Abstract
Antibiotics are particular type of drugs that are able to interfere in different ways to the metabolic pathways of bacteria. This causes also morphostructural alterations of cell wall and surface appendages (flagella, fimbriae or pili, and filaments).Atomic force microscopy (AFM) is extremely useful for analyzing the three-dimensional structure of the surface of biological specimens, particularly bacteria. A step-by-step AFM methodology to be applied to different type of bacteria is reported and visual examples of the action of antibiotics are shown. Although scanning electron microscopy is still frequently used, the introduction of the AFM technique offers substantial benefits in real quantitative data acquisition in three dimensions, minimal sample preparation times, flexibility in ambient operating conditions (i.e., no vacuum is necessary), and effective three-dimensional magnification at submicron level.
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References
Heckl, W.M. (1995) Scanning the thread of life, in The human genome (Fisher, E.P., Klose, S.), R. Piper GmbH & Co. KG, Munchen, pp. 99–146.
Braga, P.C. and Ricci, D.(1998) Atomic Force Microscopy: Application to investigation of Escherichia coli morphology before and after exposure to cefodizime. Antimicrob. Agents Chemother. 42, 18–22.
Strausser, Y.E. and Heaton, M.G. (1994) Scanning probe microscopy technology and recent innovations. American Laboratory, May 1–7.
Binning, G., Quate, C.F. and Gerber, C. (1986) Atomic force microscope. Whys. Rev. Lett. 12, 930–933.
Binnig, G.and Rohrer, H. (1982) Scanning tunnelling microscopy. Helv. Phys. Acta. 55, 726–735.
Mc Donnel, L. and Phelan, M. (1998) The scanned cantilever AFM: a versatile tool for industrial application. Microscopy and Analysis (European ed.) 52, 25–27.
Ratneshwar, L. and Scott, A.J. (1994) Biological applications of atomic force microscopy. Am. J. Physiol. 266, C1–C21.
Campbell, P.A., Gordon, R. and Walmsley, D.G. (1998) Active surface modification by scanning tunnelling microscopy. Microscopy and Analysis (European ed.) 56, 25–27.
Lorian, V. (1986) Effect of low antibiotic concentrations on bacteria: effects on ultrastructure, their virulence and susceptibility to immunodefenses, in Antibiotics in Laboratory Medicine (Lorian, V.), The Williams & Wilkins Co., Baltimore, pp. 596668.
Lorian, V., Atkinson, B., Walushacka, A. and Kim, Y. (1982) Ultrastructure, in vitro and in vivo, of staphylococci exposed to antibiotics. Curr. Microbiol., 7, 301–304.
Braga, P.C. and Ricci, D. (2000) Atomic Force Microscopy: detection of rokitamycin induced morphological alterations in Helicobacter pylori. Chemotherapy, 46, 15–22.
Braga, P.C. and Ricci, D. (2002) Differences in the susceptibility of Streptococcus pyogenes to rokitamycin and erythromycin revealed by morphostructural atomic force microscopy investigation. J. Antimicrob. Chemother. 50, 475–460.
Braga, P.C., Ricci, D., Dal Sasso, M. and Thorne, G. (2002) Bacillus cereus morphostructural damage by daptomycin: atomic force microscopy investigation. J. Chemother. 14, 336–341.
Nagao, E. and Dvorak, J.A. (1999) Developing the atomic force microscope for studies of living cells. Intern. Lab., January, 21–23.
Schaus, S.S. and Henderson, E.R. (1997) Cell viability and probe-cell membrane interactions of XR1 glial cells imaged by atomic force microscopy. Biophys. J., 73, 1205–1214.
Howland, R. and Benatar, L. (1997) A practical guide to scanning probe microscopy. Park Scientific Instrument Ed., pp. 1–73.
Acknowledgments
We would like to thank M. Dal Sasso for preparing the bacterial samples. This study was partially supported by a grant from MIUR (60%).
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Braga, P.C., Ricci, D. (2011). Imaging Bacterial Shape, Surface, and Appendages Before and After Treatment with Antibiotics. In: Braga, P., Ricci, D. (eds) Atomic Force Microscopy in Biomedical Research. Methods in Molecular Biology, vol 736. Humana Press. https://doi.org/10.1007/978-1-61779-105-5_23
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DOI: https://doi.org/10.1007/978-1-61779-105-5_23
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