Alpha Amylase 1%, Aqueous
Ready to-use glycogen digestion solution for aiding in further identification of mucosubstances used in conjunction with the Periodic Acid Schiff (PAS) Stain procedure.
Store at 2-8 C.
Shelf-life is 18 months from date of manufacture.
|Alpha Amylase 1%, Aqueous||Part 1905B|
|Periodic Acid Schiff (PAS) Glycogen Control Slides||Part 4540|
|Periodic Acid 0.5%, Aqueous||Part 13308|
|Schiff Reagent, McManus||Part 1371|
|Hematoxylin Stain, Harris||Part 12013|
|Acid Alcohol 1%||Part 10011|
|Lithium Carbonate, Saturated Aqueous||Part 12215|
|Xylene, ACS||Part 1445|
|Alcohol, Ethyl Denatured, 100%||Part 10841|
|Alcohol, Ethyl Denatured, 95%||Part 10842|
|Coplin Jar, Plastic||Part 5184 (for glycogen digestion microwave modification)|
For storage requirements and expiration date refer to individual product labels.
Newcomer Supply Alpha Amylase 1%, Aqueous is a convenient ready-to-use glycogen digestion solution for aiding in further identification of mucosubstances used in conjunction with the Periodic Acid Schiff (PAS) Stain procedure.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090)
Technique: Paraffin sections cut at 4 microns
- If necessary, heat dry tissue sections/slides in oven.
- 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.
- Digestion Step: See Procedure Note #3.
- Two control slides and two patient slides are needed.
- Label one control slide and one patient slide “with”.
- Label the other control slide and patient slide “without”.
- Place slides labeled “without” in separate Coplin jar of distilled water; hold for Step #5.
- Apply Alpha Amylase 1%, Aqueous to slides labeled “with” for 30 minutes at room temperature.
- Proceed to Step #5.
- Digestion Microwave Modification: See Procedure Note #4.
- Follow Steps #3a through #3d.
- Place slides labeled “with” in a plastic Coplin jar containing Alpha Amylase 1%, Aqueous and microwave for 1 minute at 37°C. Let sit in warm solution for an additional minute.
- Combine all slides for remaining steps; wash in running tap water for 1 minute, rinse in distilled water.
- Place in Periodic Acid 0.5%, Aqueous (Part 13308) for 10 minutes.
- Wash in three changes of tap water; rinse in distilled water.
- Place in Schiff Reagent, McManus (Part 1371) for 20 minutes.
- Wash in lukewarm tap water for 5 minutes.
- Stain with Hematoxylin Stain, Harris (Part 12013), 1-5 minutes, depending on preference of nuclear stain intensity.
- Wash in tap water for 2-3 minutes.
- Differentiate in Acid Alcohol 1% (Part 10011); 1-2 quick dips.
- Wash in tap water for 1 minute.
- Blue in Lithium Carbonate, Saturated Aqueous (Part 12215); 3-4 dips.
- Wash in several changes of tap water; rinse in distilled water.
- Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
|Glycogen digestion||Absence of magenta|
|Acid & neutral epithelial mucin||Magenta|
- Drain slides after each step to prevent solution carry over.
- Do not allow sections to dry out at any point during procedure.
- Slides labeled “with” will be treated with amylase digestion, slides labeled “without” will not be treated for digestion.
- The suggested microwave procedure has been tested at Newcomer Supply. This procedure is a guideline and techniques should be developed for use in your laboratory.
- If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.
- Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 168-171, 180.
- Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009.137-141.
- Sheehan, Dezna C., and Barbara B.Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 164-168.
- Modifications developed by Newcomer Supply Laboratory.