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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 15, Iss. 10 — May. 14, 2007
  • pp: 6420–6430

Three-dimensional laser microsurgery in light-sheet based microscopy (SPIM)

Christoph J. Engelbrecht, Klaus Greger, Emmanuel G. Reynaud, Uroš Kržic, Julien Colombelli, and Ernst H. K. Stelzer  »View Author Affiliations


Optics Express, Vol. 15, Issue 10, pp. 6420-6430 (2007)
http://dx.doi.org/10.1364/OE.15.006420


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Abstract

Advances in the life sciences rely on the ability to observe dynamic processes in live systems and in environments that mimic in-vivo situations. Therefore, new methodological developments have to provide environments that resemble physiologically and clinically relevant conditions as closely as possible. In this work, plasma-induced laser nanosurgery for three-dimensional sample manipulation and sample perturbation is combined with optically sectioning light-sheet based fluorescence microscopy (SPIM) and applied to three-dimensional biological model systems. This means: a) working with a biological system that is not confined to essentially two dimensions like cell cultures on cover glasses, b) gaining intrinsic optical sectioning capabilities by an efficient three-dimensional fluorescence imaging system, and c) using arbitrarily-shaped three-dimensional ablation-patterns by a plasma-induced laser ablation system that prevent damage to surrounding tissues. Spatial levels in our biological applications range from sub-microns during delicate ablation of single microtubules over the confined disruption of cell membranes in an MDCK-cyst to the macroscopic cutting of a millimeter-sized Zebrafish caudal fin with arbitrary three-dimensional ablation patterns. Dynamic processes like laser-induced hemocyte migration can be studied with our SPIM-microscalpel in intact, live embryos.

© 2007 Optical Society of America

OCIS Codes
(000.1430) General : Biology and medicine
(110.0180) Imaging systems : Microscopy
(170.1020) Medical optics and biotechnology : Ablation of tissue
(170.1420) Medical optics and biotechnology : Biology
(170.1530) Medical optics and biotechnology : Cell analysis
(170.2520) Medical optics and biotechnology : Fluorescence microscopy

ToC Category:
Medical Optics and Biotechnology

History
Original Manuscript: April 5, 2007
Revised Manuscript: May 3, 2007
Manuscript Accepted: May 4, 2007
Published: May 10, 2007

Virtual Issues
Vol. 2, Iss. 6 Virtual Journal for Biomedical Optics

Citation
Christoph J. Engelbrecht, Klaus Greger, Emmanuel G. Reynaud, Uroš Kržic, Julien Colombelli, and Ernst H. Stelzer, "Three-dimensional laser microsurgery in light-sheet based microscopy (SPIM)," Opt. Express 15, 6420-6430 (2007)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-15-10-6420


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