Aim:- It is a mix of hematoxylin and phosphotungstic acid. Muscle cross  striations, fibrin and glial fibers can be demonstrated by this stain.

Principle: The phosphotungstic acid all of the available hematin to form a blue lake pigment. This lake stains the muscle cross striations, fibrin and nuclei. The rest of the phosphotungstic acid stains red brown, components like collagen.

Control:- Skeletal muscle, cardiac muscle, fibrin, cerebral cortex for glial fibers.

Reagents:-

Hematoxylin                                           1gm
Phosphotungstic acid                          20gm
Distilled water                                       100ml

Dissolve the solid ingredients in separate portions of the water, the hematoxylin with aid of gentle heat. When cool mix it. Allow the mixture to ripen or add 177gm of potassium permagnate for immediate use. Properly ripened stain is rich purple in color and opaque.

0.25% potassium permagnate:-

Potassium permagnate                     0.25gm
Distilled water                                    100ml 

5% oxalic acid:-

Oxalic acid                                         5gm
Distilled water                                  100ml

Procedure:- 

1. Dehydrate and bring section to water.
2. Oxidize in potassium permagnate for 5 to 10 minutes. Discard the solution.
3. Wash in water.
4. Bleach in oxalic acid for 5 minutes or until the sections are colorless.
5. Wash thoroughly. Rinse in distilled water.
6. Put PTAH stain for 12 to 24 hour.
7. Transfer the section into 95% alcohol, followed by absolute alcohol.
8. Dehydrate, clear in xyline and mount in DPX.

Result:-

Striated muscle fibers                          blue 
Astrocytes                                              blue
Fibrin                                                       blue
Nuclei                                                      blue
Cytoplasm                                              brown red
Collagen                                                 brown pink