Tracking of single proteins in living cells.
© 2012 EPFL
Multicolor Single Molecule Tracking of Stochastically Active Synthetic Dyes.
The group of Suliana Manley(Laboratory of Experimental Biophysics) describe a new and general approach to high-density tracking of single proteins in living cells. They overcome the limitations of current methods by using genetically encoded tags to target synthetic dyes to specific proteins, and inducing them to cycle through long-lived dark states. This cycling through fluorescent states, previously exploited for super-resolution PALM/STORM imaging, allows them to readily track hundreds of molecules in a single cell. Using this combination of technologies, they are able to maintain flexibility of labeling targets while taking advantage of the superior photon yields of synthetic dyes over fluorescent proteins. As they demonstrate, it is then possible to track proteins both on the surface and within intracellular compartments, with multiple colors.