Dynamic superresolution imaging in living cells

The cortical actin cytoskeleton is a complex, highly mobile network that supports the critical cell function of trafficking, immobilizing, and localizing membrane proteins. Conventional diffraction-limited microscopy cannot resolve the characteristic distances between actin bundles. In our group, we use photoactivated localization microscopy (PALM) to image actin filaments with superresolution. We are investigating the dynamics of the cortical actin and its role in the segregation of membrane proteins into specific microdomains. To date, we have successfully performed simultaneous actin superresolution imaging and single particle tracking of plasma membrane proteins in live human embrionic kidney (HEK).

We use image processing techniques to outline the compartments created by the actin cytoskeleton on the plasma membrane. Using this information, the effect of the compartments on individual membrane proteins can be investigated by combining single particle tracking of membrane proteins with tracks of the individual compartments.

 

Group members involved

Sanaz Sadegh, Xinran Xu, Robert Turner, Patrick Mannion

Collaborators

Mike Tamkun, Pablo Visconti, Mariano Buffone, Dario Krapf