Details
| Original language | English |
|---|---|
| Title of host publication | Therapeutic Laser Applications and Laser-Tissue Interactions IV |
| Publication status | Published - 17 Jul 2009 |
| Externally published | Yes |
| Event | Therapeutic Laser Applications and Laser-Tissue Interactions IV - Munich, Germany Duration: 17 Jun 2009 → 18 Jun 2009 |
Publication series
| Name | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
|---|---|
| Volume | 7373 |
| ISSN (Print) | 1605-7422 |
Abstract
Cloning of several mammalian species has been achieved by somatic cell nuclear transfer (SCNT) in recent years. However, this method still results in very low efficiencies around 1% which originate from suboptimal culture conditions and highly invasive techniques for oocyte enucleation and injection of the donor cell using micromanipulators. In this paper, we present a new minimal invasive method for oocyte imaging and enucleation based on the application of femtosecond (fs) laser pulses. After imaging of the oocyte with multiphoton microscopy, ultrashort pulses are focused onto the metaphase plate of MII-oocytes in order to ablate the DNA molecules. We show that fs laser based enucleation of porcine oocytes completely inhibits the first mitotic cleavage after parthenogenetic activation while maintaining intact oocyte morphology in most cases. In contrast, control groups without previous irradiation of the metaphase plate are able to develop to the blastocyst stage. Further experiments have to clarify the suitability of fs laser based enucleated oocytes for SCNT.
Keywords
- Cell surgery, Femtosecond laser, Oocyte enucleation, SCNT, Somatic cell nuclear transfer
ASJC Scopus subject areas
- Materials Science(all)
- Electronic, Optical and Magnetic Materials
- Materials Science(all)
- Biomaterials
- Physics and Astronomy(all)
- Atomic and Molecular Physics, and Optics
- Medicine(all)
- Radiology Nuclear Medicine and imaging
Cite this
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- Harvard
- Apa
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- BibTeX
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Therapeutic Laser Applications and Laser-Tissue Interactions IV. 2009. 73730C (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 7373).
Research output: Chapter in book/report/conference proceeding › Conference contribution › Research › peer review
}
TY - GEN
T1 - Femtosecond laser based enucleation of porcine oocytes for somatic cell nuclear transfer
AU - Kuetemeyer, Kai
AU - Lucas-Hahn, Andrea
AU - Petersen, Bjoern
AU - Hassel, Petra
AU - Lemme, Erika
AU - Niemann, Heiner
AU - Heisterkamp, Alexander
PY - 2009/7/17
Y1 - 2009/7/17
N2 - Cloning of several mammalian species has been achieved by somatic cell nuclear transfer (SCNT) in recent years. However, this method still results in very low efficiencies around 1% which originate from suboptimal culture conditions and highly invasive techniques for oocyte enucleation and injection of the donor cell using micromanipulators. In this paper, we present a new minimal invasive method for oocyte imaging and enucleation based on the application of femtosecond (fs) laser pulses. After imaging of the oocyte with multiphoton microscopy, ultrashort pulses are focused onto the metaphase plate of MII-oocytes in order to ablate the DNA molecules. We show that fs laser based enucleation of porcine oocytes completely inhibits the first mitotic cleavage after parthenogenetic activation while maintaining intact oocyte morphology in most cases. In contrast, control groups without previous irradiation of the metaphase plate are able to develop to the blastocyst stage. Further experiments have to clarify the suitability of fs laser based enucleated oocytes for SCNT.
AB - Cloning of several mammalian species has been achieved by somatic cell nuclear transfer (SCNT) in recent years. However, this method still results in very low efficiencies around 1% which originate from suboptimal culture conditions and highly invasive techniques for oocyte enucleation and injection of the donor cell using micromanipulators. In this paper, we present a new minimal invasive method for oocyte imaging and enucleation based on the application of femtosecond (fs) laser pulses. After imaging of the oocyte with multiphoton microscopy, ultrashort pulses are focused onto the metaphase plate of MII-oocytes in order to ablate the DNA molecules. We show that fs laser based enucleation of porcine oocytes completely inhibits the first mitotic cleavage after parthenogenetic activation while maintaining intact oocyte morphology in most cases. In contrast, control groups without previous irradiation of the metaphase plate are able to develop to the blastocyst stage. Further experiments have to clarify the suitability of fs laser based enucleated oocytes for SCNT.
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=70349993378&partnerID=8YFLogxK
U2 - 10.1117/12.831880
DO - 10.1117/12.831880
M3 - Conference contribution
AN - SCOPUS:70349993378
SN - 9780819476494
T3 - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
BT - Therapeutic Laser Applications and Laser-Tissue Interactions IV
T2 - Therapeutic Laser Applications and Laser-Tissue Interactions IV
Y2 - 17 June 2009 through 18 June 2009
ER -