There is no single protocol for applying a mounting medium that’s ideal for all experiments. You will need to modify the general guideline below to better suit your specific requirements: 

1. Remove the mounting solution from the refrigerator and warm the bottle to room temperature. Make sure it is protected from light to minimize photobleaching. 
2. Remove excess liquid from your sample and apply a small drop of the mounting solution to the sample. If the sample is on a tissue culture dish or slide, take care to avoid air bubbles by carefully placing a coverslip on the drop. If the sample is on a coverslip, invert the coverslip on a clean glass slide. 
3. Remove any excess anti-fading agent.
4. Prepare samples for imaging by first incubating the mounting solution for about two hours to overnight. Use a plastic sealant or nail polish to seal the coverslip to the slide for long-term storage. Store mounted slides in the dark at 4 °C for optimum sample longevity. This will allow fluorescence imaging to remain stable for several weeks. Samples are ready to be imaged immediately after mounting.