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Nat Commun. 2017 Mar 22;8:14843. doi: 10.1038/ncomms14843.

Quantitative volumetric Raman imaging of three dimensional cell cultures.

Kallepitis C1,2,3, Bergholt MS1,2,3, Mazo MM1,2,3, Leonardo V1,2,3, Skaalure SC1,2,3, Maynard SA1,2,3, Stevens MM1,2,3.

Author information

1
Department of Materials, Imperial College London, London SW7 2AZ, UK.
2
Department of Bioengineering, Imperial College London, London SW7 2AZ, UK.
3
Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK.

Abstract

The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell-material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy.

PMID:
28327660
PMCID:
PMC5364421
DOI:
10.1038/ncomms14843
[Indexed for MEDLINE]
Free PMC Article

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