Unraveling nanotopography of cell surface receptors

Christian Franke, Tomáš Chum, Zuzana Kvíčalová, Daniela Glatzová, Alvaro Rodriguez, Dominic A. Helmerich, Otakar Frank, Tomáš Brdička, Sebastian van de Linde, Marek Cebecauer

Research output: Working paper

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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 languageEnglish
Place of PublicationCold Spring Harbor, NY
Number of pages55
DOIs
Publication statusPublished - 30 Jul 2020

Keywords

  • nanotopography
  • cell surface receptors
  • cells
  • nanoscopic receptor organization
  • cell surface morphology
  • CD4
  • CD45
  • metazoan cells

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