Muscle fibers are a component of muscle tissue and are composed of muscle cells. According to their morphological and functional characteristics, muscle fibers can be classified into smooth muscle (also known as non-striated muscle), skeletal muscle, and cardiac muscle. There are various staining methods for muscle fibers, such as the Ponceau method, Aniline Blue method, Tungsten Phosphotungstic Acid Hematoxylin method, etc. The Tannic acid-Azo Phloxine method achieves differential staining through the sequential action of two acidic dyes.
Muscle Fiber Staining Solution (Puchtler Tannic Acid-Azo Phloxine method) is mainly composed of Hematoxylin Staining Solution, Tannic Acid Differentiation Solution, Phosphomolybdic Acid Differentiation Solution, and Azo Phloxine Staining Solution. Its staining principle is that tannic acid readily penetrates collagen fibers with high permeability, staining them yellow; Azo Phloxine easily enters muscle fibers with lower permeability, staining them red. This staining solution is used to distinguish muscle fibers from collagen fibers with clear contrast and resistance to fading. It can also demonstrate myoepithelial cells and can be used in the diagnosis of myoepithelial tumors of the breast, skin, and other tissues. This reagent is for research use only. Not for use in clinical diagnosis or other purposes.
Product Components and Storage Conditions:
M1518566
| Component
| 5×50mL
| Storage
|
M1518566A
| Hematoxylin Staining Solution
| 50mL
| RT
|
M1518566B
| Tannic Acid Differentiation Solution
| 50mL
| RT
|
M1518566C
| Phosphomolybdic Acid Differentiation Solution
| 50mL
| RT . Store in the dark.
|
M1518566D
| Azo Phloxine Staining Solution
| 50mL
| 2-8℃. Store in the dark.
|
M1518566E
| Acid Alcohol Differentiation Solution
| 50mL
| RT |
Materials Required:
1. Bouin fixative, Carnoy fixative, neutral formalin, etc.
2. Distilled water, graded ethanol, xylene or eco-friendly dewaxing & clearing agent, neutral balsam
Protocol (For Reference Only):
1. Fix routine tissue specimens in Bouin fixative (Carnoy fixative or neutral formalin is also acceptable) for 2–3 hours.
2. Transfer directly into 95% ethanol for routine dehydration and embedding.
3. Prepare paraffin sections at 4 μm thickness; dewax with xylene or dewaxing & clearing agent and rehydrate to water.
4. Stain in hematoxylin staining solution for 8–10 minutes, then rinse in running water for 10 minutes.
5. Treat with tannic acid differentiation solution for 10 minutes, then rinse in running water for 1 minute.
6. Treat with phosphomolybdic acid differentiation solution for 10 minutes.
7. Stain in azo phloxine staining solution for 10–15 minutes.
8. Differentiate in acid alcohol differentiation solution for 3–5 seconds.
9. Dehydrate repeatedly in absolute ethanol, clear in xylene or dewaxing & clearing agent, and mount with neutral balsam.
Staining Results:
Muscle fibers, myoepithelial cells: Rose red
Collagen fibers: Yellow
Nuclei: Blue
Precautions:
1. Prefer Bouin fixative for tissue fixation; other fixatives may result in weaker staining.
2. Acid alcohol differentiation solution is critical. Differentiate until excess red dye is removed and muscle fibers appear bright red.
3. Azo phloxine staining solution is a supersaturated reagent. Use only the supernatant.
4. Use reagents promptly after opening to avoid affecting experimental results.
5. For your safety and health, wear a lab coat and disposable gloves during operation.
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