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Visualization of cytoskeletal elements by the atomic force microscope.

T Berdyyeva1, C D Woodworth, I Sokolov

  • 1Department of Physics, Clarkson University, Potsdam, NY 13699, USA.

Ultramicroscopy
|January 11, 2005
PubMed
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Atomic force microscopy (AFM) now visualizes human cell cytoskeletal fibers. This technique offers detailed 3D imaging of cytoskeleton structure and volume, aiding in fiber identification.

Area of Science:

  • Cell Biology
  • Biophysics
  • Nanotechnology

Background:

  • The cytoskeleton provides structural support and is crucial for cellular functions.
  • Direct visualization of cytoskeletal fibers is challenging due to their nanoscale dimensions and cellular complexity.

Purpose of the Study:

  • To introduce and validate a novel application of atomic force microscopy (AFM) for direct visualization of cytoskeletal fibers.
  • To demonstrate the capability of AFM to provide high-resolution, 3D structural information of the cytoskeleton in human foreskin epithelial cells.

Main Methods:

  • Human foreskin epithelial cells were treated with a low concentration of Triton X-100 to selectively remove membranes, soluble proteins, and organelles.
  • The remaining cytoskeletal structures were visualized using atomic force microscopy (AFM) under both liquid and air-dried conditions.

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  • AFM imaging parameters, including tip sharpness, were optimized to achieve high lateral resolution.

    • Main Results:

    • AFM successfully visualized cytoskeletal fibers, with the smallest observed fibers approximately 50 nm in diameter.
    • Scanning in liquid revealed surface filaments, while air-dried scanning provided information on both surface and volume of the fibers.
    • The technique achieved a lateral resolution of approximately 20 nm, with potential for single-nanometer resolution.
    • AFM enabled quantitative analysis of cytoskeletal density and volume, offering 3D structural insights.

    Conclusions:

    • This novel AFM application provides a direct and high-resolution method for visualizing and quantifying cytoskeletal fibers.
    • The technique offers complementary information through liquid and air-dried scanning, enhancing structural analysis.
    • AFM's 3D imaging capability and resolution allow for detailed characterization and identification of cytoskeletal fiber types, comparable to electron microscopy.