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Interferometric scattering microscopy reveals microsecond nanoscopic protein motion on a live cell membrane

View ORCID ProfileRichard W. Taylor, Reza Gholami Mahmoodabadi, Verena Rauschenberger, Andreas Giessl, View ORCID ProfileAlexandra Schambony, View ORCID ProfileVahid Sandoghdar
doi: https://doi.org/10.1101/401133
Richard W. Taylor
1Max-Planck-Zentrum für Physik und Medizin, 91058 Erlangen, Germany.
2Max Planck Institute for the Science of Light, 91058 Erlangen, Germany.
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  • ORCID record for Richard W. Taylor
Reza Gholami Mahmoodabadi
1Max-Planck-Zentrum für Physik und Medizin, 91058 Erlangen, Germany.
2Max Planck Institute for the Science of Light, 91058 Erlangen, Germany.
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Verena Rauschenberger
3Developmental Biology, Friedrich Alexander University Erlangen-Nuremberg, 91058 Erlangen, Germany.
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Andreas Giessl
4Animal Physiology, Friedrich Alexander University Erlangen-Nuremberg, 91058 Erlangen, Germany.
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Alexandra Schambony
3Developmental Biology, Friedrich Alexander University Erlangen-Nuremberg, 91058 Erlangen, Germany.
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Vahid Sandoghdar
1Max-Planck-Zentrum für Physik und Medizin, 91058 Erlangen, Germany.
2Max Planck Institute for the Science of Light, 91058 Erlangen, Germany.
5Department of Physics, Friedrich Alexander University Erlangen-Nuremberg, 91058 Erlangen, Germany.
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  • For correspondence: vahid.sandoghdar@mpl.mpg.de
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Abstract

Much of the biological functions of a cell are dictated by the intricate motion of proteins within its membrane over a spatial range of nanometers to tens of micrometers and time intervals of microseconds to minutes. While this rich parameter space is not accessible to fluorescence microscopy, it can be within reach of interferometric scattering (iSCAT) particle tracking. Being sensitive even to single unlabeled proteins, however, iSCAT is easily accompanied by a large speckle-like background, which poses a substantial challenge for its application to cellular imaging. Here, we show that these difficulties can be overcome and demonstrate tracking of transmembrane epidermal growth factor receptors (EGFR) with nanometer precision in all three dimensions at up to microsecond speeds and tens of minutes duration. We provide unprecedented examples of nanoscale motion and confinement in ubiquitous processes such as diffusion in the plasma membrane, transport on filopodia, and endocytosis.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted August 27, 2018.
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Interferometric scattering microscopy reveals microsecond nanoscopic protein motion on a live cell membrane
Richard W. Taylor, Reza Gholami Mahmoodabadi, Verena Rauschenberger, Andreas Giessl, Alexandra Schambony, Vahid Sandoghdar
bioRxiv 401133; doi: https://doi.org/10.1101/401133
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Interferometric scattering microscopy reveals microsecond nanoscopic protein motion on a live cell membrane
Richard W. Taylor, Reza Gholami Mahmoodabadi, Verena Rauschenberger, Andreas Giessl, Alexandra Schambony, Vahid Sandoghdar
bioRxiv 401133; doi: https://doi.org/10.1101/401133

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