Slidders' Orange-Fuchsin-Green (OFG)
For Pituitary Cells
Materials
- Lendrum’s celestine blue
- A progressive hemalum, such as Mayer’s
- Solution A
Material Amount Orange G 0.05 g Ethanol, 95% 100 mL Phosphotungstic acid 0.5 g - Solution B
Material Amount Acid fuchsin 0.5 g Acetic acid, glacial 0.5 mL Distilled water 100 mL - Solution C
Material Amount Phosphotungstic acid 1 g Distilled water 100 mL - Solution D
Material Amount Light green SF yellowish 1.5 g Acetic acid, glacial 1.5 mL Distilled water 100 mL
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.
Protocol
- Bring sections to water via xylene and ethanol.
- Remove mercury pigment.
- Stain nuclei with Lendrum’s celestine blue-hemalum.
- Rinse with 95% ethanol.
- Place into solution A for 2 minutes.
- Rinse well with distilled water
- Place into solution B for 2-5 minutes, until basophiles are red.
- Rinse with distilled water.
- Differentiate with solution C for 5 minutes.
- Rinse with distilled water.
- Place into solution D for 2 minutes
- Rinse well with distilled water.
- Dehydrate with ethanols.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Best results are obtained if staining times are controlled visually.
Notes
- 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.
References
- Putt, F. A.
Manual of Histopathological Staining Methods
John Wiley & Sons, New York, NY., USA
Citing:
Slidders, W. J., (1961)
Journal of Pathology and Bacteriology, v. 82, p. 532
London, England.
