Ohtani Toshio, Shichirii Motoharu, Fukushi Daisuke, Sugiyama Shigeru, Yoshino Tomoyuki, Kobori Toshiro, Hagiwara Shoji, Ushiki Tatsuo
National Food Research Institute, Tsukuba, Ibaraki, Japan.
Arch Histol Cytol. 2002 Dec;65(5):425-34. doi: 10.1679/aohc.65.425.
Topographic and fluorescent images of whole barley chromosomes stained with YOYO-1 were observed simultaneously by scanning near-field optical/ atomic force microscopy (SNOM/AFM). The chromosome was relatively smooth and flat in the topographic images and no significant difference in height was present between regions of high fluorescent and low fluorescent intensity in the chromosomes. The telomeric region, labeled by fluorescence in situ hybridization (FISH) method, was also observed by SNOM/AFM at high resolution, and fluorescent signals of the telomeric region were clearly defined on the topographic image of chromatin fibers on the chromosome at the nano-meter scale level. Although the telomeric signals were usually visualized as a single fluorescent region at the end of sister chromatids by conventional light microscopy, they were observed separately as two fluorescent regions, less than 100-200 nm distance, using the SNOM/AFM. The SNOM/AFM offers great potential in identifying particular single gene location on chromosomes in the near future.
利用扫描近场光学/原子力显微镜（SNOM/AFM）同时观察了用YOYO-1染色的大麦全染色体的形貌和荧光图像。在形貌图像中，染色体相对平滑且平坦，染色体上高荧光强度区域和低荧光强度区域之间的高度没有显著差异。通过荧光原位杂交（FISH）方法标记的端粒区域也通过SNOM/AFM在高分辨率下进行了观察，并且在纳米尺度水平上，端粒区域的荧光信号在染色体上染色质纤维的形貌图像上清晰可辨。尽管通过传统光学显微镜观察时，端粒信号通常在姐妹染色单体末端呈现为单个荧光区域，但使用SNOM/AFM时，它们被观察为两个距离小于100 - 200 nm的荧光区域。SNOM/AFM在不久的将来识别染色体上特定单基因位置方面具有巨大潜力。
