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

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

doi:10.1117/12.138753

13 January 1993 Application of x rays to the analysis of DNA packaging in mammalian sperm

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

Author Affiliations +

Proceedings Volume 1741, Soft X-Ray Microscopy; (1993) https://doi.org/10.1117/12.138753
Event: San Diego '92, 1992, San Diego, CA, United States

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 lense. 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 X 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.

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Rod Balhorn, Michele Corzett, Michael J. Allen, Catherine S. Lee, Troy W. Barbee Jr., Jeffrey A. Koch, Brian J. MacGowan, Dennis L. Matthews, Stanley Mrowka, James E. Trebes, Ian McNulty, Luiz Barroca Da Silva, Joe W. Gray, Erik H. Anderson, D. Kern, and David T. Attwood Jr. "Application of x rays to the analysis of DNA packaging in mammalian sperm", Proc. SPIE 1741, Soft X-Ray Microscopy, (13 January 1993); https://doi.org/10.1117/12.138753

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