Application of x rays to the analysis of DNA packaging in mammalian sperm

Rod Balhorn, Michele Corzett, Michael J. Allen, Catherine Lee, Troy W. Barbee, Jeffrey A. Koch, Brian J. MacGowan, Dennis L Matthews, Stan J. Mrowka, James E. Trebes, Ian McNulty, Luiz B. Da Silva, Joe W. Gray, Erik H. Anderson, D. Kern, David T. Attwood

Research output: Chapter in Book/Report/Conference proceedingConference contribution

7 Scopus citations

Abstract

X-ray microscopy has been performed on unlabeled and gold labeled rat sperm nuclei using a tantalum x-ray laser (44.83 angstrom) and an x-ray zone plate lens. Transmission images of nuclei labeled with gold using an antibody to protamine 1 show large clusters of gold as well as individual 400 angstrom gold particles coating the surface of the chromatin. Images of the same nuclei obtained with a scanning x-ray microscope demonstrate that the initial exposure of the sperm nucleus to a photon intensity of 3.0 × 1014 W/cm2 for a duration of 500 ps did not destroy or grossly distort their structure. Other nuclei labeled with an antibody to protamine 2 were partially decondensed during the labeling process and contained large vacuole-like regions. Images were also obtained of unlabeled sperm nuclei. Data obtained from one unlabeled nucleus imaged with both the atomic force and x-ray laser microscopes was used to determine the thickness of the chromatin and estimate the amount of water that must be associated with the DNA-protamine complex in air-dried nuclei. These results suggest that even air-dried sperm nuclei are extensively hydrated and that tightly bound water may comprise as much as one-third of the volume of the dried sperm nucleus.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
Place of PublicationBellingham, WA, United States
PublisherPubl by Int Soc for Optical Engineering
Pages374-385
Number of pages12
Volume1741
ISBN (Print)0819409146
StatePublished - 1993
Externally publishedYes
EventSoft X-Ray Microscopy - San Diego, CA, USA
Duration: Jul 19 1992Jul 21 1992

Other

OtherSoft X-Ray Microscopy
CitySan Diego, CA, USA
Period7/19/927/21/92

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

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