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Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging

Recent advances in serial block-face imaging using scanning electron microscopy (SEM) have enabled the rapid and efficient acquisition of 3-dimensional (3D) ultrastructural information from a large volume of biological specimens including brain tissues. However, volume imaging under SEM is often ham...

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Autores principales: Nguyen, Huy Bang, Thai, Truc Quynh, Saitoh, Sei, Wu, Bao, Saitoh, Yurika, Shimo, Satoshi, Fujitani, Hiroshi, Otobe, Hirohide, Ohno, Nobuhiko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4810419/
https://www.ncbi.nlm.nih.gov/pubmed/27020327
http://dx.doi.org/10.1038/srep23721
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author Nguyen, Huy Bang
Thai, Truc Quynh
Saitoh, Sei
Wu, Bao
Saitoh, Yurika
Shimo, Satoshi
Fujitani, Hiroshi
Otobe, Hirohide
Ohno, Nobuhiko
author_facet Nguyen, Huy Bang
Thai, Truc Quynh
Saitoh, Sei
Wu, Bao
Saitoh, Yurika
Shimo, Satoshi
Fujitani, Hiroshi
Otobe, Hirohide
Ohno, Nobuhiko
author_sort Nguyen, Huy Bang
collection PubMed
description Recent advances in serial block-face imaging using scanning electron microscopy (SEM) have enabled the rapid and efficient acquisition of 3-dimensional (3D) ultrastructural information from a large volume of biological specimens including brain tissues. However, volume imaging under SEM is often hampered by sample charging, and typically requires specific sample preparation to reduce charging and increase image contrast. In the present study, we introduced carbon-based conductive resins for 3D analyses of subcellular ultrastructures, using serial block-face SEM (SBF-SEM) to image samples. Conductive resins were produced by adding the carbon black filler, Ketjen black, to resins commonly used for electron microscopic observations of biological specimens. Carbon black mostly localized around tissues and did not penetrate cells, whereas the conductive resins significantly reduced the charging of samples during SBF-SEM imaging. When serial images were acquired, embedding into the conductive resins improved the resolution of images by facilitating the successful cutting of samples in SBF-SEM. These results suggest that improving the conductivities of resins with a carbon black filler is a simple and useful option for reducing charging and enhancing the resolution of images obtained for volume imaging with SEM.
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spelling pubmed-48104192016-04-04 Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging Nguyen, Huy Bang Thai, Truc Quynh Saitoh, Sei Wu, Bao Saitoh, Yurika Shimo, Satoshi Fujitani, Hiroshi Otobe, Hirohide Ohno, Nobuhiko Sci Rep Article Recent advances in serial block-face imaging using scanning electron microscopy (SEM) have enabled the rapid and efficient acquisition of 3-dimensional (3D) ultrastructural information from a large volume of biological specimens including brain tissues. However, volume imaging under SEM is often hampered by sample charging, and typically requires specific sample preparation to reduce charging and increase image contrast. In the present study, we introduced carbon-based conductive resins for 3D analyses of subcellular ultrastructures, using serial block-face SEM (SBF-SEM) to image samples. Conductive resins were produced by adding the carbon black filler, Ketjen black, to resins commonly used for electron microscopic observations of biological specimens. Carbon black mostly localized around tissues and did not penetrate cells, whereas the conductive resins significantly reduced the charging of samples during SBF-SEM imaging. When serial images were acquired, embedding into the conductive resins improved the resolution of images by facilitating the successful cutting of samples in SBF-SEM. These results suggest that improving the conductivities of resins with a carbon black filler is a simple and useful option for reducing charging and enhancing the resolution of images obtained for volume imaging with SEM. Nature Publishing Group 2016-03-29 /pmc/articles/PMC4810419/ /pubmed/27020327 http://dx.doi.org/10.1038/srep23721 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Nguyen, Huy Bang
Thai, Truc Quynh
Saitoh, Sei
Wu, Bao
Saitoh, Yurika
Shimo, Satoshi
Fujitani, Hiroshi
Otobe, Hirohide
Ohno, Nobuhiko
Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging
title Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging
title_full Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging
title_fullStr Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging
title_full_unstemmed Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging
title_short Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging
title_sort conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4810419/
https://www.ncbi.nlm.nih.gov/pubmed/27020327
http://dx.doi.org/10.1038/srep23721
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