Details
| Original language | English |
|---|---|
| Title of host publication | Frontiers in Ultrafast Optics |
| Subtitle of host publication | Biomedical, Scientific, and Industrial Applications X |
| Publication status | Published - 25 Feb 2010 |
| Externally published | Yes |
| Event | Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications X - San Francisco, CA, United States Duration: 24 Jan 2010 → 26 Jan 2010 |
Publication series
| Name | Proceedings of SPIE - The International Society for Optical Engineering |
|---|---|
| Volume | 7589 |
| ISSN (Print) | 0277-786X |
Abstract
Cloning of several mammalian species has been achieved by somatic cell nuclear transfer over the last decade. However, this method still results in very low efficiencies originating from biological and technical aspects. The highly-invasive mechanical enucleation belongs to the technical aspects and requires considerable micromanipulation skill. In this paper, we present a novel non-invasive method for combined oocyte imaging and automated functional enucleation using femtosecond (fs) laser pulses. After three-dimensional imaging of Hoechst-labeled porcine oocytes by multiphoton microscopy, our self-developed software automatically determined the metaphase plate position and shape. Subsequent irradiation of this volume with the very same laser at higher pulse energies in the low-density-plasma regime was used for metaphase plate ablation. We show that functional fs laser-based enucleation of porcine oocytes completely inhibited further embryonic development while maintaining intact oocyte morphology. In contrast, non-irradiated oocytes were able to develop to the blastocyst stage without significant differences to control oocytes. Our results indicate that fs laser systems offer great potential for oocyte imaging and enucleation as a fast, easy to use and reliable tool which may improve the efficiency of somatic cell clone production.
Keywords
- Cell surgery, Femtosecond laser, Oocyte enucleation, SCNT, Somatic cell nuclear transfer
ASJC Scopus subject areas
- Materials Science(all)
- Electronic, Optical and Magnetic Materials
- Physics and Astronomy(all)
- Condensed Matter Physics
- Computer Science(all)
- Computer Science Applications
- Mathematics(all)
- Applied Mathematics
- Engineering(all)
- Electrical and Electronic Engineering
Cite this
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- Apa
- Vancouver
- BibTeX
- RIS
Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications X. 2010. 75890A (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 7589).
Research output: Chapter in book/report/conference proceeding › Conference contribution › Research › peer review
}
TY - GEN
T1 - Functional enucleation of porcine oocytes for somatic cell nuclear transfer using femtosecond laser pulses
AU - Kuetemeyer, K.
AU - Lucas-Hahn, A.
AU - Petersen, Bjoern
AU - Hassel, Petra
AU - Lemme, Erika
AU - Niemann, Heiner
AU - Heisterkamp, Alexander
PY - 2010/2/25
Y1 - 2010/2/25
N2 - Cloning of several mammalian species has been achieved by somatic cell nuclear transfer over the last decade. However, this method still results in very low efficiencies originating from biological and technical aspects. The highly-invasive mechanical enucleation belongs to the technical aspects and requires considerable micromanipulation skill. In this paper, we present a novel non-invasive method for combined oocyte imaging and automated functional enucleation using femtosecond (fs) laser pulses. After three-dimensional imaging of Hoechst-labeled porcine oocytes by multiphoton microscopy, our self-developed software automatically determined the metaphase plate position and shape. Subsequent irradiation of this volume with the very same laser at higher pulse energies in the low-density-plasma regime was used for metaphase plate ablation. We show that functional fs laser-based enucleation of porcine oocytes completely inhibited further embryonic development while maintaining intact oocyte morphology. In contrast, non-irradiated oocytes were able to develop to the blastocyst stage without significant differences to control oocytes. Our results indicate that fs laser systems offer great potential for oocyte imaging and enucleation as a fast, easy to use and reliable tool which may improve the efficiency of somatic cell clone production.
AB - Cloning of several mammalian species has been achieved by somatic cell nuclear transfer over the last decade. However, this method still results in very low efficiencies originating from biological and technical aspects. The highly-invasive mechanical enucleation belongs to the technical aspects and requires considerable micromanipulation skill. In this paper, we present a novel non-invasive method for combined oocyte imaging and automated functional enucleation using femtosecond (fs) laser pulses. After three-dimensional imaging of Hoechst-labeled porcine oocytes by multiphoton microscopy, our self-developed software automatically determined the metaphase plate position and shape. Subsequent irradiation of this volume with the very same laser at higher pulse energies in the low-density-plasma regime was used for metaphase plate ablation. We show that functional fs laser-based enucleation of porcine oocytes completely inhibited further embryonic development while maintaining intact oocyte morphology. In contrast, non-irradiated oocytes were able to develop to the blastocyst stage without significant differences to control oocytes. Our results indicate that fs laser systems offer great potential for oocyte imaging and enucleation as a fast, easy to use and reliable tool which may improve the efficiency of somatic cell clone production.
KW - Cell surgery
KW - Femtosecond laser
KW - Oocyte enucleation
KW - SCNT
KW - Somatic cell nuclear transfer
UR - http://www.scopus.com/inward/record.url?scp=77951686043&partnerID=8YFLogxK
U2 - 10.1117/12.840045
DO - 10.1117/12.840045
M3 - Conference contribution
AN - SCOPUS:77951686043
SN - 9780819479853
T3 - Proceedings of SPIE - The International Society for Optical Engineering
BT - Frontiers in Ultrafast Optics
T2 - Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications X
Y2 - 24 January 2010 through 26 January 2010
ER -