Monitoring cellular behaviour using Raman spectros

Authors: A R Boyd G A Burke B J Meenan

Publish Date: 2009/12/18

Volume: 21, Issue: 8, Pages: 2317-2324

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Abstract

Raman spectroscopy has been used to determine the chemical composition of materials for over 70 years Recent spectacular advances in laser and CCD camera technology creating instruments with higher sensitivity and lower cost have initiated a strong resurgence in the technique ranging from fundamental research to process control methodology One such area of increased potential is in tissue engineering and regenerative medicine TERM where autologous cell culture stem cell biology and growth of human cells on biomaterial scaffolds are of high importance Traditional techniques for the in vitro analysis of biochemical cell processes involves cell techniques such as fixation lysis or the use of radioactive or chemical labels which are time consuming and can involve the perpetuation of artefacts Several studies have already shown the potential of Raman spectroscopy to provide useful information on key biochemical markers within cells however many of these studies have utilised micro or confocal Raman to do this which are not suited to the rapid and noninvasive monitoring of cells For this study a versatile fitforpurpose Raman spectrometer was used employing a macrosampling optical platform laser spot size 100 μm at focus on the sample to discriminate between different TERM relevant cell types and viable and nonviable cells The results clearly show that the technique is capable of obtaining Raman spectra from live cells in a nondestructive rapid and noninvasive manner however in these experiments it was not possible to discriminate between different cell lines Despite this notable differences were observed in the spectra obtained from viable and nonviable cells showing significant changes in the spectral profiles of protein DNA/RNA and lipid cell constituents after cell death It is evident that the method employed here shows significant potential for further utilisation in TERM providing data directly from live cells that fits within a quality assurance framework and provides the opportunity to analyse cells in a nondestructive manner

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