Live spheroid formation recorded with light sheet-based fluorescence microscopy

We provide a detailed protocol for a three-dimensional long-term live imaging of cellular spheroids with light sheet-based fl uorescence microscopy. The protocol allows the recording of all phases of spheroid formation in three dimensions, including cell proliferation, aggregation, and compaction. We employ the human hepatic cell line HepaRG transfected with the fusion protein H2B-GFP, i.e., a fluorescing histone. The protocol allows monitoring the effect of drugs or toxicants. © Springer Science+Business Media New York 2015.