Use of picosecond infrared laser for micromanipulation of early mammalian embryos.
A high repetition rate (80 MHz) picosecond pulse (approximately 2 psec) infrared laser was used for the inactivation (functional enucleation) of oocytes and two-cell mouse embryos and also for the fusion of blastomeres of two-cell mouse embryos. The laser inactivation of both blastomeres of two-cell mouse embryos by irradiation of nucleoli completely blocked further development of the embryo. The inactivation of one blastomere, however, did not affect the ability of the second intact blastomere to develop into a blastocyst after treatment. Laser inactivation of oocytes at Metaphase II (MII) stage and parthenogenetically activated pronuclear oocytes also completely blocked their ability for further development. Suitable doses of irradiation in cytoplasm region did not affect the ability of embryos and activated oocytes to development. The efficiency of laser induced fusion for blastomeres of two-cell embryos was 66.7% and all the tetraploid embryos developed successfully into blastocysts in culture. Our results demonstrate unique opportunities of the applications of a suitable infrared periodic pulse laser as a universal microsurgery tool for individual living cells.
Karmenyan
Artashes V
AV
Institute of Biophotonics, National Yang-Ming University, Taipei, Taiwan. artashes@ym.edu.tw
Shakhbazyan
Avetik K
AK
Sviridova-Chailakhyan
Tatiana A
TA
Krivokharchenko
Alexander S
AS
Chiou
Arthur E
AE
Chailakhyan
Levon M
LM
D000818:Animals; D001757:Blastomeres / Q000502:physiology*; D018929:Cell Culture Techniques; D002467:Cell Nucleus / Q000502:physiology; D004622:Embryo, Mammalian / Q000502:physiology*; D004626:Embryology / Q000379:methods*; D005260:Female; D007259:Infrared Rays; D007834:Lasers; D008677:Metaphase; D051379:Mice; D008810:Mice, Inbred C57BL; D008866:Microsurgery; D009865:Oocytes / Q000502:physiology*