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Putting Molecules in the Picture: Using Correlated Light Microscopy and Soft X-ray Tomography to Study Cells

Synchrotron Light Sources and Free-Electron Lasers

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Abstract

Transmission soft x-ray microscopy (SXM) is an important new technique for visualizing and quantifying biological specimens, especially individual cells. In SXM the specimen is illuminated with “water-window” soft x-rays (i.e., x-ray photons with energies in the range 284–543 eV). As the illuminating light passes through the specimen, it is attenuated more strongly by carbon- and nitrogen-containing biomolecules than by water. Consequently, even subtle differences in molecular composition produce SXM images with quantitative contrast. In this chapter, we will discuss the basic concepts behind SXM and describe how 2D SXM data is used to calculate high fidelity 3D soft x-ray tomographic (SXT) reconstructions of the specimen. SXT offers many advantages over other high-resolution imaging methods. A particular strength is the capacity to image intact, fully hydrated cells, including eukaryotes such as yeast or mammalian cells. In addition to standalone use, SXT data can be integrated with data from other imaging modalities. The recent development of cryogenic confocal fluorescence tomography (CFT) has permitted 3D SXT reconstructions to be combined directly with 3D molecular localization data measured from the same cryopreserved specimen. With the advent of correlated CFT-SXT imaging, cell biologists can now accurately locate the position of specific molecules in the context of a high-resolution reconstruction of a near-native state cell.

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    Visit http://www.nature.com/milestones/milelight/timeline.html for the major milestones in light microscopy.

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Acknowledgements

This work was supported by the US Department of Energy, Office of Biological and Environmental Research (DE-AC02-05CH11231), the National Center for Research Resources of the National Institutes of Health (P41RR019664) and the National Institutes of General Medicine of the National Institutes of Health (P41GM103445), and the Gordon and Betty Moore Foundation.

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  1. Department of Anatomy, University of California, 1550 4th St., 238B Rock Hall, 94143-2722, San Francisco,CA, USA

    Carolyn A.Larabell

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  1. Mark A. Le Gros
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  2. Jian-Hua Chen
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  3. ∎ MyanDo
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  4. ∎ GerryMcDermott
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  5. Carolyn A.Larabell
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Correspondence to Carolyn A.Larabell .

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Editors and Affiliations

  1. Helmholtz-Zentrum Berlin, Berlin, Germany

    Eberhard Jaeschke

  2. Zentrum für Synchrotronstrahlung, TU Dortmund, Dortmund, Germany

    Shaukat Khan

  3. FS-CFEL-1, DESY, Hamburg, Germany

    Jochen R. Schneider

  4. SLAC National Accelerator Laboratory Linac Coherent Light Source, Stanford University, Menlo Park, California, USA

    Jerome B. Hastings

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Gros, M., Chen, JH., MyanDo, ∎., GerryMcDermott, ∎., A.Larabell, C. (2015). Putting Molecules in the Picture: Using Correlated Light Microscopy and Soft X-ray Tomography to Study Cells. In: Jaeschke, E., Khan, S., Schneider, J., Hastings, J. (eds) Synchrotron Light Sources and Free-Electron Lasers. Springer, Cham. https://doi.org/10.1007/978-3-319-04507-8_43-1

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  • DOI: https://doi.org/10.1007/978-3-319-04507-8_43-1

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  1. Latest

    Putting Molecules in the Picture: Using Correlated Light Microscopy and Soft X-Ray Tomography to Study Cells
    Published:
    06 July 2019

    DOI: https://doi.org/10.1007/978-3-319-04507-8_43-3

  2. Putting Molecules in the Picture: Using Correlated Light Microscopy and Soft X-Ray Tomography to Study Cells
    Published:
    07 March 2019

    DOI: https://doi.org/10.1007/978-3-319-04507-8_43-2

  3. Original

    Putting Molecules in the Picture: Using Correlated Light Microscopy and Soft X-ray Tomography to Study Cells
    Published:
    28 July 2015

    DOI: https://doi.org/10.1007/978-3-319-04507-8_43-1

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