Prolong Gold Antifade Reagent from Invitrogen

Immunofluorescent experiments require enormous attention to detail. When these experiments require fixed samples, the investigator must properly fix, stain, wash, counterstain, and mount each experiment. Only after this can the samples be imaged. Optimizing each step is essential to obtaining publication quality images. Although the procedure required for each sample is unique, mounting the samples onto a slide is one common concern for any investigator.

There are quite a few options of mounting solutions on the market today. The most important property of these solutions is their ability to delay photobleaching. The real strength behind the Prolong® Gold is the ability to slow photobleaching throughout the visible spectrum and into the Far Red spectrum. Although alternatives are competitive with slowing photobleaching in the visible spectrum, none match Prolong Gold's ability to delay photobleaching in the Far Red spectrum.

Immunoflourescent assays are a staple in our lab. We work with live and fixed cells, and occasionally tissue sections. Although some experiments only require observing a few proteins of interest, recently we have been using four fluorophores (blue, green, red, and far red). This requires the use of the Far Red spectrum, and the signal from these fluorophores was noticeably bleaching during imaging. Eventually, we discovered that the mounting solution we were using could not significantly slow photobleaching in the Far Red spectrum. Since we switched to Prolong Gold, we have observed a significant improvement.

There are two versions of Prolong Gold, either containing DAPI or not. Our lab has one bottle of each, clearly labeled, in case an experiment does not specifically require DAPI. Another important consideration is the fact that Prolong solidifies and holds the coverslip to the slide making that bottle of clear nail polish a thing of the past. Overall, this is a versatile mounting solution that makes immunofluorescent experiments more robust.