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环境扫描电子显微镜中的超薄切片技术,一种适用于材料科学和生命科学的通用方法。

Ultramicrotomy in the ESEM, a versatile method for materials and life sciences.

作者信息

Zankel A, Kraus B, Poelt P, Schaffer M, Ingolic E

机构信息

Institute for Electron Microscopy, Graz University of Technology, and Center for Electron Microscopy, Graz, Steyrerg. 17, A-8010 Graz, Austria.

出版信息

J Microsc. 2009 Jan;233(1):140-8. doi: 10.1111/j.1365-2818.2008.03104.x.

Abstract

We here present the results of the first materials science analyses obtained with the prototype of a serial block-face sectioning and imaging tool, 3Viewtrade mark of Gatan, Inc (Pleasanton, CA, U.S.A.). It is a specially designed ultramicrotome operating in situ within an environmental scanning electron microscope originally developed for life science research. The microtome removes thin slices from the sample and the environmental scanning electron microscope images each new block surface of the specimen (serial block-face scanning electron microscopy). The Schottky emitter (FEG) of the microscope delivers high spatial resolution and has the advantage of stable performance and high durability. The slice thickness can typically be selected between 50 and 100 nm. It is possible to cut hundreds of slices and simultaneously acquire images with Digital Micrographtrade mark Model 700 (Gatan, Inc.). This article outlines the set-up and describes the automated process. The preparation of specimens for in situ ultramicrotomy is explained and the parameters for good image quality are discussed. In addition, special operative and analytic features of the controlling software are presented. Three different technical materials and one botanical specimen were analyzed delivering first results of this method for materials science and for botany.

摘要

我们在此展示了使用一种连续块面切片与成像工具的原型所获得的首批材料科学分析结果,该工具为美国加利福尼亚州普莱森顿市加坦公司(Gatan, Inc)的3View商标产品。它是一种经过特殊设计的超薄切片机,可在最初为生命科学研究而开发的环境扫描电子显微镜内原位操作。该超薄切片机从样品上切下薄片,环境扫描电子显微镜对标本的每个新块面进行成像(连续块面扫描电子显微镜术)。显微镜的肖特基发射极(场发射枪)提供高空间分辨率,具有性能稳定和耐用性高的优点。切片厚度通常可在50至100纳米之间选择。可以切割数百个切片,并同时使用型号700的Digital Micrograph商标软件(加坦公司)获取图像。本文概述了设置过程并描述了自动化流程。解释了原位超薄切片术标本的制备方法,并讨论了获得良好图像质量的参数。此外,还介绍了控制软件的特殊操作和分析功能。对三种不同的技术材料和一种植物标本进行了分析,得出了该方法在材料科学和植物学方面的首批结果。

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