Angermüller S, Fahimi H D
Department of Anatomy, University of Heidelberg, Federal Republic of Germany.
J Histochem Cytochem. 1988 Jan;36(1):23-8. doi: 10.1177/36.1.2891744.
The reaction product of cerium used for localization of peroxisomal oxidases is highly electron-dense but lacks sufficient contrast at the light microscopic level. We describe two methods for converting the reaction product of cerium to colored compounds visible by light microscopy. The first method is based on 3,3'-diaminobenzidine (DAB) amplification of transition metal compounds, of which cerium is one. Sections of glutaraldehyde-fixed rat liver or kidney are incubated first in media for various oxidases containing CeCl3, followed by treatment with DAB in Na acetate buffer, pH 5.3. To prevent any interference by the peroxidatic activity of catalase, NaN3 or Na pyruvate is added to the DAB amplification medium. Staining with DAB can be further intensified with NiCl2 or CoCl2. The second method is based on the conversion of the cerium reaction product with alkaline lead citrate and the final visualization of the lead compound with ammonium sulfide. These methods allow the evaluation of large sections for peroxisomal oxidases by light microscopy, making close correlation between light and electron microscopy possible.
用于过氧化物酶体氧化酶定位的铈反应产物具有很高的电子密度，但在光学显微镜水平上缺乏足够的对比度。我们描述了两种将铈反应产物转化为光学显微镜下可见的有色化合物的方法。第一种方法基于过渡金属化合物（铈是其中之一）的3,3'-二氨基联苯胺（DAB）放大。将戊二醛固定的大鼠肝脏或肾脏切片首先在含有CeCl3的各种氧化酶培养基中孵育，然后在pH 5.3的醋酸钠缓冲液中用DAB处理。为了防止过氧化氢酶的过氧化物酶活性产生任何干扰，在DAB放大培养基中加入NaN3或丙酮酸钠。用NiCl2或CoCl2可以进一步增强DAB染色。第二种方法基于用碱性柠檬酸铅转化铈反应产物，并用硫化铵最终使铅化合物可视化。这些方法允许通过光学显微镜评估过氧化物酶体氧化酶的大片段，使得光学显微镜和电子显微镜之间的紧密关联成为可能。
