利用电子光谱成像技术在电子显微切片中选择性检测金颗粒的重金属对比度增强

Ansgar Haking, Helmut Troester, Karsten Richter, Annett Burzlaff, Herbert Spring, Michael F Trendelenburg
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引用次数: 1

摘要

在电子显微镜的超薄切片中,如果胶体金标记非常小,并且底层结构显示由常规铀/铅染色引起的强烈重金属对比,则免疫细胞化学或原位杂交技术生成的图像的评估可能会变得困难。通常情况下,这个问题可以通过在更高的放大倍数下分析样品,或者通过一种叫做银增强的二次过程来放大金颗粒来解决。提出了一种利用电子能谱成像(ESI)可视化小金颗粒的方法。通过数字图像处理,对能量损失分别为0、45和120 eV的三幅能量滤波图像进行分析,使au标记物与图像背景进行光学分离。该协议可适用于与标签自动计数相关的应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Heavy metal contrast enhancement for the selective detection of gold particles in electron microscopical sections using electron spectroscopic imaging

In ultrathin sections for electron microscopy the evaluation of images generated with immunocytochemical or in situ hybridization techniques can become difficult if the colloidal gold label is very small and the underlying structure shows a strong heavy metal contrast caused by conventional uranium/lead staining. Normally, the problem is overcome by analyzing specimens at higher magnifications or by enlarging the gold grains in a secondary process called silver enhancement. We present a method to visualize the small gold particles with electron spectroscopic imaging (ESI). Three energy-filtered images acquired at energy losses of 0, 45 and 120 eV are analyzed by digital image treatment so as to allow the optical separation of the Au-markers from the image background. This protocol can be adapted for applications related to the automated counting of the label.

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