OBJECTIVE: To introduce the use of excimer lasers for penetration of the zona pellucida for micromanipulation purposes. DESIGN: Cryopreserved two-cell mouse embryos were thawed and exposed to the 248-nm line of a krypton fluoride excimer laser (Lambda Physik EMG 202, Goettingen, Germany) creating a 2 to 4-micron opening in the zona pellucida. SETTING: The Laser Ablation Laboratory at DuPont and the in Vitro Fertilization Laboratory at The Medical Center. INTERVENTIONS: The embryos were exposed in either phosphate-buffered solution (PBS) or modified human tubal fluid (HTF) with the laser power varying from 1 to 2 J/cm2 and cultured in Ham's F-10 medium (GIBCO, Grand Island, NY) with 0.4% bovine serum albumin. MAIN OUTCOME MEASURES: The outcome of each experiment was measured by blastocyst formation of laser-exposed embryos as compared with a set of unexposed control embryos handled in a similar fashion. RESULTS: Successful laser penetration of the zona pellucida was achieved using the 248-nm line of a krypton fluoride excimer laser. A higher blastocyst formation was found for embryos exposed in PBS. The higher optical absorption of the modified HTF partially inhibited embryo development. The blastocyst statistics increased 2.5-fold times by reducing the exposure of the embryos to ablation by-products. CONCLUSIONS: The use of a krypton fluoride excimer laser was introduced as a new method to open the zona pellucida of two-cell mouse embryos without interrupting blastocyst formation.
OBJECTIVE: To introduce the use of excimer lasers for penetration of the zona pellucida for micromanipulation purposes. DESIGN: Cryopreserved two-cell mouse embryos were thawed and exposed to the 248-nm line of a krypton fluoride excimer laser (Lambda Physik EMG 202, Goettingen, Germany) creating a 2 to 4-micron opening in the zona pellucida. SETTING: The Laser Ablation Laboratory at DuPont and the in Vitro Fertilization Laboratory at The Medical Center. INTERVENTIONS: The embryos were exposed in either phosphate-buffered solution (PBS) or modified human tubal fluid (HTF) with the laser power varying from 1 to 2 J/cm2 and cultured in Ham's F-10 medium (GIBCO, Grand Island, NY) with 0.4% bovine serum albumin. MAIN OUTCOME MEASURES: The outcome of each experiment was measured by blastocyst formation of laser-exposed embryos as compared with a set of unexposed control embryos handled in a similar fashion. RESULTS: Successful laser penetration of the zona pellucida was achieved using the 248-nm line of a krypton fluoride excimer laser. A higher blastocyst formation was found for embryos exposed in PBS. The higher optical absorption of the modified HTF partially inhibited embryo development. The blastocyst statistics increased 2.5-fold times by reducing the exposure of the embryos to ablation by-products. CONCLUSIONS: The use of a krypton fluoride excimer laser was introduced as a new method to open the zona pellucida of two-cell mouse embryos without interrupting blastocyst formation.
Authors: Tyl H Taylor; Janice W Gilchrist; Susan V Hallowell; Kelly K Hanshew; John J Orris; Michael J Glassner; J David Wininger Journal: J Assist Reprod Genet Date: 2010-08-05 Impact factor: 3.412