Safranin O / Fast green stain
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This method is used for the detection of cartilage, mucin, and mast cell granules on formalin-fixed, paraffin-embedded tissue sections, and may be used for frozen sections as well. The cartilage and mucin will be stained orange to red, and the nuclei will be stained black. The background is stained bluish green.
Fixation: Formalin fixed, paraffin embedded sections.
1) Deparaffinize and hydrate slides to distilled water.
2) Stain with Weigert’s iron hematoxylin working solution for 10 minutes.
3) Wash in running tap water for 10 minutes.
4) Stain with fast green (FCF) solution for 5 minutes.
5) Rinse quickly with 1% acetic acid solution for no more than 10 –15 seconds.
6) Stain in 0.1% safranin O solution for 5 minutes.
7) Dehydrate and clear with 95% ethyl alcohol, absolute ethyl alcohol, and xylene, using 2 changes each, 2 minutes each.
8) Mount using resinous medium.
Cytoplasm: bluish green
Cartilage, mucin, mast cell granules: orange to red