Modified Safranine O-Fast Green Cartilage Staining Kit 

Modified Safranine O-Fast Green Cartilage Staining Kit can be used for cartilage staining. Its staining principle lies in the fact that basophilic cartilage binds to the basic dye Safranine O, turning red, while eosinophilic bone binds to the acidic dye Fast Green, turning green or blue. This contrasts sharply with the red cartilage, thus distinguishing cartilage tissue from bone tissue.

Guidelines (The following are our recommended protocols. This protocol is for reference only and should be modified according to your specific needs.)
1. Specimen preparation: Fix in 10% formalin, decalcify, section in paraffin, and dewax to water using xylene or dewaxing clearing solution.
2. Stain with freshly prepared Weigert stain for 3-5 min, then rinse with water.
3. Differentiate with acidic ethanol differentiation solution for 15 s, then wash with distilled water for 5-10 min.
4. Immerse with Fast Green staining solution for 1-5 min.
5. Quickly wash sections with acetic acid solution for 10-15 s to remove residual Fast Green, then air dry (alternatively, quickly differentiate with acidic ethanol differentiation solution for 10-15 s, then rinse briefly with tap water).
6. Stain with Safranin O staining solution for 2-5 min.
7. Rapidly dehydrate four times with anhydrous ethanol, 3-5 seconds each time. After the fourth dehydration, examine under a microscope until the cartilage is red and the background is colorless.
8. Clear with xylene or environmentally friendly dewaxing and clearing solution, mount with neutral resin, examine under a microscope, and acquire and analyze the image.

Staining Results:
Cartilage matrix appears red, orange-red, or dark red;
Chondrocyte nuclei appear blue; chondrocyte cytoplasm appears red;
Cytoplasm, muscle, collagen fibers, and osteoblastic tissue appear grayish-green;
Cell nuclei appear blue-purple to dark blue.

Notes:
1. When cell nuclei need to be visualized, use iron hematoxylin staining whenever possible, as it has strong staining power and a rich hue. Regular hematoxylin has weak staining power.
2. Weigert staining solution should not be prepared in advance and left to stand. It generally loses its staining power after 24 hours.
3. If a large quantity of acetic acid solution is needed, a 0.05%-0.1% acetic acid aqueous solution can be prepared in-house.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

[Modified Safranine O-Fast Green Cartilage Staining Kit]

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.