Laboratory-Based Cryogenic Soft X-ray Tomography with Correlative Cryo-Light and Electron Microscopy

Journal Article · Fri Feb 01 04:00:00 EST 2013 · Microscopy and Microanalysis, 19(1):22-29

DOI:https://doi.org/10.1017/S1431927612013827· OSTI ID:1063698

Carlson, David B.; Gelb, Jeff; Palshin, Vadim; Evans, James E.

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 sub-cellular structures of Saccharomyces cerevisiae cells. The microscope is shown to achieve better than 50 nm spatial resolution with a Siemens star test sample. For whole biological cells, the microscope can image specimens up to 5 micrometers thick. Structures as small as 90 nm can be detected in tomographic reconstructions at roughly 70 nm spatial resolution 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 therefore enable greater access to three-dimensional ultrastructure determination of biological whole cells without chemical fixation or physical sectioning.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (US)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 1063698
• Report Number(s): PNNL-SA-85707
• Journal Information: Microscopy and Microanalysis, 19(1):22-29, Journal Name: Microscopy and Microanalysis, 19(1):22-29
• Country of Publication: United States
• Language: English

Related Subjects

Compact light source
Laboratory light source
correlative microscopy
cryogenic soft x-ray microscope
x-ray tomography