Grecchi S, Malatesta M
University of Verona.
Eur J Histochem. 2014 Oct 22;58(4):2449. doi: 10.4081/ejh.2014.2449.
The endocytotic pathway involves a complex, dynamic and interacting system of intracellular compartments. PKH26 is a fluorescent dye specific for long-lasting cell membrane labelling which has been successfully used for investigating cell internalization processes, at either flow cytometry or fluorescence microscopy. In the present work, diaminobenzidine photo-oxidation was tested as a procedure to detect PKH26 dye at transmission electron microscopy. Our results demonstrated that DAB-photo-oxidation is a suitable technique to specifically visualise this fluorescent dye at the ultrastructural level: the distribution of the granular dark reaction product perfectly matches the pattern of the fluorescence staining, and the electron density of the fine precipitates makes the signal evident and precisely detectable on the different subcellular compartments involved in the plasma membrane internalization routes.
内吞途径涉及细胞内区室的一个复杂、动态且相互作用的系统。PKH26是一种用于持久细胞膜标记的荧光染料，已成功用于在流式细胞术或荧光显微镜下研究细胞内化过程。在本研究中，测试了二氨基联苯胺光氧化作为在透射电子显微镜下检测PKH26染料的方法。我们的结果表明，DAB光氧化是一种在超微结构水平上特异性可视化这种荧光染料的合适技术：颗粒状暗反应产物的分布与荧光染色模式完美匹配，并且细沉淀物的电子密度使得信号在参与质膜内化途径的不同亚细胞区室上明显且可精确检测。
