PTAH STAIN (Principle and Diagnostic Application)

Principle

The PTAH staining method relies on acid-base chemistry to stain collagen and muscle fibers.
These fibers are demonstrated using a tungsten mordant provided by the phosphotungstic acid.

This mordant binds hematin and stains selective tissue components blue, while the
phosphotungstic acid is believed to stain other tissue components a red-brown color.

Tissue should preferably be fixed in Zenkers, which is believed to intensify staining reactions.
Additionally, tissues fixed in formalin are often post-fixed in Zenkers solution. A section of
skeletal muscle tissue can be used for quality control.

The amount of phosphotungstic acid in the staining solution is far greater than the amount of
hematein and it is believed that tungsten binds all available components blue, while the
phosphotungstic acid is thought to stain the red-brown components. This stain has been referred
to as a polychrome stain because one solution gives two major colors. The components colored
red-brown will lose this color with water or prolonged alcohol washes. Therefore, dehydration of
the tissue section following staining must be rapid.

Diagnostic application

1. Phosphotungstic Acid-Hematoxylin (PTAH) demonstrates and differentiates collagen and

muscle. It is especially preferred for the demonstration of cross-striations of skeletal muscle,
which may be lost in certain muscle diseases. These muscle cross-striations are also found in
tumors of muscle origin and rhabdomyosarcomas.
2. The identification of astrocytes is important in neuropathology as they react promptly to local
tissue injury, e.g. by increase in size in response to cerebral edema, or to metabolic
disturbances such as liver failure. Such cells are termed ‘reactive astrocytes’. Astrocytes
proliferate following damage to the central nervous system and will fill defects left by loss of
specialized nervous tissue with glial fibers. This is termed astrocytic gliosis and its identification
is important in analysis of disease processes in the CNS. Astrocyte stains may be performed to
demonstrate normal astrocytes, and reactive or gliotic astrocytes, but are frequently used to
demonstrate neoplastic astrocytes within an astrocytic tumor, i.e. astrocytoma, anaplastic
astrocytoma, glioblastoma multiforme, or mixed tumors such as
oligoastrocytoma, ganglioglioma, and others.
3. This stain highlighted the fibrin thrombi as hematoxyphilic blue showing numerous glomerular
fibrin thrombi in the glomerular capillaries in acute renal failure.
4. This stain highlighted the fibrin thrombi as hematoxyphilic blue showing fibrin
microthrombi in the vasculature in disseminated intravascular coagulation (DIC) and
inflammatory gastrointestinal disorders.