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. 2020 Jul 27;10:12504. doi: 10.1038/s41598-020-68892-5

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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3D BALM imaging of DNA. (a) A cartoon depiction of a DNA-coated bead immobilized on a glass surface through biotin-streptavidin linkage. (b, c) 3D BALM images (b) and the corresponding cross sections of two DNA-coated microspheres at different axial levels (c) with color-coded height map. The schematics above c illustrate different z levels. (d)–(f) 3D BALM images of 10 kb linear dsDNA tethered to the surface of the glass at both ends (d), tethered to the surface at one end and to a 1-µm diameter bead at the other end (e), tethered to a 1-µm diameter bead at one end and to a 0.4-µm diameter bead at the other end (f). (g) Line profile for determination of lateral and axial FWHM (representing dsDNA, tethered to a 1-µm diameter bead from one end and to a 0.4-µm diameter bead from the other end). Solids lines are fits to a Gaussian.