Picro Sirius Red Staining Kit

Protocol Overview

1. Heat slides in an oven at 60°C for 45 min 2. During the heating process, prepare picro-sirius red stain solution (PSR). The recipe is as follows: 0.1% direct red 80 plus 0.1% fast green FCF dissolved in saturated aqueous picric acid (1.2% picric acid in water), the operation needs about 20 min, so it can be finished before deparaffinization steps. 2.1 First prepare saturated aqueous picric acid solution as follows (200ml): Weight about 2.4g picric acid solid to a bronze bottle, added with water to 200ml, shake for 5-10mins (Picric acid is stored in 36% water, so pull out water before weighing, remember to return the water back to the bottle after weighing). 2.2 Weigh 0.2g direct red 80 and 0.2g fast green FCF powder, respectively, and then added to above solution, continuous shaking for 5-10 min.3. Deparaffinize/hydrate simialr to normal IHC staining. 4. Apply adequate picro-sirius red solution prepared above to completely cover tissue section and incubate for 60 minutes. 5. Resin slide quickly in 10 dips of H2O. 6.Quickly dehydrate and mount in xylene. 7. Coverslip slide with clean mount Mounting medium.