Quantitative 3-D imaging of eukaryotic cells using soft X-ray tomography

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TitleQuantitative 3-D imaging of eukaryotic cells using soft X-ray tomography
Publication TypeJournal Article
Year of Publication2008
AuthorsParkinson DY, McDermott G, Etkin LD, Le Gros MA, Larabell CA
Journal TitleJournal of Structural Biology
Volume162
Pages380-386
Journal DateJun
ISBN Number1047-8477
Accession NumberISI:000256684800003
Keywordsapoptosis, cellular imaging, division, Electron tomography, fission yeast, Microscopy, microtubules, mitochondrial dynamics, organelle characterization, reconstruction, resolution, schizosaccharomyces pombe, schizosaccharomyces-pombe, segmentation, soft x-ray tomography, yeast
Abstract

Imaging has long been one of the principal techniques used in biological and biomedical research. Indeed, the field of cell biology grew out of the first electron microscopy images of organelles in a cell. Since this landmark event, much work has been carried out to image and classify the organelles in eukaryotic cells using electron microscopy. Fluorescently labeled organelles can now be tracked in live cells, and recently, powerful light microscope techniques have pushed the limit of optical resolution to image single molecules. In this paper, we describe the use of soft X-ray tomography, a new tool for quantitative imaging of organelle structure and distribution in whole, fully hydrated eukaryotic Schizosaccharomyces pombe cells. In addition to imaging intact cells, soft X-ray tomography has the advantage of not requiring the use of any staining or fixation protocols-cells are simply transferred from their growth environment to a sample holder and immediately cryofixed. In this way the cells can be imaged in a near native state. Soft X-ray tomography is also capable of imaging relatively large numbers of cells in a short period of time, and is therefore a technique that has the potential to produce information on organelle morphology from statistically significant numbers of cells. (c) 2008 Elsevier Inc. All rights reserved.

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