Benjamin B. Bouchet

The ability of cells to migrate through a 3D matrix is associated with cancer invasion and metastasis. While it is recognized that the actin cytoskeleton plays a key role in interstitial migration, the role of microtubules in this process is still poorly understood. Most of the 3D cell culture setups consist in embedding cells in thick collagen I-based gels. In this type of cell culture system, the vast majority of optics available for confocal fluorescence imaging cannot be used for high-resolution observation and recording of cytoskeleton dynamics. By combining spinning disk-based confocal illumination with a new generation of high numerical aperture long working distance objective, we were able to perform high-resolution fluorescent imaging of microtubules in live cells that were fully embedded in a 3D matrix. The observations made with this new imaging setup revealed aspects of microtubule dynamics in physiologically relevant cell migration that are absent in conventional 2D cell culture systems.