Quantitative volumetric Raman imaging of three dimensional cell cultures.
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.
Department of Materials, Imperial College London, London SW7 2AZ, UK.|Department of Bioengineering, Imperial College London, London SW7 2AZ, UK.|Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK.
Charalambos
C
Kallepitis
Department of Materials, Imperial College London, London SW7 2AZ, UK.|Department of Bioengineering, Imperial College London, London SW7 2AZ, UK.|Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK.
Mads S
MS
Bergholt
Department of Materials, Imperial College London, London SW7 2AZ, UK.|Department of Bioengineering, Imperial College London, London SW7 2AZ, UK.|Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK.
Manuel M
MM
Mazo
Department of Materials, Imperial College London, London SW7 2AZ, UK.|Department of Bioengineering, Imperial College London, London SW7 2AZ, UK.|Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK.
Vincent
V
Leonardo
Department of Materials, Imperial College London, London SW7 2AZ, UK.|Department of Bioengineering, Imperial College London, London SW7 2AZ, UK.|Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK.
Stacey C
SC
Skaalure
Department of Materials, Imperial College London, London SW7 2AZ, UK.|Department of Bioengineering, Imperial College London, London SW7 2AZ, UK.|Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK.
Stephanie A
SA
Maynard
Department of Materials, Imperial College London, London SW7 2AZ, UK.|Department of Bioengineering, Imperial College London, London SW7 2AZ, UK.|Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK.
Molly M
0000-0002-7335-266X
MM
Stevens
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D006801:Humans; D021621:Imaging, Three-Dimensional; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D008055:Lipids / Q000032:analysis; D008264:Macrophages / Q000166:cytology; D009000:Monocytes / Q000166:cytology; D032383:Myocytes, Cardiac / Q000166:cytology; D017207:Rats, Sprague-Dawley; D013059:Spectrum Analysis, Raman
D008055:Lipids