Issue 12, 2010

Imaging live cells grown on a three dimensional collagen matrix using Raman microspectroscopy

Abstract

Three dimensional collagen gels have been used as matrices for the imaging of live cells by Raman spectroscopy. The study is conducted on a human lung adenocarcinoma (A549) and a spontaneously immortalized human epithelial keratinocyte (HaCaT) cell line. The lateral resolution of the system has been estimated to be <1.5 μm making it possible to access the subcellular organization. Using K-means clustering analysis, it is shown that the different subcellular compartments of individual cells can be indentified and differentiated. The biochemical specificity of the information contained in the Raman spectra allows the visualization of differences in the molecular signature of the different sub-cellular structures. Furthermore, to enhance the chemical information obtained from the spectra, principal component analysis has been employed, allowing the identification of spectral windows with a high variability. The comparison between the loadings calculated and spectra from pure biochemical compounds enables the correlation of the variations observed with the molecular content of the different cellular compartments.

Graphical abstract: Imaging live cells grown on a three dimensional collagen matrix using Raman microspectroscopy

Article information

Article type
Paper
Submitted
19 Jul 2010
Accepted
28 Sep 2010
First published
13 Oct 2010

Analyst, 2010,135, 3169-3177

Imaging live cells grown on a three dimensional collagen matrix using Raman microspectroscopy

F. Bonnier, P. Knief, B. Lim, A. D. Meade, J. Dorney, K. Bhattacharya, F. M. Lyng and H. J. Byrne, Analyst, 2010, 135, 3169 DOI: 10.1039/C0AN00539H

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