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Proceedings Paper

Nanotomography of labeled cryogenic cells
Author(s): Gerd Schneider; Christian Knoechel; Stefan Vogt; Daniel Weiss; Erik H. Anderson
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Paper Abstract

By employing the natural absorption contrast of organic matter in water at 0.5 keV photon energy, X-ray microcopy has resolved 30 nm structures in animal cells. To protect the cells from radiation damage caused by x-rays, imaging of the samples was performed at cryogenic temperatures, which makes it possible to take multiple images of a single cell. Due to the small numerical aperture of zone plates, X-ray objectives have a depth of focus on the order of several microns. By treating the X-ray microscopic images as projections of the sample absorption, computed tomography (CT) can be performed. Since cryogenic biological samples are resistant to radiation damage, it is possible to reconstruct frozen-hydrated cells imaged with a full-field X-ray microscope. This approach is used to obtain three- dimensional information about the location of specific proteins in cells. To localize proteins in cells, immunolabelling with strongly X-ray absorbing nanoparticles was performed. With the new tomography apparatus developed for the X-ray microscope XM-1 installed at the ALS, we have performed tomography of immunolabelled frozen-hydrated cells to detect protein distributions in all three dimensions inside of cells. As a first example, the distribution of the nuclear protein, male specific lethal 1 (MSL-1) in the Drosophila melanogaster cell was studied.

Paper Details

Date Published: 7 January 2002
PDF: 10 pages
Proc. SPIE 4503, Developments in X-Ray Tomography III, (7 January 2002); doi: 10.1117/12.452839
Show Author Affiliations
Gerd Schneider, Lawrence Berkeley National Lab. (United States)
Christian Knoechel, Univ. Goettingen (Germany)
Stefan Vogt, Univ. Goettingen (Germany)
Daniel Weiss, Univ. Goettingen (Germany)
Erik H. Anderson, Lawrence Berkeley National Lab. (United States)

Published in SPIE Proceedings Vol. 4503:
Developments in X-Ray Tomography III
Ulrich Bonse, Editor(s)

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