Single-cell Raman microscopy of microengineered cell scaffolds

Studying cells in a three-dimensional (3D) environment has great potential in understanding cell behaviours such as morphology, proliferation, differentiation, and migration. Microengineered 3D cell scaffolds with precise defined geometries have offered a new approach to study cell behaviour and its interactions with scaffolds. The use of Raman spectroscopy to characterise biomolecules is a rapidly expanding area and has been implemented in numerous fields including pharmacology, microbiology, toxicology, and single-cell studies. However, one area where it remains unexploited despite the vast potential of the technique is in the investigation of 3D cell scaffolds. A combination of Raman microscopy and chemometric approaches have employed to investigate the structure and biochemistry of nanofabricated scaffolds and a cell-scaffold complex. The 3D Raman mapping combined with the use of nanofabricated 3D scaffolds offers a unique opportunity to assess the influence of scaffold architecture on cell body and cell nuclei morphology and biochemistry. For the first time, we have cultured a human epithelial colorectal adenocarcinoma cell line on OrmoComp scaffolds and determined the structure and biochemistry of nanofabricated scaffolds and a cell-scaffold complex with the use of Raman microscopy combined with appropriate data analysis protocols. The results demonstrate the potential of 3D Raman mapping for identifying biochemical and physical variation within single cells as they grow and adhere to 3D scaffolds.