title: Novel image cytometric method for detection of physiological and metabolic changes in Saccharomyces cerevisiae.
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.
authors.affiliation: Department of Technology R&D, Nexcelom Bioscience LLC, Lawrence, MA 01843, USA. lchan@nexcelom.com
authors.forename: Leo L
authors.initials: LL
authors.lastname: Chan
authors.forename: Alexandria
authors.initials: A
authors.lastname: Kury
authors.forename: Alisha
authors.initials: A
authors.lastname: Wilkinson
authors.forename: Charlotte
authors.initials: C
authors.lastname: Berkes
authors.forename: Alnoor
authors.initials: A
authors.lastname: Pirani
mesh_terms: D005285:Fermentation; D005434:Flow Cytometry; D006003:Glycogen / Q000032:analysis / Q000378:metabolism; D019044:Image Cytometry / Q000295:instrumentation* / Q000379:methods*; D007700:Kinetics; D008055:Lipids / Q000032:analysis; D050296:Microbial Viability; D008856:Microscopy, Fluorescence; D012441:Saccharomyces cerevisiae / Q000378:metabolism* / Q000502:physiology; D014199:Trehalose / Q000032:analysis / Q000378:metabolism
chemical_list: D008055:Lipids; D006003:Glycogen; D014199:Trehalose