Herovici's Stain
for Young and Mature Collagen

Solutions
Weigert's iron hematoxylin or the Celestine blue hemalum sequence

Solution A
Picric acid, sat. aqu.	100	mL
Acid fuchsin, 1% aqu.	10	mL

Staining solution
Solution A	50	mL
Methyl blue, 0.05% aqu.	50	mL
Glycerol	10	mL
Lithium carbonate, sat. aqu.	0.5	mL

Tissue sample
Paraffin sections at 5µ of formol-acetic-ethanol (10:5:85) fixed tissue was recommended.

Method

1. Bring sections to water via xylene and ethanol.
2. Stain nuclei with the acid resistant nuclear stain.
3. Rinse well with tap water.
4. Place into the staining solution for 2 minutes.
5. Wash with 1% acetic acid for 2 minutes.
6. Dehydrate with ethanol.
7. Clear with xylene and mount with a resinous medium.

Expected results

• Young collagen and reticulin - blue
• Mature collagen - red
• Cytoplasm - yellow
• Nuclei - black

Notes

• Solution A is van Gieson's solution.
• Cook notes that the original included a final step with metanil yellow for cytoplasmic staining, which he omitted.

Reference
Cook, H. C., (1974)
Manual of Histological Demonstration Techniques
Butterworths, London, UK.
  Citing:–
    Herovici, c., (1963)
    Stain Technology, v. 38, p. 204

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