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Abstract:
Here we describe a two-photon microscope and laser
ablation setup combined with optical tweezers. We
tested the setup on the fission yeast Schizosaccharomyces
pombe, a commonly used model organism. We
show that long-term imaging can be achieved without
significant photo-bleaching or damage of the sample.
The setup can precisely ablate sub-micrometer structures,
such as microtubules and mitotic spindles, inside
living cells, which remain viable after the manipulation.
Longer exposure times lead to ablation, while shorter
exposures lead to photo-bleaching of the target structure.
We used optical tweezers to trap intracellular particles
and to displace the cell nucleus. Two-photon fluorescence
imaging of the manipulated cell can be performed
simultaneously with trapping. The combination of techniques
described here may help to solve a variety of
problems in cell biology, such as positioning of organelles
and the forces exerted by the cytoskeleton.
