Laser nanosurgery and manipulation in living cells

We present a combination of nonlinear microscopy, laser nanosurgery and optical trapping applied to the 3D imaging and manipulation of intracellular structures in live cells. We use Titanium-sapphire laser pulses for a combined nonlinear microscopy and nanosurgery on microtubules tagged with green fluorescent protein (GFP) in fission yeast. The same laser source is also used to trap small round lipid droplets naturally present in the cell. The trapped droplets are used as handles to exert a pushing force on the nucleus, allowing for a displacement of the nucleus away from its normal position in the center of the cell. We show that nonlinear nanosurgery and optical manipulation can be performed with sub-micrometer precision and without visible collateral damage to the cell. We present this combination as an important tool in cell biology for the manipulation of specific structures in alternative to genetic methods or chemical agents. This technique can be applied to several fundamental problems in cell biology, including the study of dynamics processes in cell division.