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TopoStats – a program for automated tracing of biomolecules from AFM images

View ORCID ProfileJoseph G. Beton, View ORCID ProfileRobert Moorehead, View ORCID ProfileLuzie Helfmann, View ORCID ProfileRobert Gray, View ORCID ProfileBart W. Hoogenboom, View ORCID ProfileAgnel Praveen Joseph, View ORCID ProfileMaya Topf, View ORCID ProfileAlice L. B. Pyne
doi: https://doi.org/10.1101/2020.09.23.309609
Joseph G. Beton
1Institute of Structural and Molecular Biology, Birkbeck, University of London, London WC1E 7HX, UK
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  • ORCID record for Joseph G. Beton
Robert Moorehead
2Department of Materials Science and Engineering, University of Sheffield, Sheffield S1 3JD, UK
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Luzie Helfmann
1Institute of Structural and Molecular Biology, Birkbeck, University of London, London WC1E 7HX, UK
3London Centre for Nanotechnology, University College London, London, WC1H 0AH, UK
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Robert Gray
1Institute of Structural and Molecular Biology, Birkbeck, University of London, London WC1E 7HX, UK
3London Centre for Nanotechnology, University College London, London, WC1H 0AH, UK
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Bart W. Hoogenboom
3London Centre for Nanotechnology, University College London, London, WC1H 0AH, UK
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  • ORCID record for Bart W. Hoogenboom
Agnel Praveen Joseph
1Institute of Structural and Molecular Biology, Birkbeck, University of London, London WC1E 7HX, UK
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  • ORCID record for Agnel Praveen Joseph
Maya Topf
1Institute of Structural and Molecular Biology, Birkbeck, University of London, London WC1E 7HX, UK
4Centre for Structural Systems Biology, Heinrich-Pette-Institut, Leibniz-Institut für Experimentelle Virologie and Universitätsklinikum Hamburg Eppendorf, Hamburg, Germany
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Alice L. B. Pyne
2Department of Materials Science and Engineering, University of Sheffield, Sheffield S1 3JD, UK
3London Centre for Nanotechnology, University College London, London, WC1H 0AH, UK
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  • For correspondence: a.l.pyne@sheffield.ac.uk
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Abstract

We present TopoStats, a Python toolkit for automated editing and analysis of Atomic Force Microscopy images. The program automates identification and tracing of individual molecules in circular and linear conformations without user input. TopoStats was able to identify and trace a range of molecules within AFM images, finding, on average, 90% of all individual molecules and molecular assemblies within a wide field of view, and without the need for prior processing. DNA minicircles of varying size, DNA origami rings and pore forming proteins were identified and accurately traced with contour lengths of traces typically within 10 nm of the predicted contour length. TopoStats was also able to reliably identify and trace linear and enclosed circular molecules within a mixed population. The program is freely available via GitHub (https://github.com/afm-spm/TopoStats) and is intended to be modified and adapted for use if required.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • Updated following review

  • https://github.com/AFM-SPM/TopoStats

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted January 05, 2021.
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TopoStats – a program for automated tracing of biomolecules from AFM images
Joseph G. Beton, Robert Moorehead, Luzie Helfmann, Robert Gray, Bart W. Hoogenboom, Agnel Praveen Joseph, Maya Topf, Alice L. B. Pyne
bioRxiv 2020.09.23.309609; doi: https://doi.org/10.1101/2020.09.23.309609
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TopoStats – a program for automated tracing of biomolecules from AFM images
Joseph G. Beton, Robert Moorehead, Luzie Helfmann, Robert Gray, Bart W. Hoogenboom, Agnel Praveen Joseph, Maya Topf, Alice L. B. Pyne
bioRxiv 2020.09.23.309609; doi: https://doi.org/10.1101/2020.09.23.309609

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