Vanderbilt Stain
for Gastrointestinal Biopsies


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.


  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


  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.


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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