Scherfeld D, Schneider G, Guttmann P, Osborn M
Institute of X-ray Physics, University of Goettingen, Geiststrasse 11, Goettingen, 37073, Germany.
J Struct Biol. 1998 Sep;123(1):72-82. doi: 10.1006/jsbi.1998.4017.
Transmission X-ray microscopy has been used to study the arrangement of cytoskeletal filaments in interphase PtK2 cells. Extraction of the soluble proteins and of some organelles with the nonionic detergent Triton X-100 was important in obtaining sufficient image contrast between the insoluble cytoskeletal filaments and the surrounding cytoplasm. If this step is not performed cytoskeletal filaments are not visualized and transmission X-ray micrographs of the cytoplasm instead show predominantly membrane-bound organelles such as vesicles and the endoplasmic reticulum. Transmission X-ray micrographs of the cytoskeletal filaments and endoplasmic reticulum in air-dried specimens, as well as in specimens examined in the wet state, can be directly compared with transmission electron micrographs of cytoskeletons prepared in the same way. The profiles seen with the two techniques are similar, although, currently, transmission X-ray micrographs have a limit of resolution of approximately 50 nm. Transmission X-ray micrographs appear to show some substructure in interphase nuclei in cells fixed either with glutaraldehyde or by cryofixation and examined in a hydrated condition.
透射X射线显微镜已被用于研究间期PtK2细胞中细胞骨架丝的排列。用非离子去污剂Triton X-100提取可溶性蛋白质和一些细胞器，对于在不溶性细胞骨架丝与周围细胞质之间获得足够的图像对比度非常重要。如果不进行这一步骤，细胞骨架丝就无法可视化，细胞质的透射X射线显微照片反而主要显示膜结合细胞器，如囊泡和内质网。空气干燥标本以及湿态检查标本中细胞骨架丝和内质网的透射X射线显微照片，可以直接与以相同方式制备的细胞骨架的透射电子显微照片进行比较。尽管目前透射X射线显微照片的分辨率极限约为50纳米，但两种技术看到的轮廓相似。在用水合状态检查的、用戊二醛固定或冷冻固定的细胞的间期核中，透射X射线显微照片似乎显示出一些亚结构。
