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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 axonal initial segments in Vectashield (VS) and GLOX BME. (a,b) Mouse cortical neurons (MCN) labelled with anti-ankyrin G (AnkG) antibody. (c,d) MCN labelled with anti-pan voltage-gated sodium channel (panNav) antibody. Left panels show TIRF images, acquired with 647 nm laser illumination prior to 3D dSTORM imaging. To ensure the quenched AF(+)647 in axonal initial segments in VS is visible, an autoscale look-up table (LUT) was used. LUT intensity scale bars show minimum and maximum grey values. Middle panels show corresponding 3D dSTORM images of AnkG (a,b) and panNav (c,d). Right panels show magnification of the boxed regions shown in the middle panels. Despite the quenching effect of VS, periodic patterns of AnkG and voltage-gated sodium channels can still be resolved. 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.