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Versatile laser‐based cell manipulator
Author(s) -
Maghelli Nicola,
TolićNørrelykke Iva M.
Publication year - 2008
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.200810026
Subject(s) - optical tweezers , microscope , organelle , biophysics , two photon excitation microscopy , schizosaccharomyces pombe , laser , microtubule , fluorescence microscope , microscopy , centrosome , nanotechnology , optics , fluorescence , chemistry , cell , materials science , yeast , microbiology and biotechnology , physics , biology , saccharomyces cerevisiae , biochemistry , cell cycle
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. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)