Switchable resolution in soft x-ray tomography of single cells

Journal Article · 23 January 2020 · PLoS ONE

DOI: https://doi.org/10.1371/journal.pone.0227601 · OSTI ID:1615292

Weinhardt, Venera  ^[1]; Chen, Jian-Hua ^[1]; Ekman, Axel A. ^[1]; Guo, Jessica ^[2]; Remesh, Soumya G. ^[3]; Hammel, Michal ^[3]; McDermott, Gerry ^[1]; Chao, Weilun ^[3]; Oh, Sharon ^[3]; Le Gros, Mark A. ^[1]; Larabell, Carolyn A. ^[1]

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1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, San Francisco, CA (United States)
2. Univ. of California, San Francisco, CA (United States)
3. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

The diversity of living cells, in both size and internal complexity, calls for imaging methods with adaptable spatial resolution. Soft x-ray tomography (SXT) is a three-dimensional imaging technique ideally suited to visualizing and quantifying the internal organization of single cells of varying sizes in a near-native state. The achievable resolution of the soft x-ray microscope is largely determined by the objective lens, but switching between objectives is extremely time-consuming and typically undertaken only during microscope maintenance procedures. Since the resolution of the optic is inversely proportional to the depth of focus, an optic capable of imaging the thickest cells is routinely selected. This unnecessarily limits the achievable resolution in smaller cells and eliminates the ability to obtain high-resolution images of regions of interest in larger cells. Here, we describe developments to overcome this shortfall and allow selection of microscope optics best suited to the specimen characteristics and data requirements. We demonstrate that switchable objective capability advances the flexibility of SXT to enable imaging cells ranging in size from bacteria to yeast and mammalian cells without physically modifying the microscope, and we demonstrate the use of this technology to image the same specimen with both optics.

• Research Organization: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
• Sponsoring Organization: German Research Foundation (DFG); National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
• Grant/Contract Number: AC02-05CH11231
• OSTI ID: 1615292
• Journal Information: PLoS ONE, Journal Name: PLoS ONE Journal Issue: 1 Vol. 15; ISSN 1932-6203
• Publisher: Public Library of ScienceCopyright Statement
• Country of Publication: United States
• Language: English