Sugiyama Shigeru, Yoshino Tomoyuki, Kanahara Hiroko, Shichiri Motoharu, Fukushi Daisuke, Ohtani Toshio
National Food Research Institute, Kannondai 2-1-12, Tsukuba, Ibaraki 305-8642, Japan.
Anal Biochem. 2004 Jan 1;324(1):39-44. doi: 10.1016/j.ab.2003.09.026.
Acetic acid treatment has been frequently used to remove cellular contaminants from plant chromosome samples for structural analyses by scanning electron microscopy and atomic force microscopy (AFM). We evaluated the effects of various concentrations of acetic acid treatments on barley chromosome structures by using AFM. The long-term 45% acetic acid treatment significantly damaged the chromosome structures, although the treatment effectively removed the cellular contaminants. On the other hand, the treatment with 15% acetic acid could not obtain sufficiently clean chromosome samples and the chromosome surface structures could not be observed. In contrast, we obtained clean chromosome preparation without severe damage by using an intermediate concentration (30%) of acetic acid treatment. In the centromeric region, we could observe fiber structures with a width of 100 nm, which were composed of ca. 50-nm granules and aligned to the axes of chromosomes. Thus, AFM analysis of chromosomes appropriately treated with acetic acid will provide important insights into the organization of higher-order structures of plant chromosomes.
醋酸处理常用于通过扫描电子显微镜和原子力显微镜（AFM）从植物染色体样本中去除细胞污染物，以进行结构分析。我们使用AFM评估了不同浓度醋酸处理对大麦染色体结构的影响。长期45%醋酸处理显著破坏了染色体结构，尽管该处理有效地去除了细胞污染物。另一方面，15%醋酸处理无法获得足够干净的染色体样本，且无法观察到染色体表面结构。相比之下，我们通过使用中等浓度（30%）的醋酸处理获得了干净的染色体标本，且没有严重损伤。在着丝粒区域，我们可以观察到宽度为100 nm的纤维结构，其由约50 nm的颗粒组成，并与染色体轴对齐。因此，对经醋酸适当处理的染色体进行AFM分析将为植物染色体高阶结构的组织提供重要见解。
