Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, 3584 CM Utrecht, the Netherlands.
Theriogenology. 2011 Jan 15;75(2):386-91. doi: 10.1016/j.theriogenology.2010.08.005. Epub 2010 Sep 27.
Manipulation of mammalian oocytes at the molecular level is hampered by low transcriptional activity and the presence of large stores of mRNA and protein. Microinjection of interfering macromolecules has become an important tool in studying oocyte maturation, although injection success, final concentrations of injected substances and viability after injection remain difficult to assess with current techniques. To address these problems, we developed an epifluorescence microscopy based technique to evaluate oocytes directly after (co-)injection of green fluorescent protein (GFP).
哺乳动物卵母细胞的分子水平操作受到转录活性低和大量 mRNA 和蛋白质储存的阻碍。尽管微注射干扰大分子已成为研究卵母细胞成熟的重要工具,但目前的技术仍然难以评估注射成功率、注射物质的最终浓度和注射后的活力。为了解决这些问题,我们开发了一种基于荧光显微镜的技术,用于在(共)注射绿色荧光蛋白 (GFP) 后直接评估卵母细胞。