Laser micromanipulation in the mouse embryo: a novel approach to zona drilling

• Published in: Fertility and sterility Vol. 57; no. 6; pp. 1337 - 1341
• Main Authors: Blanchet, Graciela B., Russell, Jeffrey B., Fincher, Curtis R., Portmann, Marc
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01.06.1992; Elsevier Science
• Subjects: Animals; Biological and medical sciences; Birth control; Blastocyst - physiology; Body Fluids; Buffers; Culture Media; Embryo, Mammalian - surgery; Embryonic and Fetal Development; Fallopian Tubes - metabolism; Female; Gynecology. Andrology. Obstetrics; Laser micromanipulation; Laser Therapy; Medical sciences; Mice; Microsurgery; Phosphates; Sodium Chloride; Sterility. Assisted procreation; Zona Pellucida; Laser micromanipulation; Drilling; Embryonic development; Embryo; Rodentia; Fecundation; In vitro; Zona pellucida; Vertebrata; Mammalia; Mouse; Animal; Excimer; Laser; Micromanipulation
• ISSN: 0015-0282; 1556-5653
• DOI: 10.1016/S0015-0282(16)55097-3

To introduce the use of excimer lasers for penetration of the zona pellucida for micromanipulation purposes. 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-μm opening in the zona pellucida. The Laser Ablation Laboratory at DuPont and the in Vitro Fertilization Laboratory at The Medical Center. The embryos were exposed in either phosphate-buffered solution (PBS) or modified human tubal fluid (HTF) with the laser power varying from 1 to 2J/cm2 and cultured in Ham’s F-10 medium (GIBCO, Grand Island, NY) with 0.4% bovine serum albumin. 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. 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. 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.