Staining Procedure:

1. Dewax the tissue section by immersing the tissue section slide in dimethylbenzene for 10 mins. Repeat this step again with fresh dimethylbenzene.
2. Immerse the tissue section slide in 100%, 95% and 75% ethanol for 1 min, respectively. Rinse the slide with water.
3. Add the Fast Green Staining Solution onto the tissue section on the slide and wait for 15-20 mins. During this time the bone tissue will be stained dark green, while the cartilage will be colorless. 
4. Wash the slide slightly and carefully with water. Do not wash for longer time to avoid washing off the Fast Green Stain.
5. Add the Safranin Staining Solution onto the tissue section on the slide and wait for 1-3 mins until the cartilage turns bright red.
6. Wash the extra Staining Solution from slide with water.
7. Immerse the tissue section slide into the color seperation solution (Anhydrous Ethanol) for 10 sec-10 mins. Observe the section carefully under the microscope, to control the degree of Green and Red stain.
8. Dry the section in room temperature and seal with neutral gum or using a cover slip.