Alcian Blue pH 2.5, Multi-Tissue
Validation Stain: Alcian Blue 1%, pH 2.5
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PRODUCT SPECIFICATIONS:
Tissue: Positive staining umbilical cord and positive staining small intestine.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain: Alcian Blue pH 2.5 quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.
Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.
CONTROL SLIDE VALIDATION:
With Alcian Blue 1%, pH 2.5 Stain Kit: | Part 9102A/B | Individual Stain Solution | |
Solution A: | Acetic Acid 3%, Aqueous | 250/500 ml | Part 10017 |
Solution B: | Alcian Blue Stain 1%, pH 2.5 Aqueous | 250/500 ml | Part 1003 |
Solution C: | Nuclear Fast Red Stain, Kernechtrot | 250/500 ml | Part 1255 |
APPLICATION:
Newcomer Supply Alcian Blue pH 2.5, Multi-Tissue Control Slides use a combination of tissue sources for the positive histochemical staining of acid epithelial mucins (sialomucin, sulfomucin) as well as stromal (mesenchymal) mucin.
NEWCOMER SUPPLY VALIDATION PROCEDURE:
- Heat dry sections in oven according to your laboratory protocol.
- Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each. Wash well with distilled water.
- See Procedure Notes #1 and #2.
- Place slides in Solution A: Acetic Acid 3%, Aqueous for 3 minutes.
- Move slides directly into Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous. Stain for 30 minutes at room temperature or for 15 minutes in a 37°C water bath.
- Wash in running tap water for 10 minutes; rinse in distilled water.
- See Procedure Note #3.
- Counterstain in Solution C: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
- Shake solution well before use; do not filter.
- Rinse well in distilled water.
- See Procedure Note #4.
- Dehydrate quickly through two changes of 95% ethyl alcohol and two changes of 100% ethyl alcohol. Clear in three xylene changes, 10 dips each; coverslip with compatible mounting medium.
RESULTS:
Acid epithelial mucins | Blue |
Stromal (mesenchymal) mucin | Blue |
Nuclei | Pink-red |
Cytoplasm | Pale pink |
PROCEDURE NOTES:
- Drain slides after each step to prevent solution carry over.
- Do not allow sections to dry out at any point during procedure.
- A brief dip in Solution A: Acetic Acid 3%, Aqueous from Step #3 can be added before water rinses to remove excess Alcian Blue Solution if needed.
- Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in dehydration steps.
- If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.
REFERENCES:
- Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 145-148.
- Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 172-175.
- Modifications developed by Newcomer Supply Laboratory.