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A simple colorimetric method for detecting cell viability in cultures of eukaryotic microorganisms

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Abstract

The dye methylene blue can be taken up by dead or severely damaged cells, but not by living cells. Based on this fact, a method was devised which permits quantitative determinations of injured cells in populations of microorganisms such asSaccharomyces cerevisiae, Rhodotorula glutinis, andEuglena gracilis. The percentage of damaged cells was determined by measuring, at 664 nm, the optical density of cell suspensions pretreated with 0.15 mM methylene blue for 6 min, a condition that does not affect cell integrity as determined by oxygen consumption and release of potassium ions. This technique is faster and simpler than the classical dye-exclusion and plate-counting methods.

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Literature Cited

  1. Bonora, A., Vannini, G. L., Mares, D. 1980. Further observations about the effects of Phosfon D onRhodotorula glutinis. Congresso Nazionale Società Botanica Italiana, Trieste 1–4 ottobre, p. 69–70.

  2. Collins, C. H. 1964. Microbiological methods. London: Butterworths Ltd.

    Google Scholar 

  3. Dominguez, A., Villanueva, J. R., Sentandreu, R. 1978. Biosynthesis ofSaccharomyces cerevisiae glycoproteins: Nature of some participating glycolipids. Antonie van Leeuwenhoek Journal of Microbiology and Serology44:183–192.

    Google Scholar 

  4. Fasulo, M. P., Vannini, G. L., Bruni, A., Mares, D. 1976. Physiological and ultrastructural changes induced in lightgrown cells ofEuglena gracilis by myomycin treatment. Zeitschrift für Pflanzenphysiologie80:407–416.

    Google Scholar 

  5. Ganter, P., Jolles, G. 1970. Histochimie, vol. 2. Paris: Gauthier-Villars.

    Google Scholar 

  6. Haugen, G. R., Hardwick, E. R. 1963. Ionic association in aqueous solutions of thionine. Journal of Physical Chemistry67:725–731.

    Google Scholar 

  7. Honsell, E. 1965. I coloranti vitali e i problemi della loro permeazione ed accumulo nella cellula vegetale. Giornale Botanico Italiano72:287–302.

    Google Scholar 

  8. Kuypers, G. A. J., Roomans, G. M. 1979. Mercury-induced loss of K+ from yeast cells investigated by electron probe X-ray microanalysis. Journal of General Microbiology115:13–18.

    PubMed  Google Scholar 

  9. Macfarlane, M. G. 1936. Phosphorylation in living yeast. Biochemical Journal30:1369–1379.

    Google Scholar 

  10. Michaelis, L. 1950. Reversible polymerization and molecular aggregation. Journal of Physical Chemistry54:1–17.

    Google Scholar 

  11. Novelli, A. 1962. Amethyst violet as a stain for distinguishing cells with a damaged membrane from normal cells. Experientia18:295–296.

    PubMed  Google Scholar 

  12. Parker, R. C. 1961. Methods of tissue culture, 3rd edition. New York: Paul B. Hoeber, Inc.

    Google Scholar 

  13. Passow, H., Rothstein, A., Loewenstein, B. 1959. An all-or-none response in the release of potassium by yeast cells treated with methylene blue and other basic redox dyes. Journal of General Physiology43:97–107.

    PubMed  Google Scholar 

  14. Rogentine, G. N., Jr., Plocinik, B. A. 1967. Application of the51Cr cytotoxicity technique to the analysis of human lymphocyte isoantigens. Transplantation5:1323–1333.

    PubMed  Google Scholar 

  15. Rotman, B., Papermaster, B. W. (1966). Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters. Proceedings of the National Academy of Sciences of the United States of America55:134–141.

    PubMed  Google Scholar 

  16. Scharff, T. G., Maupin, W. C. 1960. Correlation of the metabolic effects of benzalkonium chloride with its membrane effects in yeast. Biochemical Pharmacology5:79–86.

    PubMed  Google Scholar 

  17. Schrek, R. 1936. A method for counting the viable cells in normal and in malignant cell suspensions. American Journal of Cancer28:389–392.

    Google Scholar 

  18. Shirai, M., Nagatsuka, T., Tanaka, M. 1977. Interaction between dyes and polyelectrolytes. 2. Structural effect of polyanions on the methylene blue binding. Makromolekulare Chemie178:37–46.

    Google Scholar 

  19. Shirai, M., Nagaoka, Y., Tanaka, M. 1977. Interaction between dyes and polyelectrolytes. 5. Effect of organic and inorganic salts on metachromasy of methylene blue. Makromolekulare Chemie178:1633–1639.

    Google Scholar 

  20. Vannini, G. L., Dall'Olio, G., Donini, A. 1978. Fungal toxicity of Phosfon D and its reversion by lipids. Antimicrobial Agents and Chemotherapy13:699–700.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Institute of Pharmacology, University of Ferrara, Via Fossato di Mortara 64, I-44100, Ferrara, Italy

    Angelo Bonora

  2. Institute of Botany, University of Ferrara, Corso Porta Mare 2, I-44100, Ferrara, Italy

    Donatella Mares

Authors
  1. Angelo Bonora
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  2. Donatella Mares
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Bonora, A., Mares, D. A simple colorimetric method for detecting cell viability in cultures of eukaryotic microorganisms. Current Microbiology 7, 217–221 (1982). https://doi.org/10.1007/BF01568802

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  • DOI: https://doi.org/10.1007/BF01568802

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