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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Related Experiment Video

Updated: May 8, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
09:45

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Improving STED-Based Super-resolution-Imaging with Fluorescence Lifetimes.

Vasileios Moysidis1, Marina Mikhaylova1, Daniela Hacker2

  • 1AG Optobiology, Institute of Biology, Humboldt-Universität zu Berlin, Berlin, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|May 6, 2026
PubMed
Summary

Advanced microscopy techniques like tauSTED (Stimulated Emission Depletion) microscopy are crucial for visualizing neuronal structures. This chapter details sample preparation and imaging parameters for high-resolution tauSTED nanoscopy.

Keywords:
CytoskeletonDissociated culturesFLIMHippocampusSTEDSynapsestauSTED

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Last Updated: May 8, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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Area of Science:

  • Neuroscience
  • Cell Biology
  • Microscopy

Background:

  • The neuronal cytoskeleton and synaptic organization are vital for information processing.
  • Cytoskeletal filaments and synaptic proteins exhibit posttranslational modifications and nanodomain organization, respectively.
  • Understanding their spatial organization necessitates high-resolution imaging techniques.

Purpose of the Study:

  • To provide a comprehensive guide to tauSTED nanoscopy for biological imaging.
  • To detail critical aspects of sample preparation and imaging parameter selection for STED microscopy.
  • To highlight the advantages of tauSTED in enhancing resolution for visualizing neuronal components.

Main Methods:

  • Utilizing Stimulated Emission Depletion (STED) microscopy, a super-resolution technique.
  • Employing tauSTED, which leverages fluorescence lifetime information to improve resolution.
  • Focusing on advanced sample preparation protocols tailored for super-resolution imaging.

Main Results:

  • STED microscopy overcomes the diffraction limit of conventional fluorescence microscopy.
  • tauSTED further enhances lateral, axial, and temporal resolution compared to standard STED.
  • Successful application requires careful planning and optimized imaging parameters.

Conclusions:

  • tauSTED nanoscopy offers significant advantages for high-resolution imaging of neuronal structures.
  • Proper sample preparation and parameter selection are critical for high-quality STED imaging.
  • This chapter serves as a practical guide for researchers using tauSTED microscopy.