Vankelecom Hugo
Cold Spring Harb Protoc. 2009 Oct;2009(10):pdb.prot5298. doi: 10.1101/pdb.prot5298.
Green fluorescent protein (GFP) and its variants are used in many applications, including protein localization, high-throughput screening, and gene discovery. Transgenes that use GFP-based proteins as a reporter provide a convenient way to follow protein expression over the course of development and disease progression in the mouse. The basic principle of visualizing fluorescent proteins is the same as for the fluorophores used in fluorescent microscopy. This protocol describes a method for fixation and paraffin-embedding of fresh mouse embryos or tissue for use in GFP microscopy.
绿色荧光蛋白(GFP)及其变体被应用于许多领域,包括蛋白质定位、高通量筛选和基因发现。使用基于GFP的蛋白质作为报告基因的转基因提供了一种便捷的方法,可在小鼠发育和疾病进展过程中追踪蛋白质表达。可视化荧光蛋白的基本原理与荧光显微镜中使用的荧光团相同。本方案描述了一种用于新鲜小鼠胚胎或组织的固定和石蜡包埋方法,以用于GFP显微镜检查。