Slidders OFG
Orange-Fuchsin-Green for Pituitary

Solutions

Lendrum’s celestine blue

A progressive hemalum, such as Mayer’s

Solution A
Orange G	0.5	g
Ethanol, 95%	100	mL
Phosphotungstic acid	0.5	g

Solution B
Acid fuchsin	0.5	g
Acetic acid, glacial	0.5	mL
Distilled water	100	mL

Solution C
Phosphotungstic acid	1	g
Distilled water	100	mL

Solution D
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.

Method

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

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

Notes

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

 

Reference

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.