人类中期染色体的扫描电镜。

Scanning electron microscopy Pub Date : 1986-01-01
T D Allen, E M Jack, C J Harrison, D Claugher
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引用次数: 0

摘要

染色体扫描电镜的制备方法取决于材料的原始来源。通过分离缓冲液从未固定的中期细胞中提取的染色体往往表现出地形和表面形态,这可能是由分离缓冲液本身的选择引起的。此外,这种类型的制备通常排除任何染色体鉴定,因为许多中期已经合并,而且这些制备的染色体不适合临床细胞遗传学中经常使用的显带技术。我们自己的方法是使用标准的细胞遗传学方法,从甲醇-乙酸固定,风干中期扩散开始,既可以识别单个染色体,也可以进行各种带带程序,如G带和C带。随后,使用补液、戊二醛固定和硫代碳肼(TCH)的锇浸渍,染色体被“重新准备”用于扫描电镜。这种方法产生的染色体可以在没有金属涂层的情况下以高分辨率检查其形貌,表面形态和染色质组织,以及通过带技术产生的变化,这些变化引起结构改变,从而在光学显微镜下产生差异染色。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Scanning electron microscopy of human metaphase chromosomes.

Preparative methods for scanning electron microscopy of chromosomes are dependent on the original source of material. Chromosomes extracted from unfixed metaphase cells via isolation buffers tend to show topography and surface morphology which may have been induced by the choice of isolation buffer itself. Furthermore, this type of preparation often precludes any chromosome identification, as many metaphases have been pooled, and also the chromosomes from these preparations are not suitable for the banding techniques regularly used in clinical cytogenetics. Our own approach has been to use the standard cytogenetic approach, starting with methanol-acetic acid fixed, air dried metaphase spreads, allowing both identification of individual chromosomes, and also the facility for various banding procedures such as G and C banding to be performed. Chromosomes are subsequently "reprepared" for SEM, using rehydration, glutaraldehyde fixation, and osmium impregnation using Thiocarbohydrazide (TCH). This method produces chromosomes which can be examined at high resolution, without metallic coating, for their topography, surface morphology and chromatin organisation, and the changes produced by banding techniques which give rise to a structural alterations resulting in differential staining in the light microscope.

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