Vanderbilt Stain
for Gastrointestinal Biopsies

Solutions

An alum hematoxylin

An aqueous or alcoholic eosin Y solution

Solution A
Acetic acid, glacial	3	mL
Distilled water	97	mL

Solution B
Alcian blue 8GN	1	g
Acetic acid, glacial	3	mL
Distilled water	97	mL

Solution C
Lithium carbonate	0.5	g
Distilled water	100	mL

Solution D
Saffron	1.5	g
Acetic acid, glacial	1	mL
Ethanol, absolute	99	mL

Tissue sample
Hollande's fiixative is recommended. 5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.

Method

1. Bring sections to water via xylene and ethanol.
2. Place in solution A for 3 minutes.
3. Place into solution B for 5 minutes.
4. Wash well with water.
5. Place into solution C for 1 minute.
6. Wash well with water.
7. Do a standard H&E.
8. Rinse with 95% ethanol, then two changes of absolute ethanol of 1 minute each.
9. Place into solution D for 30 seconds.
10. Rinse with ethanol, clear with xylene and mount in a resinous medium

Expected results

• Nuclei  –  blue
• Mucins  –  blue
• Cytoplasm  –  pink
• Collagen  –  yellow
• Smooth muscle  –  salmon

Notes

1. If overstained with saffron (solution D), destain with ammonia water and restain from the eosin step of the H&E.
2. The saffron solution may be replaced with a tartrazine solution.

 

Reference
Staples, T. C., 1996.
Alcian blue-H&E-saffron stain for gastrointestinal biopsies
Histo-Logic, v. 26, N°1, March, 1996.

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