期刊
MICROSCOPY AND MICROANALYSIS
卷 19, 期 1, 页码 22-29出版社
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S1431927612013827
关键词
compact light source; laboratory light source; X-ray tomography; correlative microscopy; cryogenic soft X-ray microscope
资金
- National Institutes of Health [5RC1GM091755]
- U.S. Department of Energy [DE-AC05-76RL01830]
- [2R44RR022488-04]
Here we present a novel laboratory-based cryogenic soft X-ray microscope for whole cell tomography of frozen hydrated samples. We demonstrate the capabilities of this compact cryogenic microscope by visualizing internal subcellular structures of Saccharomyces cerevisiae cells. The microscope is shown to achieve better than 50 nm half-pitch spatial resolution with a Siemens star test sample. For whole biological cells, the microscope can image specimens up to 5 mu m thick. Structures as small as 90 nm can be detected in tomographic reconstructions following a low cumulative radiation dose of only 7.2 MGy. Furthermore, the design of the specimen chamber utilizes a standard sample support that permits multimodal correlative imaging of the exact same unstained yeast cell via cryo-fluorescence light microscopy, cryo-soft X-ray microscopy, and cryo-transmission electron microscopy. This completely laboratory-based cryogenic soft X-ray microscope will enable greater access to three-dimensional ultrastructure determination of biological whole cells without chemical fixation or physical sectioning.
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