Van Noorden C J, Frederiks W M
Laboratory of Cell Biology, University of Amsterdam, The Netherlands.
J Microsc. 1993 Jul;171(Pt 1):3-16. doi: 10.1111/j.1365-2818.1993.tb03354.x.
Methods based on the use of cerium to detect the activity of oxidases and phosphatases are rapidly expanding. Both classes of enzymes can be demonstrated with cerium at the electron and light microscopical level. The in situ detection of H2O2 production with cerium is another application that has great potential, particularly in experimental pathological research. The fine precipitate of the cerium-containing final reaction product, cerium perhydroxide or cerium phosphate, enables a very precise localization. With such techniques, important advances have been made in cell biology, such as the discovery of new organelles, functional subcompartmentization of peroxisomes, tubular lysosomes and the elucidation of the function of extracellular ATPases. At the light microscopical level, the activity of enzymes can be quantified in situ because the production of final reaction product in the cerium methods is proportional to enzyme activity in tissue sections or cells. Cerium precipitates have strong reflectance properties and this enables their use in confocal scanning laser microscopy. In the present review, the principles of cerium methods are outlined and applications in cell biology and pathology are discussed.
基于使用铈来检测氧化酶和磷酸酶活性的方法正在迅速发展。这两类酶都可以在电子显微镜和光学显微镜水平上用铈进行显示。用铈原位检测过氧化氢的产生是另一个具有巨大潜力的应用,特别是在实验病理学研究中。含铈最终反应产物(过氢氧化铈或磷酸铈)的细微沉淀能够实现非常精确的定位。通过这些技术,细胞生物学取得了重要进展,例如发现新的细胞器、过氧化物酶体、管状溶酶体的功能亚区隔以及阐明细胞外ATP酶的功能。在光学显微镜水平上,可以对酶的活性进行原位定量,因为铈法中最终反应产物的产生与组织切片或细胞中的酶活性成正比。铈沉淀具有很强的反射特性,这使得它们能够用于共聚焦扫描激光显微镜。在本综述中,概述了铈法的原理,并讨论了其在细胞生物学和病理学中的应用。