Reference: Achilles J, et al. (2006) Analysis of living S. cerevisiae cell states--a three color approach. Cytometry A 69(3):173-7

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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 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.

Reference Type
Journal Article | Research Support, Non-U.S. Gov't
Authors
Achilles J, Harms H, Müller S
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