Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, The Netherlands.
J Physiol Pharmacol. 2009 Oct;60 Suppl 4:125-9.
Imaging of reporter molecules such as fluorescent proteins in intact animals, tissues and cells has become an indispensable tool in cell biology. Imaging activity of enzymes, which is called metabolic mapping, provides information on subcellular localisation in combination with functions of the enzymes. The principle of metabolic mapping is imaging of the formation of a reaction product that is fluorescent or coloured by conversion of either a fluorogenic or chromogenic substrate or a fluorescent substrate with different spectral characteristics. Chromogenic and fluorogenic staining methods are discussed here in the context of metabolic mapping in living animals, unfixed cryostat sections of tissues and living cells.
在完整的动物、组织和细胞中对报告分子(如荧光蛋白)进行成像已成为细胞生物学中不可或缺的工具。对酶活性的成像,称为代谢映射,提供了有关亚细胞定位的信息,同时还提供了酶功能的信息。代谢映射的原理是通过转化荧光或显色的荧光底物或具有不同光谱特征的荧光底物来成像反应产物的形成。本文讨论了在活体动物、未经固定的冰冻切片组织和活细胞中进行代谢映射时,显色和荧光染色方法。