Abstract
Separation and transport of defined populations of living cells grown on a thin membrane can be achieved by laser microdissection LMD of the sample of interest, followed by a laser-induced forward transport process laser pressure "catapulting" LPC of the dissected cell cluster. We investigate the dynamics of LMD and LPC with focused and defocused UV-A laser pulses by means of timeresolved photography. Catapulting is driven by plasma formation when tightly focused pulses are used, and by confined thermal ablation at the bottom of the sample for defocused catapulting. With both modalities, the initial specimen velocity amounts to about 50 to 60 m/ s. Time-resolved photography of live cell catapulting reveals that in defocused catapulting, strong shear forces arise when the sample is accelerated out of the culture medium covering the cells. By contrast, pulses focused at the periphery of the specimen cause a fast rotational movement that minimizes the flow of culture medium parallel to the sample surface, and thus the resulting shear stresses. Therefore, the recultivation rate of catapulted cells is much higher when focused pulses are used. Compared to collateral damage by mechanical forces, side effects by heat and UV exposure of the cells play only a minor role.
| Original language | English |
|---|---|
| Article number | 054016 |
| Journal | Journal of Biomedical Optics |
| Volume | 12 |
| Issue number | 5 |
| ISSN | 1083-3668 |
| DOIs | |
| Publication status | Published - 09.2007 |
Funding
This work was sponsored by the German Bundesministerium für Bildung und Forschung (BMBF) under grant number 13N8461. Palm Microlaser Technologies provided the microbeam system used. We appreciate stimulating discussions with Karin Schütze, Bernd Sägmüller, and Yilmaz Niyaz of Palm-Microlaser Technologies, and with Heyke Diddens (Institute of Biomedical Optics, University of Lübeck), who also provided the CHO cell line used for live cell catapulting. The suggestion to use oil to prevent evaporation of the culture medium shortly before catapulting was made by one of the reviewers of this work. We thank Sebastian Freidank, Florian Wölbeling, Helge Meyer, and Reinhard Schulz of the Institute of Biomedical Optics, University of Lübeck, for technical assistance.
UN SDGs
This output contributes to the following UN Sustainable Development Goals (SDGs)
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SDG 9 Industry, Innovation, and Infrastructure
Research Areas and Centers
- Academic Focus: Biomedical Engineering
Fingerprint
Dive into the research topics of 'Principles of laser-induced separation and transport of living cells'. Together they form a unique fingerprint.Projects
- 1 Finished
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New methods for laser microdissection and non-contact transportation of tissue samples and living cells for molecular biological analyses
Gebert, A. (Principal Investigator (PI)), Klages , C.-P. (Principal Investigator (PI)) & Vogel, A. (Principal Investigator (PI))
01.01.06 → 31.12.10
Project: DFG Individual Projects
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