Quantifying the Autophagy-Triggering Effects of Drugs in Cell Spheroids with Live Fluorescence Microscopy

• DOI: 10.1007/978-1-4939-0856-1_3
• Publication Date: 2014-05-17T05:29:41Z
• Authors: Nariman Ansari; Stefanie Hardung; Katharina Hötte; Stefanie Rakel; Patrick Antonietti; Donat Kögel; Ernst H. K. Stelzer; Francesco Pampaloni
• Keywords: Spheroid; Live cell imaging

We present a 3D assay for the quantification of the autophagic flux in live cell spheroids by using the fluorescent reporter mRFP-GFP-LC3. The protocol describes the formation of the spheroids from the astrocytoma cell line U343, live long-term 3D fluorescence imaging of drug-treated spheroids, and the image processing workflow required to extract quantitative data on the autophagic flux.