Vanderbilt Stain
for Gastrointestinal Biopsies
Materials
- An alum hematoxylin
- An aqueous or alcoholic eosin Y solution
- Solution A
Material Amount Acetic acid, glacial 3 mL Distilled water 97 mL - Solution B
Material Amount Alcian blue 8GN 1 g Acetic acid, glacial 3 mL Distilled water 97 mL - Solution C
Material Amount Lithium carbonate 0.5 g Distilled water 100 mL - Solution D
Material Amount Saffron 1.5 g Acetic acid, glacial 1 mL Ethanol, absolute 99 mL
Tissue Sample
Hollande’s fixative is recommended. 5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.
Protocol
- Bring sections to water via xylene and ethanol.
- Place in solution A for 3 minutes.
- Place into solution B for 5 minutes.
- Wash well with water.
- Place into solution C for 1 minute.
- Wash well with water.
- Do a standard H&E.
- Rinse with 95% ethanol, then two changes of absolute ethanol of 1 minute each.
- Place into solution D for 30 seconds.
- 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
- If overstained with saffron (solution D), destain with ammonia water and restain from the eosin step of the H&E.
- The saffron solution may be replaced with a tartrazine solution.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Staples, T. C., 1996.
Alcian blue-H&E-saffron stain for gastrointestinal biopsies
Histo-Logic, v. 26, N°1, March, 1996.
