Collagen Staining by Picro-Sirius Red

Protocol After Paraffin Block Sectioning

1. Rehydration (Xylene 2x5 minutes, ethanol 100% 2 minutes, ethanol 95% 2x2 minutes, ethanol 70% 2 minutes, deionized water 2 minutes) 
2. Picro-Sirius Red staining 1 hours then wash deionized water 10 dips
3. Gill Hematoxylin for 5 minutes then wash in tap water for 5 minutes  
4. Dehydration (ethanol 70% 10 seconds, ethanol 95% 20 seconds, ethanol 100% 10 seconds, xylene 2x10 seconds) 
5. Mounting in Xylene based medium

Result

Recepie for Picro-Sirius Red:

1. Weight 2.4 gr 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. Add 0.2 gr Direct Red 80 to above solution, then shaking/mixing for 5 minutes

Recommended Reagents:

1. Picric acid (Sigma Aldrich) NOTE: Becareful picric acid is toxic and potentially explosive when dry)
2. Direct Red 80 (Sigma Aldrich)