Nanotomography of labeled cryogenic cells

Gerd Schneider; Christian Knoechel; Stefan Vogt; Daniel Weiss; Erik H. Anderson

doi:10.1117/12.452839

7 January 2002 Nanotomography of labeled cryogenic cells

Gerd Schneider, Christian Knoechel, Stefan Vogt, Daniel Weiss, Erik H. Anderson

Proceedings Volume 4503, Developments in X-Ray Tomography III; (2002) https://doi.org/10.1117/12.452839
Event: International Symposium on Optical Science and Technology, 2001, San Diego, CA, United States

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.

Citation Download Citation

Gerd Schneider, Christian Knoechel, Stefan Vogt, Daniel Weiss, and Erik H. Anderson "Nanotomography of labeled cryogenic cells", Proc. SPIE 4503, Developments in X-Ray Tomography III, (7 January 2002); https://doi.org/10.1117/12.452839

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