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Special Staining (I)

Special Staining (I). Carbohydrates. 1- PERIODIC ACID SCHIFF'S ( PAS ). Purpose: Glycogen is present in liver, kidney, skeletal and cardiac muscle.

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Special Staining (I)

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  1. Special Staining (I)

  2. Carbohydrates

  3. 1- PERIODIC ACID SCHIFF'S (PAS ) • Purpose: Glycogen is present in liver, kidney, skeletal and cardiac muscle. • The PAS stain is used to demonstrate neutral polysaccharides that are present in basement membranes, and mucus substances secreted from the epithelia of various organs. • A routine stain for: liver and kidney biopsies.

  4. Principle: The PAS stain is a histochemical reaction in that the periodic acid oxidizes the carbon to carbon bond forming aldehydes which react to the fuchsin-sulfurous acid which form the magenta color. • Results: Glycogen: magenta (red)

  5. 2- Alcian Blue pH 2.5 Acid Mucopolysaccharides PURPOSE: Alcian blue stains acid mucus substances and acidic mucins. • Excessive amounts of non-sulfated acidic mucus substances are seen in mesotheliomas, certain amounts occur normally in blood vessel walls but increase in early lesions of atherosclerosis.

  6. PRINCIPLE: Alcian blue is a group of polyvalent basic dyes that are water soluble. The blue color is due to the presence of copper in the molecule. - Alcian blue stains both sulfated and carboxylated acid mucopolysaccharides and sulfated and carboxylated glycoproteins. - It is believed to form salt linkages with the acid groups of acid mucopolysaccharides. • RESULTS: Acid mucins/mucus substances: blue

  7. 3- Alcian Blue-PAS (PAb) • Purpose: To differentiate between neutral and acidic mucus substances. • Routine stain for G.I. biopsies. • Results: Acid mucus substances: blue Neutral polysaccharides: magenta

  8. Acid mucus substances: blue Neutral polysaccharides: magenta

  9. 4- Mucicarmine Stain - Mucin PURPOSE: To stain acid mucopolysaccharides (mucin), which is a secretion produced by a variety of epithelial cells and connective tissue cells. • Excess mucin is secreted by epithelial cells in certain inflammations and in certain intestinal carcinomas. • The mucicarmine technique is also useful in determining the site of a primary tumor in that finding mucin positive tumor cells.

  10. Principle: aluminum is believed to form a chelation complex with the carmine, changing the molecule to a positive charge allowing it to bind with the acid substrates of low density such as mucins. • Results: Mucin: deep rose Nuclei: black Other tissue elements: yellow

  11. Mucin: deep rose Nuclei: black Other tissue elements: yellow

  12. Nucleic Acids

  13. Methyl Green Pyronin Stain • PURPOSE: The nuclear chromatin in the nucleus is composed of nucleoproteins. • The two types of nucleic acids are deoxyribonucleic acid (DNA), found in the nucleus, and ribonucleic acid (RNA), found in the nucleolus and cytoplasm. • DNA functions in cell heredity, and synthesis of RNA. • RNA functions principally in protein synthesis.

  14. Principle: This stain demonstrates RNA and DNA. DNA stains with methyl green, RNA is colored red with pyronine. • RESULTS: DNA: blue-green to green RNA: pink to red

  15. Methyl Green Pyronin Stain DNA: blue-green to green RNA: pink to red

  16. Feulgen stain • A selective cytochemical reaction for DNA in which sections or cells are first hydrolyzed with hydrochloric acid to produce apurinic acid and then are stained with Schiff reagent to produce magenta-stained nuclei. • Generally the concentration of DNA in nucleoli and mitochondria is too low to permit detection by this stain

  17. Feulgen stain

  18. Lipids

  19. Oil Red O • Purpose: To demonstrate fat or lipids in fresh frozen sections. • Fat occurring in an abnormal place, such as fatty emboli, and tumors arising from fat cells (liposarcomas) can be differentiated from other types of tumors. • Principle: Staining with oil-soluble dyes is based on the greater solubility of the dye in the lipoid substances than in the usual hydroalcoholic dye solvents. • Results: Fat: red Nuclei: blue

  20. Oil Red O Fat: red Nuclei: blue

  21. Sudan Black • Purpose: For the demonstration of fat using fresh frozen sections (cryostat sections). • Principle: Sudan Black is slightly basic dye and will combine with acidic groups in compound lipids, thus staining phospholipids also. • Results: Fat: blue-black Nuclei: red

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