COMPUTED TOMOGRAPHY OF CRYOGENIC CELLS

G. Schneider, E. Anderson, S. Vogt, C. Knöchel, D. Weiss, M. Legros and C. Larabell
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G. Schneider: Center for X-ray Optics, Lawrence Berkeley National Laboratory, One Cyclotron Road MS 2-400, Berkeley, CA 94720, USA
E. Anderson: Center for X-ray Optics, Lawrence Berkeley National Laboratory, One Cyclotron Road MS 2-400, Berkeley, CA 94720, USA
S. Vogt: Institut für Röntgenphysik, Universität Göttingen, Geiststraße. 11, D-37073 Göttingen, Germany
C. Knöchel: Institut für Röntgenphysik, Universität Göttingen, Geiststraße. 11, D-37073 Göttingen, Germany
D. Weiss: Institut für Röntgenphysik, Universität Göttingen, Geiststraße. 11, D-37073 Göttingen, Germany
M. Legros: Life Sciences, Lawrence Berkeley National Laboratory, One Cyclotron Road MS 6-2100, Berkeley, CA 94720, USA
C. Larabell: Life Sciences, Lawrence Berkeley National Laboratory, One Cyclotron Road MS 6-2100, Berkeley, CA 94720, USA

Surface Review and Letters (SRL), 2002, vol. 09, issue 01, 177-183

Abstract: Soft X-ray microscopy has resolved 30 nm structures in biological cells. To protect the cells from radiation damage caused by X-rays, imaging of the samples has to be 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, immunolabeling with strongly X-ray absorbing nanoparticles was performed. With the new tomography setup developed for the X-ray microscope XM-1 installed at the ALS, we have performed tomography of immunolabeled frozen-hydrated cells to detect protein distributions inside of cells. As a first example, the distribution of the nuclear protein male-specific lethal 1 (MSL-1) in theDrosophila melanogastercell was studied.

Date: 2002
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DOI: 10.1142/S0218625X02001914

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