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.
Highlights
The plasma membrane is the interface through which cells interact with their environment
We aimed to determine whether fluorescent magnetic nanoparticles (FMNPs) were compatible with single-particle tracking (SPT) and magnetic manipulation in the focal plane of a fluorescence microscope in aqueous conditions
We made use of particles consisting of a 100 nm diameter ferromagnetic core and a polymer shell conjugated with a fluorescent dye and streptavidin
Summary
The plasma membrane is the interface through which cells interact with their environment. 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. Local accumulation or separation of specific membrane molecules is crucial for efficient execution and regulation of fundamental biological processes, such as (synaptic) signaling[3], cellular secretion[4] and internalization of nutrients[5], cellular movement[6], and tissue formation[7] Imaging techniques such as super-resolution imaging, SPT, and fluorescence correlation spectroscopy have been instrumental to describe the local membrane composition and dynamics[8]. This allows for the exact correlation of membrane protein location with cellular events and structures
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