Safranin O-Fast Green stain can well demonstrate cartilage layer and tidal line structures, excellently display structures of cartilago articularis, subchondral bone and bone tissues. This product stain cartilage matrix with homogenous red and green for bone with sharp contrast.
1.Deparafiinize and rehydrate to distilled water.
2.Staining with Fast-Green solution 5-10 min, wash to remove the excess stain solution until cartilage is colorless. Rinse in hydrochloric acid alcohol slight to differentiate, wash in tap water.
3.Staining in Safranin O solution for 13-30 s, rapid dehydration by 3 cylinders absolute alcohol.
4.Clear in xylene 5 min, mount with resin mounting medium.
5.Microscopy determination and analysis.
Cartilage is red or orange in green background.
1.If staining is unsuccessful, wash in water to clear fast-green and differentiate in hydrochloric acid alcohol to eliminate Safranin O. when all colors are cleared, restain slides starting from step 2.
2.Staining time has a great impact on coloring degree. If the cartilages show significant green after staining in Fast-Green, differentiate in water is necessary to decolorize to light green or colorless. On the other way, if bone tissues show light color after differentiate, restain in Fast-Green and re-differentiate with water if necessary. The final aim is to show light green or colorless in cartilage while sharp green in bone tissues.
3.Precipitations may occur in Safranin O when over-used, result low efficiency to stain. Please change fresh Safranin O solution in time.