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

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 proceeding › Conference contribution

8 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 language	English (US)
Title of host publication	Proceedings of SPIE - The International Society for Optical Engineering
Publisher	Publ by Int Soc for Optical Engineering
Pages	374-385
Number of pages	12
ISBN (Print)	0819409146
State	Published - 1993
Externally published	Yes
Event	Soft X-Ray Microscopy - San Diego, CA, USA Duration: Jul 19 1992 → Jul 21 1992

Publication series

Name	Proceedings of SPIE - The International Society for Optical Engineering
Volume	1741
ISSN (Print)	0277-786X

Other

Other	Soft X-Ray Microscopy
City	San Diego, CA, USA
Period	7/19/92 → 7/21/92

ASJC Scopus subject areas

Electronic, Optical and Magnetic Materials
Condensed Matter Physics
Computer Science Applications
Applied Mathematics
Electrical and Electronic Engineering

Cite this

Balhorn, R., Corzett, M., Allen, M. J., Lee, C., Barbee, T. W., Koch, J. A., MacGowan, B. J., Matthews, D. L., Mrowka, S. J., Trebes, J. E., McNulty, I., Da Silva, L. B., Gray, J. W., Anderson, E. H., Kern, D., & Attwood, D. T. (1993). Application of x rays to the analysis of DNA packaging in mammalian sperm. In Proceedings of SPIE - The International Society for Optical Engineering (pp. 374-385). (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 1741). Publ by Int Soc for Optical Engineering.

Application of x rays to the analysis of DNA packaging in mammalian sperm. / Balhorn, Rod; Corzett, Michele; Allen, Michael J. et al.
Proceedings of SPIE - The International Society for Optical Engineering. Publ by Int Soc for Optical Engineering, 1993. p. 374-385 (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 1741).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Balhorn, R, Corzett, M, Allen, MJ, Lee, C, Barbee, TW, Koch, JA, MacGowan, BJ, Matthews, DL, Mrowka, SJ, Trebes, JE, McNulty, I, Da Silva, LB, Gray, JW, Anderson, EH, Kern, D & Attwood, DT 1993, Application of x rays to the analysis of DNA packaging in mammalian sperm. in Proceedings of SPIE - The International Society for Optical Engineering. Proceedings of SPIE - The International Society for Optical Engineering, vol. 1741, Publ by Int Soc for Optical Engineering, pp. 374-385, Soft X-Ray Microscopy, San Diego, CA, USA, 7/19/92.

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title = "Application of x rays to the analysis of DNA packaging in mammalian sperm",

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.",

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

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pages = "374--385",

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AU - Balhorn, Rod

AU - Corzett, Michele

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AU - Lee, Catherine

AU - Barbee, Troy W.

AU - Koch, Jeffrey A.

AU - MacGowan, Brian J.

AU - Matthews, Dennis L.

AU - Mrowka, Stan J.

AU - Trebes, James E.

AU - McNulty, Ian

AU - Da Silva, Luiz B.

AU - Gray, Joe W.

AU - Anderson, Erik H.

AU - Kern, D.

AU - Attwood, David T.

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N2 - 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.

AB - 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.

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

Abstract

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Other

ASJC Scopus subject areas

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