Muscle fibers are components of muscle tissue and are composed of muscle cells. According to their morphological and functional characteristics, muscle fibers can be classified into smooth muscle, skeletal muscle, and cardiac muscle. There are many staining methods for muscle fibers, such as Ponceau staining, Aniline Blue staining, and Phosphotungstic Acid Hematoxylin staining. When phosphotungstic acid hematoxylin staining solution was first developed, Mallory’s PTAH method included several variants of phosphotungstic acid hematoxylin staining. Around 1900, Mallory combined aqueous phosphotungstic acid solution with hematoxylin staining solution and found this method produced excellent staining of muscle fibers. The widely used formulation today is Mallory Phosphotungstic Acid Hematoxylin Staining Solution (PTAH, natural oxidation method). Hematoxylin can also be prepared by the PTAH chemical oxidation method, but it has a shorter shelf life and tends to lose staining potency. Although natural oxidation is time‑consuming, the resulting hematoxylin solution can be stored for more than 2 years with stable staining capacity, making it an ideal staining solution. It is suitable for tissues of the central nervous system, general histological structures, and tissues fixed with all standard fixatives. Staining time varies depending on the preparation method, fixative used, and the histological structures to be demonstrated.
Mallory Phosphotungstic Acid Hematoxylin Staining Solution (PTAH Natural Oxidation Method) mainly consists of PTAH oxidant, oxalic acid solution, and Mallory PTAH staining solution. The Mallory PTAH staining solution is naturally ripened, with good staining strength and long storage life. It is mostly used to demonstrate cross‑striations in striated muscle and is clinically applied in the diagnosis of rhabdomyosarcoma. Rhabdomyosarcoma shows diverse histological morphology and is difficult to distinguish from undifferentiated mesenchymal tumors. After phosphotungstic acid hematoxylin staining, the identification of blue cross‑striations in the cytoplasm of tumor cells confirms striated muscle differentiation. This staining solution can also be used to stain fibrin in inflammatory exudates, fibrin in capillaries in DIC, and in neuropathological examinations. This reagent is for research use only and not for clinical diagnosis or other purposes.
Product Components and Storage Conditions:
M1518515
| Component
| 3×100mL
| Storage
|
M1518515A
| PTAH Oxidant A
| 50mL
| RT. Store in the dark.
|
M1518515B
| PTAH Oxidant B
| 50mL
| RT
|
M1518515C
| Oxalic Acid Solution
| 100mL
| RT
|
M1518515D
| Mallory PTAH Staining Solution (Natural Oxidation Method)
| 100mL
| RT. Store in the dark.
|
Immediately before use, mix equal volumes of A and B to prepare PTAH Oxidant. Prepare and use immediately.
Materials Required (User-supplied):
1. 10% Formalin fixative, distilled water, graded ethanol, xylene or eco-friendly dewaxing clearing agent, neutral balsam
2. Light microscope
Protocol (For Reference Only):
1. Fix tissues in 10% formalin, followed by routine dehydration and embedding.
2. Prepare 4 μm paraffin sections, dewax with xylene or dewaxing clearing agent, and hydrate to water.
3. Oxidize in freshly prepared PTAH Oxidant for 5 min, then rinse briefly with water.
4. Bleach in oxalic acid solution for 1–2 min, wash in running water for 2 min, then rinse once with distilled water.
5. Stain in Mallory PTAH Staining Solution (covered) for 24–48 h.
6. Take out sections and quickly wash off excess stain with 95% ethanol.
7. Perform routine dehydration, clear with xylene or dewaxing clearing agent, and mount with neutral balsam.
Staining Results:
Cross-striations of striated muscle, fibrin, nuclei, erythrocytes, glial fibers: Deep blue
Collagen fibers, cartilage matrix: Reddish-brown
Coarse elastic fibers: Purple
Precautions:
1. If the blue color of cross-striations is weak or appears bright red during staining, it indicates insufficient or excessive oxidation. Replace the staining solution or prepare a fresh one.
2. Do not wash with water after Mallory PTAH staining, and rinse quickly with 95% ethanol. Prolonged washing with water or ethanol will remove the phosphotungstic acid hematoxylin stain.
3. Mallory PTAH Staining Solution is a progressive stain; avoid overstaining. After 24 h of staining, sections can be examined under a microscope to check staining intensity.
4. For your safety and health, wear a lab coat and disposable gloves during operation.
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