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Quantitative volumetric Raman imaging of three dimensional cell cultures

Research output: Contribution to journalArticlepeer-review

Charalambos Kallepitis, Mads S Bergholt, Manuel M Mazo, Vincent Leonardo, Stacey C Skaalure, Stephanie A Maynard, Molly M Stevens

Original languageEnglish
Article number14843
Number of pages9
JournalNature Communications
Volume8
Issue number1
Early online date22 Mar 2017
DOIs
Accepted/In press3 Feb 2017
E-pub ahead of print22 Mar 2017
Published22 Mar 2017

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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.

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