Switchable resolution in soft x-ray tomography of single cells

Autoři: Venera Weinhardt aff001;  Jian-Hua Chen aff001;  Axel A. Ekman aff001;  Jessica Guo aff002;  Soumya G. Remesh aff001;  Michal Hammel aff001;  Gerry McDermott aff001;  Weilun Chao aff003;  Sharon Oh aff003;  Mark A. Le Gros aff001;  Carolyn A. Larabell aff001
Působiště autorů: Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, California, United States of America aff001;  Department of Anatomy, University of California San Francisco, San Francisco, California, United States of America aff002;  Center for X-ray Optics, Material Science Division, Lawrence Berkeley National Laboratory, Berkeley, California, United States of America aff003
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0227601


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.

Klíčová slova:

B cells – Light microscopy – Optical lenses – Schizosaccharomyces pombe – Tomography – X-ray radiography – Yeast – X-ray microscopy


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