Parton Richard M, Vallés Ana Maria, Dobbie Ian M, Davis Ilan
Cold Spring Harb Protoc. 2010 Apr;2010(4):pdb.prot5402. doi: 10.1101/pdb.prot5402.
The fruit fly Drosophila melanogaster is an important model for basic research into the molecular mechanisms underlying cell function and development, as well as a major biomedical research tool. A significant advantage of Drosophila is the ability to apply live cell imaging to a variety of living tissues that can be dissected and imaged in vivo, ex vivo, or in vitro. Drosophila egg chambers, for example, have proven to be a useful model system for studying border cell migration, Golgi unit transport, the rapid movement of mRNA and protein particles, and the role of microtubules in meiosis and oocyte differentiation. A crucial first step before imaging is preparation of the experimental material to ensure physiological relevance and to achieve the best conditions for image quality. Early- to mid-stage egg chambers cannot be mounted in an aqueous-based medium, because this causes a change in microtubule organization and follicle cell morphology. Such egg chambers survive better in Halocarbon oil, which allows free diffusion of oxygen, has low viscosity, and thus prevents dehydration and hypoxia. With a refractive index similar to glycerol, Halocarbon oil also has good optical properties for imaging. It also provides a good environment for injection and is particularly useful for long-term imaging of embryos. However, unlike with aqueous solutions, changes in the medium are not possible. This protocol describes the isolation of Drosophila egg chambers.
果蝇（Drosophila melanogaster）是研究细胞功能和发育潜在分子机制基础研究的重要模型，也是一种主要的生物医学研究工具。果蝇的一个显著优势是能够将活细胞成像应用于多种可在体内、体外或离体状态下进行解剖和成像的活组织。例如，果蝇卵室已被证明是研究边缘细胞迁移、高尔基体单位运输、mRNA和蛋白质颗粒的快速移动以及微管在减数分裂和卵母细胞分化中的作用的有用模型系统。成像前至关重要的第一步是制备实验材料，以确保生理相关性并获得最佳图像质量条件。早期至中期的卵室不能安装在水基介质中，因为这会导致微管组织和卵泡细胞形态发生变化。这种卵室在全氟烃油中存活得更好，全氟烃油能使氧气自由扩散，粘度低，从而防止脱水和缺氧。由于其折射率与甘油相似，全氟烃油也具有良好的成像光学特性。它还为注射提供了良好的环境，对胚胎的长期成像特别有用。然而，与水溶液不同的是，介质无法更换。本方案描述了果蝇卵室的分离方法。
