Informative three-dimensional survey of cell/tissue architectures in thick paraffin sections by simple low-vacuum scanning electron microscopy

Scientific Reports
Akira SawaguchiYujiro Asada

Abstract

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...Continue Reading

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Citations

Aug 31, 2019·Laboratory Investigation; a Journal of Technical Methods and Pathology·Hideya KawasakiTakahiko Hariyama
Apr 22, 2020·Journal of the European Academy of Dermatology and Venereology : JEADV·Toru Kawai TRiichiro Abe
Jun 12, 2021·Communications Biology·Akira SawaguchiYoshitaka Hishikawa
Jul 4, 2021·International Journal of Pharmaceutics·Nidhi RavalRakesh K Tekade
Oct 31, 2021·Nature Protocols·Tariq Sami HaddadIris Nagtegaal

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Methods Mentioned

BETA
electron microscopy
light microscopy
xenograft
xenografts
transmission electron microscopy
fluorescence microscopy
light

Software Mentioned

Hitachi map 3D

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