Preparation of biological tissue sections for correlative ion, electron, and light microscopy
Preparation of biological tissue sections for correlative ion, electron, and light microscopy
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用于相关离子、电子和光学显微镜的生物组织切片的制备
DOI：
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
J. Shelburne
中科院分区：
文献类型：
--
作者：
K. Kupke;J. P. Pickett;P. Ingram;D. Griffis;R. W. Linton;P. Burger;J. Shelburne
为了正确解释使用离子显微镜（IM）获得的化学图像，将它们与常规光学显微镜（LM），二次电子成像（SEI），背散射电子成像（BEI）和电子探针显微分析（EPMA）提供的信息相关联是有用的。因此，我们设计了一种标本制备技术，该技术允许将几种不同的微量分析技术应用于安装在同一基底上的单个组织切片。将切片切割到聚酯塑料盖玻片上（使用EPMA，没有原子序数> 9的任何元素的峰）并通过LM研究。在用碳轻旋转涂层（以防止充电）后，可以通过SEI，BEI和EPMA检查切片。可以使用安装在物镜上的针式组织微定位器或通过在感兴趣的特征上放置切割成特定几何形状的小块金属箔来标记特定区域以进行IM研究。在用铂涂覆样品后，使用典型IM样品室上可用的低功率反射光显微镜作为离子束放置的指南，可以看到无金属的阴影。导电涂层还最大限度地减少了IM期间的试样充电。IM后光学显微镜，SEI和BEI用于确认IM实验中探测的特定区域的位置，并提供有关差分离子溅射伪影和组织污染物的信息。这种新的相关技术应允许更好地理解这些不同的仪器获得的图像。
In order to correctly interpret the chemical images obtained using ion microscopy (IM), it is useful to correlate them with the information provided by conventional light microscopy (LM), secondary electron imaging (SEI), backscattered electron imaging (BEI), and electron probe microanalysis (EPMA). Accordingly, we have devised a technique of specimen preparation which allows for the application of several different microanalytical techniques to a single histologic section mounted on the same substrate. Sections are cut onto polyester plastic coverslips (devoid of peaks for any element with atomic number > 9 using EPMA) and studied by LM. After a light rotary coating with carbon (to prevent charging), the section can then be examined by SEI, BEI, and EPMA. Specific areas can be marked for IM study either with an objective-mounted pin tissue microlocater, or by placing small pieces of metal foil, cut in specific geometric shapes, over features of interest. After sputter-coating the sample with platinum, metal-free shadows are visible using a low-power reflected light microscope available on a typical IM sample chamber as a guide for ion beam placement. The conductive coatings also minimize specimen charging during IM. Post-IM light microscopy, SEI, and BEI are used to confirm the location of specific areas probed in the IM experiments and to provide information on differential ion-sputtering artifacts and tissue contaminants. This new correlative technique should permit better understanding of the images obtained with these diverse instruments.
影响因子：
56.9
作者：
PERL, DP;GAJDUSEK, DC;GIBBS, CJ
通讯作者：
GIBBS, CJ