What Are the Limitations of Using β Galactosidase in Histology?

Despite its widespread use, there are limitations to using β Galactosidase in histology:

Endogenous Activity: Some tissues may have endogenous β Galactosidase activity, which can lead to background staining and false-positive results.
Enzyme Stability: The enzyme can be sensitive to fixation and processing conditions, potentially leading to loss of activity.
Substrate Diffusion: Diffusion of chromogenic or fluorogenic substrates can sometimes blur the boundaries of enzyme activity, complicating the interpretation of results.