Abstract
We provide a detailed protocol for a three-dimensional long-term live imaging of cellular spheroids with light sheet-based fluorescence 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.
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Acknowledgments
The authors thank the Deutsche Forschungsgemeinschaft (DFG) and the German Federal Ministry of Education and Research (BMBF) for financial support (project ProMEBS). We thank Berit Langer for her outstanding support.
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Pampaloni, F., Richa, R., Ansari, N., Stelzer, E.H.K. (2015). Live Spheroid Formation Recorded with Light Sheet-Based Fluorescence Microscopy. In: Verveer, P. (eds) Advanced Fluorescence Microscopy. Methods in Molecular Biology, vol 1251. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2080-8_3
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DOI: https://doi.org/10.1007/978-1-4939-2080-8_3
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2079-2
Online ISBN: 978-1-4939-2080-8
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