Informative three-dimensional survey of cell/tissue architectures in thick paraffin sections by simple low-vacuum scanning electron microscopy.
Informative three-dimensional survey of cell/tissue architectures in thick paraffin sections by simple low-vacuum scanning electron microscopy.
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DOI：
10.1038/s41598-018-25840-8
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发表时间：
2018-05-10
影响因子：
4.6
通讯作者：
Asada Y
中科院分区：
文献类型：
--
作者：
Sawaguchi A;Kamimura T;Yamashita A;Takahashi N;Ichikawa K;Aoyama F;Asada Y
生物医学研究的最新进展，例如从干细胞产生再生器官，需要对细胞/组织结构进行三维分析。电子显微镜的高分辨率成像是阐明复杂细胞/组织结构的最佳方法，但传统方法需要熟练和耗时的准备。本研究开发了一种三维测量方法，通过利用低真空扫描电子显微镜中的背散射电子成像来评估30 μ m厚石蜡切片中的细胞/组织结构。结果，在肾脏中，清楚地观察到足细胞及其突起覆盖肾小球。30 µm的厚度便于在上皮和内皮的正面（而不是切片）图像上进行研究，这在传统的薄切片中很少见。在睾丸中，已分化的精子在曲细精管的中部三维组装。进一步应用于血管损伤血栓形成揭示了纤维蛋白纤维和血小板的独特网络，将红细胞捕获到血栓中。四段BSE探测器提供了地形鸟瞰图像，允许在电子显微镜水平上对细胞/组织结构进行三维理解。在这里，我们描述了这种成像方法的精确程序，并提供了正常大鼠器官，实验血栓形成和三维培养的肿瘤细胞的代表性电子显微照片。
Recent advances in bio-medical research, such as the production of regenerative organs from stem cells, require three-dimensional analysis of cell/tissue architectures. High-resolution imaging by electron microscopy is the best way to elucidate complex cell/tissue architectures, but the conventional method requires a skillful and time-consuming preparation. The present study developed a three-dimensional survey method for assessing cell/tissue architectures in 30-µm-thick paraffin sections by taking advantage of backscattered electron imaging in a low-vacuum scanning electron microscope. As a result, in the kidney, the podocytes and their processes were clearly observed to cover the glomerulus. The 30 µm thickness facilitated an investigation on face-side (instead of sectioned) images of the epithelium and endothelium, which are rarely seen within conventional thin sections. In the testis, differentiated spermatozoa were three-dimensionally assembled in the middle of the seminiferous tubule. Further application to vascular-injury thrombus formation revealed the distinctive networks of fibrin fibres and platelets, capturing the erythrocytes into the thrombus. The four-segmented BSE detector provided topographic bird’s-eye images that allowed a three-dimensional understanding of the cell/tissue architectures at the electron-microscopic level. Here, we describe the precise procedures of this imaging method and provide representative electron micrographs of normal rat organs, experimental thrombus formation, and three-dimensionally cultured tumour cells.
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影响因子：
1.8
作者：
Yano, Tetsuya;Soejima, Yurie;Sawabe, Motoji
通讯作者：
Sawabe, Motoji
影响因子：
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作者：
Inaga, Sumire;Hirashima, Sayuri;Naguro, Tomonori
通讯作者：
Naguro, Tomonori
影响因子：
2.8
作者：
Lou JJ;Mirsadraei L;Sanchez DE;Wilson RW;Shabihkhani M;Lucey GM;Wei B;Singer EJ;Mareninov S;Yong WH
通讯作者：
Yong WH
影响因子：
1.6
作者：
Kokkat, Theresa J.;Patel, Miral S.;Baloch, Zubair W.
通讯作者：
Baloch, Zubair W.
影响因子：
4.6
作者：
Creech, Matthew K.;Wang, Jing;Gibbs, Summer L.
通讯作者：
Gibbs, Summer L.