Operetta imaging platform

A549 cells in black, micro-clear bottom 96-well plates were imaged using an Operetta Imaging platform (Perkin Elmer; Waltham, MA) with the following settings: four fields per well, 20X, long-working distance objective, and in wide-field mode. The cell plates were exposed to varying excitation spectra depending on the dyes in each well, and the emission spectrum for each dye was detected. Each dye was added to untreated cells (controls) to adjust for background staining and to assess dye interference from each color overlay. Cells intersecting the border of each image were excluded from the data analysis. Each image contains a reference bar which describes the level of spatial resolution.

OVCAR8 and 786-O cells were treated with pre-prepared nanoparticles at a concentration of 20 μM one day before treatment with ML210 (concentrations ranging from 20 μM to 0 μM). Cells were imaged at day 2, 3 and 6 of ML210 treatment on an Incucyte at 20x magnification and at day 6 with Hoechst DNA stain at 1:1000 on an Operetta imaging platform (PerkinElmer) for cell counting.