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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Use of energy-filtering transmission electron microscopy for routine ultrastructural analysis of high-pressure-frozen or chemically fixed plant cells
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Use of energy-filtering transmission electron microscopy for routine ultrastructural analysis of high-pressure-frozen or chemically fixed plant cells

机译：使用能量过滤透射电子显微镜对高压冷冻或化学固定的植物细胞进行常规超微结构分析

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摘要

In the present study energy-filtering transmission electron microscopy by use of an in-column spectrometer is employed as a powerful tool for ultrastructural analysis of plant cells. Images of unstained very thin (50 nm) and thick (140 nm) sections of the unicellular green alga Micrasterias denticulata, as a model system for a growing plant cell, taken by conventional transmission electron microscopy are compared to those obtained from filtering at zero energy loss (elastic bright field) and to those generated by energy filtering below the carbon-specific absorption edge at about 250 eV. The results show that the high-contrast images produced by the latter technique are distinctly superior in contrast and information content to micrographs taken at conventional transmission electron microscopy mode or at elastic bright field. Post- or en bloc staining with heavy metals, which is indispensable for conventional bright-field transmission electron microscopy, can be completely omitted. Delicate structural details such as membranous or filamentous connections between organelles, organelle interactions, or vesicle and vacuole contents are clearly outlined against the cytoplasmic background. Also. immunoelectron microscopic localization of macromolecules benefits from energy-filtering transmission electron microscopy by a better and more accurate assignment of antigens and structures and by facilitating the detection of immunomarkers without renunciation of contrast.
机译：在本研究中，通过使用柱内光谱仪的能量过滤透射电子显微镜被用作植物细胞超微结构分析的有力工具。将通过常规透射电子显微镜拍摄的单细胞绿藻Micrasterias denticulata的未染色非常薄（50 nm）和厚（140 nm）切片的图像作为生长植物细胞的模型系统，与通过零能量过滤获得的图像进行比较损耗（弹性明场）以及通过能量过滤在约250 eV的碳比吸收边缘以下产生的能量损失。结果表明，由后一种技术产生的高对比度图像在对比度和信息含量方面明显优于常规透射电子显微镜模式或弹性明场下拍摄的显微照片。重金属的后染色或整块染色是常规明场透射电子显微镜必不可少的。微妙的结构细节，例如细胞器之间的膜状或丝状连接，细胞器相互作用或囊泡和液泡内容物，均针对细胞质背景进行了概述。也。大分子的免疫电子显微镜定位得益于能量过滤透射电子显微镜，它可以更好，更准确地分配抗原和结构，并有助于在不放弃对比度的情况下检测免疫标记。

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