O Safranin Staining (Cartilage staining):
Stains, cartilage, mucin and also mast cells.
Is used for Frozen section or paraffined tissue sections,
Materials and Solutions:
Weigert's Iron Hematoxylin:
50ml hematoxilyn, 1% Feric Chlorid (in ddH2O), 0.5ml Hcl (concentrated).
make to 100ml (store up to 1 month)
Mix equal parts of stock solution A and B. This working solution is stable for about 4 weeks.
0.1% Safranin O
0.001% Fast Green
1% Acetic Acid
Section should be hydrated to ddH20
1. Stain with for 10min with Weigerts iron hematoxylin.
2. Wash in running tap water for 15 minutes (use coplin jar).
3. Stain with 0.001%fast green, 5 minutes (no more then 5min).
4. Rinse twice quickly with 1% acetic acid.
5. Stain with 0.1% safranin O for 5 minutes.
6. Dehydrate using these steps (ecry step twice for 2min each): 1- 95% ethyl alcohol, 2- absolute ethyl alcohol, 3- xylene
Cartilage, mucin, mast cell will be Red, Nuclei black, Cytoplasm green.
1. Kahveci Z, Minbay FZ, Cavusoglu L (2000) Safranin O staining using a microwave oven. Biotech Histochem. 75(6):264-8.
2. Camplejohn KL, Allard SA. Limitations of safranin 'O' staining in proteoglycan-depleted cartilage demonstrated with monoclonal antibodies. Histochemistry. 1988;89(2):185-8.