The lag phase and G1 phase of a single yeast cell monitored by Raman microspectroscopy

We optically trapped a single yeast cell for up to 3 h and monitored the changes in the Raman spectra during the lag phase of its growth and the G1 phase of its cell cycle. A non-budding cell (corresponding either to the G0 or G1 phase) was chosen for each experiment. During the lag phase, the cell synthesises new proteins and lipids and the observed behaviour of the peaks corresponding to these constituents as well as those of RNA served as a sensitive indicator of the adaptation of the cell to its changed environment. Temporal behaviour of the Raman peaks observed was different in the lag phase as compared to the late lag phase. Two different laser wavelengths were applied to study the effect of long-term optical trapping on the living cells. Yeast cells killed either by boiling or by a chemical protocol were also trapped for a long time in a single beam optical trap to understand the effect of optical trapping on the behaviour of observed Raman peaks. 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. Copyright © 2006 John Wiley & Sons, Ltd.

[1]  A. Ashkin,et al.  Optical trapping and manipulation of single cells using infrared laser beams , 1987, Nature.

[2]  I. Tolic-Nørrelykke,et al.  Anomalous diffusion in living yeast cells. , 2004, Physical review letters.

[3]  D. Grier A revolution in optical manipulation , 2003, Nature.

[4]  Gajendra P. Singh,et al.  Dual wavelength optical tweezers for confocal Raman spectroscopy , 2005 .

[5]  A. Nasim,et al.  Molecular biology of the fission yeast , 1989 .

[6]  L L Hench,et al.  Spectroscopic study of human lung epithelial cells (A549) in culture: living cells versus dead cells. , 2003, Biopolymers.

[7]  Yong-qing Li,et al.  Raman sorting and identification of single living micro-organisms with optical tweezers. , 2005, Optics letters.

[8]  Yasuaki Naito,et al.  In vivo time‐resolved Raman imaging of a spontaneous death process of a single budding yeast cell , 2005 .

[9]  Wei Tang,et al.  Study of dynamical process of heat denaturation in optically trapped single microorganisms by near-infrared Raman spectroscopy , 2003 .

[10]  Keiichi Torimitsu,et al.  Laser trapping and Raman spectroscopy of single cellular organelles in the nanometer range. , 2002, Lab on a chip.

[11]  Peter F. Stanbury,et al.  Principles of Fermentation Technology , 1984 .

[12]  W. L. Fangman,et al.  Cell cycle phases in the unequal mother/daughter cell cycles of Saccharomyces cerevisiae , 1984, Molecular and cellular biology.

[13]  G. Fink,et al.  Methods in enzymology vol 194 guide to yeast genetics and molecular biology , 1991 .

[14]  J. Warner,et al.  Coordinate control of syntheses of ribosomal ribonucleic acid and ribosomal proteins during nutritional shift-up in Saccharomyces cerevisiae , 1981, Molecular and cellular biology.

[15]  L. Jespersen,et al.  Protein expression during lag phase and growth initiation in Saccharomyces cerevisiae. , 2002, International journal of food microbiology.

[16]  J. Conboy,et al.  Optical-trapping Raman microscopy detection of single unilamellar lipid vesicles. , 2003, Analytical chemistry.

[17]  Juergen Popp,et al.  Raman and Fluorescence Spectra of Single Optically Trapped Microdroplets in Emulsions , 1994 .

[18]  K. O. Greulich,et al.  Micromanipulation by light in biology and medicine : the laser microbeam and optical tweezers , 1999 .

[19]  Hiro-o Hamaguchi,et al.  Raman spectroscopic signature of life in a living yeast cell , 2004 .

[20]  K. Torimitsu,et al.  Detection of glutamate in optically trapped single nerve terminals by Raman spectroscopy. , 2004, Analytical chemistry.

[21]  Douglas E. Bassett,et al.  Yeast genes and human disease , 1996, Nature.

[22]  Hiro-o Hamaguchi,et al.  Molecular‐level pursuit of yeast mitosis by time‐ and space‐resolved Raman spectroscopy , 2003 .

[23]  J Greve,et al.  Nonresonant confocal Raman imaging of DNA and protein distribution in apoptotic cells. , 2003, Biophysical journal.

[24]  H. Barr,et al.  Raman spectroscopy for identification of epithelial cancers. , 2004, Faraday discussions.

[25]  K. Crawford,et al.  Raman Vibrational Evidence for the Presence of Conjugated Regions in Individual Micron Diameter Polystyrene Particles Irradiated with Visible Radiation , 1998 .

[26]  A. Ward,et al.  Laser tweezers Raman study of optically trapped aerosol droplets of seawater and oleic acid reacting with ozone: implications for cloud-droplet properties. , 2004, Journal of the American Chemical Society.

[27]  L. Jespersen,et al.  Genome-wide transcriptional changes during the lag phase of Saccharomyces cerevisiae , 2003, Archives of Microbiology.

[28]  Giovanni Volpe,et al.  Raman imaging of floating cells. , 2005, Optics express.

[29]  J. Greve,et al.  Studying single living cells and chromosomes by confocal Raman microspectroscopy , 1990, Nature.

[30]  S. J. Pirt,et al.  Principles of microbe and cell cultivation , 1975 .

[31]  Yong-qing Li,et al.  Confocal micro-Raman spectroscopy of single biological cells using optical trapping and shifted excitation difference techniques , 2003 .

[32]  Prof. Dr. Karl Otto Greulich Micromanipulation by Light in Biology and Medicine , 1999, Methods in Bioengineering.

[33]  P. Gemperline,et al.  Identification of single bacterial cells in aqueous solution using confocal laser tweezers Raman spectroscopy. , 2005, Analytical chemistry.

[34]  Giovanni Volpe,et al.  Real-time detection of hyperosmotic stress response in optically trapped single yeast cells using Raman microspectroscopy. , 2005, Analytical chemistry.

[35]  Gerald R. Fink,et al.  Guide to yeast genetics and molecular biology , 1993 .

[36]  James P Freyer,et al.  Raman spectroscopy detects biochemical changes due to proliferation in mammalian cell cultures. , 2005, Biophysical journal.

[37]  S. Lane,et al.  Reagentless identification of single bacterial spores in aqueous solution by confocal laser tweezers Raman spectroscopy. , 2004, Analytical chemistry.

[38]  Ioan Notingher,et al.  In situ spectral monitoring of mRNA translation in embryonic stem cells during differentiation in vitro. , 2004, Analytical chemistry.

[39]  M. Houlne,et al.  Confocal Raman microscopy for monitoring chemical reactions on single optically trapped, solid-phase support particles. , 2002, Analytical chemistry.

[40]  B. Wood,et al.  A portable Raman acoustic levitation spectroscopic system for the identification and environmental monitoring of algal cells. , 2005, Analytical chemistry.

[41]  Wolfgang Kiefer,et al.  Raman-Microsampling Technique Applying Optical Levitation by Radiation Pressure , 1984 .

[42]  Jürgen Popp,et al.  Raman spectroscopic identification of single yeast cells , 2005 .

[43]  G. Cooper The Cell: A Molecular Approach , 1996 .

[44]  Yong-qing Li,et al.  Real-time Raman spectroscopy of optically trapped living cells and organelles. , 2004, Optics express.