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Proceedings Paper

Quantitative single-molecule localization microscopy reports on protein numbers in signaling protein complexes
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Paper Abstract

Knowledge of how proteins organize into functional complexes is essential to understand their biological function. Optical super-resolution techniques provide the spatial resolution necessary to visualize and to investigate individual protein complexes in the context of their cellular environment. Single-molecule localization microscopy (SMLM) builds on the detection of single fluorophore labels, which next to the generation of high-resolution images provides access to quantitative molecular information. We developed various tools for quantitative SMLM (qSMLM), an imaging method that both super-resolves individual protein clusters and reports on molecular numbers by analyzing the kinetics of single emitter blinking. This method is compatible with both fluorescent proteins and organic fluorophores. With qSMLM, we quantify protein copy numbers in single clusters, and we study how changes in the stoichiometry of protein complexes translates into function.

Paper Details

Date Published: 13 February 2020
PDF: 7 pages
Proc. SPIE 11246, Single Molecule Spectroscopy and Superresolution Imaging XIII, 112460N (13 February 2020); doi: 10.1117/12.2550635
Show Author Affiliations
Christos Karathanasis, Goethe-Univ. Frankfurt am Main (Germany)
Tim N. Baldering, Goethe-Univ. Frankfurt am Main (Germany)
Carolin Boeger, Goethe-Univ. Frankfurt am Main (Germany)
Marie-Lena I. E. Harwardt, Goethe-Univ. Frankfurt am Main (Germany)
Yunqing Li, Goethe-Univ. Frankfurt am Main (Germany)
Mark S. Schröder, Goethe-Univ. Frankfurt am Main (Germany)
Nina S. Deußner-Helfmann, Goethe-Univ. Frankfurt am Main (Germany)
Mathilda Glaesmann, Goethe-Univ. Frankfurt am Main (Germany)
Sebastian Malkusch, Goethe-Univ. Frankfurt am Main (Germany)
Marina S. Dietz, Goethe-Univ. Frankfurt am Main (Germany)
Anne-Sophie Hafner, Max Planck Institute for Brain Research (Germany)
Erin Schuman, Max Planck Institute for Brain Research (Germany)
Gerhard Hummer, Max Planck Institute of Biophysics (Germany)
Mike Heilemann, Goethe-Univ. Frankfurt am Main (Germany)

Published in SPIE Proceedings Vol. 11246:
Single Molecule Spectroscopy and Superresolution Imaging XIII
Ingo Gregor; Felix Koberling; Rainer Erdmann, Editor(s)

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