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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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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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Atomic Force Microscopy01:08

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Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
The AFM Probe
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Updated: Jan 7, 2026

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip
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Offsetting ROS-Mediated Arrest of Endothelial Fenestration Dynamics Permits Long-Term Optical Super-Resolution

Annika Kiel1, Marcin Luty2,3, Angela Kralemann-Köhler1

  • 1Department of General- and Visceral Surgery - Liver- and Tumor Biology, Medical Faculty OWL, Bielefeld University, Bielefeld 33615, Germany.

ACS Applied Materials & Interfaces
|January 5, 2026
PubMed
Summary

Photoactivated reactive oxygen species (ROS) arrest fenestrations in live-cell imaging. Supplementing with N-acetylcysteine (NAC) preserves fenestration dynamics during super-resolution microscopy.

Keywords:
atomic force microscopy (AFM)fenestrationslive-cell imagingliver sinusoidal endothelial cells (LSEC)reactive oxygen species (ROS)structured illumination microscopy (SIM)super-resolution microscopy

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Area of Science:

  • Cell Biology
  • Microscopy
  • Biophysics

Background:

  • Live-cell super-resolution fluorescence imaging is crucial for studying subcellular structures.
  • Phototoxicity limits live-cell imaging, causing artifacts and biased interpretations.
  • Liver Sinusoidal Endothelial Cells (LSECs) present unique challenges due to dynamic fenestrations.

Purpose of the Study:

  • Identify the cause of fenestration arrest during fluorescence microscopy.
  • Optimize live-cell super-resolution imaging conditions for LSECs.
  • Develop a framework to mitigate imaging artifacts in sensitive cell types.

Main Methods:

  • Three-dimensional super-resolution structured illumination microscopy (3D SR-SIM).
  • Systematic evaluation of fluorophores and reactive oxygen species (ROS) scavengers.
  • Atomic force microscopy (AFM) for validation and nanomechanical analysis.

Main Results:

  • Photoactivation-generated ROS were identified as the primary cause of fenestration arrest.
  • Combining BioTracker staining with N-acetylcysteine (NAC) and CO2-independent medium preserved fenestration dynamics.
  • AFM confirmed ROS-mediated fenestration impairment and revealed nanomechanical changes.

Conclusions:

  • ROS generated during imaging significantly impact LSEC fenestration dynamics.
  • NAC supplementation effectively prevents imaging-induced artifacts in LSECs.
  • The developed imaging strategy provides a broadly applicable framework for live-cell super-resolution microscopy.