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. 2023 Apr 1;13:5366. doi: 10.1038/s41598-023-32524-5

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Pre-expansion labeling of presynaptic markers with different signal amplification systems in X10ht. (A) Schematic visualization of the different amplification systems. See the main text for more details. Final used fluorophores are Alexa Fluor488 (AF488) or Abberior Star635P (AS635P). (B) Representative confocal images of expanded neurons immunostained with either the antibody-AF488-based amplification system for SHANK2, or (C) with the different nanobody-based amplification systems detecting VGLUT1. (D) Quantitative analysis of SHANK2 with N = 7 images for 0. Ab w/o amplification, and N = 22 images for 2. Ab-AF-Ab from 3 experiments; N = 18 images for 0. Nb w/o amplification (VGLUT1) and 1. Nb-AF-Ab, N = 13 images for 3. Nb-BT-Ab, and N = 14 images for 4. NbALFA-SpaMo-Ab all to detect VGLUT1, with several ROIs analyzed per image generated from 2 experiments. Data are presented as single data points, mean ± SD.