Abstract

The plasma membrane is the interface through which cells interact with their environment. Membrane proteins are embedded in the lipid bilayer of the plasma membrane and their function in this context is often linked to their specific location and dynamics within the membrane. However, few methods are available to manipulate membrane protein location at the single-molecule level. Here, we use fluorescent magnetic nanoparticles (FMNPs) to track membrane molecules and to control their movement. FMNPs allow single-particle tracking (SPT) at 10 nm and 5 ms spatiotemporal resolution, and using a magnetic needle, we pull membrane components laterally with femtonewton-range forces. In this way, we drag membrane proteins over the surface of living cells. Doing so, we detect barriers which we could localize to the submembrane actin cytoskeleton by super-resolution microscopy. We present here a versatile approach to probe membrane processes in live cells via the magnetic control of membrane protein motion. Membrane proteins are embedded in the lipid bilayer of the plasma membrane and their function in this context is often linked to their specific location and dynamics within the membrane. Here authors report the use of fluorescent magnetic nanoparticles to track membrane molecules and to manipulate their movement and pull membrane components laterally through the membrane with femtonewton-range forces.

Highlights

  • 1531-Pos Bayesian Grouping of Localizations, Sub-nanometer Precision, Counting and Resolution Doubling Mohamadreza Fazel1, Sebastian Restrepo Cruz2, Jennifer Gillette2, Bernd Rieger3, Ralf Jungmann4, Keith A

  • We describe a method for Bayesian Grouping of Localizations (BaGoL) that combines localizations from multiple blinking/binding events

  • BaGoL can improve localization precision to better than 1 nm, can be used for inference on the number of fluorophores in small clusters, and can be applied to any SMLM data set for improved resolution

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Summary

Introduction

1531-Pos Bayesian Grouping of Localizations, Sub-nanometer Precision, Counting and Resolution Doubling Mohamadreza Fazel1, Sebastian Restrepo Cruz2, Jennifer Gillette2, Bernd Rieger3, Ralf Jungmann4, Keith A. Since SMLM relies on localizing a sparse subset of single photoactivated fluorophores in space and time, long data acquisition times are required to thoroughly sample the labeled biomolecules. During this data acquisition time, cells must be exposed to the high-energy photons of the photoactivating laser.

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