Fast Red Super

To prepare the reagent, add five drops of buffer reagent B to 4.9 milliliters of distilled or deionized water in a test tube and mix well. Add a Fast Red tablet and allow it to stand in the solution for 5 to 10 minutes, then vortex for 3 to 5 minutes until the tablet is completely dissolved. Add 5 drops of substrate reagent S and mix. For IHC and ICC applications, following the alkaline phosphatase reaction, wash the slide with buffer and distilled or deionized water. Apply several drops of the prepared Fast Red reagent and incubate at room temperature or 37°C for 5 to 10 minutes. Wash the slides two to five times with buffer, followed by a rinse with distilled or deionized water. Apply a counterstain compatible with red color, such as hematoxylin, for 30 to 60 seconds, then wash with tap water or a buffer with a pH over 7 to transition the color to blue. The slide may be mounted with an aqueous mounting medium after a final rinse with distilled or deionized water, or it may be mounted with an organic mounting medium following dehydration through increasing concentrations of alcohol and clearing with limonene, toluene, or xylene.