Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging.
Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging.
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DOI：
10.1038/srep23721
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发表时间：
2016-03-29
影响因子：
4.6
通讯作者：
Ohno N
中科院分区：
文献类型：
--
作者：
Nguyen HB;Thai TQ;Saitoh S;Wu B;Saitoh Y;Shimo S;Fujitani H;Otobe H;Ohno N
使用扫描电子显微镜（SEM）的连续块面成像的最新进展已经使得能够从包括脑组织的大量生物标本中快速有效地获取三维（3D）超微结构信息。然而，在SEM下的体积成像通常受到样品充电的阻碍，并且通常需要特定的样品制备以减少充电并增加图像对比度。在本研究中，我们介绍了碳基导电树脂的亚细胞超微结构的三维分析，使用连续块面扫描电镜（SBF-SEM）图像样品。导电树脂是通过将炭黑填料Ketjen黑添加到通常用于生物样品的电子显微镜观察的树脂中来生产的。炭黑主要局限于组织周围，并没有渗透细胞，而导电树脂显着减少样品在SBF-SEM成像过程中的充电。当获得连续图像时，嵌入到导电树脂中通过促进样品在SBF-SEM中的成功切割来提高图像的分辨率。这些结果表明，提高树脂的导电性与炭黑填料是一个简单而有用的选择，减少充电和提高分辨率的体积成像与SEM获得的图像。
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.