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Imaging soft materials with scanning tunneling microscopy

J T Woodward¹, J A Zasadzinski

¹Department of Physics, University of California, Santa Barbara 93106-5080, USA.

Scanning Microscopy. Supplement

|January 1, 1996

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Researchers developed a scanning tunneling microscope (STM) technique using modified freeze-fracture replication to image soft biomaterials in 3D. This method achieves high resolution for hydrated lipid and protein samples.

Area of Science:

Biophysics
Materials Science
Microscopy

Background:

Standard electron microscopy (EM) techniques have limitations for imaging soft, non-conductive biomaterials.
Freeze-fracture replication is a common EM sample preparation method.
Scanning tunneling microscopy (STM) offers high-resolution imaging but requires conductive or rigid samples.

Purpose of the Study:

To adapt freeze-fracture replication for scanning tunneling microscopy (STM).
To enable high-resolution 3D imaging of soft, non-conductive biomaterials.
To characterize the ripple structure of dimyristoylphosphatidyl-choline (DMPC) lipid bilayers.

Main Methods:

Modified freeze-fracture replication with a silver backing layer and mounting on silver filters for enhanced rigidity.

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Imaging in a dry nitrogen atmosphere to prevent tip-sample coupling.

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Image analysis using cross-correlation averaging for noise reduction.

Main Results:

The modified STM/replica technique successfully imaged soft biomaterials in 3D at high resolution.
Rigidity enhancements and dry atmosphere imaging minimized height amplification artifacts.
Analyzed ripple shape and amplitude in the P beta phase of DMPC.
Achieved lateral resolution of approximately 1 nm and vertical resolution of approximately 0.3 nm.

Conclusions:

The developed STM/replica technique is a versatile method for high-resolution 3D imaging of hydrated lipid and protein samples.
This technique overcomes limitations of traditional EM and STM for soft biological materials.
It provides valuable insights into the structural details of biomolecular systems.