Department of Cell Biology and Histology, Academic Medical Center, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands.
J Histochem Cytochem. 2010 Jun;58(6):481-97. doi: 10.1369/jhc.2010.955518. Epub 2010 Feb 1.
For the understanding of functions of proteins in biological and pathological processes, reporter molecules such as fluorescent proteins have become indispensable tools for visualizing the location of these proteins in intact animals, tissues, and cells. For enzymes, imaging their activity also provides information on their function or functions, which does not necessarily correlate with their location. Metabolic mapping enables imaging of activity of enzymes. The enzyme under study forms a reaction product that is fluorescent or colored by conversion of either a fluorogenic or chromogenic substrate or a fluorescent substrate with different spectral characteristics. Most chromogenic staining methods were developed in the latter half of the twentieth century but still find new applications in modern cell biology and pathology. Fluorescence methods have rapidly evolved during the last decade. This review critically evaluates the methods that are available at present for metabolic mapping in living animals, unfixed cryostat sections of tissues, and living cells, and refers to protocols of the methods of choice.
为了理解蛋白质在生物和病理过程中的功能，报告分子（如荧光蛋白）已成为在完整动物、组织和细胞中可视化这些蛋白质位置的不可或缺的工具。对于酶来说，对其活性的成像也提供了关于其功能或功能的信息，这些信息不一定与它们的位置相关。代谢映射使酶活性的成像成为可能。所研究的酶形成反应产物，该产物通过荧光或显色转化为荧光或显色底物，或者转化为具有不同光谱特征的荧光底物。大多数显色染色方法是在 20 世纪后半叶开发的，但在现代细胞生物学和病理学中仍有新的应用。荧光方法在过去十年中迅速发展。本综述批判性地评估了目前可用于活体动物、未固定的冰冻切片组织和活细胞中代谢映射的方法，并提到了所选方法的方案。
