High-Precision Protein-Tracking With Interferometric Scattering Microscopy

Richard W Taylor; Cornelia Holler; Reza Gholami Mahmoodabadi; Michelle Küppers; Houman Mirzaalian Dastjerdi; Vasily Zaburdaev; Alexandra Schambony; Vahid Sandoghdar

doi:10.3389/fcell.2020.590158

High-Precision Protein-Tracking With Interferometric Scattering Microscopy

Front Cell Dev Biol. 2020 Nov 3:8:590158. doi: 10.3389/fcell.2020.590158. eCollection 2020.

Authors

Richard W Taylor^{1

2}, Cornelia Holler^{1

2}, Reza Gholami Mahmoodabadi^{1

2}, Michelle Küppers^{1

2

3}, Houman Mirzaalian Dastjerdi^{1

2

4}, Vasily Zaburdaev^{2

5}, Alexandra Schambony^{1

2

5}, Vahid Sandoghdar^{1

2

3}

Affiliations

¹ Max Planck Institute for the Science of Light, Erlangen, Germany.
² Max-Planck-Zentrum für Physik und Medizin, Erlangen, Germany.
³ Department of Physics, Friedrich Alexander University Erlangen-Nuremberg, Erlangen, Germany.
⁴ Department of Computer Science, Friedrich Alexander University Erlangen-Nuremberg, Erlangen, Germany.
⁵ Department of Biology, Friedrich Alexander University Erlangen-Nuremberg, Erlangen, Germany.

Abstract

The mobility of proteins and lipids within the cell, sculpted oftentimes by the organization of the membrane, reveals a great wealth of information on the function and interaction of these molecules as well as the membrane itself. Single particle tracking has proven to be a vital tool to study the mobility of individual molecules and unravel details of their behavior. Interferometric scattering (iSCAT) microscopy is an emerging technique well-suited for visualizing the diffusion of gold nanoparticle-labeled membrane proteins to a spatial and temporal resolution beyond the means of traditional fluorescent labels. We discuss the applicability of interferometric single particle tracking (iSPT) microscopy to investigate the minutia in the motion of a protein through measurements visualizing the mobility of the epidermal growth factor receptor in various biological scenarios on the live cell.

Keywords: epidermal growth factor receptor; iSCAT; iSPT; interferometric scattering microscopy; live cell imaging; membrane organization; single particle tracking.