96 well ultra low attachment treated spheroid microplates

A549 cells were seeded at the cell density of 2.5 × 10³ cells/well into Corning^™ 96-Well Ultra-Low Attachment Treated Spheroid Microplates and incubated for 72 h at 37 °C and 5% CO₂. Post 72 h, the spheroids were treated with FITC labeled BN-OH and BN/NAVI nanoparticles and 980 nm laser for 6 min were applied to evaluate the effect of laser on penetration efficiency. After 6 min of laser exposure, the formulations were incubated for 3 h and after that, the spheroids were washed with phosphate buffer saline and observed at 10× magnification under a confocal microscope at different focal lengths (Z-stack) and images were captured.

A549 cells were seeded at the cell density of 2.5 × 10³ cells/well in Corning^™ 96-Well Ultra-Low Attachment Treated Spheroid Microplates and incubated for 72 h at 37 °C and 5% CO₂. DCFH-DA dye was used as a fluorescent probe to measure ROS production in 3D spheroids. The spheroids were treated with BN-OH and BN/NAVI (at IC50 concentrations: 20 μM) and 980 nm laser was applied for 6 min. After 6 min, the spheroids were further incubated with the above formulations for 3 h. Then, the dye H₂DCFDA (5 μM) was added and incubated at 37 °C for 20 min and photographed using a confocal microscope in the green channel (Ex/Em: 492–495/517–527 nm).