In drug discovery programs, multicellular spheroids have emerged as powerful tools to bridge the gap between in vitro cell culture models and in vivo tissues. However, one of the greatest challenges in higher throughput 3D imaging is the acquisition of images of solid spheroids, owing to the reduced light penetration.
One solution is to use optical clearing techniques, which can enhance the imaging depth in spheroids by removing lipid and protein molecules.
In this technical note, we compare different optical clearing strategies for 3D spheroids and quantify the clearing effectiveness and alterations in spheroid morphology, and demonstrate how to increase imaging depth in 3D spheroids by a factor of four.