Genta Robason and Graham Stain for Simultaneous Visualization of Helicobacter Pylori and Gastric Morphology
EMS Catalog #:26116-series
Fixation:
Formalin fixed
Sections:
Paraffin @ 4 um sections
Staining:
- Deparaffinize and hydrate sections to distilled water.
- Sensitize sections by placing them in Uranium Nitrate. 1%, Aqueous (#26116-01) at room temperature for 3 minutes and transfer them to distilled water.
- Rinse slides in distilled water until the possibility of cross contamination is eliminated.
- Place sections in Silver Nitrate, 1% (#26116-02) at room temperature and then heat then in a microwave oven to just below the boiling point. DO NOT BOIL!
- Rinse slides in 3 changes of distilled water.
- Rinse in 2 changes of 95% alcohol.
- Rinse in 2 changes of 100% alcohol.
- Place slides in 2.5% Gum Mastic (#26116-03) for 5 minutes.
- Air dry sections for 1 minute
- Rinse in 2 changes of distilled water.
- Reduce in reducing solution in a 450°C water bath for 10 to 15 minutes or until sections have developed satisfactorily with black or dark brown H-Pylori and a light yellow background. Check periodically under microscope.
To prepare reducing solution
- Gum Mastic, 2.5 % (#26116-03) - 10 ml
- Hydroquinone, 2% (#26116-04) - 25 ml
- Absolute Alcohol (#15055) - 5 ml
- Silver Nitrate, 0.04% (#26116-02) - 2.5 ml
Mix together - DO NOT FILTER!!
- Rinse sections in water to stop reduction.
- Stain in Alcian Blue, pH 2.5 (#26116-06) for 10 minutes.
- Rinse in running tap water.
- Stain sections in Harris' Hematoxylin (#26116-07) for 5 minutes.
- Rinse in running tap water.
- Quickly dip in Acid Alcohol (#26116-08).
- Rinse in running tap water.
- Rinse in Ammonia Water, 0.25% (#26116-09) until blue.
- Rinse in running tap water.
- Stain sections in Eosin Y, 1% (#26116-10) for 5 minutes.
- Quickly rinse in tap water.
- Rinse twice in 95% alcohol.
- Rinse three times in absolute alcohol.
- Rinse three times in Xylene (#23400) and then coverslip.
Staining Results:
Mucus cells |
Brownish yellow |
Lamina propria ground substance |
Light yellowish or greyish |
Smooth muscle fibers |
Light pink |
Cytoplasm of neutrophils |
Pale pink |
Eosinophils |
Intense red |
Nucleus |
Dark brown |
Intestinal metaplasma |
Bright blue |
H-pylori are easily recognized in the mucus, on the cell surface, and within the gastric pits.
References:
Genta PM., Robason GO, Graham DY- Simultaneous Visualization of Helicobactor Pylon and Gastric Morphology A New Stain Hum Pathol 25: 22 1-226, 1994.
Product Information
Genta. Robason and Graham Stain for Simultanious Visualization of Helicobacter Pylori and Gastric Morphology