3D Ultrastructural Organization of Whole Chlamydomonas reinhardtii Cells Studied by Nanoscale Soft X-Ray Tomography |
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| Authors: | Eric Hummel Peter Guttmann Stephan Werner Basel Tarek Gerd Schneider Michael Kunz Achilleas S Frangakis Benedikt Westermann |
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| Institution: | 1. Institut für Zellbiologie, Universität Bayreuth, Bayreuth, Germany.; 2. Helmholtz-Zentrum für Materialien und Energie GmbH, Institute for Soft Matter and Functional Materials, Berlin, Germany.; 3. Frankfurt Institute for Molecular Life Sciences and Institute of Biophysics, Goethe University Frankfurt, Frankfurt am Main, Germany.; University of Hyderabad, India, |
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| Abstract: | The complex architecture of their structural elements and compartments is a hallmark of eukaryotic cells. The creation of high resolution models of whole cells has been limited by the relatively low resolution of conventional light microscopes and the requirement for ultrathin sections in transmission electron microscopy. We used soft x-ray tomography to study the 3D ultrastructural organization of whole cells of the unicellular green alga Chlamydomonas reinhardtii at unprecedented spatial resolution. Intact frozen hydrated cells were imaged using the natural x-ray absorption contrast of the sample without any staining. We applied different fiducial-based and fiducial-less alignment procedures for the 3D reconstructions. The reconstructed 3D volumes of the cells show features down to 30 nm in size. The whole cell tomograms reveal ultrastructural details such as nuclear envelope membranes, thylakoids, basal apparatus, and flagellar microtubule doublets. In addition, the x-ray tomograms provide quantitative data from the cell architecture. Therefore, nanoscale soft x-ray tomography is a new valuable tool for numerous qualitative and quantitative applications in plant cell biology. |
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