Raman spectroscopy and microscopy of individual cells and cellular components

Raman spectroscopy provides the unique opportunity to nondestructively analyze chemical concentrations in individual cells on the submicrometer length scale without the need for optical labels. This enables the rapid assessment of cellular biochemistry inside living cells, and it allows for their continued analysis. Here, we review recent developments in the analysis of single cells, subcellular compartments, and chemical imaging based on Raman spectroscopy. Spontaneous Raman spectroscopy provides for the full spectral assessment of cellular biochemistry, while coherent Raman techniques, such as coherent anti-Stokes Raman scattering is primarily used as an imaging tool comparable to confocal fluorescence microscopy. These techniques are complemented by surface-enhanced Raman spectroscopy, which provides higher sensitivity and local specificity, and also extends the techniques to chemical indicators, i.e. pH sensing. We review the strengths and weaknesses of each technique, demonstrate some of their applications and discuss their potential for future research in cell biology and biomedicine.

[1]  D. Naumann,et al.  Identification of medically relevant microorganisms by vibrational spectroscopy. , 2002, Journal of microbiological methods.

[2]  K. S. Krishnan,et al.  A New Type of Secondary Radiation , 1928, Nature.

[3]  Pavel Matousek,et al.  Fluorescence suppression in resonance Raman spectroscopy using a high-performance picosecond Kerr gate , 2001 .

[4]  J. Molloy.  Muscle contraction: actin filaments enter the fray. , 2005, Biophysical journal.

[5]  K. Rowlen,et al.  Flashy Silver Nanoparticles , 2004 .

[6]  Shuming Nie,et al.  Efficient Raman enhancement and intermittent light emission observed in single gold nanocrystals , 1999 .

[7]  R. Dasari,et al.  Ultrasensitive chemical analysis by Raman spectroscopy. , 1999, Chemical reviews.

[8]  Keith T. Carron,et al.  Determination of the Distance Dependence and Experimental Effects for Modified SERS Substrates Based on Self-Assembled Monolayers Formed Using Alkanethiols , 1999 .

[9]  Marion Peter,et al.  Imaging molecular interactions by multiphoton FLIM , 2004, Biology of the cell.

[10]  Marcus Motzkus,et al.  Time-resolved two color single-beam CARS employing supercontinuum and femtosecond pulse shaping , 2006 .

[11]  M. Ishikawa,et al.  Thermal Activation of Blinking in SERS Signal , 2004 .

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

[13]  S. Wachsmann-Hogiu,et al.  Simultaneous forward and epi-CARS microscopy with a single detector by time-correlated single photon counting. , 2008, Optics express.

[14]  W. R. Wiley,et al.  Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering , 1999 .

[15]  Rainer Erdmann,et al.  Time-resolved laser scanning microscopy with FLIM and advanced FCS capability , 2005, SPIE BiOS.

[16]  Shuming Nie,et al.  Re-examining the origins of spectral blinking in single-molecule and single-nanoparticle SERS. , 2006, Faraday discussions.

[17]  J. Greve,et al.  Line-Scan Raman Microspectrometry for Biological Applications , 1997 .

[18]  H. Bruining,et al.  Rapid Identification of Candida Species by Confocal Raman Microspectroscopy , 2002, Journal of Clinical Microbiology.

[19]  D. A. Stuart,et al.  In vivo glucose measurement by surface-enhanced Raman spectroscopy. , 2006, Analytical chemistry.

[20]  Conor L Evans,et al.  Heterodyne coherent anti-Stokes Raman scattering (CARS) imaging. , 2006, Optics letters.

[21]  K. König,et al.  Fluorescence lifetime imaging by time‐correlated single‐photon counting , 2004, Microscopy research and technique.

[22]  Michael S. Feld,et al.  Surface-Enhanced Raman Spectroscopy in Single Living Cells Using Gold Nanoparticles , 2002 .

[23]  Mattias Goksör,et al.  Resonance Raman spectroscopy of optically trapped functional erythrocytes. , 2004, Journal of biomedical optics.

[24]  B. Wood,et al.  Resonance Raman spectroscopy provides evidence of heme ordering within the functional erythrocyte , 2005 .

[25]  C. Mirkin,et al.  Nanoparticles with Raman spectroscopic fingerprints for DNA and RNA detection. , 2002, Science.

