STRUCTURAL DETERMINATION OF INTERPHASE CHROMOSOMES

新开发的仪器和软件现在允许 三维重建，在一般的方式，亚细胞 结构，在光和高压电子显微镜 程度. 18个核已重建和解释在一个 量化的时尚 六个细胞核的细胞遗传学位点与 三维结构显示，许多染色体特征， 三维确定。 建议在三维上确定许多相邻的多线 在确定的发育阶段和组织中， 可以用定量的方式来观察结构关系。 提出了许多计算方法来处理图像， 结构，定位带，因此基因的三维， 来确定原子核的精细结构。 这些研究还将利用 单克隆抗体定位活性基因位点及其关闭 与另一个和核膜的联系， 结构到功能。 此外，二倍体细胞核将在2000年进行三维重建。 完整果蝇胚胎的最高光学显微镜分辨率。 与间期3D模式差异相关的问题 染色体作为发育的函数，细胞周期将 研究了 实验室将继续重建有丝分裂染色体， 来自果蝇多线染色体的interband/bands 三维光线使分辨率提高成为可能 显微镜和高得多的分辨率的高压电子 显微镜利用系统的高倾斜约70度， 三维结构。 三维图像处理的新计算方法是 正在继续。 用于光学显微镜的新型图像采集硬件 和高压电子显微镜。

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.