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Related Experiment Video

Updated: Mar 12, 2026

Use of Image Cytometry for Quantification of Pathogenic Fungi in Association with Host Cells
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A novel concentration and viability detection method for Brettanomyces using the Cellometer image cytometry.

Brian Martyniak1, Jason Bolton1, Dmitry Kuksin2

  • 1Cooperative Extension and the School of Food and Agriculture, University of Maine, Orono, ME, 04469, USA.

Journal of Industrial Microbiology & Biotechnology
|November 14, 2016
PubMed
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Image cytometry offers a more accurate and efficient method for counting Brettanomyces yeast cells, overcoming limitations of traditional manual and spectrophotometer techniques. This improved cell enumeration can enhance the quality of fermented beverages.

Area of Science:

  • Microbiology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Brettanomyces yeasts exhibit complex cell morphologies, including pseudohyphae and budding, complicating accurate cell enumeration.
  • Traditional methods like manual counting and optical density measurements suffer from subjectivity, operator variability, and lack of precision.
  • Accurate monitoring of Brettanomyces cell populations is crucial for controlling fermentation processes and ensuring product quality in beverages.

Purpose of the Study:

  • To evaluate the efficacy of an image cytometer for precise enumeration of Brettanomyces cell concentrations, viabilities, and pseudohyphae formation.
  • To compare the performance of image cytometry with traditional cell counting methods for Brettanomyces spp.
  • To demonstrate the application of image cytometry in monitoring Brettanomyces growth dynamics during fermentation.
Keywords:
BrettanomycesBuddingCell countingCellometerFermentationImage cytometryPropagationPseudohyphae

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Main Methods:

  • Utilized an image cytometer with fluorescence viability dyes (acridine orange and propidium iodide) for direct counting of individual cell nuclei.
  • Optimized software counting parameters for Brettanomyces clausenii, Brettanomyces bruxellensis, and Brettanomyces lambicus during propagation.
  • Conducted a fermentation experiment to monitor B. lambicus and B. clausenii growth patterns in beer.

Main Results:

  • Image cytometry accurately enumerated cells within pseudohyphae chains, distinguishing individual nuclei.
  • The method demonstrated varying growth patterns, viabilities, and pseudohyphae percentages across different Brettanomyces species.
  • Results showed improved accuracy and efficiency compared to manual counting, with reduced operator-dependent variations.

Conclusions:

  • Image cytometry provides a robust, efficient, and objective method for quantifying Brettanomyces yeast cell characteristics.
  • This technique overcomes the limitations of traditional methods, offering enhanced precision in cell enumeration.
  • The proposed method has the potential to significantly improve process control and quality assurance in the production of beverages utilizing Brettanomyces yeasts.