1. Fixation: Tissue samples or cell cultures are fixed using agents like formaldehyde to preserve cellular structure and enzyme activity. 2. Incubation with X-gal: The fixed tissues are incubated with a solution containing X-gal, a substrate that β-galactosidase cleaves to produce a blue dye. 3. Washing: After incubation, the samples are washed to remove excess substrate. 4. Visualization: The samples are then examined under a microscope to identify regions of blue staining, which correspond to areas of β-galactosidase activity.
