Imaging of fine structures of cellular organelles in hydrated biological cells by a soft x-ray microscope combined with a fluorescence microscope

Abstract

We have proposed and developed a new hybrid microscopy system using a soft x-ray microscope and a fluorescence microscope imaging the same biological cells at the nearly same time. Combining the powerful advantages such as high spatial resolution of the soft x-ray microscope and the accurate organelle identification of the fluorescence microscope, we can observe fine structures of the cellular organelles in live hydrated biological cells in situ. Staining the cells with several fluorescent dyes such as Mito-tracker, Phalloidin, and DAPI, the soft x-ray images of the cells have been directly compared with the fluorescent images and the cellular organelles such as mitochondria, actin filaments, and chromosomes in the soft x-ray images have been clearly identified. Since the soft x-ray microscope has higher spatial resolution than that of the fluorescence microscope, not only shape of the cellular organelles but also the fine structures of the cellular organelles of the live biological cells have been clearly observed for the first time.