Skip to main content
SpringerLink
Log in

Novel image cytometric method for detection of physiological and metabolic changes in Saccharomyces cerevisiae

  • Biotechnology Methods
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The studying and monitoring of physiological and metabolic changes in Saccharomyces cerevisiae (S. cerevisiae) has been a key research area for the brewing, baking, and biofuels industries, which rely on these economically important yeasts to produce their products. Specifically for breweries, physiological and metabolic parameters such as viability, vitality, glycogen, neutral lipid, and trehalose content can be measured to better understand the status of S. cerevisiae during fermentation. Traditionally, these physiological and metabolic changes can be qualitatively observed using fluorescence microscopy or flow cytometry for quantitative fluorescence analysis of fluorescently labeled cellular components associated with each parameter. However, both methods pose known challenges to the end-users. Specifically, conventional fluorescent microscopes lack automation and fluorescence analysis capabilities to quantitatively analyze large numbers of cells. Although flow cytometry is suitable for quantitative analysis of tens of thousands of fluorescently labeled cells, the instruments require a considerable amount of maintenance, highly trained technicians, and the system is relatively expensive to both purchase and maintain. In this work, we demonstrate the first use of Cellometer Vision for the kinetic detection and analysis of vitality, glycogen, neutral lipid, and trehalose content of S. cerevisiae. This method provides an important research tool for large and small breweries to study and monitor these physiological behaviors during production, which can improve fermentation conditions to produce consistent and higher-quality products.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Arlorio M, Coïsson JD, Martelli A (1999) Identification of Saccharomyces cerevisiae in bakery products by PCR amplification of the ITS region of ribosomal DNA. Eur Food Res Technol 209:185–191

    Article  CAS  Google Scholar 

  2. Ciani M, Mannazzu I, Marinangeli P, Clementi F, Martini A (2004) Contribution of winery-resident Saccharomyces cerevisiae strains to spontaneous grape must fermentation. Antonie Van Leeuwenhoek 85:159–164

    Article  PubMed  CAS  Google Scholar 

  3. Novak J, Basarova G, Teixeira JA, Vicente AA (2007) Monitoring of brewing yeast propagation under aerobic and anaerobic conditions employing flow cytometry. J Inst Brew 113:249–255

    Article  CAS  Google Scholar 

  4. Bauer A, Kölling R (1996) Characterization of the SAC3 gene of Saccharomyces cerevisiae. Yeast 12:965–975

    Article  PubMed  CAS  Google Scholar 

  5. Hernlem B, Hua S-S (2010) Dual fluorochrome flow cytometric assessment of yeast viability. Curr Microbiol (published online)

  6. Millbank JW (1962) The action of acriflavine on yeast protoplasts. Antonie Van Leeuwenhoek 28:215–220

    Article  Google Scholar 

  7. Miura Y, Wada N, Nishida Y, Mori H, Kobayashi K (2003) Chemoenzymatically synthesized glycoconjugate polymers. Biomacromolecules 4:410–415

    Article  PubMed  CAS  Google Scholar 

  8. Raschke D, Knorr D (2009) Rapid monitoring of cell size, vitality and lipid droplet development in the oleaginous yeast Waltomyces lipofer. J Microbiol Methods 79:178–183

    Article  PubMed  CAS  Google Scholar 

  9. Schlee C, Miedl M, Leiper KA, Stewart GG (2006) The potential of confocal imaging for measuring physiological changes in brewer’s yeast. J Inst Brew 112:134–147

    Article  CAS  Google Scholar 

  10. Slaughter JC, Minabe M (1994) Fatty acid-containing lipids of the yeast Saccharomyces cerevisiae during post-fermentation decline in viability. J Sci Food Agric 65:497–501

    Article  CAS  Google Scholar 

  11. Slaughter JC, Nomura T (1992) Intracellular glycogen and trehalose contents as predictors of yeast viability. Enzyme Microb Technol 14:64–67

    Article  CAS  Google Scholar 

  12. Rodríguez-Porrata B, Novo M, Guillamón J, Rozès N, Mas A, Otero RC (2008) Vitality enhancement of the rehydrated active dry wine yeast. Int J Food Microbiol 126:116–122

    Article  PubMed  Google Scholar 

  13. Henry-Stanley MJ, Garni RM, Wells CL (2004) Adaptation of FUN-1 and Calcofluor white stains to assess the ability of viable and nonviable yeast to adhere to and be internalized by cultured mammalian cells. J Microbiol Methods 59:289–292

    Article  PubMed  CAS  Google Scholar 

  14. Zhang T, Fang HHP (2004) Quantification of Saccharomyces cerevisiae viability using BacLight. Biotechnol Lett 26:989–992

    Article  PubMed  CAS  Google Scholar 

  15. Hsu SYL, Hsu HF, Isacson P, Cheng HF (1977) Schistosoma mansoni and S. japonicum: Methylene Blue Test for the viability of Schitosomula in vitro. Exp Parasitol 41:329–334

    Article  PubMed  CAS  Google Scholar 

  16. Boyd AR, Gunasekera TS, Attfield PV, Simic K, Vincent SF, Veal DA (2003) A flow-cytometric method for determination of yeast viability and cell number in a brewery. FEMS Yeast Res 3:11–16

