Imaging growth of neurites in conditioned hydrogel by coherent anti-Stokes Raman scattering microscopy

Cultured DRGs in different gel scaffolds were analyzed using CARS microscopy to determine its possible use as a label-free imaging option for tracking cellular growth in a gel scaffold. This study demonstrates for the first time the applicability of CARS microscopy to the imaging of live neuronal cells in GAG hydrogels. By tuning the laser beating frequency, ωp ? ωs, to match the vibration of C-H bonds in the cell membrane, the CARS signal yields detailed, high-quality images of neurites with single membrane detection sensitivity. The results demonstrate that CARS imaging allows monitoring of cellular growth in a tissue scaffold over time, with a contrast that shows comparable cellular structures to those obtained using standard fluorescent staining techniques. These findings show the potential of CARS microscopy to assist in the understanding of organogenesis processes in a tissue scaffold.