[26]  Nicole J. Crane,et al.  Raman imaging demonstrates FGF2-induced craniosynostosis in mouse calvaria. , 2005, Journal of biomedical optics.

[27]  Peter J. Verveer,et al.  Spectral Imaging in a Programmable Array Microscope by Hadamard Transform Fluorescence Spectroscopy , 1999 .

[28]  G. Puppels,et al.  In situ investigation of the chemical composition of ceroid in human atherosclerosis by Raman spectroscopy , 2002 .

[29]  Olga Lyandres,et al.  Real-time glucose sensing by surface-enhanced Raman spectroscopy in bovine plasma facilitated by a mixed decanethiol/mercaptohexanol partition layer. , 2005, Analytical chemistry.

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

[31]  Tammy Y. Olson,et al.  Synthesis, characterization, and tunable optical properties of hollow gold nanospheres. , 2006, The journal of physical chemistry. B.

[32]  Eric O. Potma,et al.  Cover Picture: Direct Visualization of Lipid Phase Segregation in Single Lipid Bilayers with Coherent Anti‐Stokes Raman Scattering Microscopy (ChemPhysChem 1/2005) , 2005 .

[33]  C. A. Murray,et al.  Measurement of the molecule–silver separation dependence of surface enhanced Raman scattering in multilayered structures , 1982 .

[34]  D. L. Jeanmaire,et al.  Surface raman spectroelectrochemistry: Part I. Heterocyclic, aromatic, and aliphatic amines adsorbed on the anodized silver electrode , 1977 .

[35]  Martin Tjahjono,et al.  Remote Monitoring of a Multi-Component Liquid-Phase Organic Synthesis by Infrared Emission Spectroscopy: The Recovery of Pure Component Emissivities by Band-Target Entropy Minimization , 2007, Applied spectroscopy.

[36]  Thomas Huser,et al.  Improving nanoprobes using surface-enhanced Raman scattering from 30-nm hollow gold particles. , 2006, Analytical chemistry.

[37]  Sang Bok Lee,et al.  Surface‐enhanced Raman scattering of o‐mercaptobenzoic acid in silver sol , 1991 .

[38]  R. Dasari,et al.  Surface-enhanced Raman scattering and biophysics , 2001 .

[39]  X. Xie,et al.  Nonperturbative chemical imaging of organelle transport in living cells with coherent anti-stokes Raman scattering microscopy. , 2006, Biophysical journal.

[40]  Yong-qing Li,et al.  Near-infrared Raman spectroscopy of single optically trapped biological cells. , 2002, Optics letters.

[41]  A. Ashkin,et al.  Applications of laser radiation pressure. , 1980, Science.

[42]  K. Shi,et al.  SUPERCONTINUUM CARS TWEEZERS , 2007 .

[43]  S. Lane,et al.  Micro-Raman spectroscopy detects individual neoplastic and normal hematopoietic cells. , 2006, Biophysical journal.

[44]  Alberto Diaspro,et al.  Rapid dissemination of two‐photon excitation microscopy prompts new applications , 2004, Microscopy research and technique.

[45]  Marc Dellian,et al.  Acid production in glycolysis-impaired tumors provides new insights into tumor metabolism. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[46]  Ji-Xin Cheng,et al.  Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy. , 2007, Journal of biomedical optics.

[47]  B Willekens,et al.  Nonresonant Raman imaging of protein distribution in single human cells. , 2003, Biopolymers.

[48]  J. Sweedler,et al.  Quantitative submonolayer spatial mapping of Arg-Gly-Asp-containing peptide organomercaptan gradients on gold with matrix-assisted laser desorption/ionization mass spectrometry. , 2004, Analytical chemistry.

[49]  Steven R. Emory,et al.  Probing Single Molecules and Single Nanoparticles by Surface-Enhanced Raman Scattering , 1997, Science.

[50]  T. Molitor,et al.  Gammadelta T-lymphocyte cytotoxic activity against Mycobacterium bovis analyzed by flow cytometry. , 2005, Journal of immunological methods.

[51]  S. Ly,et al.  Time-gated single photon counting enables separation of CARS microscopy data from multiphoton-excited tissue autofluorescence. , 2007, Optics express.

[52]  B. R. Johnson,et al.  All-optical nanoscale pH meter. , 2006, Nano letters.

[53]  J. Parys,et al.  Subcellular distribution of the inositol 1,4,5‐trisphosphate receptors: functional relevance and molecular determinants , 2004, Biology of the cell.

