高锰酸钾/铅染色可在低真空扫描电子显微镜下对组织结构进行无铀成像。
KMnO/Pb staining allows uranium free imaging of tissue architectures in low vacuum scanning electron microscopy.
作者信息
Sawaguchi Akira, Kamimura Takeshi, Kitagawa Kyoko, Nagashima Yoko, Takahashi Nobuyasu
机构信息
Division of Ultrastructural Cell Biology, Department of Anatomy, Faculty of Medicine, University of Miyazaki, Miyazaki, 889-1692, Japan.
Hitachi High-Tech Corporation, Tokyo, 105-6409, Japan.
出版信息
Npj Imaging. 2024 Oct 1;2(1):40. doi: 10.1038/s44303-024-00045-z.
Scanning electron microscopy under low-vacuum conditions allows high-resolution imaging of complex cell/tissue architectures in nonconductive specimens. However, the conventional methods for metal staining of biological specimens require harmful uranium compounds, which hampers the applications of electron microscopy. Here, we introduce a uranium-free KMnO/Pb metal staining protocol that allows multiscale imaging of extensive cell/tissue architectures to intensive subcellular ultrastructures. The obtained image contrast was equivalent to that of Ur/Pb staining and sufficient for ultrastructural observation, showing the fine processes of podocytes in the glomerulus, which were invisible by light microscopy. The stainability in the elastic tissue indicated that the distinct histochemical properties of KMnO oxidation led to Pb deposition and BSE signal enhancement superior to Ur staining. Elemental analysis clarified that the determinant of the backscattered electron signal intensity was the amount of Pb deposition enhanced by KMnO oxidation. This user-friendly method is anticipated to create a new approach for biomedical electron microscopy.
在低真空条件下进行扫描电子显微镜检查可对非导电标本中的复杂细胞/组织结构进行高分辨率成像。然而,用于生物标本金属染色的传统方法需要使用有害的铀化合物,这阻碍了电子显微镜的应用。在此,我们介绍一种无铀的KMnO/Pb金属染色方案,该方案可对广泛的细胞/组织结构至精细的亚细胞超微结构进行多尺度成像。所获得的图像对比度与Ur/Pb染色相当,足以进行超微结构观察,显示出肾小球中足细胞的精细突起,而这些在光学显微镜下是不可见的。弹性组织中的可染色性表明,KMnO氧化独特的组织化学性质导致Pb沉积和背散射电子(BSE)信号增强,优于Ur染色。元素分析表明,背散射电子信号强度的决定因素是KMnO氧化增强的Pb沉积量。这种用户友好型方法有望为生物医学电子显微镜创造一种新方法。
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