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Approach to map nanotopography of cell surface receptors

C. Franke, T. Chum, Z. Kvíčalová, D. Glatzová, GJ. Gentsch, A. Rodriguez, DA. Helmerich, L. Herdly, H. Mavila, O. Frank, T. Brdička, S. van de Linde, M. Cebecauer

. 2022 ; 5 (1) : 218. [pub] 20220309

Jazyk angličtina Země Velká Británie

Typ dokumentu časopisecké články, práce podpořená grantem

Perzistentní odkaz   https://www.medvik.cz/link/bmc22019207

Grantová podpora
SBF003\1163 Wellcome Trust (Wellcome)
Wellcome Trust - United Kingdom
19-0704S Grantová Agentura České Republiky (Grant Agency of the Czech Republic)
EP/N509760/1 RCUK | Engineering and Physical Sciences Research Council (EPSRC)
2031229 RCUK | Engineering and Physical Sciences Research Council (EPSRC)
British Heart Foundation - United Kingdom

Cells communicate with their environment via surface receptors, but nanoscopic receptor organization with respect to complex cell surface morphology remains unclear. This is mainly due to a lack of accessible, robust and high-resolution methods. Here, we present an approach for mapping the topography of receptors at the cell surface with nanometer precision. The method involves coating glass coverslips with glycine, which preserves the fine membrane morphology while allowing immobilized cells to be positioned close to the optical surface. We developed an advanced and simplified algorithm for the analysis of single-molecule localization data acquired in a biplane detection scheme. These advancements enable direct and quantitative mapping of protein distribution on ruffled plasma membranes with near isotropic 3D nanometer resolution. As demonstrated successfully for CD4 and CD45 receptors, the described workflow is a straightforward quantitative technique to study molecules and their interactions at the complex surface nanomorphology of differentiated metazoan cells.

Citace poskytuje Crossref.org

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$a Chum, Tomáš $u Department of Biophysical Chemistry, J. Heyrovsky Institute of Physical Chemistry, Czech Academy of Sciences, Prague, Czech Republic
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