Issue 6, 2009
From the journal:

Analyst

Impact of fixation on in vitro cell culture lines monitored with Raman spectroscopy†

Melissa M. Mariani,a   Peter Lampen,a   Jürgen Popp,bc   Bayden R. Woodd  and  Volker Deckert*ae  

* Corresponding authors

a ISAS-Institute for Analytical Sciences, Bunsen-Kirchhoff-Str. 11–13, Dortmund 44139, Germany
E-mail: deckert@isas.de
Fax: +49 231 1392 120
Tel: +49 231 139 2263

b IPHT-Institute for Photonic Technology, Albert-Einstein-Str. 9, 07745 Jena, Germany

c FSU Jena, Institute for Physical Chemistry, Helmholtzweg 4, 07743 Jena, Germany
E-mail: Juergen.Popp@uni-jena.de

d Monash University, Clayton, Australia
E-mail: Bayden.Wood@sci.monash.edu.au
Fax: +61 3 9905 4597
Tel: +61 3 9905 5721

e Technische Universität Dortmund, Otto-Hahn-Str. 6, 44227 Dortmund, Germany

Abstract

Raman spectroscopy provides chemical-rich information about the composition of analytes and is a powerful tool for biological studies. With the ability to investigate specific cellular components or image whole cells, compatible methods of sample preservation must be implemented for accurate spectra to be collected. Unfortunately, the effects of many commonly used sample preservation methods have not been explored with cultured cells. In this study, two human cell lineages of varying phenotypes were used to investigate the effects of sample preservation methods. Cells were cultured directly onto quartz substrates and either formalin-fixed, desiccated or air dried. The results indicate that the methodology applied to cell cultures for Raman analysis significantly influences the quality and reproducibility of the resulting spectral data. Formalin fixation was not found to be as universally efficient as anticipated for a commonly used fixative. This was due largely to the inconsistency in sample preservation between cell lines and loss of signal intensity. Sample air-drying was found to be largely inconsistent in terms of spectral reproducibility. Our study shows that sample desiccation displayed good spectral reproducibility and resulted in a good signal-to-noise ratio. Lipid and protein content in both activated and inactivated cells were maintained and provided a more controlled method compared with air-drying, revealing that the speed of drying is important for sample preservation

Graphical abstract: Impact of fixation on in vitro cell culture lines monitored with Raman spectroscopy

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Article information

DOI
https://doi.org/10.1039/B822408K
Article type
Paper
Submitted
15 Dec 2008
Accepted
06 Apr 2009
First published
28 Apr 2009

Analyst, 2009,134, 1154-1161

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Impact of fixation on in vitro cell culture lines monitored with Raman spectroscopy

M. M. Mariani, P. Lampen, J. Popp, B. R. Wood and V. Deckert, Analyst, 2009, 134, 1154 DOI: 10.1039/B822408K

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