Biotek cytation 3 automated fluorescence microscope

To visualize the effects of CBD treatment on Salmonella cells, fluorescence microscopy was utilized. Salmonella cells were fixed and stained using DAPI (4′,6-diamidino-2-phenylindole) to determine cell membrane integrity. DAPI stain binds exclusively to dsDNA, which is only accessible as a result of compromised membrane integrity and expulsion of the cytoplasmic material including the nucleus out of the cell. Visualization of DAPI fluorescence under the fluorescence microscope (Biotek Cytation™ 3 Automated Fluorescence Microscope) (Agilent Technologies Inc., Santa Clara, CA, USA) confirmed cell membrane damage and hence cell death. Salmonella cells were treated with CBD dilutions (1.25, 0.125, or 0.0125 μg/mL) or left untreated (control). Following treatment, cells were fixed at either 5 min or 30 min and assessed using fluorescence microscopy [27 (link)].

To investigate the effect of capsaicin on Vero cells, the cells were cultured in 96-tissue well plates containing DMEM media supplemented with 10% FBS. Capsaicin at 1000 µg/mL, 100 µg/mL, 10 µg/mL, 1 µg/mL, 0.1 µg/mL, 0.01 µg/mL and 0.001 µg/mL was mixed with the culture media and incubated with capsaicin for 8 h. Control cultures did not receive capsaicin treatment. At the 8 h time point, AlamarBlue at 10% was added to the growing culture and allowed to incubate for an additional 4 h. Absorbance at 570 nm was read using Cytation 3 imaging/plate reader (Biotek Cytation™ 3 Automated Fluorescence Microscope) (Agilent Technologies Inc., Santa Clara, CA, USA) plotted on a bar chart to evaluate the percentage of cell viability.