Dmi8 automated

To visually compare the effects of untreated, and Cis, AuNCs, US, US + Cis, US + AuNCs, and US + AuNCs + Cis treatments in A2780/A2780cis ovarian cancer cells, calcein-based cell viability assay, which interacts with living cells to emit green fluorescence, was performed according to the previously published protocol [27] (link). The treatments were carried out as previously described. After 48 h, RPMI-1640 medium was replaced with 3 µM of calcein (green) (Thermo Fisher Scientific, USA) which was dissolved in dimethyl sulfoxide (DMSO, Sigma, USA) and diluted with RPMI-1640. The cells were incubated for 2 h and the dye solution was carefully replaced with the culture medium. The images were captured using an inverted fluorescent microscope (Leica, DMI8 Automated, Germany). The green fluorescence intensities of the images were calculated with ImageJ.

Formalin fixed paraffin embedded tumor tissue from mice treated for 2 days were sliced manually with a manual microtome (Leica, RM2155) to 5 μm depth. H&E staining was preformed using Gill's II Hematoxylin (Sigma-Aldrich, GHS216) and Eosin Y (VWR, 95057-848). For IHC, slides were stained with cleaved caspase 3 antibody (Cell Signaling Technology, 9661), and counterstained with Gill's II Hematoxylin. Slides were visualized using a Leica DMi8 automated microscope and also scanned by HistoWiz for low-power visualization.