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Related Experiment Videos

Immobilizing and imaging microtubules by atomic force microscopy

A Vinckier1, I Heyvaert, A D'Hoore

  • 1Department of Chemistry, University of Leuven, Heverlee, Belgium.

Ultramicroscopy
|March 1, 1995
PubMed
Summary

Researchers immobilized pig brain microtubules on silicon wafers for atomic force microscopy (AFM) studies. This method ensures stable microtubule positioning, enabling reproducible imaging and detailed surface analysis.

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Area of Science:

  • Biophysics
  • Materials Science
  • Microscopy

Background:

  • Microtubules are essential cytoskeletal components crucial for cellular structure and transport.
  • Developing stable immobilization techniques for microtubules is vital for high-resolution imaging and biophysical studies.
  • Atomic Force Microscopy (AFM) offers potential for nanoscale imaging of biological samples like microtubules.

Purpose of the Study:

  • To develop and validate a method for immobilizing microtubules from pig brains onto inorganic substrates for AFM analysis.
  • To investigate the structural and mechanical properties of immobilized microtubules using AFM and Scanning Tunneling Microscopy (STM).
  • To compare AFM/STM imaging results with traditional electron microscopy techniques.

Main Methods:

  • Microtubules were isolated from pig brains and immobilized on silicon wafers using 4-aminobutyldimethylmethoxysilane and glutaraldehyde.

Related Experiment Videos

  • Atomic Force Microscopy (AFM) in tapping mode was employed for high-resolution imaging.
  • Scanning Tunneling Microscopy (STM) was used to image gold-coated microtubules on silicon and tubules on graphite.

    • Main Results:

    • Stable covalent immobilization of microtubules on silicon wafers was achieved, allowing reproducible AFM experiments.
    • AFM revealed apparent microtubule heights of 10 nm in buffer and 15 nm in air, smaller than transmission electron microscopy (25 nm).
    • Microtubule surface softness was greater in buffer solution than in air; highest resolution (10 nm) achieved with tapping mode AFM.

    Conclusions:

    • The developed immobilization technique provides a stable platform for studying microtubule structure and mechanics using scanning probe microscopy.
    • AFM and STM offer valuable insights into microtubule morphology and surface properties, complementing electron microscopy.
    • Careful substrate selection and artifact identification are crucial for accurate interpretation of STM images of biological samples.