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Optics Express

Optics Express

  • Editor: Michael Duncan
  • Vol. 13, Iss. 10 — May. 16, 2005
  • pp: 3690–3696

Pulse energy dependence of subcellular dissection by femtosecond laser pulses

A. Heisterkamp, I. Z. Maxwell, E. Mazur, J. M. Underwood, J. A. Nickerson, S. Kumar, and D. E. Ingber  »View Author Affiliations


Optics Express, Vol. 13, Issue 10, pp. 3690-3696 (2005)
http://dx.doi.org/10.1364/OPEX.13.003690


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Abstract

Precise dissection of cells with ultrashort laser pulses requires a clear understanding of how the onset and extent of ablation (i.e., the removal of material) depends on pulse energy. We carried out a systematic study of the energy dependence of the plasma-mediated ablation of fluorescently-labeled subcellular structures in the cytoskeleton and nuclei of fixed endothelial cells using femtosecond, near-infrared laser pulses focused through a high-numerical aperture objective lens (1.4 NA). We find that the energy threshold for photobleaching lies between 0.9 and 1.7 nJ. By comparing the changes in fluorescence with the actual material loss determined by electron microscopy, we find that the threshold for true material ablation is about 20% higher than the photobleaching threshold. This information makes it possible to use the fluorescence to determine the onset of true material ablation without resorting to electron microscopy. We confirm the precision of this technique by severing a single microtubule without disrupting the neighboring microtubules, less than 1 µm away.

© 2005 Optical Society of America

OCIS Codes
(140.7090) Lasers and laser optics : Ultrafast lasers
(170.0180) Medical optics and biotechnology : Microscopy
(180.2520) Microscopy : Fluorescence microscopy

ToC Category:
Research Papers

History
Original Manuscript: February 28, 2005
Revised Manuscript: May 1, 2005
Published: May 16, 2005

Citation
A. Heisterkamp, I. Z. Maxwell, E. Mazur, J. M. Underwood, J. A. Nickerson, S. Kumar, and D. E. Ingber, "Pulse energy dependence of subcellular dissection by femtosecond laser pulses," Opt. Express 13, 3690-3696 (2005)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-13-10-3690


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