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Living up to Life Special Stain Kit Alcian Blue

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Living up to Life Special Stain Kit Alcian Blue Living up to Life Special Stain Kit Alcian Blue Stain Kit Catalog No: 38016SS3 Intended Use Staining Protocol (Microwave) For In Vitro Diagnostic Use. For Laboratory Use. Exercise caution when using the microwave to heat any solution or reagent. The microwave The reagents in this kit are intended for In Vitro use only. Alcian Blue, when used with the must be properly ventilated to prevent the accumulation of fumes in the laboratory. Microwave appropriate histological staining protocol, may be useful for the demonstration of acidic mucins transparent coplin jars and caps should be used during the staining process. The caps should in tissue sections. The pH 2.5 Alcian Blue provided in this kit reacts with both carboxylated and be loosely applied to prevent spills. Caps with ventilation holes also may be used. sulfated mucins. All microwaves should be used in accordance with the manufacturer’s instructions. The procedures described here were performed using an Energy Beam Sciences H2250 laboratory Probable Mode of Action microwave. All microwaved steps were conducted at a power setting of 800 watts unless The Alcian Blue molecule is a large conjugated dye molecule that contains a central copper otherwise noted. Because of differences in microwave power and frequencies among various containing phthalocyanine ring and four isothiouronium groups. The isothiouronium groups models, it may be necessary to adjust power levels or times to achieve optimal results. are basic and impart an overall positive charge to the Alcian Blue molecule. The cationic 1. Deparaffinize with xylene or a xylene substitute and rehydrate though graded alcohols to isothiouronium groups likely bind via electrostatic interactions to the anionic sulfate and deionized water.a carboxylate groups located within the carbohydrate moieties of mucin. 2. Place slides in a plastic Coplin jar with the Alcian Blue solution (40–50 mL) and microwave Reagents Provided for 30 seconds. Alcian Blue pH 2.5 (Item No. 38016SS3A, 500 mL) 3. Gently mix the Alcian blue solution by swirling and allow the solution to stand for 1 minute. Nuclear Fast Red (Item No. 38016SS3B, 500 mL) 4. Wash the slides in running tap water for 10 minutes. Please see reverse side for hazardous ingredients and warning symbols. 5. Rinse briefly in deionized water.a 6. Place sections in a plastic Coplin jar with the Nuclear Fast Red (40–50 mL) and microwave Storage and Stability for 20 seconds.b Store at room temperature (15-30 °C). Do not use after the expiration date. 7. Gently mix the Nuclear Fast Red by swirling and allow the solutions to stand for 2 minutes. Specimen Preparations 8. Rinse in running tap water for 1 minute. Fixation 9. Dehydrate through two changes of 95% alcohol and 100% alcohol two minutes each. Any general fixative including but not limited to neutral buffered formalin, alcoholic formalin and 10. Clear in two changes of xylene (two minutes each) and coverslip using a mounting medium Bouin’s may be used. miscible in xylene. Paraffin Sections Technical Notes Following processing and paraffin embedding, cut sections at 4–6 microns. a. Distilled water may be substituted for deionized water. Preparation of Weigert’s Hematoxylin Working Solution b. Filter Nuclear Fast Red before use. Mix equal parts of Weigert’s Hematoxylin Solutions A and B. Weigert’s working solution is Expected Results useable for 10 days. Acid mucins, proteoglycans and hyaluronic acid — Blue Staining Protocol (Conventional, Room Temperature) Nuclei — Red/Pink 1. Deparaffinize with xylene or a xylene substitute and rehydrate through graded alcohols to Cytoplasm – Pink deionized water.a 2. Stain in Alcian Blue solution for 30 minutes. Recommended Controls 3. Wash in running tap water for 10 minutes. Gastrointestinal epitlelium of the small intestine, appendix or colon. 4. Rinse briefly in deionized water.a References 5. Stain with Nuclear Fast Red for 5 minutes.b 1. Scott J.E., Quintarelli G. and Dellovo M.C. (1964) The chemical and histochemical 6. Rinse in running tap water for 1 minute. properties of alcian blue. I. The mechanisms of alcian blue staining. Histochemie 4:73-85 7. Dehydrate through two changes of 95% alcohol and 100% alcohol two minutes each. 2. Quintarelli G., Scott J.E. and Dellovo M.C. (1964) The chemical and histochemical 8. Clear in two changes of xylene (two minutes each) and coverslip with a mounting medium properties of alcian blue. II. Dye binding polyanions. Histochemie 4: 86-98 that is miscible with xylene. 3. Mowry R.W. (1956) Alcian blue techniques for the histochemical study of acidic carbohydrates. J Histochem and Cytochem 4:403-407 Date of Issue July 2010 Leica Biosystems Richmond, Inc. 5205 Route 12 Richmond, IL 60071 USA ( 1-800-225-3035 www.leica-microsystems.com Living up to Life Special Stain Kit Alcian Blue Stain Kit Catalog No: 38016SS3 Alcian Blue pH 2.5 (Item No. 38016SS3A, 500 mL) Hazardous Ingredient % wt. CAS No. Alcian Blue <2 75881-23-1 Buffer <0.5 Propriety Buffer Salts <0.5 Propriety NFPA 0 1 0 Nuclear Fast Red (Item No. 38016SS3B, 500 mL) Hazardous Ingredient % wt. CAS No. Aluminum Sulfate <10 10043-01-3 Nuclear Fast Red <1 64009-77-4 Ethylene Glycol <10 107-21-1 Proclin 300 <0.1 96118-96-6 Water >85 7732-18-5 NFPA 0 1 0 Leica Biosystems Richmond, Inc. 5205 Route 12 Richmond, IL 60071 USA ( 1-800-225-3035 www.leica-microsystems.com © Leica Microsystems GmbH • HRB 5187 • 95.9623 Rev B • RM 3816SS3.
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