Table 1.

Troubleshooting.

Issue	Solution
Higher Sytox intensity in stimulated cells than with Triton X-100 leading to cell death higher than 100%	Resuspend more vigorously and repeat after 1 h Increase Triton X-100 concentration
Trypsinization incomplete for D10 spheroids	Include an incubation step with collagenase I instead of using trypsin
Spheroids disaggregate after 1 day during ferroptosis assay when using adherent plates	Use suspension plate instead of adherent plates to avoid cells adhering to plate
Intensity too high: overestimated	Use lower gains Optimize gain settings for new cell lines