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The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
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Related Experiment Video

Updated: Dec 13, 2025

Engineering Fibrin-based Tissue Constructs from Myofibroblasts and Application of Constraints and Strain to Induce Cell and Collagen Reorganization
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Controlling Fibronectin Fibrillogenesis Using Visible Light.

Tetyana Gudzenko1, Clemens M Franz1,2

  • 1DFG-Center for Functional Nanostructures, Karlsruhe Institute of Technology, Karlsruhe, Germany.

Frontiers in Molecular Biosciences
|August 1, 2020
PubMed
Summary
This summary is machine-generated.

Visible light inhibits fibronectin (FN) fibrillogenesis, a process crucial for cell adhesion and signaling. This photosensitivity impacts live-cell imaging and requires specific conditions for accurate study of FN mechanics.

Keywords:
AFMfibrillogenesisfibronectinmatrix nanomechanicsvisible light

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Area of Science:

  • Cell biology
  • Biophysics
  • Materials science

Background:

  • Fibronectin (FN) fibrillogenesis is vital for cell adhesion and mechanosensitive signaling.
  • Previous imaging attempts using fluorescently labeled FN failed under standard light microscopy.
  • This failure suggested an unknown sensitivity of the FN fibrillogenesis process.

Purpose of the Study:

  • To investigate the photo sensitivity of cell-induced plasma fibronectin fibrillogenesis.
  • To identify optimal conditions for imaging FN fibrillogenesis using live-cell microscopy.
  • To understand the impact of light exposure on FN mechanics and cellular responses.

Main Methods:

  • Timelapse atomic force microscopy (AFM) to image FN fibrillogenesis.
  • Live-cell fluorescence microscopy with varying illumination wavelengths.
  • Cell seeding on pre-illuminated FN substrates.
  • Measurement of reactive oxygen species (ROS) levels.
  • Analysis of cell migration, focal adhesion, and signaling pathway phosphorylation (FAK, paxillin).

Main Results:

  • Visible light illumination, particularly below 560 nm, significantly inhibited FN fibrillogenesis.
  • This photoinhibition was independent of phototoxicity to cells but enhanced by fluorophore labeling.
  • Using long-wavelength dyes (Alexa Fluor 633) and ROS scavengers enabled successful imaging.
  • Light-exposed FN showed increased mechanical stiffness, affecting cell migration and signaling.

Conclusions:

  • Fibronectin fibrillogenesis is photosensitive, posing challenges for live-cell fluorescence imaging.
  • Optimized imaging protocols using specific dyes and ROS scavengers are necessary.
  • Light-induced changes in FN mechanics influence cell behavior and mechanosensitive signaling.
  • Careful consideration of light exposure is crucial when interpreting results from FN-coated substrates.