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. 2020 Apr 15;10:6441. doi: 10.1038/s41598-020-63418-5

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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 dSTORM super-resolution imaging of neurofilament light chain (NfL) in Vectashield (VS). (a) Widefield images of ND7/23 cells immunolabelled with anti-NfL primary antibody, followed by AF(+)647-conjugated secondary antibody. The same field of view is shown in PBS and in VS. To compare the effect of two different imaging media, look-up table (LUT) intensity scale bars were used. Brightness and contrast were linearly adjusted to show the same display range in both PBS and VS conditions. (b) TIRF image of the boxed regions in panel (a). An image was acquired in VS, with 647 nm laser illumination prior to dSTORM imaging. To ensure the quenched AF(+)647-labelled cell is visible, an auto-scale LUT was used. LUT intensity scale bars show minimum and maximum grey values. The corresponding 3D dSTORM image is shown on the right. The z positions in the 3D dSTORM images are colour-coded according to the height maps shown on right. Height maps contain minimal, maximal and focal (F) z position values.