[54]  Quantum Langevin Equation , 1981 .

[55]  D. Palanker,et al.  Wide-field coherent anti-Stokes Raman scattering microscopy with non-phase-matching illumination. , 2007, Optics letters.

[56]  N. Xia,et al.  SERS Titration of 4-Mercaptopyridine Self-Assembled Monolayers at Aqueous Buffer/Gold Interfaces. , 1999, Analytical chemistry.

[57]  H. Winhold,et al.  Optical trapping and coherent anti-Stokes Raman scattering (CARS) spectroscopy of submicron-size particles , 2005, IEEE Journal of Selected Topics in Quantum Electronics.

[58]  M D Duncan,et al.  Scanning coherent anti-Stokes Raman microscope. , 1982, Optics letters.

[59]  S. Kawata,et al.  Molecular vibration imaging in the fingerprint region by use of coherent anti-Stokes Raman scattering microscopy with a collinear configuration. , 2000, Optics letters.

[60]  H. Hamaguchi,et al.  Vibrationally resonant imaging of a single living cell by supercontinuum-based multiplex coherent anti-Stokes Raman scattering microspectroscopy. , 2005, Optics express.

[61]  Ziding Feng,et al.  Ultrasensitive detection and characterization of posttranslational modifications using surface-enhanced Raman spectroscopy. , 2006, Analytical chemistry.

[62]  P. Hamilton,et al.  Raman microscopy for the chemometric analysis of tumor cells. , 2006, The journal of physical chemistry. B.

[63]  Annika Enejder,et al.  Dual-pump coherent anti-Stokes-Raman scattering microscopy. , 2006, Optics letters.

[64]  N J Halas,et al.  Surface-enhanced Raman scattering on tunable plasmonic nanoparticle substrates , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[65]  B. Malissen,et al.  CD38 Is Associated with Lipid Rafts and upon Receptor Stimulation Leads to Akt/Protein Kinase B and Erk Activation in the Absence of the CD3-ζ Immune Receptor Tyrosine-based Activation Motifs* , 2002, The Journal of Biological Chemistry.

[66]  R. Dasari,et al.  Single Molecule Detection Using Surface-Enhanced Raman Scattering (SERS) , 1997 .

[67]  R R Duncan,et al.  Multi‐dimensional time‐correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) to detect FRET in cells , 2004, Journal of microscopy.

[68]  Tuan Vo-Dinh,et al.  Surface-enhanced Raman scattering detection of the breast cancer susceptibility gene BRCA1 using a silver-coated microarray platform , 2002 .

[69]  D. Renker.  New trends on photodetectors , 2007 .

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

[71]  Y. Kokubun,et al.  Fabrication technologies for vertically coupled microring resonator with multilevel crossing busline and ultracompact-ring radius , 2005, IEEE Journal of Selected Topics in Quantum Electronics.

[72]  M Mazilu,et al.  Dual beam fibre trap for Raman micro-spectroscopy of single cells. , 2006, Optics express.

[73]  Ramasamy Manoharan,et al.  Detection and identification of a single DNA base molecule using surface-enhanced Raman scattering (SERS) , 1998 .

[74]  R. Levin,et al.  Discrimination of viable Vibrio vulnificus cells from dead cells in real-time PCR. , 2006, Journal of microbiological methods.

[75]  Jan Greve,et al.  Applications of Micro-Raman Imaging in Biomedical Research , 1997 .

[76]  Karen Ross.  Drugs in the water. , 2006, Analytical chemistry.

[77]  C. Soeller,et al.  A mode of thought in excitation-contraction coupling. , 2002, Biophysical journal.

[78]  J. Bukowska,et al.  Surface‐enhanced Raman scattering (SERS) of 4‐mercaptobenzoic acid on silver and gold substrates , 2003 .

[79]  M. Hearn,et al.  Open tubular capillary electrochromatography of synthetic peptides on etched chemically modified columns. , 2004, Analytical chemistry.

[80]  Surface critical phenomena and scaling in the eight-vertex model. , 1996 .

[81]  R. Shi,et al.  Second harmonic and sum frequency generation imaging of fibrous astroglial filaments in ex vivo spinal tissues. , 2007, Biophysical journal.

[82]  R. V. Van Duyne,et al.  A glucose biosensor based on surface-enhanced Raman scattering: improved partition layer, temporal stability, reversibility, and resistance to serum protein interference. , 2004, Analytical chemistry.

