Sayeed's Stain
for Helicobacter
Materials
- Schiff’s reagent
- 0.5% periodic acid
- Mayer’s hemalum
- Stock methylene blue
Material Amount Methylene blue 1.4 g Ethanol, 95% 100 mL - Working methylene blue
Material Amount Stock methylene blue 10 mL Tap water 90 mL
Tissue Sample
5µ paraffin sections of formalin fixed tissue are suitable.
Protocol
- Bring sections to water via xylene and ethanol.
- Rinse with distilled water.
- Place in periodic acid for 1 minute.
- Rinse well with tap water.
- Rinse with distilled water.
- Place in Schiff’s reagent for 1 minute (see note).
- Wash with tap water for 2 minutes.
- Rinse with distilled water.
- Place in Mayer’s hemalum for 2 minutes.
- Rinse well with tap water
- Place in working methylene blue for 1 minute.
- Rinse with distilled water.
- Dehydrate, clear and mount with a resinous medium.
Expected Results
- Mucin – pink
- Helicobacter – blue
Notes
- The time in Schiff’s reagent is an estimate and should be altered to produce the desired effect.
- The authors specified Coleman’s Schiff reagent. Others would likely be satisfactory.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- 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.