    PubMed  CAS  Google Scholar 

  17. Oh K-B, Matsuoka H (2002) Rapid viability assessment of yeast cells using vital staining with 2-NBDG, a fluorescent derivative of glucose. Int J Food Microbiol 76:47–53

    Article  PubMed  CAS  Google Scholar 

  18. Anton-Leberre V, Haanappel E, Marsaud N, Trouilh L, Benbadis L, Boucherie H, Massou S, François JM (2010) Exposure to high static or pulsed magnetic fields does not affect cellular processes in the yeast Saccharomyces cerevisiae. Bio Electro Magn 31:28–38

    CAS  Google Scholar 

  19. Bouchez JC, Cornu M, Danzart M, Leveau JY, Duchiron F, Bouix M (2004) Physiological significance of the cytometric distribution of fluorescent yeasts after viability staining. Biotechnol Bioeng 86:520–530

    Article  PubMed  CAS  Google Scholar 

  20. Chan LL, Lyettefi EJ, Pirani A, Smith T, Qiu J, Lin B (2010) Direct concentration and viability measurement of yeast in corn mash using a novel imaging cytometry method. J Ind Microbiol Biotechnol 38:1109–1115

    Article  PubMed  Google Scholar 

  21. McCaig R (1990) Evaluation of the fluorescent dye 1-Anilino-8-Naphthalene sulfonic acid for yeast viability determination. J Am Soc Brew Chem 48:22–25

    CAS  Google Scholar 

  22. Zandycke SMV, Simal O, Gualdoni S, Smart KA (2003) Determination of yeast viability using fluorophores. J Am Soc Brew Chem 61:15–22

    Google Scholar 

  23. Cahill G, Walsh PK, Donnelly D (2000) Determination of yeast glycogen content by individual cell spectroscopy using image analysis. Biotechnol Bioeng 69:312–322

    Article  PubMed  CAS  Google Scholar 

  24. Paulillo SCDL, Yokoya F, Basso LC (2003) Mobilization of endogenous glycogen and trehalose of industrial yeasts. Brazilian J Microbiol 34:249–254

    Article  Google Scholar 

  25. Nikolova M, Savova I, Marinov M (2002) An optimised method for investigation of the yeast viability by means of fluorescent microscopy. J Cult Collect 3:66–71

    Google Scholar 

  26. King LM, Schisler DO, Ruocco JJ (1981) Epifluorescent method for detection of nonviable yeast. J Am Soc Brew Chem 39:52–54

    CAS  Google Scholar 

  27. Chang WL, Heyde H C v d, Klein BS (1998) Flow cytometric quantitation of yeast a novel technique for use in animal model work and in vitro immunologic assays. J Immunol Methods 211:51–63

    Article  PubMed  CAS  Google Scholar 

  28. Malacrinó P, Zapparoli G, Torriani S, Dellaglio F (2001) Rapid detection of viable yeasts and bacteria in wine by flow cytometry. J Microbiol Methods 45:127–134

    Article  PubMed  Google Scholar 

  29. Deere D, Shen J, Vesey G, Bell P, Bissinger P, Veal D (1998) Flow cytometry and cell sorting for yeast viability assessment and cell selection. Yeast 14:147–160

    Article  PubMed  CAS  Google Scholar 

  30. Chan LL, Zhong X, Pirani A, Lin B (2012) A novel method for kinetic measurements of rare cell proliferation using Cellometer image-based cytometry. J Immunol Methods 377:8–14

    Article  PubMed  CAS  Google Scholar 

  31. Chan LL, Zhong X, Qiu J, Li PY, Lin B (2011) Cellometer Vision as an alternative to flow cytometry for cell cycle analysis, mitochondrial potential, and immunophenotyping. Cytometry Part A 79A:507–517

    Article  Google Scholar 

  32. Chan LL-Y, Lai N, Wang E, Smith T, Yang X, Lin B (2011) A rapid detection method for apoptosis and necrosis measurement using the Cellometer imaging cytometry. Apoptosis 16:1295–1303

    Article  PubMed  Google Scholar 

Download references

Conflict of interest

The authors, LLC, and AP declare competing financial interests, and the work performed in this manuscript is for reporting on product performance for Nexcelom Bioscience, LLC. The performance of the instrumentation has been compared to standard approaches currently used in biomedical research institutions.

Author information

Authors and Affiliations

  1. Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA

    Leo L. Chan, Alexandria Kury & Alisha Wilkinson

  2. Department of Applications, Nexcelom Bioscience LLC, Lawrence, MA, 01843, USA

    Alnoor Pirani

  3. Department of Biology, Merrimack College, North Andover, MA, 01845, USA

    Alexandria Kury, Alisha Wilkinson & Charlotte Berkes

  4. Center for Biotechnology and Biomedical Sciences, Merrimack College, North Andover, MA, 01845, USA

    Leo L. Chan, Alexandria Kury, Alisha Wilkinson & Charlotte Berkes

Authors
  1. Leo L. Chan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alexandria Kury
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alisha Wilkinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Charlotte Berkes
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alnoor Pirani
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Leo L. Chan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chan, L.L., Kury, A., Wilkinson, A. et al. Novel image cytometric method for detection of physiological and metabolic changes in Saccharomyces cerevisiae . J Ind Microbiol Biotechnol 39, 1615–1623 (2012). https://doi.org/10.1007/s10295-012-1177-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10295-012-1177-y

Keywords

Search

Navigation