Biological specimen viability analysis by hybrid microscope combined optical microscope and Environmental-SEM

M. Nakajima, H. Akimoto, T. Hirano, M. Kojima, N. Hisamoto, M. Homma, T. Fukuda
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引用次数: 5

Abstract

This paper presents a novel biological specimen analysis system by hybrid microscope, which is combined optical and environmental-scanning electron microscope (E-SEM) based on nanorobotic manipulation system. As previous works, we present single cell analysis by E-SEM nanorobotic manipulation system. It can be realized that the nano-scale manipulation and evaluation by nano-tools using the nano-scale image of E-SEM. The E-SEM provides us the high resolution image of specimen with water containing condition by specially built detector and temperature controlling stage. However the E-SEM image, especially SEM image, is surface image, hence it is impossible to obtain the inner structural imaging which is important for biological analysis. In this work, we combine optical microscope (OM) and E-SEM to realize biological specimen analysis by optical microscope image including fluorescent imaging, and nano-scale manipulation by E-SEM imaging. We observe C. elegans as biological specimen analysis, because it is one of important model organism for various diseases analysis. The viability analysis has been done from the eGFP fluorescent intensity of C. elegans under E-SEM observation and electron beam irradiation. This system will be applied as future nano-surgery system based on nanomanipulation system for biological specimen.
光学显微镜与环境扫描电镜相结合的混合显微镜分析生物标本活力
提出了一种基于纳米机器人操作系统的光学与环境扫描电镜(E-SEM)相结合的混合显微镜生物标本分析系统。与之前的工作一样,我们提出了利用E-SEM纳米机器人操作系统进行单细胞分析。利用电子扫描电镜的纳米尺度图像,可以实现纳米工具的纳米尺度操作和评价。E-SEM通过特制的探测器和控温台,为我们提供了含水条件下样品的高分辨率图像。然而,电子扫描电镜图像,特别是电子扫描电镜图像是表面图像,因此不可能获得对生物分析至关重要的内部结构成像。本研究将光学显微镜(OM)与电子扫描电镜(E-SEM)相结合,通过包括荧光成像在内的光学显微镜图像对生物标本进行分析,并通过电子扫描电镜成像进行纳米级操作。我们将秀丽隐杆线虫作为生物标本进行分析,因为它是各种疾病分析的重要模式生物之一。通过电子扫描电镜观察和电子束辐照下秀丽隐杆线虫的eGFP荧光强度,对其活力进行了分析。该系统将在生物标本纳米操作系统的基础上作为未来的纳米手术系统。
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