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High Resolution Imaging of Immunoglobulin G Antibodies and Other Biomolecules Using Amplitude Modulation Atomic Force Microscopy in Air

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Atomic Force Microscopy in Biomedical Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 736))

Abstract

The atomic force microscope (AFM) is a very versatile tool for studying biological samples at ­nanometre-scale resolution. The resolution one achieves depends on many factors, including the sample properties, the imaging environment, the AFM tip and cantilever probe characteristics, and the signal detection and feedback control mechanism, to name a few. This chapter describes how to routinely achieve the highest possible spatial resolution on isolated protein molecules on mica surfaces. This is illustrated with Immunoglobulin G antibodies but the methods apply equally well to any other globular multi-subunit protein, as well as other biomolecules. Double-stranded DNA is used as a model sample to illustrate the effects of the force regime in amplitude modulation atomic force microscopy (AM AFM) on the image resolution and contrast. AM control is a widely used technique in biological AFM for reasons which are discussed.

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Acknowledgements

We thank Anthony Maxwell for kindly providing us with monoclonal IgG antibodies against the A-subunit of DNA gyrase. SS is funded through a Doctoral Training Grant of the BBSRC. We acknowledge the University of Leeds for strategic investment in AFM infrastructure.

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Authors and Affiliations

  1. School of Physics and Astronomy, University of Leeds, Leeds, UK

    Neil H. Thomson

Authors
  1. Sergio Santos
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  2. Neil H. Thomson
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Corresponding author

Correspondence to Neil H. Thomson .

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Editors and Affiliations

  1. School of Medicine, Department of Pharmacology, University of Milan, Via Vanvitelli 32, Milano, 20129, Italy

    Pier Carlo Braga

  2. , Robotics, Brain & Cognitive Sciences, Italian Institute of Technology, Via Morego 30, Genova, 16163, Italy

    Davide Ricci

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Santos, S., Thomson, N.H. (2011). High Resolution Imaging of Immunoglobulin G Antibodies and Other Biomolecules Using Amplitude Modulation Atomic Force Microscopy in Air. In: Braga, P., Ricci, D. (eds) Atomic Force Microscopy in Biomedical Research. Methods in Molecular Biology, vol 736. Humana Press. https://doi.org/10.1007/978-1-61779-105-5_5

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  • DOI: https://doi.org/10.1007/978-1-61779-105-5_5

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-104-8

  • Online ISBN: 978-1-61779-105-5

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