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

Macromolecular-scale resolution in biological fluorescence microscopy.

Gerald Donnert1, Jan Keller, Rebecca Medda

  • 1Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37070 Göttingen, Germany.

Proceedings of the National Academy of Sciences of the United States of America
|July 26, 2006
PubMed
Summary

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

K-Ras controls asymmetric cell divisions from the primary cilium.

Cell death & disease·2026
Same author

Direct visualization of native GSDMD pores reveals lipid-driven stabilization during pyroptosis.

Science advances·2026
Same author

Daily relationship between air pollution and physical activity in an industrial region.

Applied psychology. Health and well-being·2026
Same author

Ethanol-driven structural transitions of human serum albumin and surface-assisted 2D network formation on Ti-6Al-4V: A kinetic-morphological map.

Colloids and surfaces. B, Biointerfaces·2026
Same author

Smart event-triggered MINFLUX microscopy to catch and follow rare events.

Nature communications·2026
Same author

Protocol for microfluidic-based high-precision general polarization fluorescence microscopy of lipid packing in membrane vesicles.

STAR protocols·2026

We achieved 15-20 nm resolution in fluorescence microscopy by preventing molecular triplet states, significantly reducing photobleaching and increasing signal. This breakthrough enables detailed nanoscale imaging of biological structures.

Area of Science:

  • Biophysics
  • Cell Biology
  • Optical Microscopy

Background:

  • Conventional fluorescence microscopy is limited by the diffraction barrier, restricting resolution.
  • Photobleaching of fluorescent dyes, particularly due to molecular triplet state excitation, hinders high-resolution imaging.
  • Stimulated emission depletion (STED) microscopy offers improved resolution but can be limited by photobleaching.

Purpose of the Study:

  • To demonstrate far-field fluorescence microscopy with significantly enhanced resolution below the diffraction limit.
  • To overcome photobleaching limitations in STED microscopy by managing molecular triplet states.
  • To enable detailed nanoscale visualization of biological structures using conventional optics.

Main Methods:

  • Eliminating molecular triplet state excitation during excitation-depletion cycles in STED microscopy.

Related Experiment Videos

  • Implementing triplet-state relaxation by adjusting laser repetition rates or scanning speeds.
  • Utilizing far-field fluorescence microscopy with optimized illumination schemes.

    • Main Results:

    • Achieved a focal-plane resolution of 15-20 nm in biological samples, a 10- to 12-fold increase below the diffraction barrier.
    • Reduced photobleaching by managing triplet states, leading to up to a 30-fold increase in total fluorescence signal.
    • Decreased the effective focal spot area by up to approximately 140-fold compared to diffraction limits.
    • Successfully visualized nanoscale protein patterns, intermediate filament structures, and nuclear speckles in mammalian cells.

    Conclusions:

    • The developed method provides diffraction-unlimited fluorescence microscopy with unprecedented resolution.
    • This technique overcomes major photobleaching limitations in super-resolution microscopy.
    • Opens new avenues for investigating fundamental biological questions at the nanoscale using conventional optical setups.