Live/Dead staining
Following the protocol of the LIVE/DEAD® Viability/Cytotoxicity Assay
Kit (Invitrogen, America), a working solution was prepared by combining
20 µL of EthD-1 stock solution and 5 µL of calcein AM stock solution in
10 mL of D-PBS (Meilunbio, China). This working solution was applied to
cells that had undergone two washes with D-PBS. After a 30-minute
incubation, fluorescence microscopy was employed to capture images.