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
Mike Tamkun, Pablo Visconti, Maria Gracia Gervasi, Mariano Buffone, Dario Krapf
S. Sadegh, J.L. Higgins, P.C. Mannion, M.M. Tamkun, and D. Krapf, “Plasma Membrane is Compartmentalized by a Self-Similar Cortical Actin Meshwork”, Phys. Rev. X 7, 011031 (2017) pdf
A. Alvau et al., “The tyrosine kinase FER is responsible for the capacitation-associated increase in tyrosine phosphorylation in murine sperm”, Development 143, 2325 (2016) pdf
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