APPLICATION
Mammalian Tissue
DESIGNED TO SHOW
Helicobacter Pylori
SPECIMEN REQUIREMENTS
Fixation: Tissue; Any well-fixed tissue, however, it is suggested to use formalin fixed tissue.
Sections: Paraffin sections cut at 5 microns.
COMPONENTS AND REAGENTS
1. Periodic Acid Solution - 4oz. /120 ml Refrigerate Corrosive; Oxidizer
2. Sodium Metabisulfite Solution - 4oz. /120 ml Refrigerate Corrosive
3. Alcian Yellow Stain Solution - 4oz. /120 ml Refrigerate
4. Toluidine Blue Solution - 4oz. /120 ml Refrigerate
5 Sodium Hydroxide
6 Pap Jars
1 Stained and 3 unstained helicobacter Pylori control slides.
*WORKING TOLUIDINE BLUE
Add 10 drops Sodium Hydroxide to every 25ml of Toluidine Blue Solution before use. Solution is good for one week
STAINING PROCEDURE
1. Deparaffinize sections and hydrate to deionized water.
2. Oxidize sections in Periodic Acid Solution for 10 minutes.
3. Wash sections well under deionized water for 2 minutes.
4. Treat sections in Sodium Metabisulfite Solution for 5 minutes.
5. Wash sections well under deionized water for 2 minutes.
6. Stain sections in Alcian Yellow Stain Solution for 5 minutes.
7. Wash sections well under deionized water for 1-2 minutes.
8. Stain sections in working Toluidine Blue Solution for 3 minutes.
9. Wash sections well under deionized water for 1-2 minutes.
10. Blot dry.
11. Dehydrate through three changes of absolute Reagent Alcohols, Clear and mount.
RESULTS
1. Helicobacter Pylori - Blue
2. Mucin - Yellow
3. Remaining tissue background - pale blue
NOTES
1. Distilled water or Tap water may be used for rinsing and hydration. Always check the pH of your tap water and chlorine levels before proceeding with any type of biological tissue staining.
2. Always increase your time in deparaffinization when incorporating the use of Xylene substitutes.
3. IMEB provides control tissue with all kits. Use an IMEB control tissue when staining along with patient tissue. Additional control packs can be ordered from IMEB Inc.
REFERENCES
1. Leung K., & Gibbon, K.J., A rapid staining method for helicobacter pylori in gastric biopsies; Journal of Histotechnology, Vol 19, No. 2, June 1996
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