This review summarises progress in Raman spectroscopy and its application in diagnostics, toxicological testing and tissue engineering. Applications of Raman spectroscopy in cell biology are in the early stages of development, however, recent publications have demonstrated its utilisation as a diagnostic and development tool with the key advantage that investigations of living cells can be performed non-invasively.Some of the research highlighted here demonstrates the ability of Raman spectroscopy to accurately characterise cancer cells and distinguish between similar cell types. Many groups have used Raman spectroscopy to study tissues, but recently increased effort has gone into single cell analysis of cell lines; the advantages being that cell lines offer ease of handling and increased reproducibility over tissue studies and primary cells. The main goals of bio-Raman spectroscopy at this stage are twofold. Firstly, the aim is to further develop the diagnostic ability of Raman spectroscopy so it can be implemented in a clinical environment, producing accurate and rapid diagnoses. Secondly, the aim is to optimise the technique as a research tool for the non-invasive real time investigation of cell/material interactions in the fields of tissue engineering and toxicology testing.
"Processes and kinetic changes on the surfaces and in the structures of these molecules can be automatically observed in a very short time with a Raman analytic system (Owen, Notingher, Hill, Stevens, & Hench, 2006; Smith & Dent, 2005). The other advantage of Raman over other analytical techniques is its ability to work on water-rich samples such as food (Numata, Yoshiyuki, & Tanaka, 2011; Owen et al., 2006). Well-designed databases and chemometric algorithms are available for identification of Raman spectra, so both qualitative and quantitative results can be obtained through Raman (Dörfer, Schumacher, Tarcea, Schmitt, & Popp, 2010). "
[Show abstract][Hide abstract] ABSTRACT: In this study, the utility of Raman spectroscopy (RS) with chemometric methods for quantification of multiple components in the fermentation process was investigated. Vinegar, the product of a two stage fermentation, was used as a model and glucose and fructose consumption, ethanol production and consumption and acetic acid production were followed using RS and the partial least squares (PLS) method. Calibration of the PLS method was performed using model solutions. The prediction capability of the method was then investigated with both model and real samples. HPLC was used as a reference method. The results from comparing RS-PLS and HPLC with each other showed good correlations were obtained between predicted and actual sample values for glucose (R(2)=0.973), fructose (R(2)=0.988), ethanol (R(2)=0.996) and acetic acid (R(2)=0.983). In conclusion, a combination of RS with chemometric methods can be applied to monitor multiple components of the fermentation process from start to finish with a single measurement in a short time.
"Raman spectroscopy allows the study of living cells under physiological conditions, without labels and fixation (Notingher et al., 2005). Based on inelastic scattering of incident laser light that is characteristic of the composition of chemicals within living cells (Ling et al., 2002), it has been used for clinical diagnostics, tissue engineering and toxicological testing (Owen et al., 2006a). Due to its sensitivity to chemical variations in biomolecules, this method has been utilized to investigate toxic effects of pharmaceuticals on living cells in vitro (Owen et al., 2006b), interactions of drugs with DNA or protein (Morari and Muntean, 2003; Nakamura et al., 1997; Stanicova et al., 1999; Sukhanova et al., 2002) and to discover real-time biochemical alterations in living cells in order to identify specific toxic agents (Notingher et al., 2004a,b). "
[Show abstract][Hide abstract] ABSTRACT: In this study, confocal Raman spectroscopy, atomic force microscope (AFM) and multiplex ELISA were applied to analyze the biophysical responses (biomechanics and biospectroscopy) of normal human primary small airway epithelial cells (SAECs) and human lung carcinoma epithelial A549 cells to in vitro short term DEP exposure (up to 2 h). Raman spectra revealed the specific cellular biomolecular changes in cells induced by DEP compared to unexposed control cells. Principal component analysis was successfully applied to analyze spectral differences between control and treated groups from multiple individual cells, and indicated that cell nuclei are more sensitive than other cell locations. AFM measurements indicated that 2 h of DEP exposure induced a significant decrease in cell elasticity and a dramatic change in membrane surface adhesion force. Cytokine and chemokine production measured by multiplex ELISA demonstrated DEP-induced inflammatory responses in both cell types.
"A second disadvantage is that Raman scattering in endogenous compounds produces relatively weak signals that require long measurement times (seconds). Still, there is significant interest in using these approaches to develop diagnostic tools for cancer and other diseases (Kendall et al., 2009)(Nijssen et al., 2009)(Owen et al., 2006). "
[Show abstract][Hide abstract] ABSTRACT: Significant advances have been made in the preparation and applications of surface-enhanced Raman scattering (SERS)-active materials for biomolecular analysis. Bright signals, photostability, and narrow spectral features of SERS-active materials offer attractive advantages for cytometric analyses. However, SERS cytometry is still in an early stage of development, and advances in both instrumentation and reagents will be necessary to realize its full potential. In this chapter, we discuss the challenges of expanding the numbers of fluorescent labels that can be measured in cytometry, and introduce SERS tags with extremely narrow spectral peaks as an approach to make more efficient use of the optical spectrum and increase the number of parameters in cytometry.
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