Approach to map nanotopography of cell surface receptors

Christian Franke; Tomáš Chum; Zuzana Kvíčalová; Daniela Glatzová; Gregor Jörg Gentsch; Alvaro Rodriguez; Dominic A. Helmerich; Lucas Herdly; Harsha Mavila; Otakar Frank; Tomáš Brdička; Sebastian van de Linde; Marek Cebecauer

doi:10.1038/s42003-022-03152-y

Approach to map nanotopography of cell surface receptors

Christian Franke^*, Tomáš Chum, Zuzana Kvíčalová, Daniela Glatzová, Gregor Jörg Gentsch, Alvaro Rodriguez, Dominic A. Helmerich, Lucas Herdly, Harsha Mavila, Otakar Frank, Tomáš Brdička, Sebastian van de Linde, Marek Cebecauer

^*Corresponding author for this work

Physics

Research output: Contribution to journal › Article › peer-review

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Abstract

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.

Original language	English
Article number	218
Number of pages	16
Journal	Communications Biology
Volume	5
Issue number	1
DOIs	https://doi.org/10.1038/s42003-022-03152-y
Publication status	Published - 9 Mar 2022

Funding

We would like to thank Lilliana Barbieri (University of Oxford), Peter Kapusta, Silke Kerruth and Harsha Mavila for technical assistance and professional advice. We are grateful to Markus Sauer (University of Würzburg) for initial support of the project. C.F. would like to thank Laure Plantard and Jan Peychl from the MPI-CBG LMF for technical support. M.C. acknowledges funding from Czech Science Foundation (19-0704S), S.v.d.L. and L.H. acknowledge funding from Academy of Medical Sciences/the British Heart Foundation/the Government Department of Business, Energy and Industrial Strategy/the Wellcome Trust Springboard Award (SBF003\1163) and from the EPSRC through a doctoral fellowship (EPSRC studentship 2031229 and EPSRC Doctoral Training Partnership (DTP) Grant EP/N509760/1). The measurements at the Imaging Methods Core Facility in BIOCEV, Vestec, Czech Republic were supported by MEYS CR grant Z.02.1.01/0.0/0.0/16_013/0001775. We would like to thank Lilliana Barbieri (University of Oxford), Peter Kapusta, Silke Kerruth and Harsha Mavila for technical assistance and professional advice. We are grateful to Markus Sauer (University of W?rzburg) for initial support of the project. C.F. would like to thank Laure Plantard and Jan Peychl from the MPI-CBG LMF for technical support. M.C. acknowledges funding from Czech Science Foundation (19-0704S), S.v.d.L. and L.H. acknowledge funding from Academy of Medical Sciences/the British Heart Foundation/the Government Department of Business, Energy and Industrial Strategy/the Wellcome Trust Springboard Award (SBF003\1163) and from the EPSRC through a doctoral fellowship (EPSRC studentship 2031229 and EPSRC Doctoral Training Partnership (DTP) Grant EP/N509760/1). The measurements at the Imaging Methods Core Facility in BIOCEV, Vestec, Czech Republic were supported by MEYS CR grant Z.02.1.01/0.0/0.0/16_013/0001775.

Keywords

map
nanotopography
cell surface receptors
cell communication
morphology

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10.1038/s42003-022-03152-yLicence: CC BY 4.0

Franke-etal-CB-2022-Approach-to-map-nanotopography-of-cell-surface-receptorsFinal published version, 6.72 MBLicence: CC BY 4.0

Cite this

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AU - Gentsch, Gregor Jörg

AU - Rodriguez, Alvaro

AU - Helmerich, Dominic A.

AU - Herdly, Lucas

AU - Mavila, Harsha

AU - Frank, Otakar

AU - Brdička, Tomáš

AU - van de Linde, Sebastian

AU - Cebecauer, Marek

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AB - 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.

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Approach to map nanotopography of cell surface receptors

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Tuneable photoswitches for chromatic aberration-free multicolour super-resolution imaging

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Approach to map nanotopography of cell surface receptors

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Tuneable photoswitches for chromatic aberration-free multicolour super-resolution imaging

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