@article{ad69782931084eacb662801045f754a7,
title = "Fix your membrane receptor imaging: Actin cytoskeleton and CD4 membrane organization disruption by chemical fixation",
abstract = "Single-molecule localization microscopy (SMLM) techniques allow near molecular scale resolution (~ 20 nm) as well as precise and robust analysis of protein organization at different scales. SMLM hardware, analytics and probes have been the focus of a variety of studies and are now commonly used in laboratories across the world. Protocol reliability and artifact identification are increasingly seen as important aspects of super-resolution microscopy. The reliability of these approaches thus requires in-depth evaluation so that biological findings are based on solid foundations. Here we explore how different fixation approaches that disrupt or preserve the actin cytoskeleton affect membrane protein organization. Using CD4 as a model, we show that fixation-mediated disruption of the actin cytoskeleton correlates with changes in CD4 membrane organization. We highlight how these artifacts are easy to overlook and how careful sample preparation is essential for extracting meaningful results from super-resolution microscopy.",
keywords = "Actin cortex, Artefact analysis, CD4, Fixation, Super-resolution imaging",
author = "Pereira, {Pedro M.} and David Albrecht and Si{\^a}n Culley and Caron Jacobs and Mark Marsh and Jason Mercer and Ricardo Henriques",
note = "Funding Information: This work was funded by grants from the UK Biotechnology and Biological Sciences Research Council (BB/M022374/1; BB/P027431/1; BB/R000697/1; BB/S507532/1) (RH, PP, and CJ); The UK Medical Research Council (MR/K015826/1) (RH, JM, and MM); The Wellcome Trust (203276/Z/16/Z) (RH); the MRC Programme Grant (MC_UU12018/7) (JM); The European Research Council (649101-UbiProPox) (JM); The MRC Programme Grant (MC_U12016/1) (MM); DA has received funding from the European Union{\textquoteright}s Horizon 2020 research and innovation programme under the Marie Sk{\l}odowska-Curie grant agreement No. 750673. CJ was funded by a Commonwealth scholarship, funded by the UK government. Publisher Copyright: {\textcopyright} 2019 Pereira, Albrecht, Culley, Jacobs, Marsh, Mercer and Henriques. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.",
year = "2019",
doi = "10.3389/fimmu.2019.00675",
language = "English",
volume = "10",
journal = "Frontiers in Immunology",
issn = "1664-3224",
publisher = "Frontiers Media",
number = "APR",
}