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A Novel Preparation Method for High Resolution AFM Introduced With 2d-Streptavidin Crystals Grown on a Biotinlipid Monolayer

Published online by Cambridge University Press:  02 July 2020

Simon Scheuring
Affiliation:
M.E.Müller-Institut for Microscopy, Biozentrum Universität Basel, Klingelbergstrasse 70, CH-4056, Basel, Switzerland.
Daniel J. Müller
Affiliation:
M.E.Müller-Institut for Microscopy, Biozentrum Universität Basel, Klingelbergstrasse 70, CH-4056, Basel, Switzerland.
Philippe Ringler
Affiliation:
M.E.Müller-Institut for Microscopy, Biozentrum Universität Basel, Klingelbergstrasse 70, CH-4056, Basel, Switzerland.
J. Bernard Heymann
Affiliation:
M.E.Müller-Institut for Microscopy, Biozentrum Universität Basel, Klingelbergstrasse 70, CH-4056, Basel, Switzerland.
Andreas Engel
Affiliation:
M.E.Müller-Institut for Microscopy, Biozentrum Universität Basel, Klingelbergstrasse 70, CH-4056, Basel, Switzerland.
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Extract

The AFM (atomic force microscopy) requires a stable and flat substrate on which a biological sample is readily immobilized. For instance, membrane protein 2D crystals in lipid bilayers have been adsorbed to silanized glass with covalently attachment using a photocrosslinker or to mica by physisorption under appropriate ionic conditions. For the immobilization of 2D crystals of water soluble proteins formed on lipid monolayers, a different approach is required to attach the highly hydrophobic tails of the lipid molecules. To find a suitably hydrophobic substrate, the hydrophobicity of different materials was determined by the sitting drop method. Briefly, a 10 μ1 droplet of deionized and filtered water was deposited on the substrate surface and the diameter measured. Table 1 shows a decrease in droplet diameter with increasing hydrophobicity, indicating that substrates such as silanized glass and HOPG (highly ordered pyrolytic graphite) may be suitable for adsorbing lipid monolayers.

Type
Scanned Probe Microscopy: Much More Than Just Beautiful Images
Copyright
Copyright © Microscopy Society of America

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