Preparing NFR involves dissolving the dye in a suitable solvent, usually water or an aqueous medium, and then applying it to tissue sections. Here is a general protocol for using NFR:
Deparaffinize and rehydrate tissue sections if they are embedded in paraffin. Stain the sections with the primary stain (e.g., hematoxylin) as required. Rinse the sections in water to remove excess primary stain. Immerse the sections in NFR solution for a specified duration, usually a few minutes. Rinse the sections again to remove excess NFR. Dehydrate, clear, and mount the sections for microscopic examination.
