Abstract
The practical use of flow cytometry is shown in several microbial assays. Recent technical improvements in the optics and electronics of flow cytometric systems as well as in staining techniques permit the measurements of minute cellular components such as the cellular DNA and the protein content of bacteria, algae, moulds and yeasts. Single cell ingredients can be measured by this assay according to their specific stainability. The cell DNA was stained by propidium iodide while the cell protein was fluorochromed by fluorescein-iso-thiocyanate. The DNA synthesis of Saccharomyces cerevisiae and Saccharomyces pastorianus runs discontinuously while the protein content increases continuously during the vegetative growth. The different stages of DNA synthesis of yeast cells can be divided into two ‘gap’ phases, a synthesis and a mitosis period, corresponding to Howard and Pelc's model of DNA synthesis. Living and dead cells can be counted differentially after staining with Erythrosine B. The red fluorescence of the chlorophyll in algae can readily be used to determine the chlorophyll content of these cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Berkhan, E. 1972. DNS-Messung von Zellen aus Vaginalabstrichen. — Ärztl. Lab. 18: 77–79.
Darzynkiewicz, Z., Traganos, F., Sharpless, T. and Melamed, M. D. 1975. Conformation of RNA in situ as studied by acridine orange staining and automated cytofluorometry. — Exptl. Cell Res. 95: 143–153.
van Dilla, M. A., Trujillo, T. T., Mullaney, P. F. and Coulter, J. R. 1969. Cellmicrofluorometry: A method for rapid fluorescence measurement. — Science 163: 1213–1214.
van Dilla, M. A., Gray, J. W., Carrano, A. V., Minkler, J. L. and Steinmetz, L. L. 1976. New directions for flow cytometry: Chromosome analysis. pp. 57–62. In W. Göhde, J. Schumann, Th. Bücher (eds), 2nd International Symposium Pulse-Cytophotometry. — European Press, Ghent, Belgium.
Dittrich, W. und Göhde, W. 1969. Impulscytophotometrie bei Einzelzellen in Suspension. — Z. Naturforschg. 24b: 360–361.
Duffus, J. H. 1971. The cell cycle in yeast. — J. Inst. Brew. 77: 500–508.
Göhde, W. und Dittrich, W. 1970. Simultane Impulscytophotometrie des DNS-und Proteingehaltes von Tumorzellen. — Z. Anal. Chem. 252: 328–330.
Gurr, E. 1971. (ed.). Anionic dyes, wholly acid. p. 197–198. In Synthetic dyes in biology, medicine and chemistry. — Academic Press, London & New York.
Halvorson, H. O., Carter, B. L. A. and Tauro, P. 1971. Synthesis of enzymes during the cell cycle. — Adv. Microbiol. Physiol. 6: 47–106.
Hartwell, L. H. 1974. Saccharomyces cerevisiae cell cycle. — Bacteriol. Rev. 38: 164–198.
Howard, A. and Pelc, S. R. 1953. Synthesis of deoxyribonucleic acid in normal and irradiated cells and its relation to chromosome breakage. — Heredity 6: (Suppl.); 261–273.
Hutter, K.-J., Göhde, W. und Emeis, C.-C. 1975. Untersuchungen über die DNS-, RNS- und Proteinsynthese ausgewählter Mikroorganismenpopulationen mit Hilfe der Zytophotometrie und der Impulscytophotometrie. I. Mitteilung: Methodische Untersuchungen über geeignete Färbeverfahren. — Chem. Mikrobiol. Technol. Lebensm. 4: 29–32.
Hutter, K.-J. and Eipel, H. E. 1978a. DNA determination of yeast by flow cytometry. — FEMS Microbiol. Lett. 3: 35–38.
Hutter, K.-J. and Eipel, H. E. 1978b. Protein distribution in population of baker's yeast. — Europ. J. Appl. Microbiol. Biotechnol. 5: 203–206.
