Raman spectroscopy of a single living cell in environmentally stressed conditions

Gajendra P. Singh; Caitriona Creely; Giovanni Volpe; Helga Grotsch; Dmitri Petrov

doi:10.1117/12.613166

26 August 2005 Raman spectroscopy of a single living cell in environmentally stressed conditions

Gajendra P. Singh, Caitriona Creely, Giovanni Volpe, Helga Grotsch, Dmitri Petrov

Proceedings Volume 5930, Optical Trapping and Optical Micromanipulation II; 593009 (2005) https://doi.org/10.1117/12.613166
Event: Optics and Photonics 2005, 2005, San Diego, California, United States

Abstract

Living cells initiate a stress response in order to survive environmentally stressful conditions. We monitored changes in the Raman spectra of an optically trapped Saccharomyces cerevisiae yeast cell under normal and hyperosmotic stress conditions. When the yeast cells were challenged with a high concentration of glucose so as to exert hyperosmotic stress, it was shown that two chemical substances - glycerol and ethanol - could be monitored in real time in a single cell. The volume of the detection area of our confocal microspectrometer is approximately 1 fL. The average quantities of detected glycerol and ethanol are about 300 attomol and 700 attomol respectively. This amounts to the detection of approximately 10⁸ glycerol molecules and 4 X 10⁸ ethanol molecules after 36 min of hyper osmotic stress. Besides this, we also optically trapped a single yeast cell for up to three hours under normal conditions and monitored the changes in the Raman spectra during the lag phase of its growth and the G₁ phase of its cell cycle. During the lag phase the cell synthesises new proteins and the observed behavior of the peaks corresponding to these proteins as well as those of RNA served as a sensitive indicator of the adaptation of the cell to its changed environment. The changes observed in the Raman spectra of a trapped yeast cell in the late G₁ phase or the beginning of S phase corresponded to the growth of a bud.

Citation Download Citation

Gajendra P. Singh, Caitriona Creely, Giovanni Volpe, Helga Grotsch, and Dmitri Petrov "Raman spectroscopy of a single living cell in environmentally stressed conditions", Proc. SPIE 5930, Optical Trapping and Optical Micromanipulation II, 593009 (26 August 2005); https://doi.org/10.1117/12.613166

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