Garvey's Stain
for Elastic, Fibrin & Collagen
Materials
Stock Verhoeff A
| Material | Amount | |
|---|---|---|
| Hematoxylin | 3 | g |
| Ethanol, absolute | 100 | mL |
Stock Verhoeff B
| Material | Amount | |
|---|---|---|
| Ferric chloride | 2.3 | g |
| Distilled water | 100 | mL |
Stock Verhoeff C
| Material | Amount | |
|---|---|---|
| Potassium iodide | 4 | g |
| Iodine | 2 | g |
| Distilled water | 100 | mL |
Working Verhoeff
| Material | Amount | |
|---|---|---|
| Stock solution A | 3 | volumes |
| Stock solution B | 2 | volumes |
| Stock solution C | 1 | volume |
Polyacid
| Material | Amount | |
|---|---|---|
| Phosphotungstic acid | 2 | g |
| Distilled water | 100 | mL |
Acetic water
| Material | Amount | |
|---|---|---|
| Acetic acid, glacial | 0.5 | mL |
| Distilled water | 100 | mL |
Picro-mercuric chloride
| Material | Amount | |
|---|---|---|
| Picric acid, sat. aqu. | 100 | mL |
| Mercuric chloride | 5 | g |
| Distilled water | 100 | mL |
Erythrocyte stain
| Material | Amount | |
|---|---|---|
| Lissamine fast yellow | 1 | g |
| Acetic acid, glacial | 0.5 | mL |
| Distilled water | 100 | mL |
Plasma stain
| Material | Amount | |
|---|---|---|
| Biebrich scarlet | 0.75 | g |
| Acid fuchsin | 0.75 | g |
| Ponceau 2R | 0.75 | g |
| Acetic acid, glacial | 0.5 | mL |
| Distilled water | 100 | mL |
| Material | Amount for Var I | Amount for Var II | ||
|---|---|---|---|---|
| Aniline blue | 0.5 | g | – | |
| Light green SF yellowish | – | 2 | g | |
| Acetic acid, glacial | 1 | mL | 2 | mL |
| Distilled water | 100 | mL | 100 | mL |
Tissue Sample
Formol sublimate fixation is preferred, although 10% formalin variants are acceptable. Paraffin sections at 3µ are preferred.
Protocol
- Bring sections to water via xylene and ethanol.
- Optionally, place into picro-mercuric chloride for 1 hour.
- Wash well with warm water to remove picric acid.
- Place in verhoeff’s solution for 9 minutes.
- Wash with warm tap water for 5 minutes.
- Place in lissamine fast yellow for 2 minutes.
- Rinse with 0.5% acetic acid.
- Place in the plasma stain for 5 minutes.
- Rinse with distilled water.
- Place in polyacid for 10 minutes.
- Rinse with distilled water.
- Place in a fiber stain variant for 2 minutes.
- Rinse with 0.5% acetic acid.
- Dehydrate, clear and mount with a resinous medium.
Expected Results
- Nuclei – black
- Elastic fibres – black
- Fibrin – young – yellow
- Fibrin – mature – red
- Fibrin – old – blue or green
- Erythrocytes – yellow
- Muscle – red
- Collagen – blue or green (some may be red)
Notes
- Sections should be treated with picro-mercuric chloride if formol sublimate or similar fixation was not used initially.
- The ferric chloride should be the hexahydrate crystals.
- Although the iodine content of the Verhoeff elastic stain component may remove any mercury pigment, in methods such as this, some technologists prefer to apply the iodine-thiosulphate sequence before staining.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Garvey W., et. al., (1987),
A combined elastic, fibrin and collagen stain
Stain Technology, V. 62, No 6, pp 365-367.
