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Synchrotron-Based Chemical Nano-Tomography of Microbial Cell-Mineral Aggregates in their Natural, Hydrated State

Published online by Cambridge University Press:  19 February 2014

Gregor Schmid ,
Fabian Zeitvogel ,
Likai Hao ,
Pablo Ingino ,
Wolfgang Kuerner ,
James J. Dynes ,
Chithra Karunakaran ,
Jian Wang ,
Yingshen Lu  and
Travis Ayers
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Gregor Schmid
Affiliation:
Department of Geosciences, Center for Applied Geoscience, University of Tuebingen, Hoelderlinstr. 12, 72074 Tuebingen, Germany
Fabian Zeitvogel
Affiliation:
Department of Geosciences, Center for Applied Geoscience, University of Tuebingen, Hoelderlinstr. 12, 72074 Tuebingen, Germany
Likai Hao
Affiliation:
Department of Geosciences, Center for Applied Geoscience, University of Tuebingen, Hoelderlinstr. 12, 72074 Tuebingen, Germany
Pablo Ingino
Affiliation:
Department of Geosciences, Center for Applied Geoscience, University of Tuebingen, Hoelderlinstr. 12, 72074 Tuebingen, Germany
Wolfgang Kuerner
Affiliation:
Department of Geosciences, Center for Applied Geoscience, University of Tuebingen, Hoelderlinstr. 12, 72074 Tuebingen, Germany
James J. Dynes
Affiliation:
Canadian Light Source Inc., Saskatoon, Canada
Chithra Karunakaran
Affiliation:
Canadian Light Source Inc., Saskatoon, Canada
Jian Wang
Affiliation:
Canadian Light Source Inc., Saskatoon, Canada
Yingshen Lu
Affiliation:
Canadian Light Source Inc., Saskatoon, Canada
Travis Ayers
Affiliation:
Luxel Corp., Friday Harbor, USA
Chuck Schietinger
Affiliation:
Luxel Corp., Friday Harbor, USA
Adam P. Hitchcock
Affiliation:
McMaster University, Hamilton, Canada
Martin Obst*
Affiliation:
Department of Geosciences, Center for Applied Geoscience, University of Tuebingen, Hoelderlinstr. 12, 72074 Tuebingen, Germany
*
*Corresponding author. [email protected]
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Abstract

Chemical nano-tomography of microbial cells in their natural, hydrated state provides direct evidence of metabolic and chemical processes. Cells of the nitrate-reducing Acidovorax sp. strain BoFeN1 were cultured in the presence of ferrous iron. Bacterial reduction of nitrate causes precipitation of Fe(III)-(oxyhydr)oxides in the periplasm and in direct vicinity of the cells. Nanoliter aliquots of cell-suspension were injected into custom-designed sample holders wherein polyimide membranes collapse around the cells by capillary forces. The immobilized, hydrated cells were analyzed by synchrotron-based scanning transmission X-ray microscopy in combination with angle-scan tomography. This approach provides three-dimensional (3D) maps of the chemical species in the sample by employing their intrinsic near-edge X-ray absorption properties. The cells were scanned through the focus of a monochromatic soft X-ray beam at different, chemically specific X-ray energies to acquire projection images of their corresponding X-ray absorbance. Based on these images, chemical composition maps were then calculated. Acquiring projections at different tilt angles allowed for 3D reconstruction of the chemical composition. Our approach allows for 3D chemical mapping of hydrated samples and thus provides direct evidence for the localization of metabolic and chemical processes in situ.

Keywords

chemical tomographyhydratedbacteriaSTXMsoft X-rays3D
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 20 , Issue 2 , April 2014 , pp. 531 - 536
Copyright
© Microscopy Society of America 2014 

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