Jump to main content
Jump to site search

Issue 8, 2017
Previous Article Next Article

Probing glycosaminoglycan spectral signatures in live cells and their conditioned media by Raman microspectroscopy

Author affiliations

Abstract

Spectroscopic markers characteristic of reference glycosaminoglycan molecules were identified previously based on their vibrational signatures. Infrared spectral signatures of glycosaminoglycans in fixed cells were also recently demonstrated but probing live cells still remains challenging. Raman microspectroscopy is potentially interesting to perform studies under physiological conditions. The aim of the present work was to identify the Raman spectral signatures of GAGs in fixed and live cells and in their conditioned media. Biochemical and Raman analyses were performed on five cell types: chondrocytes, dermal fibroblasts, melanoma (SK-MEL-28), wild type CHO, and glycosaminoglycan-defective mutant CHO-745 cells. The biochemical assay of sulfated GAGs in conditioned media was only possible for chondrocytes, dermal fibroblasts, and wild type CHO due to the detection limit of the test. In contrast, Raman microspectroscopy allowed probing total glycosaminoglycan content in conditioned media, fixed and live cells and the data were analysed by principal component analysis. Our results showed that the Raman technique is sensitive enough to identify spectral markers of glycosaminoglycans that were useful to characterise the conditioned media of the five cell types. The results were confirmed at the single cell level on both live and fixed cells with a good differentiation between the cell types. Furthermore, the principal component loadings revealed prominent glycosaminoglycan-related spectral information. Raman microspectroscopy allows monitoring of the glycosaminoglycan profiles of single live cells and could therefore be developed for cell screening purposes and holds promise for identifying glycosaminoglycan signatures as a marker of cancer progression in tissues.

Graphical abstract: Probing glycosaminoglycan spectral signatures in live cells and their conditioned media by Raman microspectroscopy

Back to tab navigation

Supplementary files

Publication details

The article was received on 31 Aug 2016, accepted on 19 Mar 2017 and first published on 20 Mar 2017


Article type: Paper
DOI: 10.1039/C6AN01951J
Citation: Analyst, 2017,142, 1333-1341
  • Open access: Creative Commons BY-NC license
  •   Request permissions

    Probing glycosaminoglycan spectral signatures in live cells and their conditioned media by Raman microspectroscopy

    S. Brézillon, V. Untereiner, H. T. Mohamed, J. Hodin, A. Chatron-Colliet, F.-X. Maquart and G. D. Sockalingum, Analyst, 2017, 142, 1333
    DOI: 10.1039/C6AN01951J

    This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material and it is not used for commercial purposes.

    Reproduced material should be attributed as follows:

    • For reproduction of material from NJC:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the Centre National de la Recherche Scientifique (CNRS) and the RSC.
    • For reproduction of material from PCCP:
      [Original citation] - Published by the PCCP Owner Societies.
    • For reproduction of material from PPS:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC.
    • For reproduction of material from all other RSC journals:
      [Original citation] - Published by The Royal Society of Chemistry.

    Information about reproducing material from RSC articles with different licences is available on our Permission Requests page.

Search articles by author

Spotlight

Advertisements