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Living up to Life Special Stain Kit Periodic Acid Schiff (PAS) Stain

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Living up to Life Special Stain Kit Periodic Acid Schiff (PAS) Stain Living up to Life Special Stain Kit Periodic Acid Schiff (PAS) Stain Kit Catalog No: 38016SS4 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. The PAS kit, when used with the must be properly ventilated to prevent the accumulation of fumes in the laboratory. Microwave appropriate histological procedures, may be used for the detection of specific carbohydrate transparent coplin jars and caps should be used during the staining process. The caps should containing molecules found in tissue sections. be loosely applied to prevent spills. Caps with ventilation holes also may be used. All microwaves should be used in accordance with the manufacturer’s instructions. The Probable Mode of Action procedures described here were performed using an Energy Beam Sciences H2250 laboratory Carbohydrates that contain adjacent hydroxyl (1, 2–glycol) groups are found in glycogen, microwave. All microwaved steps were conducted at a power setting of 800 watts unless neutral mucins as well as specific glycoproteins. The 1, 2-glycol groups are oxidized by periodic otherwise noted. Because of differences in microwave power and frequencies among various acid to produce adjacent aldehyde groups. The aldehyde groups react with the Schiff reagent models, it may be necessary to adjust power levels or times to achieve optimal results. which subsequently produces a bright magenta coloration in areas of tissue containing these 1. Deparaffinize with xylene or a xylene substitute and rehydrate through graded alcohols to carbohydrates. deionized water.a Reagents Provided 2. Place sections in a plastic Coplin jar containing Periodic Acid (40–50 mL) and microwave Periodic Acid 0.5% (Item No. 38016SS4A, 500 mL) for 10 seconds. Schiff Reagent (Item No. 38016SS4B, 500 mL) 3. Gently mix the solution by swirling and allow to stand for 1 minute. Gill II Hematoxylin (Item No. 38016SS4C, 500 mL) 4. Rinse in several changes of deionized water. Please see reverse side for hazardous ingredients and warning symbols. 5. Add the Schiff Reagent (40–50 mL) to a plastic Coplin jar and microwave for 15 seconds.a 6. Gently mix the Schiff Reagent by swirling and allow to stand for 1 minute. Storage and Stability 7. Wash in lukewarm tap water for 5 minutes. Periodic acid and Gill II Hematoxylin may be stored at room temperature. The Schiff Reagent 8. Counterstain with Gill II Hematoxylin for 3–4 minutes. should be stored at 2–8 °C. Do not use after the expiration date. 9. Rinse in running tap water for 5 minutes. Specimen Preparations 10. Dehydrate through two changes of 95% alcohol and 100% alcohol, two minutes each. Fixation 11. Clear in two changes of xylene (two minutes each) and mount with a mounting media that Any general fixative including but not limited to neutral buffered formalin, alcoholic formalin and is miscible with xylene. Bouin’s may be used. Fixatives containing dialdehyde (glutaraldehyde) should be avoided, as free aldehyde groups may bind the Schiff reagent and produce non-specific staining. Technical Notes a. Distilled water may be used in place of deionized water. Paraffin Sections Following processing and paraffin embedding, cut sections at 4–6 microns. Expected Results Staining Protocol (Conventional, Room Temperature) Many tissues and cell types will demonstrate a bright magenta coloration indicative of a positive PAS reaction. This includes, but is not limited to: glycogen containing hepatocytes, mucin in 1. Deparaffinize with xylene or a xylene substitute and rehydrate through graded alcohols to a the gastrointestinal tract, as well as basement membranes in various sites. Nuclei should stain deionized water. purple/blue with hematoxylin. 2. Place in Periodic Acid solution for 5 minutes. 3. Rinse in several changes of deionized water.a Recommended Controls 4. Place slides in Schiff Reagent for 15 minutes. Liver or gastrointestinal epithelium (small intestine, appendix, colon.) 5. Wash in lukewarm tap water for 10 minutes. References 6. Counterstain with Gill II Hematoxylin for 3–4 minutes. 2. McManus J.F.A. (1946) Histological demonstration of mucin after periodic acid. Nature 7. Rinse in running tap water for 5 minutes. (London) 158:202- 8. Dehydrate through two changes of 95% alcohol and absolute alcohol two minutes each. 3. Hotchkiss R.D.A. (1948) A microchemical reaction resulting in the staining of polysaccharide 9. Clear in two changes of xylene and mount in a mounting media that is miscible with xylene. structures in fixed tissue preparations. Acrh Biochem 16:131-141. 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 Periodic Acid Schiff (PAS) Stain Kit Catalog No: 38016SS4 Periodic Acid 0.5% (Item No. 38016SS4A, 500 mL) Hazardous Ingredient % wt. CAS No. Periodic acid <0 10450-60-9 Water 99 7732-18-5 NFPA 0 2 0 Schiff Reagent (Item No. 38016SS4B, 500 mL) Hazardous Ingredient % wt. CAS No. Basic fuchsin <1 632-99-5 Sodium bisulfite <2 7757-83-7 Hydrochloric acid <1 7647-01-0 NFPA 0 2 0 Gill II Hematoxylin (Item No. 38016SS4C, 500 mL) Hazardous Ingredient % wt. CAS No. Hematoxylin powder <1 517-28-2 Ethylene Glycol 25 107-21-1 Aluminum Sulfate <5 10043-01-3 Sodium Iodate <1 7681-55-2 Acetic Acid <5 64-19-7 NFPA 1 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 • 95.9766 Rev B • RM 3816SS4.
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