Quantifying the autophagy-triggering effects of drugs in cell spheroids with live fluorescence microscopy.

Nariman Ansari,Stefanie Hardung, Katharina Hötte,Stefanie Rakel, Patrick Antonietti,Donat Kögel,Ernst H K Stelzer,Francesco Pampaloni

CANCER CELL SIGNALING: METHODS AND PROTOCOLS, SECOND EDITION（2014）

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摘要

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.

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关键词

Three-dimensional cell cultures,Live-cell assay,Glioma tumor spheroids,Autophagic flux,Drug development,Live cell imaging,Automated fluorescence microscopy,Live confocal microscopy

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