid-diffusion limit (Dtau(0)/s(2) >> 1) with the Tb(3+)-dipicolinate donor.
    • Observed a 50% energy transfer at a rhodamine B concentration of 0.67 muM, three orders of magnitude lower than in the static limit.
    • Demonstrated excellent agreement between experimental data and theoretical predictions across a wide range of diffusion coefficients.
    • Determined the distance of closest approach (a) to be 10 A for a Tb.(DPA)(3) chelate within a membrane vesicle.

    Conclusions:

    • Long-lived energy donors (millisecond lifetimes) enable the study of translational motion in membranes and other assemblies.
    • Energy transfer in the rapid-diffusion limit provides a sensitive method for measuring the distance of closest approach between molecules.
    • This technique allows for the determination of transverse chromophore locations in both model and biological membranes.