4.11. Bioluminescence Analysis Using Copper Caged-Luciferin-1

Copper content of HCT-116 and HT-29 cells maintained in different experimental conditions, supplemented with TM or CuSO₄ was further assessed by in vivo bioluminescence imaging using a specific Copper-Caged-Luciferin-1 (CCL-1) kindly provided by C.J. Chang [16]. Briefly, HCT-116 and HT-29 cells (2.5 × 10³ cells/96 multi well dishes) were supplemented with TM at different doses (5 or 50 µM) for 24 hours. Then the medium was removed and 100 µL of CCL-1 (75 µM) were added into the dishes. As control a parallel set of cells were incubated with firefly luciferase. The plates were imaged with the IVIS Lumina II equipped with the Living Image software as previously described, and the data was analyzed and quantified [67].