Chromosome Interior Observation by Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) Using Ionic Liquid Technique.
Chromosome Interior Observation by Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) Using Ionic Liquid Technique.
复制标题
使用离子液体技术通过聚焦离子束/扫描电子显微镜 (FIB/SEM) 观察染色体内部。
DOI：
10.1017/s143192761401280x
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
and Fukui K.
中科院分区：
文献类型：
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作者：
Hamano T.;Dwiranti A.;Kaneyoshi K.;Fukuda S.;Kometani R.;Nakao M.;Takata H.;Uchiyama S.;Ohmido N.;and Fukui K.
长期以来，阐明染色体结构的尝试一直难以捉摸。电子显微镜因其高分辨率和高放大倍率而在染色体结构研究中非常有用。然而，染色体等生物样品需要经过各种准备步骤，包括脱水、干燥和金属/碳涂层，这可能会导致收缩和伪影。离子液体技术是最近发展起来的，它可以在不脱水、干燥或涂覆的情况下制备样品，提供更接近自然条件的样品。与此同时，聚焦离子束/扫描电子显微镜(FIB/SEM)已经发展起来，使研究和直接分析染色体内部成为可能。在这项研究中，我们使用离子液体技术制备的植物和人类染色体，用FIB/SEM研究了染色体内部。结果，用临界点干燥法制备的大麦和人的染色体，都可以看到两种类型的染色体，有腔的和没有腔的。然而，用离子液体技术制备时，染色体内部只显示为固体结构，没有空洞。我们的结果表明，空穴的存在和大小取决于制备过程。因此，离子液体技术与FIB/SEM相结合是染色体研究的有力工具。
Attempts to elucidate chromosome structure have long remained elusive. Electron microscopy is useful for chromosome structure research because of its high resolution and magnification. However, biological samples such as chromosomes need to be subjected to various preparation steps, including dehydration, drying, and metal/carbon coating, which may induce shrinkage and artifacts. The ionic liquid technique has recently been developed and it enables sample preparation without dehydration, drying, or coating, providing a sample that is closer to the native condition. Concurrently, focused ion beam/scanning electron microscopy (FIB/SEM) has been developed, allowing the investigation and direct analysis of chromosome interiors. In this study, we investigated chromosome interiors by FIB/SEM using plant and human chromosomes prepared by the ionic liquid technique. As a result, two types of chromosomes, with and without cavities, were visualized, both for barley and human chromosomes prepared by critical point drying. However, chromosome interiors were revealed only as a solid structure, lacking cavities, when prepared by the ionic liquid technique. Our results suggest that the existence and size of cavities depend on the preparation procedures. We conclude that combination of the ionic liquid technique and FIB/SEM is a powerful tool for chromosome study.