Sayeed's Stain
for Helicobacter


Schiff's reagent
0.5% periodic acid
Mayer's hemalum
Stock methylene blue
Methylene blue 1.4 g
Ethanol, 95% 100 mL
Working methylene blue
Stock methylene blue 10 mL
Tap water 90 mL

Tissue sample
5µ paraffin sections of formalin fixed tissue are suitable.


  1. Bring sections to water via xylene and ethanol.
  2. Rinse with distilled water.
  3. Place in periodic acid for 1 minute.
  4. Rinse well with tap water.
  5. Rinse with distilled water.
  6. Place in Schiff's reagent for 1 minute (see note).
  7. Wash with tap water for 2 minutes.
  8. Rinse with distilled water.
  9. Place in Mayer's hemalum for 2 minutes.
  10. Rinse well with tap water
  11. Place in working methylene blue for 1 minute.
  12. Rinse with distilled water.
  13. Dehydrate, clear and mount with a resinous medium.

Expected results


  1. The time in Schiff's reagent is an estimate and should be altered to produce the desired effect.
  2. The authors specified Coleman's Schiff reagent. Others would likely be satisfactory.


Cohen LF, Sayeeduddin MHT, Phillips C.
A new staining method for identification of Helicobacter pylori and simultaneous visualization of gastric morphologic features.
Modern Pathology 1997; v. 10, No. 11, pp. 1160-1163.




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