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Title (Primary) Analysis of living S. cerevisiae cell states - a three color approach
Author Achilles, J.; Harms, H.; Müller, S.;
Journal Cytometry Part A
Year 2006
Department UMB;
Volume 69A
Issue 3
Language englisch;
Keywords multiparametric flow cytometry; Hoechst 33342; 2-NBD-glucose; viability; population dynamics; metabolic modes; Saccharomyces cerevisiae
Abstract Background: Biosyntheses often fluctuate with the state of the cell in the cell cycle and on the capacity of the cell to access and metabolize a carbon source, Visualization of Substrate uptake by individual cells, together with the simultaneous analysis of proliferation activity and the proportion of dead cells, facilitate reliable and quasi-online process optimization. Methods: Flow cytometry and Hoechst 33342 staining were used to follow proliferation activity of living Saccha romyces cerevisiae cells, whereas 2-NBD-glucose was employed to analyze the cells' substrate affinity. Propidium iodide was used to determine the proportion of dead cells. Calibration and verification experiments were performed with cells grown batch-wise as well as in transient state regimes. Results: A new and rapid three-color assay was developed and tested under varying microenvironmental conditions. Conclusions: Live/dead cell states and the affinity to 2-NBD-glucose vs. proliferation states were determined during respiratory and/or fermentative modes of metabolism.
ID 2441
Persistent UFZ Identifier
Achilles, J., Harms, H., Müller, S. (2006):
Analysis of living S. cerevisiae cell states - a three color approach
Cytom. Part A 69A (3), 173 - 177