The basic principle of ISH involves the use of a labeled probe that is complementary to the target sequence. The probe can be labeled with fluorescent dyes, radioactive isotopes, or enzymes for detection. The steps typically include:
Preparation: Tissue sections or cell samples are fixed to preserve morphology and nucleic acids. Hybridization: The labeled probe is applied to the sample, where it binds to its complementary sequence. Washing: Excess probe is washed away to reduce non-specific binding. Detection: The bound probe is visualized using appropriate methods, such as fluorescence microscopy or autoradiography.
