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. 2018 May 17;8:7809. doi: 10.1038/s41598-018-24119-2

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© The Author(s) 2018

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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Two-color STED nanoscopy of peroxisome membrane and matrix. Dual immunofluorescence on human skin fibroblasts. (A) Monoclonal anti-PMP70 labeled with KK114 secondary label in red, polyclonal rabbit anti-Catalase (anti-CAT1) labeled with Atto594-coupled secondary in green, and a merge of both (right). (B) Blow-up of box in (A). (C) Gaussian fit of the line scan marked in (B). (D) Anti-PMP70 labeled with KK114-coupled secondary antibody in red, polyclonal rabbit anti-acetyl-CoA acyltransferase1 (anti-ACAA1) in green, and merged images of both. (E) Inset magnification of (D). (F) Gaussian fit of the line scan marked in (E). Images were smoothed by 3 × 3 average filter and linearly scaled. Scale bars 500 nm (A,D), and 100 nm (B,E).