Department of Oral Anatomy, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi, Japan.
J Microsc. 2018 Mar;269(3):338-345. doi: 10.1111/jmi.12620. Epub 2017 Nov 10.
When polymorphonuclear leukocytes (PMNs) phagocytose opsonised zymosan particles (OPZ), free radicals and reactive oxygen species (ROS) are formed in the phagosomes. ROS production is mediated by NADPH oxidase (Nox), which transfers electrons in converting oxygen to superoxide (O ). Nox-generated O is rapidly converted to other ROS. Free radical-forming secretory vesicles containing the Nox redox center flavocytochrome b558, a membrane protein, and azurophil granules with packaged myeloperoxidase (MPO) have been described. Presuming the probable fusion of these vesicular and granular organelles with phagosomes, the translation process of the enzymes was investigated using energy-filtering and energy-dispersive spectroscopy-scanning transmission electron microscopy. In this work, the primary method for imaging cerium (Ce) ions demonstrated the localisation of H O generated by phagocytosing PMNs. The MPO activity of the same PMNs was continuously monitored using 0.1% 3,3'-diaminobenzidine-tetrahydrochloride (DAB) and 0.01% H O . A detailed view of these vesicular and granular structures was created by overlaying each electron micrograph with pseudocolors: blue for Ce and green for nitrogen (N).
当多形核白细胞 (PMN) 吞噬调理化酵母聚糖颗粒 (OPZ) 时，吞噬体中会形成自由基和活性氧物质 (ROS)。ROS 的产生是由 NADPH 氧化酶 (Nox) 介导的，它在将氧气转化为超氧阴离子 (O ) 时传递电子。Nox 产生的 O 会迅速转化为其他 ROS。已经描述了含有 Nox 氧化还原中心 flavocytochrome b558、一种膜蛋白和含有包装过的髓过氧化物酶 (MPO) 的嗜苯胺颗粒的形成自由基的分泌小泡。假定这些囊泡和颗粒细胞器与吞噬体的融合，使用能量过滤和能量分散光谱扫描透射电子显微镜研究了酶的翻译过程。在这项工作中，使用铈 (Ce) 离子的主要成像方法证明了吞噬 PMN 产生的 H O 的定位。使用 0.1% 3,3'-二氨基联苯胺四盐酸盐 (DAB) 和 0.01% H O 连续监测相同 PMN 的 MPO 活性。通过用伪色叠加每个电子显微镜图像来创建这些囊泡和颗粒结构的详细视图：蓝色表示 Ce，绿色表示氮 (N)。
