Why are the permeabilization procedures critical to in situ hybridization?

Effective permeabilization is crucial for in situ hybridization because the proteins surrounding the target DNA or RNA sequences can block the access of hybridization probes. These proteins are often extensively cross-linked, making it challenging for the probe to invade the tissue and bind to its target. Therefore, permeabilization procedures are essential for removing these issues and ensuring successful hybridization between the probe and the target nucleic acid sequences.