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Slidders’ Orange-Fuchsin-Green for Pituitary Cells

Slidders' Orange-Fuchsin-Green (OFG)

For Pituitary Cells



  • Lendrum’s celestine blue
  • A progressive hemalum, such as Mayer’s
  • Solution A
    Orange G0.05g
    Ethanol, 95%100mL
    Phosphotungstic acid0.5g
  • Solution B
    Acid fuchsin0.5g
    Acetic acid, glacial0.5mL
    Distilled water100mL
  • Solution C
    Phosphotungstic acid1g
    Distilled water100mL
  • Solution D
    Light green SF yellowish1.5g
    Acetic acid, glacial1.5mL
    Distilled water100mL

Tissue Sample

Although 5µ paraffin sections of neutral buffered formalin fixed tissue are likely adequate, superior results are obtained with fixation in formal sublimate, preferably about 48 hours. Other fixatives may be satisfactory. Formalin fixed tissues may benefit from secondary fixation of sections in Bouin’s fluid.


  1. Bring sections to water via xylene and ethanol.
  2. Remove mercury pigment.
  3. Stain nuclei with Lendrum’s celestine blue-hemalum.
  4. Rinse with 95% ethanol.
  5. Place into solution A for 2 minutes.
  6. Rinse well with distilled water
  7. Place into solution B for 2-5 minutes, until basophiles are red.
  8. Rinse with distilled water.
  9. Differentiate with solution C for 5 minutes.
  10. Rinse with distilled water.
  11. Place into solution D for 2 minutes
  12. Rinse well with distilled water.
  13. Dehydrate with ethanols.
  14. Clear with xylene and mount with a resinous medium.

Expected Results

  • Best results are obtained if staining times are controlled visually.


  • Nuclei – black
  • Erythrocytes – orange
  • Collagen – green
  • Acidophils – orange
  • Basophils – red
  • Chromophobes – pale gray-green

Safety Note

Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.


  1. Putt, F. A.
    Manual of Histopathological Staining Methods
    John Wiley & Sons, New York, NY., USA
    Slidders, W. J., (1961)
    Journal of Pathology and Bacteriology, v. 82, p. 532
    London, England.