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Implementation of subcellular water mapping by electron energy loss spectroscopy in a medium-voltage scanning transmission electron microscope

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Abstract

The water concentration in biological cells plays a predominant role in cellular life. Using electron energy loss spectroscopy, the feasibility to measure the water content in cells has already been demonstrated. In this paper, we present an upgrade of water measurement in hydrated cryosections by spectrum imaging mode in a medium-voltage scanning transmission electron microscope. The electron energy loss spectra are recorded in spectrum imaging mode in a 2n×2n pixels array. Each spectrum is processed in order to determine the water mass content in the corresponding pixel. Then a parametric image is obtained in which grey levels are related to water concentration. In this image, it is possible to recognize the different subcellular compartments. By averaging the water concentration over the relevant pixels, we can determine the water mass content in the concerned subcellular compartment. As an example, we present water mass content measurement at subcellular level in rat hepatocytes.

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Acknowledgements

The authors thank Dr. Karl Zierold (Max Planck Institut für Molekular Physiology, Dortmund, Germany) for his precious advice on cryomethods and for helpful discussions.

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Authors and Affiliations

  1. Laboratoire de Microscopie Electronique Analytique, INSERM ERM 0203, 21 rue Clément Ader, 51685 Cedex 2, Reims, France

    C. Terryn, J. Michel, D. Laurent-Maquin & G. Balossier

  2. GSMA, UMR 6089, Faculté des Sciences, BP 1039, 51687 Cedex 2, Reims , France

    X. Thomas

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  1. C. Terryn
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  2. J. Michel
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  3. X. Thomas
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  4. D. Laurent-Maquin
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  5. G. Balossier
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Correspondence to C. Terryn.

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Terryn, C., Michel, J., Thomas, X. et al. Implementation of subcellular water mapping by electron energy loss spectroscopy in a medium-voltage scanning transmission electron microscope. Eur Biophys J 33, 321–327 (2004). https://doi.org/10.1007/s00249-003-0347-2

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  • DOI: https://doi.org/10.1007/s00249-003-0347-2

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