Periodic Acid Schiff (PAS) Stain Kit
Stains glycoproteins and aids in the differential diagnosis of tumors through the detection of acid/neutral epithelial mucins and/or glycogen. Digestion steps can be employed for further identification of mucosubstances. Also useful for staining basement membranes and fungal cell walls.
PERIODIC ACID SCHIFF (PAS) STAIN KIT INCLUDES:
|Part 9162A||Part 9162B|
|Solution A:||Periodic Acid 0.5%, Aqueous||250 ml||500 ml|
|Solution B:||Schiff Reagent, McManus||250 ml||500 ml|
|Solution C:||Hematoxylin Stain, Harris||250 ml||500 ml|
|Solution D:||Acid Alcohol 1%||250 ml||500 ml|
|Solution E:||Lithium Carbonate, Saturated Aqueous||250 ml||500 ml|
COMPLIMENTARY POSITIVE CONTROL SLIDES: Enclosed are two complimentary unstained positive control slides for initial verification of staining techniques and reagents. Verification must be documented by running one Newcomer Supply complimentary positive control slide along with your current positive control slide for the first run. Retain the second complimentary control slide for further troubleshooting, if needed.
Individual stain solutions and additional control slides may be available for purchase under separate part numbers at www.newcomersupply.com.
|Xylene, ACS||Part 1445|
|Alcohol, Ethyl Denatured, 100%||Part 10841|
|Alcohol, Ethyl Denatured, 95%||Part 10842|
|Alpha Amylase 1%, Aqueous||Part 1905 (for glycogen digestion)|
|Coplin Jar, Plastic||Part 5184 (for glycogen digestion microwave modification)|
For storage requirements and expiration date refer to individual bottle labels.
Newcomer Supply Periodic Acid Schiff (PAS) Stain Kit procedure, with methods for glycogen digestion, is used for staining glycoproteins and may aid in the differential diagnosis of tumors through detection of acid/neutral epithelial mucins and/or glycogen. Digestion steps can be employed for further identification of mucosubstances. PAS can also be used for staining basement membranes and fungal cell walls.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090)
Technique: Paraffin sections cut at 4 microns
Solutions: All solutions manufactured by Newcomer Supply, Inc.
All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below. Some solutions in the kit may contain extra volumes.
- 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.
- Proceed to Step #5 if not running Digestion.
- 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 (Part 1905) 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 (1905) 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 Solution A: Periodic Acid 0.5%, Aqueous for 10 minutes.
- Wash in three changes of tap water; rinse in distilled water.
- Place in Solution B: Schiff Reagent, McManus for 20 minutes.
- Wash in lukewarm tap water for 5 minutes.
- Stain with Solution C: Hematoxylin Stain, Harris 1-5 minutes, depending on preference of nuclear stain intensity.
- Wash in tap water for 2-3 minutes.
- Differentiate in Solution D: Acid Alcohol 1%; 1-2 quick dips.
- Wash in tap water for 1 minute.
- Blue in Solution E: Lithium Carbonate, Saturated Aqueous; 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|
|Fungal cell walls||Red to purple|
|Basement membranes||Red to purple|
- 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, 245.
- Modifications developed by Newcomer Supply Laboratory.