Institute of Biophotonics, National Yang-Ming University, Taipei, Taiwan.
Mol Reprod Dev. 2009 Oct;76(10):975-83. doi: 10.1002/mrd.21045.
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.
一种高重复率(80MHz)皮秒脉冲(约 2psec)红外激光被用于卵母细胞和 2 细胞期小鼠胚胎的失活(功能去核),以及 2 细胞期小鼠胚胎的卵裂球融合。用核仁照射完全阻断了两细胞期小鼠胚胎两个卵裂球的进一步发育。然而,一个卵裂球的失活并不影响第二个完整卵裂球在处理后发育成囊胚的能力。M 期(MII)期卵母细胞的激光失活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。在细胞质区域的合适剂量照射不会影响胚胎和激活卵母细胞的发育能力。2 细胞期胚胎卵裂球的激光诱导融合效率为 66.7%,所有四倍体胚胎在培养中均成功发育成囊胚。我们的结果表明,适当的红外周期脉冲激光作为一种通用的单细胞微创手术工具,具有独特的应用机会。