Open AccessThree-dimensional laser microsurgery in light-sheet based microscopy (SPIM)
Author(s) -
Christoph J. Engelbrecht,
Klaus Greger,
Emmanuel G. Reynaud,
Uroš Kržič,
Julien Colombelli,
Ernst H. K. Stelzer
Publication year - 2007
Publication title -
optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.15.006420
Subject(s) - light sheet fluorescence microscopy , optics , materials science , microscopy , optical sectioning , laser ablation , ablation , laser , biophotonics , biophysics , nanotechnology , scanning confocal electron microscopy , physics , biology , engineering , aerospace engineering
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.