Computed tomography of cryogenic cells

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Due to the short wavelengths of X-rays and low numerical aperture of the Fresnel zone plates used as X-ray objectives, the depth of field is several microns. Within the focal depth, imaging a thick specimen is to a good approximation equivalent to projecting the specimen absorption. Therefore, computed tomography based on a tilt series of X-ray microscopic images can be used to reconstruct the local linear absorption coefficient and image the three-dimensional specimen structure. To preserve the structural integrity of biological objects during image acquisition, microscopy is performed at cryogenic temperatures. Tomography based on X-ray microscopic images was applied to ... continued below

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7 pages

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Schneider, Gerd; Anderson, E.; Vogt, S.; Knochel, C.; Weiss, D.; LeGros, M. et al. August 30, 2001.

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Due to the short wavelengths of X-rays and low numerical aperture of the Fresnel zone plates used as X-ray objectives, the depth of field is several microns. Within the focal depth, imaging a thick specimen is to a good approximation equivalent to projecting the specimen absorption. Therefore, computed tomography based on a tilt series of X-ray microscopic images can be used to reconstruct the local linear absorption coefficient and image the three-dimensional specimen structure. To preserve the structural integrity of biological objects during image acquisition, microscopy is performed at cryogenic temperatures. Tomography based on X-ray microscopic images was applied to study the distribution of male specific lethal 1 (MSL-1), a nuclear protein involved in dosage compensation in Drosophila melanogaster, which ensures that males with single X chromosome have the same amount of most X-linked gene products as females with two X chromosomes. Tomographic reconstructions of X-ray microscopic images were used to compute the local three-dimensional linear absorption coefficient revealing the arrangement of internal structures of Drosophila melanogaster cells. Combined with labelling techniques, nanotomography is a new technique to study the 3D distribution of selected proteins inside whole cells. We want to improve this technique with respect to resolution and specimen preparation. The resolution in the reconstruction can be significantly improved by reducing the angular step size to collect more viewing angles, which requires an automated data acquisition. In addition, fast-freezing with liquid ethane instead of cryogenic He gas will be applied to improve the vitrification of the hydrated samples. We also plan to apply cryo X-ray nanotomography in order to study different types of cells and their nuclear protein distributions.

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7 pages

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OSTI as DE00793803

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  • International Conference on Vacuum Ultraviolet Radiation Physics VUV-XIII, Trieste (IT), 07/23/2001--07/27/2001

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  • Report No.: LBNL--49932
  • Grant Number: AC03-76SF00098
  • Office of Scientific & Technical Information Report Number: 793803
  • Archival Resource Key: ark:/67531/metadc738176

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Office of Scientific & Technical Information Technical Reports

Reports, articles and other documents harvested from the Office of Scientific and Technical Information.

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  • August 30, 2001

Added to The UNT Digital Library

  • Oct. 19, 2015, 7:39 p.m.

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  • April 4, 2016, 6:36 p.m.

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Schneider, Gerd; Anderson, E.; Vogt, S.; Knochel, C.; Weiss, D.; LeGros, M. et al. Computed tomography of cryogenic cells, article, August 30, 2001; California. (digital.library.unt.edu/ark:/67531/metadc738176/: accessed April 24, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.