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Updated: Jul 22, 2026
Quantitative Live Cell Fluorescence-microscopy Analysis of Fission Yeast
Published on: January 23, 2012
Analysis of living S. cerevisiae cell states--a three color approach
J Achilles1, H Harms, S Müller
1Department of Environmental Microbiology, Centre for Environmental Research Leipzig/Halle in the Helmholtz association, Leipzig, Germany.
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 Saccharomyces 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.
