The location of the mAuNPs captured by cells was determined with biological transmission electron microscopy and dark field microscopy. After a 24-hour incubation with mAuNPs at a concentration equivalent to single cell uptake, trypsinized B16-F10 cells were harvested by centrifugation and fixed overnight at 4°C in 3% glutaraldehyde solution (in PBS). The fixed cells were then dehydrated, polymerized, sectioned, and finally double-stained with uranyl acetate and lead citrate. Observations were performed on a JEM-1230 transmission electron microscopy (JEOL Co., Japan) at 100 kV. For the dark field microscopy experiment, B16-F10 cells were cultured with AuNPs for 24 hours in a Ф35 mm Petri dish with a coverslip placed in advance. The coverslip was placed on a glass slide and then observed under a dark field microscope (Olympus, Japan).

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