Cartilage tissue is composed of chondrocytes and cartilage matrix. Cartilage tissue and the surrounding perichondrium constitute cartilage. There are many staining methods for cartilage, such as toluidine blue method, alcian blue method, safranin O method, etc. According to the different fibrous components in the matrix, cartilage can be divided into hyaline cartilage, elastic cartilage and fibrocartilage. Hyaline cartilage is mainly distributed in the upper respiratory tract, medial end of the ribs, articular surfaces of short bones and other sites.
Unna alkaline methylene blue, also known as polychrome methylene blue, is an oxidized mixture of methylene blue. It exhibits polychromatism including blue, purple and red after preparation by a specific process, and has unique applications in microbiology, pathology and materials science. Hyaline Cartilage Staining Solution (Unna alkaline methylene blue method) uses alizarin red combined with alkaline methylene blue for staining. Bone tissue (where calcium salts are deposited) binds to alizarin red and appears red; cartilage reacts with polychrome methylene blue and appears purple; cytoplasm is yellow and nuclei are blue. This method is the optimal staining method for fetal phalanges at 6–8 months of gestation. It is composed of potassium carbonate, methylene blue, alizarin red, etc. This reagent is for research use only and not suitable for clinical diagnosis or other purposes.
Product Components and Storage Conditions:
T1511541
| Component
| 2×50mL
| Storage
|
T1511541A
| Alizarin Red S Staining Solution
| 50mL
| RT
|
T1511541B
| Unna Alkaline Methylene Blue Staining Solution
| 50mL
| RT. Store in the dark. |
Materials Required (User-supplied):
1. Graded ethanol, distilled water, xylene or environment-friendly dewaxing solution, environment-friendly clearing solution, neutral balsam
2. Microscope
Protocol (For Reference Only):
1. Fix tissues in 90% ethanol for 12–24 hours, embed in paraffin and section. Section thickness: 3–5 μm. Dewax to water, then rinse with distilled water.
2. Stain in Alizarin Red S Staining Solution for 5–20 minutes, rinse briefly with distilled water.
3. Counterstain in Unna Alkaline Methylene Blue Staining Solution for 2–6 minutes, differentiate in 95% ethanol until staining is clear.
4. Perform routine dehydration, clear in xylene or environment-friendly clearing solution, and mount with neutral balsam.
Staining Results:
Areas of calcium deposition: red
Cartilage matrix: purple
Cytoplasm: yellow
Cell nuclei: blue
Precautions:
1. For staining insoluble calcium, the staining time of Alizarin Red S should be determined according to calcium content, and observed under a microscope until the staining is intense and non-diffuse.
2. After staining with Alizarin Red S Staining Solution, calcium deposits are birefringent.
3. Unna Alkaline Methylene Blue Staining Solution is volatile; cover the container during staining.
4. For your safety and health, please wear a lab coat and disposable gloves during operation.
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