UMMS Affiliation

Department of Cell Biology

Publication Date

2005-07-23

Document Type

Article

Subjects

Actins; Animals; Cell Nucleus; Cytoskeleton; Endothelial Cells; *Lasers; Microscopy, Electron, Transmission; Microscopy, Fluorescence; Microtubules; Radiation Dosage

Disciplines

Cell Biology

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 micrometer away.

Rights and Permissions

This paper was published in Optics Express and is made available as an electronic reprint with the permission of OSA. The paper can be found at the following URL on the OSA website: Link to article on publisher's website. Systematic or multiple reproduction or distribution to multiple locations via electronic or other means is prohibited and is subject to penalties under law.

DOI of Published Version

10.1364/OPEX.13.003690

Source

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). The paper can be found at the following URL on the OSA website: Link to article on publisher's website.

Journal/Book/Conference Title

Optics express

Related Resources

Link to Article in PubMed

PubMed ID

16035172

Included in

Cell Biology Commons

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