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

Supercritical angle fluorescence correlation spectroscopy.

Jonas Ries1, Thomas Ruckstuhl, Dorinel Verdes

  • 1Biotechnologisches Zentrum, Technical University of Dresden, Dresden, Germany.

Biophysical Journal
|September 11, 2007
PubMed
Summary
This summary is machine-generated.

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

Fluorescence Correlation Spectroscopy to Examine Protein-Lipid Interactions in Membranes.

Methods in molecular biology (Clifton, N.J.)·2026
Same author

Ion- and Temperature-Programmable Reconfiguration of Subcompartments in Synthetic Cells.

Chembiochem : a European journal of chemical biology·2026
Same author

Optimizing spatial organization of FtsZ rings for large-scale constriction in synthetic cells.

Nature communications·2026
Same author

On the nature of the earliest known lifeforms.

eLife·2026
Same author

Chlamydial membrane vesicles deliver the beta barrel outer membrane protein OmpA to mitochondria to inhibit apoptosis.

PLoS pathogens·2026
Same author

Continuum architecture dynamics of vesicle tethering in exocytosis.

Cell·2026
Same journal

Quantifying the Peripheral Surface Information Entropy from Conformational Ensembles of Globular Protein-Peptide Complexes.

Biophysical journal·2026
Same journal

Anisotropic unbinding and location-dependent hovering of a kinesin motor head over microtubule.

Biophysical journal·2026
Same journal

Kinesin-5/Cut7 C-terminal tail phosphorylation influence on motor regulation through multi-scale molecular modeling.

Biophysical journal·2026
Same journal

Dynamic conformations of fluorophores on self-labeling protein tags.

Biophysical journal·2026
Same journal

Different actions of RyR2 open and closed channel block explained by a multiscale Ca<sup>2+</sup> release model.

Biophysical journal·2026
Same journal

Membrane Environment Sets the Functional pK<sub>a</sub> of Ionizable Lipids.

Biophysical journal·2026
See all related articles

Supercritical angle (SA) microscopy improves fluorescence correlation spectroscopy (FCS) by creating a smaller detection volume. This novel SA-FCS method offers enhanced surface sensitivity and outperforms standard confocal FCS for membrane diffusion studies.

Area of Science:

  • Biophysics
  • Microscopy
  • Spectroscopy

Background:

  • Fluorescence Correlation Spectroscopy (FCS) is a powerful technique for studying molecular dynamics.
  • Standard confocal FCS has limitations in achieving high surface sensitivity and small detection volumes.
  • Developing novel microscopy objectives is crucial for advancing biophysical measurements.

Purpose of the Study:

  • To investigate the utility of a supercritical angle (SA) objective for Fluorescence Correlation Spectroscopy (FCS).
  • To demonstrate the enhanced surface sensitivity and detection volume of SA-FCS.
  • To provide a theoretical framework for quantitative analysis of SA-FCS data.

Main Methods:

  • Utilized a novel supercritical angle (SA) microscope objective combining an aspheric lens and a parabolic mirror.

Related Experiment Videos

  • Performed diffusion measurements on model membranes with excess free dye.
  • Compared SA-FCS measurements directly against standard confocal FCS.
  • Developed an extensive theoretical framework for SA-FCS data analysis.

    • Main Results:

    • The SA objective creates a significantly smaller detection volume compared to conventional methods.
    • SA-FCS exhibits excellent surface sensitivity, validated by diffusion measurements in model membranes.
    • SA-FCS demonstrated a clear advantage over standard confocal FCS, particularly for membrane diffusion studies.
    • A comprehensive theoretical framework for accurate SA-FCS correlation curve evaluation was established.

    Conclusions:

    • Supercritical angle microscopy is a promising advancement for Fluorescence Correlation Spectroscopy.
    • SA-FCS offers superior performance for studying molecular diffusion at surfaces and in membranes.
    • The developed theoretical framework enables precise quantitative analysis of SA-FCS experiments.