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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

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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...
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Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
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Photobleaching Enables Super-resolution Imaging of the FtsZ Ring in the Cyanobacterium Prochlorococcus
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Bleaching-Resistant Super-Resolution Fluorescence Microscopy.

Jiwoong Kwon1, Mohamed Saleh Elgawish2,3, Sang-Hee Shim2

  • 1Department of Biophysics and Biophysical Chemistry, Johns Hopkins University, Baltimore, MD, 21205, USA.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|January 28, 2022
PubMed
Summary
This summary is machine-generated.

Photobleaching permanently damages fluorescence, limiting super-resolution microscopy (SRM). New strategies, including exploiting on-off states, are overcoming this, enabling molecular-scale resolution and longer observations.

Keywords:
fluorophorephotobleachingphotostabilityphotoswitchingsuper-resolution fluorescence microscopy

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Area of Science:

  • Microscopy
  • Biophysics
  • Biochemistry

Background:

  • Photobleaching, the irreversible loss of fluorescence, significantly hinders super-resolution microscopy (SRM).
  • This limitation restricts both the spatial and temporal resolution achievable in SRM.
  • Overcoming photobleaching is crucial for advancing nanoscale imaging.

Purpose of the Study:

  • To review current strategies for preventing and overcoming photobleaching in SRM.
  • To highlight novel approaches that leverage fluorescence dynamics for photostabilization.
  • To discuss the potential of these strategies in enhancing SRM capabilities.

Main Methods:

  • Review of photostabilization techniques from conventional fluorescence microscopy applied to SRM.
  • Analysis of SRM-specific strategies exploiting fluorescence on-off transitions.
  • Exploration of how off-states can protect probes or facilitate replacement.

Main Results:

  • Photostabilization strategies from conventional microscopy are being adapted for SRM.
  • SRM-specific methods utilize fluorescence blinking to mitigate photobleaching.
  • Off-states offer protection from light or allow for probe exchange.

Conclusions:

  • Overcoming photobleaching in SRM is achievable through adapted and novel strategies.
  • Exploiting fluorescence on-off dynamics presents a promising route for photostabilization.
  • These advancements are expected to push SRM resolution to molecular scales and extend observation times.