The CISH procedure typically involves the following steps:
Sample preparation: Tissue sections are prepared and fixed on slides. Pre-treatment: The tissue is treated to permeabilize cell membranes and remove proteins that may interfere with hybridization. Hybridization: Labeled probes are applied to the tissue and allowed to hybridize with the target sequences. Washing: Excess probes are washed away to reduce background signals. Detection: Enzyme-conjugated antibodies are applied to bind to the labeled probes. Color development: A chromogenic substrate is added, producing a visible signal where the target sequence is present. Analysis: The stained tissue sections are examined under a bright-field microscope.
