High-speed tracking of intracellular structures: understanding the transport mechanisms in living plant cells

Zera (R) technology offers the possibility to artificially induce the formation of spherical organelles in different kinds of cells. Their large size and high density, compared to the native organelles of the cell, make them good candidates to be used as a handles for the realization of biophysical experiments inside living cells. Furthermore, they present a high signal-to-noise ratio in fluorescence microscopy and small photobleaching. This work focuses mainly on the nature of protein body motion in Nicotiana Benthamiana (tobacco) cells. The high-speed tracking of these structures reveals that they move in a stepwise mode, suggesting that myosin XI motors directly pull these organelles through the cytosol. Our results indicate that these artificially-induced structures are well integrated into the natural processes of the cell so that the technique may be advantageous for the study of the intracellular transport mechanisms. Large forces can be exerted with optical tweezers to mechanically interact with the moving organelles.