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Co<sup>2+</sup> mediated adsorption facilitates atomic force microscopy of DNA molecules at double-helix resolution

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posted on 2025-08-26, 08:33 authored by Mark PailingMark Pailing, Bart HoogenboomBart Hoogenboom
<p dir="ltr">Atomic force microscopy (AFM) has demonstrated the ability to resolve single DNA molecules in liquid at a spatial resolution that is sufficient to visualize the double helix structure and variations therein. Such variations can be due to inherent configurational flexibility and may be related to, e.g., DNA sequence, ionic screening, supercoiling, or protein binding. These AFM experiments require DNA to be adhered to a solid and preferably flat support. For high-resolution, in-liquid AFM studies so far, such adhesion has commonly been achieved using Ni<sup>2+</sup> ions to electrostatically bridge between the negatively charged DNA and a negatively charged, atomically flat mica surface, yet Ni<sup>2+</sup> ions tend to cause precipitation of salts on the surface, increasing the risk of AFM tip contamination and increasing the corrugation of the support surface, making it harder to distinguish secondary DNA structure.</p><p dir="ltr">Here, we provide the dataset that underpins a study on sample preparation protocol that, instead, relies on Co<sup>2+</sup> ions to adhere DNA to mica. While the Co<sup>2+</sup> is similarly effective as Ni<sup>2+</sup> for facilitating DNA adsorption onto mica, it leads to significantly reduced salt precipitation with the potential to provide enhanced reproducibility in high-resolution DNA imaging by AFM. We expect this to substantially facilitate high-resolution AFM studies of DNA in aqueous solutions.</p><p dir="ltr">The data are in the form of AFM files that can be opened, e.g., by open-source software as available at <a href="https://gwyddion.net/" rel="noreferrer" target="_blank">https://gwyddion.net/</a>, and numerical values in xlsx format for the quantification of the images.</p><p dir="ltr">These data belong to a study Pailing et al., accepted for publication in Nanoscale and available as a preprint at <a href="https://www.biorxiv.org/content/10.1101/2025.05.29.655787v1" rel="noreferrer" target="_blank">https://www.biorxiv.org/content/10.1101/2025.05.29.655787v1</a>.</p>

Funding

Turnkey video-rate atomic force microscopy for nanometre resolution imaging of functional biomolecules and cellular surfaces

Biotechnology and Biological Sciences Research Council

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Single molecule visualisation of PARP trapping by PARP inhibitors

Biotechnology and Biological Sciences Research Council

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Experimental equipment at UCL

Engineering and Physical Sciences Research Council

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