The ATPase staining procedure typically involves the following steps:
1. Tissue Preparation: Muscle tissue is collected and frozen to preserve enzyme activity. Cryosectioning is performed to obtain thin tissue sections. 2. Incubation: The tissue sections are incubated in a solution containing ATP at a specific pH (usually pH 4.3, 4.6, or 9.4) to activate the enzyme. 3. Color Development: After incubation, the sections are incubated in a solution containing calcium ions, which precipitates with inorganic phosphate to form a visible product. 4. Counterstaining: Optional counterstaining with dyes such as hematoxylin may be performed to provide contrast and enhance visualization. 5. Microscopy: The stained sections are examined under a microscope to identify the different types of muscle fibers based on their staining patterns.
