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Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions
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Methods for imaging DNA in liquid with lateral molecular-force microscopy.

R L Harniman1, J A Vicary, J K H Hörber

  • 1H H Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol, UK. Robert.Harniman@bristol.ac.uk

Nanotechnology
|February 2, 2012
PubMed
Summary

This study introduces a new method for imaging DNA molecules in liquid using shear force microscopy with a scattered evanescent wave (SEW) detection system and vertically oriented cantilevers (VOCs). This technique enables reliable, true non-contact imaging of biomolecules, advancing nanoscale visualization in aqueous environments.

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Area of Science:

  • Atomic Force Microscopy
  • Nanotechnology
  • Biophysics

Background:

  • Shear force microscopy (SFM) is typically used for surface imaging in air, not typically for biomolecules in liquid.
  • Imaging delicate biomolecules in liquid requires non-invasive techniques to preserve their structure and function.

Purpose of the Study:

  • To demonstrate the capability of a novel scattered evanescent wave (SEW) detection system with custom-designed vertically oriented cantilevers (VOCs) for true non-contact imaging of DNA molecules in liquid.
  • To establish optimal experimental parameters for shear force imaging of biomolecules in aqueous environments.

Main Methods:

  • Utilized a scattered evanescent wave (SEW) detection system integrated with custom-designed vertically oriented cantilevers (VOCs).
  • Determined the effective range of cantilever spring constants for successful shear force imaging (0.05–0.09 N m⁻¹).

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  • Acquired true non-contact images of lambda DNA (λ-DNA) adsorbed on mica in distilled water at high scan rates (8000 pixels s⁻¹).
  • Developed and demonstrated a constant-height force mapping mode for VOCs with sub-nanometer vertical accuracy.

    • Main Results:

    • Achieved reliable, true non-contact imaging of DNA molecules in a liquid environment.
    • Identified specific cantilever spring constants crucial for successful shear force microscopy in liquid.
    • Successfully mapped shear forces above λ-DNA molecules in liquid using the novel force mapping mode, demonstrating precise tip-sample distance control.

    Conclusions:

    • The SEW detection system combined with VOCs offers a robust method for non-contact imaging of biomolecules in liquid.
    • The developed force mapping mode provides high accuracy for controlling tip-sample interactions, essential for nanoscale force measurements.
    • This advancement opens new possibilities for high-resolution visualization and manipulation of biological samples in their native aqueous environment.