[83]  Thomas Huser,et al.  Intracellular pH sensors based on surface-enhanced raman scattering. , 2004, Analytical chemistry.

[84]  K. Kneipp,et al.  One- and two-photon excited optical ph probing for cells using surface-enhanced Raman and hyper-Raman nanosensors. , 2007, Nano letters.

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

[86]  Mark Bachman,et al.  Choosing one from the many: selection and sorting strategies for single adherent cells , 2006, Analytical and bioanalytical chemistry.

[87]  T. McConnell,et al.  Activation-dependent phases of T cells distinguished by use of optical tweezers and near infrared Raman spectroscopy. , 2005, Journal of immunological methods.

[88]  Louis E. Brus,et al.  Ag Nanocrystal Junctions as the Site for Surface-Enhanced Raman Scattering of Single Rhodamine 6G Molecules , 2000 .

[89]  X. Xie,et al.  Polarization coherent anti-Stokes Raman scattering microscopy. , 2001, Optics letters.

[90]  K. Maquelin,et al.  Discrimination between nontumor bladder tissue and tumor by Raman spectroscopy. , 2006, Analytical chemistry.

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

[92]  M. Wabuyele,et al.  Surface-enhanced Raman scattering for medical diagnostics and biological imaging , 2005 .

[93]  W. Hill,et al.  Potential- and pH-dependent surface-enhanced Raman scattering of p-mercapto aniline on silver and gold substrates , 1993 .

[94]  P. Fratzl,et al.  Raman imaging of two orthogonal planes within cortical bone. , 2007, Bone.

[95]  Y. Ozaki,et al.  Surface-Enhanced Raman Spectroscopy , 2005 .

[96]  Qi Ye,et al.  Surface-Enhanced Raman Scattering from Functionalized Self-Assembled Monolayers. 2. Distance Dependence of Enhanced Raman Scattering from an Azobenzene Terminal Group , 1997 .

[97]  Rebecca Richards-Kortum,et al.  Resonance Raman Spectroscopy at 257 nm Excitation of Normal and Malignant Cultured Breast and Cervical Cells , 1999 .

[98]  Maria Cristina Chavantes,et al.  Near-infrared Raman spectroscopy of human coronary arteries: histopathological classification based on Mahalanobis distance. , 2003, Journal of clinical laser medicine & surgery.

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

[100]  Qiuxu Wei,et al.  Study of the effect of alcohol on single human red blood cells using near‐infrared laser tweezers Raman spectroscopy , 2005 .

[101]  R. Dasari,et al.  Population pumping of excited vibrational states by spontaneous surface-enhanced Raman scattering. , 1996, Physical review letters.

[102]  D S Elson,et al.  Fluorescence lifetime system for microscopy and multiwell plate imaging with a blue picosecond diode laser. , 2002, Optics letters.

[103]  Pavel Matousek,et al.  Efficient Rejection of Fluorescence from Raman Spectra Using Picosecond Kerr Gating , 1999 .

[104]  M Al-Rubeai,et al.  Use of intracellular pH and annexin-V flow cytometric assays to monitor apoptosis and its suppression by bcl-2 over-expression in hybridoma cell culture. , 1998, Journal of immunological methods.

[105]  Tuan Vo-Dinh,et al.  Cancer gene detection using surface-enhanced Raman scattering (SERS) , 2002 .

[106]  P. French,et al.  Time-resolved fluorescence microscopy , 2005 .

[107]  Kees Maquelin,et al.  Rapid epidemiological analysis of Acinetobacter strains by Raman spectroscopy. , 2006, Journal of microbiological methods.

[108]  Thomas R Huser,et al.  Surface-enhanced Raman scattering from individual au nanoparticles and nanoparticle dimer substrates. , 2005, Nano letters.

[109]  Andreas Volkmer,et al.  Time-resolved coherent anti-Stokes Raman scattering microscopy: Imaging based on Raman free induction decay , 2002 .

[110]  Rainer Erdmann,et al.  Dead-time optimized time-correlated photon counting instrument with synchronized, independent timing channels. , 2007, The Review of scientific instruments.

[111]  M. Moskovits.  Surface-enhanced spectroscopy , 1985 .

[112]  Don McNaughton,et al.  Resonance Raman spectroscopy of red blood cells using near-infrared laser excitation , 2007, Analytical and bioanalytical chemistry.