用于定量全内反射荧光显微镜分析的发育中卵母细胞的卵质提取物。
Ooplasmic Extract from Developing Oocytes for Quantitative TIRF Microscopy Analysis.
作者信息
Gaspar Imre, Ephrussi Anne
机构信息
European Molecular Biology Laboratory (EMBL), Developmental Biology Unit, Heidelberg, Meyerhofstrasse 1, D-69117, Germany.
出版信息
Bio Protoc. 2017 Jul 5;7(13). doi: 10.21769/BioProtoc.2380.
Abstract
Understanding the dynamic behavior and the continuously changing composition of macromolecular complexes, subcellular structures and organelles is one of areas of active research in both cell and developmental biology, as these changes directly relate to function and subsequently to the development and homeostasis of the organism. Here, we demonstrate the use of the developing oocyte to study dynamics of messenger ribonucleoprotein complexes (mRNPs) with high spatiotemporal resolution. The combination of genetics with total internal reflection (TIRF) microscopy, image processing and data analysis gives insight into mRNP motility and composition dynamics with unprecedented precision.
摘要
了解大分子复合物、亚细胞结构和细胞器的动态行为及其不断变化的组成，是细胞生物学和发育生物学中活跃的研究领域之一，因为这些变化直接关系到功能，进而关系到生物体的发育和内稳态。在这里，我们展示了利用发育中的卵母细胞以高时空分辨率研究信使核糖核蛋白复合物（mRNP）的动态变化。遗传学与全内反射（TIRF）显微镜、图像处理和数据分析相结合，以前所未有的精度深入了解了mRNP的运动性和组成动态变化。
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