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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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Comparison of the effect of 25% Vectashield (25% VS) on AF647 and AF(+)647 fluorescence. (a) Mouse cortical neurons (MCN) immunolabelled with anti-pan voltage-gated sodium channels (panNav) antibody, followed by AF647-conjugated secondary antibody. (b) MCN immunolabelled with panNav, followed by AF(+)647-conjugated secondary antibody. Upper panels show widefield images of the same field of view acquired in PBS and in 25% VS. Brightness and contrast were linearly adjusted to show the same display range in both PBS and 25% VS conditions (as indicated by look-up table (LUT) intensity scale bars). Lower panels show corresponding 3D dSTORM images of the boxed regions from panels (a and b). 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.