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Updated: Aug 17, 2026
Laser-assisted Cytoplasmic Microinjection in Livestock Zygotes
Published on: October 5, 2016
Laser micromanipulation in the mouse embryo: a novel approach to zona drilling
G B Blanchet1, J B Russell, C R Fincher
1E.I. DuPont de Nemours, Wilmington, Delaware.
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.
