Development of X-Ray Microscopy for Cultured Cells

在过去的几年里，扫描透射x射线显微镜已经成为现实。该项目的目标是开发辅助设备和所需的知识基础，使这种形式的显微镜对整个培养细胞的生物学研究有用。这项工作的目的是使观察完整的、潮湿的、未染色的细胞的空间分辨率比光学显微镜精细十倍。细胞可能是活的，也可能是固定的。这种新型显微技术对细胞生物学和神经生物学，特别是细胞超微结构与功能关系的研究具有广泛的意义。我们将开发一种适合于活培养细胞成像的标本系统。然后将获得原型图像，以确定未染色标本的自然对比显示其超微结构的程度。整个固定细胞的免疫染色技术也将被描述。将进行辐射损伤效应的研究，以确定维持活细胞和固定标本短期和长期完整性的可接受照射范围。还将开发一种改进的系统，最适合成像小于整个哺乳动物细胞的标本，如细菌或肌原纤维。

During the past several years, scanning transmission x-ray microscopy has become a reality. The objective of this project is to develop auxiliary equipment, and the required knowledge base, to make this form of microscopy useful for biological studies of whole cultured cells. This work is intended to make it practical to view whole, wet, unstained cells with a spatial resolution ten times finer than that of the light microscope. The cells may be either alive or fixed. This new kind of microscopy has broad significance for researchers in cell biology and neurobiology, particularly those studying the relation of cellular ultrastructure to function. We will develop a specimen system appropriate for imaging live cultured cells. Prototype images will then be obtained to determine the extent to which the natural contrast of unstained specimens reveals their ultrastructure. Immunostaining techniques for whole fixed cells will also be characterized. Studies of radiation damage effects will be made to define the acceptable range of exposures for maintaining both short-term and long-term integrity of live cells and fixed specimens. A modified system optimal for imaging specimens smaller than whole mammalian cells, such as bacteria or myofibrils, will also be developed.