FluoroQuest™ Anti-fading Kit I *Optimized for Slide Imaging*

Protocol summary

1. Prepare Samples (slides or microplate wells)
2. Add a drop of a component and mount
3. Examine the specimen under microscope

Important notes
These ready-to-use anti-fading reagents can be applied directly on the washed specimen. Although the reagents have been tested with lots of fixed samples, their optimal anti-fading efficiencies strongly depend on the properties of your samples. We suggest that you try more than one component for your imaging samples to get the ideal component. For example, one component may be more compatible with a fluorescent labeled antibody conjugate (or an enzyme substrate or a special mounting specimens that contain lipophilic plasma membrane stains like DiI) than another one.

Table 1. Recommendations for usage

1. Thaw all the kit components at room temperature, and keep from light.
   
   
2. Remove any excess liquid from your specimen. Add a small drop of the selected component to the specimen. If the sample is on a slide or tissue culture dish, carefully place a coverslip on the drop, avoiding air bubbles. If the sample is on a coverslip, invert the coverslip on a clean glass slide. Remove any excess anti-fading component.
   
   
3. The anti-fading reagents should be incubated for 2 hours to overnight. For long-term storage, seal the coverslip to the slide with nail polish or a plastic sealant. Mounted slides should be stored at 4ºC in the dark for optimum sample longevity. The fluorescence imaging would remain stable for many weeks. Samples can be imaged immediately after mounting. A typical image is shown in Figure 1.