4 6 diamidino 2 phenylindole dihydrochloride dapi dye

In this study, we used rhodamine phalloidin dye R-415 (Thermo Fisher Scientific, Waltham, MA, USA) for cytoskeleton labeling, and then used 4′,6-diamidino- 2-phenylindole, dihydrochloride (DAPI) dye (Thermo Fisher Scientific) for nuclei labeling in ARPE-19 retinal cells. We viewed and analyzed fluorescent images of cytoskeleton and nuclei in ARPE-19 retinal cells using Leica DM IRB inverted fluorescence microscope and Leica Application Suite software (LAS) V4.12 (Wetzlar, Germany) individually. We visualized the rhodamine-labelled cytoskeleton of ARPE-19 retinal cells by red fluorescence that was excited at 540 nm and detected the dye emission at 565 nm. We visualized DAPI-labelled nuclei of ARPE-19 retinal cells labeled by blue fluorescence that were excited at 358 nm and detected the dye emission at 461 nm.

In this study, Rhodamine Phalloidin dye R-415 (Thermo Fisher Scientific, Waltham, MA, USA) was used for cytoskeleton labeling, and 4′,6-Diamidino-2- Phenylindole, Dihydrochloride (DAPI) dye (Thermo Fisher Scientific), was used for nuclei labeling in the 4T1 breast cancer cells. The prepared 4T1 breast cancer cell specimens were securely mounted with coverslips. Fluorescence images of the 4T1 breast cancer cells were viewed using a Leica DM IRB inverted fluorescence microscope and analyzed using Leica Application Suite software (LAS) V4.12 (Wetzlar, Germany). To visualize the cytoskeleton labeled by Rhodamine, the cells were irradiated at 540 nm, and the dye emission was detected at 565 nm. The blue fluorescence of DAPI-labeled nuclei was excited at 358 nm, and the emission was detected at 461 nm. In addition, dichlorofluorescein diacetate (DCFDA) (Molecular Probes, Eugene, OR, USA) was used for reactive oxygen species (ROS) labeling, and the 3,3’-dihexyloxa- carbocyanine iodide (DiOC6 (3)) method (Sigma–Aldrich Co.) was used for mitochondrial membrane potential labeling in the 4T1 breast cancer cells. The mean fluorescence intensity (FL-1) of the 4T1 breast cancer cells was analyzed using an FACS Calibur flow cytometer (Becton–Dickison, PharMingen, Lake Franklin, NJ, USA).