Garvey-Movat Pentachrome

for Elastic, Mucin & Collagen

steps

materials

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Expected Results

Movat staining showing amyloid, lipofuscin, and myocardial fibrosis in an autopsy of senile cardiac amyloidosis

fig1-movat-staining-amyloid-lipofuscin-myocardial-fibrosis-autopsy-senile-cardiac-amyloidosis-600x400

Figure 1. Movat Staining Showing Elastic Fibres in an Autopsy Specimen of Senile Cardiac Amyloidosis

Movat staining of an autopsy specimen of senile cardiac amyloidosis shows nuclei and elastic fibres (in black), collagen and reticular fibers (in yellow), ground substance and mucin (in blue), fibrin (in bright red), and muscle (in red). Amyloid is visible as extracellular muddy brown material (right of image), abundant lipofuscin as dark red granular material, and myocardial fibrosis as yellow (left of image). Cardiac amyloidosis very high mag movat by Nephron / Michael Bonert, Pathology & Molecular Medicine, McMaster University is licensed under CC BY-SA 3.0.

Nuclei – black
Elastic fibres – black
Fibrin – mature – red
Muscle – red
Collagen – yellow
Ground substance – blue-green

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

Alcian blue

Material	Amount
Alcian blue	1	g
Acetic acid, glacial	3	mL
Distilled water	100	mL

Plasma stain

Material	Amount
Biebrich scarlet, 1% aqu.	8	mL
Acid fuchsin, 1% aqu.	2	mL
Acetic acid, 1% aqu.	100	mL

Fibre stain

Material	Amount
Saffron du Gatinais	6	g
Ethanol, absolute	100	mL

Preparation

Add the saffron to the ethanol and seal the container.
Incubate at 56°C for 2 weeks. This solution must be anhydrous.

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.
1. Optionally, place into picro-mercuric chloride for 1 hour.
2. Wash well with warm water to remove picric acid.
Rinse with 3% acetic acid.
Place in alcian blue at 60°C for 10 minutes
Rinse with distilled water.
Place in working Verhoeff’s solution for 6 minutes.
Wash with warm tap water for 6 minutes.
Place in the plasma stain for 3 minutes
Rinse with distilled water.
Place in the polyacid for 15 minutes.
Rinse with 1% acetic acid.
Dehydrate thoroughly with absolute ethanol 3 changes.
Place in the fiber stain for 5-6 minutes.
Dehydrate, clear and mount with a resinous medium

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.
Sections thicker than 4µ may need the elastic stain differentiated by treating with 1% aqueous ferric chloride for 20-30 seconds, then washing well with tap water.
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., (1986),
Improved Movat pentachrome stain
Stain Technology, V. 61, No 1, pp 60-62.

Garvey-Movat Pentachrome for Elastic, Mucin & Collagen