Laser micromanipulation in the mouse embryo: a novel approach to zona drilling.
OBJECTIVE
To introduce the use of excimer lasers for penetration of the zona pellucida for micromanipulation purposes.


METHODS
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.


METHODS
The Laser Ablation Laboratory at DuPont and the in Vitro Fertilization Laboratory at The Medical Center.


METHODS
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.


METHODS
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.
Blanchet
G B
GB
E.I. DuPont de Nemours, Wilmington, Delaware.
Russell
J B
JB
Fincher
C R
CR
Portmann
M
M
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D001826:Body Fluids; D002021:Buffers; D003470:Culture Media; D004622:Embryo, Mammalian / Q000601:surgery*; D005314:Embryonic and Fetal Development; D005187:Fallopian Tubes / Q000378:metabolism; D005260:Female; D053685:Laser Therapy; D051379:Mice; D008866:Microsurgery; D010710:Phosphates; D012965:Sodium Chloride; D015044:Zona Pellucida
D002021:Buffers; D003470:Culture Media; D010710:Phosphates; D012965:Sodium Chloride