STRUCTURAL DETERMINATION OF INTERPHASE CHROMOSOMES

新开发的仪器和软件现在允许 亚细胞的普遍形式的三维重建 光电子显微镜和高压电子显微镜下的结构 级别。对18个原子核进行了重建和解释 量化时尚。六个核的细胞遗传学基因座与 显示出许多染色体特征的三维结构是 三维确定的。 提出了用三维方法确定多个相邻多线的方法。 在特定的发育阶段和组织中的细胞核，因此它将是 有可能以定量的方式看到结构关系。 提出了许多计算方法来处理图像、细化图像 这些结构，以三维方式定位条带，从而定位基因，以及 以确定原子核的精细结构。这些研究还将利用 定位活性基因位点的单抗及其近似性 彼此之间的联系和核膜，并联系 结构才能发挥作用。 此外，二倍体细胞核还将在 完整果蝇胚胎的最高光学显微镜分辨率。 与界面三维图形差异有关的问题 染色体作为发育的函数，细胞周期将是 学习。 该实验室将继续重建有丝分裂染色体和 果蝇多线染色体的带间带 分辨率-由三维光实现的重叠方式 高压电子的显微镜和更高的分辨率 利用系统高倾角约70度的显微镜 三维结构。 三维图像处理的新计算方法是 还在继续。用于光学显微镜的新型图像采集硬件 提出了高压电子显微镜的概念。

The newly developed instrumentation and software now allows three-dimensional reconstruction, in a generalized fashion, of subcellular structures, at both the light and high voltage electron microscopic levels. Eighteen nuclei have been reconstructed and interpreted in a quantitative fashion. Six nuclei have their cytogenetic loci aligned with the three-dimensional structures showing that many chromosome features are three-dimensionally determined. It is proposed to determine, three-dimensionally, many adjacent polytene nuclei in defined developmental stages and tissues so that it will be possible to see in a quantitative fashion the structural relationships. Many computational approaches are proposed to process the images, to refine the structures, to position the bands, hence genes three-dimensionally, and to determine the fine structure of nuclei. These studies will also utilize monoclonal antibodies to localize active gene loci and their close associations one with another and the nuclear envelope, and to relate structure to function. Diploid nuclei will, in addition, be three-dimensionally reconstructed at highest possible light microscope resolution in intact Drosophila embryos. Questions related to differences in the 3D patterns of the interphase chromosomes as a function of development, and the cell cycle will be studied. The laboratory will continue to reconstruct mitotic chromosomes and interband/bands from polytene chromosomes from Drosophila in a resolution-overlaping fashion made possible by the three-dimensional light microscopy and much higher resolution of the high voltage electron microscope utilizing systematic high tilt about 70 degrees for three-dimensional structure. New computational approaches to image processing in three dimensions are being continued. New image collection hardware for the light microscopy and high voltage electron microscopy are proposed.