OSA's Digital Library

Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editor: Gregory W. Faris
  • Vol. 2, Iss. 6 — Jun. 13, 2007

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)

View Full Text Article

Enhanced HTML    Acrobat PDF (1544 KB) Open Access

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



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

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

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)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. M. W. Berns, J. Aist, J. Edwards, K. Strahs, J. Girton, P. McNeill, J. B. Rattner, M. Kitzes, M. Hammer-Wilson, L. H. Liaw, A. Siemens, M. Koonce, S. Peterson, S. Brenner, J. Burt, R. Walter, P. J. Bryant, D. van Dyk, J. Coulombe, T. Cahill, and G. S. Berns, "Laser microsurgery in cell and developmental biology," Science 213, 505-513 (1981). [CrossRef] [PubMed]
  2. S. W. Grill, P. Gönczy, E. H. K. Stelzer, and A. A. Hyman, "Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo," Nature 409, 630-633 (2001). [CrossRef] [PubMed]
  3. E. L. Botvinick, V. Venugopalan, J. V. Shah, L. H. Liaw, and M. W. Berns, "Controlled ablation of microtubules using a picosecond laser," Biophys. J. 87, 4203-4212 (2004). [CrossRef] [PubMed]
  4. J. Colombelli, E. G. Reynaud, J. Rietdorf, R. Pepperkok, and E. H. K. Stelzer, "In vivo selective cytoskeleton dynamics quantification in interphase cells induced by pulsed ultraviolet laser nanosurgery," Traffic 6, 1093-1102 (2005). [CrossRef] [PubMed]
  5. S. Kumar, I. Z. Maxwell, A. Heisterkamp, T. R. Polte, T. P. Lele, M. Salanga, E. Mazur, and D. E. Ingber, "Viscoelastic retraction of single living stress fibers and its impact on cell shape, cytoskeletal organization, and extracellular matrix mechanics," Biophys. J. 90, 3762-3773 (2006). [CrossRef] [PubMed]
  6. D. P. Kiehart, C. G. Galbraith, K. A. Edwards, W. L. Rickoll, and R. A. Montague, "Multiple forces contribute to cell sheet morphogenesis for dorsal closure in Drosophila," J. Cell Biol. 149, 471-490 (2000). [CrossRef] [PubMed]
  7. M. S. Hutson, Y. Tokutake, M. Chang, J. W. Bloor, S. Venakides, D. P. Kiehart, and G. S. Edwards, "Forces for morphogenesis investigated with laser microsurgery and quantitative modeling," Science 300, 145-149 (2003). [CrossRef] [PubMed]
  8. W. Supatto, D. Débarre, B. Moulia, E. Brouzés, J. Martin, E. Farge, and E. Beaurepaire, "In vivo modulation of morphogenetic movements in Drosophila embryos with femtosecond laser pulses," Proc. Natl. Acad. Sci. U S A 102, 1047-1052 (2005). [CrossRef] [PubMed]
  9. D. B. Edelman and E. W. Keefer, "A cultural renaissance: in vitro cell biology embraces three-dimensional context," Exp. Neurol. 192, 1-6 (2005). [CrossRef] [PubMed]
  10. L. G. Griffith, and M. A. Swartz, "Capturing complex 3D tissue physiology in vitro," Nat. Rev. Mol. Cell Biol. 7, 211-224 (2006). [CrossRef] [PubMed]
  11. H. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, "Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain," Nat. Methods 4, 331-336 (2007). [CrossRef] [PubMed]
  12. J. Huisken, J. Swoger, F. D. Bene, J. Wittbrodt, and E. H. K. Stelzer, "Optical sectioning deep inside live embryos by selective plane illumination microscopy," Science 305, 1007-1009 (2004). [CrossRef] [PubMed]
  13. K. Greger, J. Swoger, and E. H. K. Stelzer, "Basic building units and properties of a fluorescence Single Plane Illumination Microscope (SPIM)," Rev. Sci. Instrum. 78, 023705 (2007). [CrossRef] [PubMed]
  14. C. J. Engelbrecht, and E. H. K. Stelzer, "Resolution enhancement in a light-sheet-based microscope (SPIM)," Opt. Lett. 31, 1477-1479 (2006). [CrossRef] [PubMed]
  15. P. J. Verveer, J. Swoger, F. Pampaloni, K. Greger, M. Marcello, and E.H.K. Stelzer, "High-resolution three-dimensional imaging of large specimens with light sheet-based microscopy," Nat. Methods 4, 311-313 (2007). [PubMed]
  16. J. Colombelli, S. W. Grill, and E. H. K. Stelzer, "UV diffraction limited nanosurgery of live biological tissues," Rev. Sci. Instrum. 75, 472-478 (2004). [CrossRef]
  17. A. Vogel, and V. Venugopalan, "Mechanisms of pulsed laser ablation of biological tissues," Chem. Rev. 103, 577-644 (2003). [CrossRef] [PubMed]
  18. S. W. Grill, and E.H. K. Stelzer, "Method to calculate lateral and axial gain factors of optical setups with a large solid angle," J. Opt. Soc. Am. A 16, 2658-2665 (1999). [CrossRef]
  19. L. E. O'Brien, M. M. P. Zegers, and K. E. Mostov, "Opinion: Building epithelial architecture: insights from three-dimensional culture models," Nat. Rev. Mol. Cell Biol. 3, 531-537 (2002). [CrossRef]
  20. B. Stramer, W. Wood, M. J. Galko, M. J. Redd, A. Jacinto, S. M. Parkhurst, and P. Martin, "Live imaging of wound inflammation in Drosophila embryos reveals key roles for small GTPases during in vivo cell migration," J. Cell Biol. 168, 567-573 (2005). [CrossRef] [PubMed]
  21. A. Nimmerjahn, F. Kirchhoff, and F. Helmchen, "Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo," Science 308, 1314-1318 (2005). [CrossRef] [PubMed]
  22. R. Farooqui, and G. Fenteany, "Multiple rows of cells behind an epithelial wound edge extend cryptic lamellipodia to collectively drive cell-sheet movement," J. Cell Sci. 118, 51-63 (2005). [CrossRef]
  23. F. Jankovics, and D. Brunner, "Transiently reorganized microtubules are essential for zippering during dorsal closure in Drosophila melanogaster," Dev. Cell 11, 375-385 (2006). [CrossRef] [PubMed]

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.

Supplementary Material

» Media 1: MOV (799 KB)     
» Media 2: MOV (592 KB)     

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited