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Related Experiment Videos

Covalent binding of biological samples to solid supports for scanning probe microscopy in buffer solution

S Karrasch1, M Dolder, F Schabert

  • 1Maurice E. Müller-Institute for High-Resolution Electron Microscopy, University of Basel, Switzerland.

Biophysical Journal
|December 1, 1993
PubMed
Summary

Researchers developed a covalent binding method for immobilizing biological samples on glass surfaces using photoactivatable cross-linkers. This technique enables repeated, high-quality imaging with scanning force microscopy without sample displacement.

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Area of Science:

  • Biophysics
  • Surface Chemistry
  • Microscopy

Background:

  • Scanning force microscopy (SFM) requires stable sample immobilization on solid supports for imaging biological molecules in native buffer solutions.
  • Displacement of biological samples by the SFM scanning tip is a common challenge, leading to image degradation.

Purpose of the Study:

  • To develop a robust method for covalently attaching biological samples to glass surfaces for SFM analysis.
  • To enable repeated, high-fidelity imaging of biological molecules in their native state.

Main Methods:

  • Chemical modification of glass coverslips with N-5-azido-2-nitrobenzoyloxysuccinimide, a photoactivatable cross-linker.
  • Immobilization of biological samples by sandwiching them between derivatized coverslips.
  • Cross-linking of samples to the glass surface via ultraviolet light irradiation.

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Main Results:

  • Successful covalent attachment of biological samples to glass surfaces.
  • Stable immobilization allowing repeated SFM imaging without sample displacement.
  • Preservation of image quality during repeated scans, unlike non-immobilized samples.

Conclusions:

  • The described method provides a reliable approach for immobilizing biological samples for SFM.
  • This technique overcomes sample displacement issues, enhancing the utility of SFM for studying biological molecules.
  • Enables long-term, high-resolution imaging of biological specimens in aqueous environments.