Fertilization of bovine oocytes induced solely with combined laser microbeam and optical tweezers |
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Authors: | Annette Clement-sengewald Karin Schütze Arthur Ashkin Gustavo A. Palma Gertrude Kerlen Gottfried Brem |
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Affiliation: | (1) Bayerische Klonierungsforschungs-GmbH & Co. KG, Hackerstr. 27, 85764 Oberschleissheim, Germany;(2) Applikatives Laserzentrum Harlaching, Sanatoriumsplatz 2, 81545 Munich, Germany;(3) AT&T Bell Laboratories, 07733 Holmdel, New Jersey;(4) Institut für den Wissenschaftlichen Film, Nonnenstieg 72, 37075 Göttingen, Germany;(5) Bosques de Reforma 1416, Casa 10B Col. Bosques de Las Lomas, 11700 México, D. F. |
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Abstract: | Purpose: Our purpose was to show that fertilization of oocytes can be obtained solely by laser light-mediated manipulation of gametes.Method: A small channel was drilled into the zona pellucida of bovine oocytes using an ultraviolet (UV)-laser microbeam. Highly diluted cattle sperm were not able to fertilize the laser drilled oocytes.Results: Fertilization was achieved only when three to five cattle sperm were trapped with optical tweezers and inserted directly through the laser drilled hole into the perivitelline space. After 20 hr, 3 of 79 (3.8%) oocytes revealed two pronuclei and a sperm tail within their cytoplasm. Cattle sperm are difficult to catch. Therefore, the gametes had to remain for about 20 min in room atmosphere, which might be the reason for the low fertilization results.Conclusions: The results indicate that a combined UV-laser microbeam and optical tweezers trap can be used successfully for noncontact microinsemination procedures.Presented in part at the IXth World Congress on In Vitro Fertilization and Alternate Assisted Reproduction, April 3–7, 1995, Vienna, Austria. |
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Keywords: | laser micromanipulation laser-assisted fertilization laser zona drilling laser-mediated subzonal sperm insertion microinsemination |
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