Can I run multiplex IHC using fluorescent detection?

Yes, you can run multiplex IHC using fluorescent detection. The wide array of fluorochromes available enables multiple targets to be detected simultaneously by emitting light at distinct wavelengths. When running multiplex IHC experiments, the key to minimizing spectral overlap is to choose your fluorochromes carefully. You must also take care to design your multiplex fluorescent experiment to reduce cross-reactivity. 

One effective strategy is to select primary antibodies from different host species, which largely mitigates issues with cross-reactivity. In such instances, species-specific secondary antibodies will specifically bind to only one primary antibody, ensuring accurate detection.