1887

Abstract

Summary

Experiments were performed to determine whether a modern flow cytometer could be used to study bacterial populations in suspension, with particular reference to their morphological characteristics and their responses to antibiotics. The FACScan, a commercial benchtop flow cytometer fitted with an air-cooled laser, designed primarily for the study of eukaryotic peripheral blood mononuclear cells, yielded reproducible data relating to bacterial shape and internal architecture. It was sensitive enough to detect changes in bacterial morphology on entry into the growth cycle and after exposure to antibiotics. Antibiotic-induced morphological changes affecting subpopulations of bacteria were sufficiently specific to allow differentiation between antibiotics with different cell-wall enzyme targets. Simultaneously, the effect of such antibiotics on the integrity of the outer cell membrane of was assessed by measurement of the association of the nucleic acid-binding dye propidium iodide with the bacteria. These experiments demonstrated complex patterns of probable cell-wall leakage, related to the modes of action of the antibiotics. The FACScan is a useful and sensitive tool for the study of the morphology and physiology of bacterial populations in suspension, and is especially applicable to the study of antibiotic action.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-39-2-147
1993-08-01
2024-12-08
Loading full text...

Full text loading...

/deliver/fulltext/jmm/39/2/medmicro-39-2-147.html?itemId=/content/journal/jmm/10.1099/00222615-39-2-147&mimeType=html&fmt=ahah

References

  1. Tuomanen E, Schwartz J. Penicillin-binding protein 7 and its relationship to lysis of nongrowing Escherichia coli. J Bacteriol 1987; 169:4912–4915
    [Google Scholar]
  2. Brown MRW, Collier PJ, Gilbert P. Influence of growth rate on susceptibility to antimicrobial agents: modification of the cell envelope and batch and continuous culture studies. Antimicrob Agents Chemother 1990; 34:1623–1628
    [Google Scholar]
  3. Tuomanen E, Cozens R, Tosch W, Zak O, Tomasz A. The rate of killing of Escherichia coli by β-lactam antibiotics is strictly proportional to the rate of growth. J Gen Microbiol 1986; 132:1297–1304
    [Google Scholar]
  4. Phillips AP, Martin KL. Limitations of flow cytometry for the specific detection of bacteria in mixed populations. J Immunol Methods 1988; 106:109–117
    [Google Scholar]
  5. Mansour JD, Robson JA, Arndt CW, Schulte TH. Detection of Escherichia coli in blood using flow cytometry. Cytometry 1985; 6:186–190
    [Google Scholar]
  6. Donnelly CW, Baigent GJ. Method for flow cytometric detection of Listeria monocytogenes in milk. Appl Environ Microbiol 1986; 52:689–695
    [Google Scholar]
  7. Tyndall RL, Hand RE, Mann RC, Evans C, Jemigan R. Application of flow cytometry to detection and characterisation of Legionella spp. Appl Environ Microbiol 1985; 49:852–857
    [Google Scholar]
  8. Monfort P, Baleux B. Comparison of flow cytometry and epifluorescence microscopy for counting bacteria in aquatic ecosystems. Cytometry 1992; 13:188–192
    [Google Scholar]
  9. Cohen CY, Sahar E. Rapid flow cytometric bacterial detection and determination of susceptibility to amikacin in body fluids and exudates. J Clin Microbiol 1989; 27:1250–1256
    [Google Scholar]
  10. Steen HB, Skarstak, Boye E. Flow cytometry of bacteria: cell cycle kinetics and effects of antibiotics. Ann NY Acad Sci 1983; 468:329–338
    [Google Scholar]
  11. Steen HB. Flow cytometric studies of microorganisms. In: Melamed Lindmo Mendelsohn. (eds) Flow cytometry and sorting New York: Wiley-Liss Inc.; 1991605
    [Google Scholar]
  12. Lorian V, Amaral A. Effects of low concentrations of antibiotics. Antibiot Newslett 1990; 7:65–70
    [Google Scholar]
  13. Martinez OV, Gratzner HG, Malinin TI, Ingram M. The effect of some β-lactam antibiotics on Escherichia coli studied by flow cytometry. Cytometry 1982; 3:129–133
    [Google Scholar]
/content/journal/jmm/10.1099/00222615-39-2-147
Loading
/content/journal/jmm/10.1099/00222615-39-2-147
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error