Hutter, K.-J., Eipel, H. E. and Hettwer, H. 1978a. Rapid determination of the purity of yeast cultures by flow cytometry. — Europ. J. Appl. Microbiol. Biotechnol. 5: 109–112.
Hutter, K.-J., Görtz, Th., Oldiges, H. und Eipel, H. E. 1978b. Durchflußzytophotometrische Bestimmung des DNS-Gehaltes von Nectria coccinea Pers. ex Fr. bei Fungizideinwirkung. —Chemosphere 1: 51–58.
Kamentsky, L. A., Melamed, M. R. and Derman, H. 1965. Spectrophotometer: New instrument for ultrarapid cell analysis. — Science 150: 630–631.
Kaplow, L. S., Dauber, H. and Lerner, E. 1976. Assessment of monocyte esterase activity by flow cytophotometry. — J. Histochem. Cytochem. 24: 363.
Kessler, E. und Czygan, F.-C. 1970. Physiologische und biochemische Beiträge zur Taxonomie der gattung Chlorella. IV. Verwertung organischer Stickstoffverbindungen. — Arch. Mikrobiol. 70: 211–216.
Mitchison, J. W. 1971. (ed.). In Biology of the cell cycle. — Cambridge University Press.
Munch, R., Beck, G. und Manojlovic, N. 1971. Untersuchung über die Reaktion in vitro gezüchteter Zellen nach Inkubation in SiO2-Staub. — Beitr. Silikoseforschg. 23: 4.
Ortho Instruments. 1977. Measurement of DNA in yeast cells. In Ortho Instr. (eds), Protocols No. 24. — Ortho Instr., Westwood, Mass., USA.
Ortho Instruments. 1978. Determination of purity of yeast cultures. In Ortho Instr., (eds), Protocols No. 26.— Ortho Instr., Westwood, Mass., USA.
Paau, A. S., Lee, D. and Cowles, J. R. 1977a. Comparison of nucleic acid content in populations of free-living and symbiotic Rhizobium meliloti by flow-microfluorometry. — J. Bacteriol. 129: 1156–1158.
Paau, A. S., Cowles, J. R. and Oro, J. 1977b. Flow-microfluorometric analysis of escherichia coli, Rhizobium meliloti, and Rhizobium japonicum at different stages of the growth cycle. — Canad. J. Microbiol. 23: 1165–1169.
Slater, M. L., Sharrow, S. O. and Gart, J. J. 1977. Cell cycle of Saccharomyces cerevisiae in populations growing at different stages. — Proc. Natl. Acad. Sci. USA. 74: 3850–3854.
Smith, J. E. and Berry, D. R. 1974. The vegetative state. p. 106–151. In J. E. Smith and D. R. Berry (eds), An introduction to biochemistry of fungal development. — Academic Press, London & New York.
Sprenger, E., Sandritter, W., Böhm, N., Schaden, M., Hilgarth, M. und Wagner, D. 1971. Durchflußfluoreszenzzytophotometrie: Ein Prescreening-Verfahren für die gynäkokogische Zytodiagnostik. — Beitr. Path. 143: 323–344.
Strugger, S. 1949. (ed.). p. 89–92 In: Fluoreszenzmikroskopie und Mikrobiologie. — Verlag M. & H. Schaper, Hannover.
Williamson, D. H. and Scopes, A. W. 1961. Protein synthesis and nitrogen uptake in synchronously dividing cultures of Saccharomyces cerevisiae. — J. Inst. Brew. 67: 39–42.
Williamson, D. H. 1965. The timing of de oxyribonucleic acid synthesis in the cell cycle of Saccharomyces cerevisiae. — J. Cell. Biol. 25: 517–528.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hutter, K.J., Eipel, H.E. Flow cytometric determinations of cellular substances in algae, bacteria, moulds and yeasts. Antonie van Leeuwenhoek 44, 269–282 (1978). https://doi.org/10.1007/BF00394305
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00394305