Open AccessSwitchable resolution in soft x-ray tomography of single cells
Author(s) -
Venera Weinhardt,
Jianhua Chen,
Axel Ekman,
Jessica Guo,
Soumya G. Remesh,
Michal Hammel,
Gerry McDermott,
Weilun Chao,
Sharon Oh,
Mark A. Le Gros,
Carolyn A. Larabell
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0227601
Subject(s) - microscope , resolution (logic) , microscopy , image resolution , lens (geology) , tomography , optics , focus (optics) , optical microscope , optical coherence tomography , flexibility (engineering) , materials science , computer science , biomedical engineering , artificial intelligence , physics , mathematics , medicine , scanning electron microscope , statistics
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.