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Spectroscopy: An International Journal
Volume 27, Article ID 746597, 5 pages
http://dx.doi.org/10.1155/2012/746597

Raman Microspectroscopy of the Yeast Vacuoles

1Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo Nám. 2, 166 10 Prague 6, Czech Republic
2Institute of Physics, Faculty of Mathematics and Physics, Charles University in Prague, Ke Karlovu 5, 121 16 Prague 2, Czech Republic

Copyright © 2012 Lucie Bednárová et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

In the present work, real ability of a confocal Raman microspectroscopy to monitor chemical composition of the vacuoles within living yeast cells was investigated and critically assessed. Simple, economical, and practical protocols of the yeast immobilization suitable for less laborious, high-throughput, and spatially resolved Raman measurements were tested for their possible impacts on physiological states and viability of the cells. We have demonstrated that, acquiring Raman spectra from statistically sound sets of immobilized cells and employing advanced multivariate methods for spectral analysis, the chemical composition of the yeast vacuoles can be reliably studied. The most easily and accurately quantifiable seems to be the concentration of polyphosphates which can be unambiguously identified due to unmistakable Raman features. Our approach can be useful for routine, label-free, and noninvasive monitoring of the chemical composition of the vacuoles of living yeasts exposed to various stress factors, the information important in biomedical research of pathogens.