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. 1997 Apr;72(4):1521–1526. doi: 10.1016/S0006-3495(97)78800-3

X-ray micrography and imaging of Escherichia coli cell shape using laser plasma pulsed point x-ray sources.

J M Rajyaguru 1, M Kado 1, M C Richardson 1, M J Muszynski 1
PMCID: PMC1184348  PMID: 9083658

Abstract

High-resolution x-ray microscopy is a relatively new technique and is performed mostly at a few large synchrotron x-ray sources that use exposure times of seconds. We utilized a bench-top source of single-shot laser (ns) plasma to generate x-rays similar to synchrotron facilities. A 5 microlitres suspension of Escherichia coli ATCC 25922 in 0.9% phosphate buffered saline was placed on polymethylmethyacrylate coated photoresist, covered with a thin (100 nm) SiN window and positioned in a vacuum chamber close to the x-ray source. The emission spectrum was tuned for optimal absorption by carbon-rich material. Atomic force microscope scans provided a surface and topographical image of differential x-ray absorption corresponding to specimen properties. By using this technique we observed a distinct layer around whole cells, possibly representing the Gram-negative envelope, darker stained areas inside the cell corresponding to chromosomal DNA as seen by thin section electron microscopy, and dent(s) midway through one cell, and 1/3- and 2/3-lengths in another cell, possibly representing one or more division septa. This quick and high resolution with depth-of-field microscopy technique is unmatched to image live hydrated ultrastructure, and has much potential for application in the study of fragile biological specimens.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arakawa H., Umemura K., Ikai A. Protein images obtained by STM, AFM and TEM. Nature. 1992 Jul 9;358(6382):171–173. doi: 10.1038/358171a0. [DOI] [PubMed] [Google Scholar]
  2. Edstrom R. D., Meinke M. H., Yang X. R., Yang R., Elings V., Evans D. F. Direct visualization of phosphorylase-phosphorylase kinase complexes by scanning tunneling and atomic force microscopy. Biophys J. 1990 Dec;58(6):1437–1448. doi: 10.1016/S0006-3495(90)82489-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Feder R., Banton V., Sayre D., Costa J., Baldini M., Kim B. Direct imaging of live human platelets by flash x-ray microscopy. Science. 1985 Jan 4;227(4682):63–64. doi: 10.1126/science.3964958. [DOI] [PubMed] [Google Scholar]
  4. Ford T. W., Stead A. D., Cotton R. A. Soft X-ray contact microscopy of biological materials. Electron Microsc Rev. 1991;4(2):269–292. doi: 10.1016/0892-0354(91)90006-x. [DOI] [PubMed] [Google Scholar]
  5. Hansma H. G., Bezanilla M., Zenhausern F., Adrian M., Sinsheimer R. L. Atomic force microscopy of DNA in aqueous solutions. Nucleic Acids Res. 1993 Feb 11;21(3):505–512. doi: 10.1093/nar/21.3.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hansma H. G., Weisenhorn A. L., Edmundson A. B., Gaub H. E., Hansma P. K. Atomic force microscopy: seeing molecules of lipid and immunoglobulin. Clin Chem. 1991 Sep;37(9):1497–1501. [PubMed] [Google Scholar]
  7. Hoh J. H., Hansma P. K. Atomic force microscopy for high-resolution imaging in cell biology. Trends Cell Biol. 1992 Jul;2(7):208–213. doi: 10.1016/0962-8924(92)90248-l. [DOI] [PubMed] [Google Scholar]
  8. Ill C. R., Keivens V. M., Hale J. E., Nakamura K. K., Jue R. A., Cheng S., Melcher E. D., Drake B., Smith M. C. A COOH-terminal peptide confers regiospecific orientation and facilitates atomic force microscopy of an IgG1. Biophys J. 1993 Mar;64(3):919–924. doi: 10.1016/S0006-3495(93)81452-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ohnesorge F., Binnig G. True atomic resolution by atomic force microscopy through repulsive and attractive forces. Science. 1993 Jun 4;260(5113):1451–1456. doi: 10.1126/science.260.5113.1451. [DOI] [PubMed] [Google Scholar]
  10. Panessa-Warren B. J., Warren J. B. Determining biological fine structure by differential absorption of soft x-rays. Ann N Y Acad Sci. 1980;342:350–367. doi: 10.1111/j.1749-6632.1980.tb47233.x. [DOI] [PubMed] [Google Scholar]
  11. Rees W. A., Keller R. W., Vesenka J. P., Yang G., Bustamante C. Evidence of DNA bending in transcription complexes imaged by scanning force microscopy. Science. 1993 Jun 11;260(5114):1646–1649. doi: 10.1126/science.8503010. [DOI] [PubMed] [Google Scholar]
  12. Schabert F. A., Henn C., Engel A. Native Escherichia coli OmpF porin surfaces probed by atomic force microscopy. Science. 1995 Apr 7;268(5207):92–94. doi: 10.1126/science.7701347. [DOI] [PubMed] [Google Scholar]
  13. Wigren R., Elwing H., Erlandsson R., Welin S., Lundström I. Structure of adsorbed fibrinogen obtained by scanning force microscopy. FEBS Lett. 1991 Mar 25;280(2):225–228. doi: 10.1016/0014-5793(91)80298-h. [DOI] [PubMed] [Google Scholar]
  14. Yang J., Takeyasu K., Shao Z. Atomic force microscopy of DNA molecules. FEBS Lett. 1992 Apr 20;301(2):173–176. doi: 10.1016/0014-5793(92)81241-d. [DOI] [PubMed] [Google Scholar]
  15. Zasadzinski J. A., Helm C. A., Longo M. L., Weisenhorn A. L., Gould S. A., Hansma P. K. Atomic force microscopy of hydrated phosphatidylethanolamine bilayers. Biophys J. 1991 Mar;59(3):755–760. doi: 10.1016/S0006-3495(91)82288-3. [DOI] [PMC free article] [PubMed] [Google Scholar]

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