Centre for Biospectroscopy, School of Chemistry, Monash University, Clayton, VIC, 3800, Australia.
Department of Microbiology, Monash University, Clayton, VIC, 3800, Australia.
Sci Rep. 2017 Aug 21;7(1):8945. doi: 10.1038/s41598-017-08973-0.
Confocal Raman spectroscopy (CRS) can provide information about oocyte competency through measurement of changes in the macromolecular architecture during oocyte development and maturation. Hitherto most spectroscopic studies have been limited to fixed oocytes due to the inherent difficulties working with live cells. Here we report the first three-dimensional images of living murine oocytes using CRS. We show that fixation induces significant changes in the macromolecular chemistry compared to living oocytes. A band at 1602 cm, assigned to a marker for mitochondria function was found in living oocytes but absent from fixed oocytes providing an in vivo marker. Fixation resulted in significant changes in protein and nucleic acid bands and the spatial distribution of organelles. Raman imaging of Metaphase I and II (MI, MII) and germinal vesicle stage oocytes showed changes in nuclear organisation and cytoplasm macromolecular architecture during these development and maturation stages related to changes in chromosome condensation, mitochondria aggregation and lipid droplet numbers.
共聚焦拉曼光谱（CRS）可以通过测量卵母细胞发育和成熟过程中大分子结构的变化，提供关于卵母细胞能力的信息。迄今为止，由于活细胞的固有困难，大多数光谱研究仅限于固定的卵母细胞。在这里，我们报告了使用 CRS 对活的小鼠卵母细胞的首次三维成像。我们发现，与活卵母细胞相比，固定会导致大分子化学发生显著变化。在活卵母细胞中发现了一个位于 1602cm 处的带，被分配给线粒体功能的标志物，但在固定的卵母细胞中不存在，这为活体内提供了一个标志物。固定导致蛋白质和核酸带以及细胞器的空间分布发生显著变化。对 MⅠ、MⅡ期和生发泡期卵母细胞的拉曼成像显示，在这些发育和成熟阶段，核组织和细胞质大分子结构发生变化，与染色体浓缩、线粒体聚集和脂滴数量的变化有关。
