Quantification methods of determining brewer's and pharmaceutical yeast cell viability: accuracy and impact of nanoparticles

For industrial processes, a fast, precise, and reliable method of determining the physiological state of yeast cells, especially viability, is essential. However, an increasing number of processes use magnetic nanoparticles (MNPs) for yeast cell manipulation, but their impact on yeast cell viability and the assay itself is unclear. This study tested the viability of Saccharomyces pastorianus ssp. carlsbergensis and Pichia pastoris by comparing traditional colourimetric, high-throughput, and growth assays with membrane fluidity. Results showed that methylene blue staining is only reliable for S. pastorianus cells with good viability, being erroneous in low viability (R-2= 0.945; (sigma) over bar= 5.78%). In comparison, the fluorescence microscopy-based assay of S. pastorianus demonstrated a coefficient of determination of R2=0.991 at a = 0 ((sigma) over bar= 2.50%) and flow cytometric viability determination using carboxyfluorescein diacetate (CFDA), enabling high-throughput analysis of representative cell numbers; R-2= 0.972 (a = 0; (sigma) over bar= 3.89%). Membrane fluidity resulted in a non-linear relationship with the viability of the yeast cells (a not equal 0 ). We also determined similar results using P. pastoris yeast. In addition, we demonstrated that MNPs affected methylene blue staining by overestimating viability. The random forest model has been shown to be a precise method for classifying nanoparticles and yeast cells and viability differentiation in flow cytometry by using CFDA. Moreover, CFDA and membrane fluidity revealed precise results for both yeasts, also in the presence of nanoparticles, enabling fast and reliable determination of viability in many experiments using MNPs for yeast cell manipulation or separation.

Automated summary

For industrial processes, it is crucial to have a fast, precise, and reliable method for determining the physiological state, particularly viability, of yeast cells. This study examined the impact of magnetic nanoparticles (MNPs) on yeast cell viability and the assay itself, testing two types of yeast cells. The results showed that traditional colorimetric staining was only reliable for cells with good viability, while fluorescence microscopy and flow cytometry-based assays demonstrated high precision and reliability in determining viability, even in the presence of MNPs. Moreover, the study revealed a non-linear relationship between membrane fluidity and yeast cell viability. The use of CFDA and membrane fluidity provided accurate results for both yeast types, allowing for efficient viability determination in experiments involving yeast cell manipulation or separation using MNPs.