Analyzing indirect secondary electron contrast of unstained bacteriophage T4 based on SEM images and Monte Carlo simulations
Analyzing indirect secondary electron contrast of unstained bacteriophage T4 based on SEM images and Monte Carlo simulations
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DOI：
10.1016/j.bbrc.2009.01.046
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发表时间：
2009-03-06
影响因子：
3.1
通讯作者：
Ogura, Toshihiko
中科院分区：
文献类型：
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作者：
Ogura, Toshihiko
扫描电子显微镜（SEM）的间接二次电子衬度（ISEC）条件产生高对比度检测，对安装在薄碳层下的未染色生物样品的损伤最小。高对比度图像是由低加速电压在碳膜下产生的二次电子信号产生的。在这里，我们表明，ISEC条件是清楚地能够检测未染色的噬菌体T4下的薄碳膜（10-15 nm），通过使用高分辨率场发射（FE）SEM。结果表明，FE-SEM比电子发射SEM具有更高的分辨率。此外，我们研究了散射电子在碳膜的ISEC条件下的区域使用Monte Carlo模拟。模拟结果表明，图像分辨率的差异与碳膜中的散射宽度和电子束光斑大小有关。在未染色的病毒样品上使用ISEC条件将产生低的电子损伤，因为电子束不直接照射样品。除常规分析外，该方法还可用于病毒、细菌和蛋白质复合物等各种生物样品的结构分析。(c)2009 Elsevier Inc.版权所有
The indirect secondary electron contrast (ISEC) condition of the scanning electron microscopy (SEM) produces high contrast detection with minimal damage of unstained biological samples mounted Under a thin carbon him. The high contrast image is created by a secondary electron signal produced under the carbon film by a low acceleration voltage. Here, we show that ISEC condition is clearly able to detect unstained bacteriophage T4 under a thin carbon film (10-15 nm) by using high-resolution field emission (FE) SEM. The results show that FE-SEM provides higher resolution than thermionic emission SEM. Furthermore, we investigated the scattered electron area within the carbon film under ISEC conditions using Monte Carlo Simulation. The Simulations indicated that the image resolution difference is related to the scattering width in the carbon film and the electron beam spot size. Using ISEC conditions on unstained virus samples would produce low electronic damage, because the electron beam does not directly irradiate the sample. In addition to the routine analysis, this method can be utilized for Structural analysis Of various biological samples like Viruses, bacteria, and protein complexes. (c) 2009 Elsevier Inc. All rights reserved