Use of Picosecond Infrared Laser for Micromanipulation of Early Mammalian Embryos
Use of Picosecond Infrared Laser for Micromanipulation of Early Mammalian Embryos
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DOI：
10.1002/mrd.21045
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发表时间：
2009-10-01
影响因子：
2.5
通讯作者：
Chailakhyan, Levon M.
中科院分区：
文献类型：
--
作者：
Karmenyan, Artashes V.;Shakhbazyan, Avetik K.;Chailakhyan, Levon M.
高重复频率（80 MHz）皮秒脉冲（类似于2 psec）红外激光用于卵母细胞和双细胞小鼠胚胎的失活（功能性去核）和双细胞小鼠胚胎的卵裂球融合。激光照射核仁使双细胞小鼠胚胎的两个卵裂球失活，完全阻断了胚胎的进一步发育。然而，一个卵裂球的失活并不影响第二个完整的卵裂球在处理后发育成囊胚的能力。激光灭活中期（MII）卵母细胞和孤雌激活的原核卵母细胞也完全阻断了它们进一步发育的能力。细胞质区适当剂量的辐照对胚胎和活化卵母细胞的发育没有影响。激光诱导两细胞胚的卵裂球融合效率为66.7%，四倍体胚均成功发育成囊胚。我们的研究结果展示了一种合适的红外周期性脉冲激光作为单个活细胞的通用显微手术工具的独特应用机会。
A high repetition rate (80 MHz) picosecond pulse (similar to 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.