PicroSirius red staining

Edited by Junyan 05/21/12

Materials

Methods

1. Heat slides in an oven at 60°C for 45 min (not similar to IHC, could done in the same day)
2. During the heating process, prepare picro-sirius red stain solution (PSR). The recipe is as following: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 could finished before deparaffinize steps.
   1. Firstly prepare saturated aqueous picric acid solution as following(200ml): Weight about 2.4g picric acid solid to a bronze bottle, added with water to 200ml, shaking for 5-10mins(Picric acid stored in 36% water, so pulled out water before weighting, remember to return back the water to the bottle after weighting).
   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 using the following series of washes: Xylenes 2X 5 min 100% EtOH 2X 2 min 95% EtOH 2X 2 min 70% EtOH 1X 2 min 50% EtOH 1X 2 min dH2O 2X 2 min Outline with PAP pen dH2O 2X 3 min
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. Quickly resin in 70% EtOH 1X 20s 100% EtOH 2X 10 dips Xylenes 2X 10dips
7. Coverslip slide with clean mount mounting medium.