The ATPase staining procedure involves several steps:
1. Sample Preparation: Muscle tissue samples are collected and frozen. The frozen sections are then cut using a cryostat. 2. Pre-incubation: The sections are incubated in a series of buffers with different pH levels (commonly pH 4.3, 4.6, and 9.4). 3. Staining: After pre-incubation, sections are incubated in a solution containing ATP. The enzyme activity leads to the formation of a precipitate that can be visualized using a specific dye, often cobalt chloride and ammonium sulfide. 4. Differentiation: Based on the pH sensitivity, different muscle fibers will exhibit varying degrees of staining intensity, allowing for their classification.
