Label-free volumetric quantitative imaging of human osteosarcoma cells by hyperspectral coherent anti-Stokes Raman scattering

Quantitative determination of the chemical composition of unstained samples, non-invasively, with high three- dimensional spatio-temporal resolution, will accelerate progress in cell biology. The current state of the art in bioimaging is dominated by either chemically non-specific or invasive methods. In this work, we demonstrate label-free, non-invasive quantitative volumetric imaging of human osteosarcoma cells using coherent anti-Stokes Raman scattering microscopy. A data analysis method developed in-house was applied to represent the chemical composition of the cells as volumetric three-dimensional images indicating water, proteins, DNAP (mixture of DNA and proteins), and lipids, and to determine the dry masses of the organic components with picogram resolution.