Canaria Christie A, Lansford Rusty
Cold Spring Harb Protoc. 2011 Nov 1;2011(11):1375-82. doi: 10.1101/pdb.prot066621.
Traditionally, our understanding of developmental biology has been based on the fixation and study of embryonic samples. Detailed microscopic scrutiny of static specimens at varying ages allowed for anatomical assessment of tissue development. The advent of confocal and two-photon excitation (2PE) microscopy enables researchers to acquire volumetric images in three dimensions (x, y, and z) plus time (t). The development of transgenic Japanese quail has provided an embryonic model system that is more easily accessible than mammalian models and more efficient to breed than the classic avian model, the chicken. This protocol describes the methods necessary to prepare transgenic quail embryos for imaging in vitro, and for acquiring, processing, and analyzing three-dimensional (3D) time-lapse images.
传统上，我们对发育生物学的理解基于对胚胎样本的固定和研究。对不同年龄的静态标本进行详细的显微镜检查，以便对组织发育进行解剖学评估。共聚焦显微镜和双光子激发（2PE）显微镜的出现，使研究人员能够获取三维（x、y和z）加时间（t）的体积图像。转基因日本鹌鹑的发展提供了一个胚胎模型系统，它比哺乳动物模型更容易获取，并且比经典的禽类模型鸡更高效地繁殖。本方案描述了为体外成像制备转基因鹌鹑胚胎，以及获取、处理和分析三维（3D）延时图像所需的方法。
