The procedure for Oil Red O staining involves several steps: 1. Fixation: Tissue samples are fixed using formalin or another appropriate fixative to preserve cellular structures. 2. Cryosectioning: Because lipids are soluble in alcohols and xylene, paraffin embedding is unsuitable. Therefore, samples are usually cryosectioned. 3. Staining: Sections are stained with Oil Red O solution, which involves dissolving the dye in a solvent like isopropanol. 4. Counterstaining: Sections may be counterstained with hematoxylin to provide contrast. 5. Mounting: Stained sections are mounted in an aqueous mounting medium to prevent dissolution of lipids.
