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Analysis of living S. cerevisiae cell states—A three color approach
Author(s) -
Achilles J.,
Harms H.,
Müller S.
Publication year - 2006
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20212
Subject(s) - flow cytometry , propidium iodide , saccharomyces cerevisiae , cell growth , yeast , cell , cytometry , substrate (aquarium) , staining , biology , microbiology and biotechnology , chemistry , biochemistry , apoptosis , programmed cell death , genetics , ecology
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. © 2006 International Society for Analytical Cytology