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Ultrastructural Analysis of Bacteria–Host Cell Interactions

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Bacterial Pathogenesis

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 431))

Summary

Electron microscopy of bacterial pathogens and interactions between bacteria and host cells and tissues provides valuable insights into structural and molecular properties and processes involved in pathogenesis. Applications for electron microscopy in bacterial pathogenesis range from discovering etiologic agents and following chronological events during infections by conventional examination of clinical samples to assessing molecular host–cell responses to infection and in situ interactions between receptors and ligands using specific immune-labeling techniques. This chapter focuses on techniques for preparing samples of bacteria and host cells for conventional transmission (TEM) and scanning electron microscopy (SEM) and use of luminescent nanocrystals or “quantum dots” as specific probes for correlative light and electron microscopy. Conventional TEM and SEM are well established tools for high resolution examination of structural effects and chronological events associated with bacterial infections. The recent development of quantum dots as physiological and immunological probes in biology has provided a powerful technique for bridging fluorescent analyses of fixed and live material with preparation and examination by TEM and SEM.

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References

  1. Hawkes, P. W., and Spence, J. C. H. (eds) (2006) Science of Microscopy. Springer, New York, NY.

    Google Scholar 

  2. Hayat, M. A. (ed.) (2000) Principles and Techniques of Electron Microscopy: Biological Applications. Cambridge University Press, Cambridge, UK.

    Google Scholar 

  3. Flegler, S. L., Heckman, J. W., and Klomparens, K. L. (eds) (1993) Scanning and Transmission Electron Microscopy: An Introduction. Oxford University Press, New York, NY.

    Google Scholar 

  4. Giberson, R. T., and Demaree, R. S. (eds) (2001) Microwave Techniques and Protocols. Humana Press, Totowa, NJ.

    Google Scholar 

  5. Ubero-Pascal, N., Fortuno, J. M., and de Los Angeles Puig, M. (2005) New application of air-drying techniques for studying Ephemoptera and Plecoptera eggs by scanning electron microscopy. Micros. Res. Tech. 68, 264–271.

    Article  Google Scholar 

  6. McIntosh, J. R. (2001) Electron microscopy of cells: a new beginning for a new century. J. Cell Biol. 153, F25–F32.

    Article  CAS  PubMed  Google Scholar 

  7. Giepmans, B. N., Deerinck, T. J., Smarr, B. L., Jones, Y. Z., and Ellisman, M. H. (2005) Correlated light and electron microscopy imaging of multiple endogenous proteins using quantum dots. Nat. Methods 2, 742–749.

    Article  Google Scholar 

  8. van Dam, G. J., Bogitsh, B. J., Fransen, J. A., Kornelis, D., van Zeyl, R. J., and Deelder, A. M. (1991) Application of the FITC-anti-FITC-gold system to ultrastructural localization of antigens. J. Histochem. Cytochem. 39, 1725–1728.

    PubMed  Google Scholar 

  9. Plowman, S. J., Muncke, C., Paxton, R. G., and Hancock, J. F. (2005) H-ras, K-ras, and inner plasma membrane raft proteins operate in nano clusters with differential dependence on the actin cytoskeleton. Proc. Nat. Acad. Sci. U.S.A. 102, 15500–15550.

    Article  CAS  Google Scholar 

  10. Gaietta, G., Deerinck, T. J., Adams S. R., Bouwer, J., Tour, O., Laird, D. W., Sosinsky, G. E., Tsien, R. Y., and Ellisman, M. H. (2002) Multicolor and electron microscopic imaging of connexin trafficking. Science 296, 503–507.

    Article  CAS  PubMed  Google Scholar 

  11. Rodriguez-Viejo, J., et al. (1997) Cathodoluminescence and photoluminescence of highly luminescent CdSe/ZnS quantum dot composites. Appl. Phys. Lett. 70, 2132–3134.

    Article  CAS  Google Scholar 

  12. Rodriguez-Viejo, J., et al. (2000) Evidence of photo-and electrodarkening of (CdSe)ZnS quantum dot composites. J. Appl. Phys. 87, 8526–8534.

    Article  CAS  Google Scholar 

  13. Leon, R., Nadeau, J., Evans, K., Paskove, T., and Monemar, B. (2004) Electron irradiation effects on nanocrystal quantum dots used in bio-sensing applications. IEEE Trans. Nucl. Sci. 51, 3186–3192.

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases.

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© 2008 Humana Press, a part of Springer Science+Business Media, LLC

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Dorward, D.W. (2008). Ultrastructural Analysis of Bacteria–Host Cell Interactions. In: DeLeo, F.R., Otto, M. (eds) Bacterial Pathogenesis. Methods in Molecular Biology™, vol 431. Humana Press. https://doi.org/10.1007/978-1-60327-032-8_14

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  • DOI: https://doi.org/10.1007/978-1-60327-032-8_14

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-740-2

  • Online ISBN: 978-1-60327-032-8

  • eBook Packages: Springer Protocols

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