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Correlative organelle fluorescence microscopy and synchrotron X-ray chemical element imaging in single cells

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Abstract

X-ray chemical element imaging has the potential to enable fundamental breakthroughs in the understanding of biological systems because chemical element interactions with organelles can be studied at the sub-cellular level. What is the distribution of trace metals in cells? Do some elements accumulate within sub-cellular organelles? What are the chemical species of the elements in these organelles? These are some of the fundamental questions that can be addressed by use of X-ray chemical element imaging with synchrotron radiation beams. For precise location of the distribution of the elements, identification of cellular organelles is required; this can be achieved, after appropriate labelling, by use of fluorescence microscopy. As will be discussed, this approach imposes some limitations on sample preparation. For example, standard immunolabelling procedures strongly modify the distribution of the elements in cells as a result of the chemical fixation and permeabilization steps. Organelle location can, however, be performed, by use of a variety of specific fluorescent dyes or fluorescent proteins, on living cells before cryogenic fixation, enabling preservation of element distribution. This article reviews the methods used for fluorescent organelle labelling and X-ray chemical element imaging and speciation of single cells. Selected cases from our work and from other research groups are presented to illustrate the potential of the combination of the two techniques.

Synchrotron X-ray fluorescence distribution maps of Ca, P and S in yeast cells. Elemental distribution maps (green color scale) were combined with the image of vacuoles labeled with Arg-CMAC (red color scale). The yellow signal of superposed images shows that Ca and P are preferentially located within the vacuole.

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Acknowledgments

The authors thank the guest editors N. Jakubowski and P. Dittrich for their kind invitation to publish in this special issue dedicated to “Single Cell Analysis”. In their research the authors have benefited from access to beamlines ID21 and ID22 at the European Synchrotron Radiation Facility (ESRF), Grenoble, France. We are sincerely grateful to M. Salomé, J. Susini, G. Veronesi, S. Bohic, R. Tucoulou, and P. Cloetens from ESRF and to G. Devès and T. Bacquart from CENBG for providing assistance during the experiments. Some of the developments reported in this review were financially supported by the ANR (Agence Nationale de la Recherche) program PIRIBIO (ANR-09-PIRI-0029-01).

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Authors and Affiliations

  1. University of Bordeaux, CNRS, CENBG, UMR 5797, 33170, Gradignan, France

    Stéphane Roudeau, Asuncion Carmona, Laura Perrin & Richard Ortega

  2. CNRS, IN2P3, CENBG, UMR 5797, 33170, Gradignan, France

    Stéphane Roudeau, Asuncion Carmona, Laura Perrin & Richard Ortega

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  1. Stéphane Roudeau
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  2. Asuncion Carmona
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  4. Richard Ortega
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Correspondence to Richard Ortega.

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Published in the topical collection Single Cell Analysis with guest editors Petra Dittrich and Norbert Jakubowski.

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Roudeau, S., Carmona, A., Perrin, L. et al. Correlative organelle fluorescence microscopy and synchrotron X-ray chemical element imaging in single cells. Anal Bioanal Chem 406, 6979–6991 (2014). https://doi.org/10.1007/s00216-014-8004-4

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  • DOI: https://doi.org/10.1007/s00216-014-8004-4

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