Schiff Reagent, McManus

Schiff Reagent, McManus

  • Store at room temperature for up to one year.

Product Options:

Part # 1371A125 ml $32.40
Part # 1371B500 ml $104.90
Part # 1371C1 L $124.70
Part # 1371D4 L $256.30

 

 

 

Technical Memo 1

Schiff Reagent, McManus for Periodic Acid Schiff (PAS) Stain 

 

 

SOLUTION:                                                                                                                                       

  125 ml 500 ml 1 Liter 4 Liters
Schiff Reagent, McManus Part 1371A Part 1371B Part 1371C Part 1371D

               

Additionally Needed For Periodic Acid Schiff (PAS) Stain:                                               

Periodic Acid Schiff (PAS) Glycogen Control Slides Part 4540
Periodic Acid 0.5%, Aqueous Part 13308
Hematoxylin Stain, Harris Modified Part 1201
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

For storage requirements and expiration date refer to individual product labels.

APPLICATION:

Newcomer Supply Schiff Reagent, McManus a crucial element in the Periodic Acid Schiff (PAS) Stain and for staining glycoproteins, epithelial mucins, basement membranes and fungal cell walls. Schiff Reagent, McManus is used in a variety of staining procedures that include:

  • PAS with and without diastase
  • Alcian Blue/PAS for mucins
  • PAS/Light Green for Fungus (See Below)
  • Feulgen Reaction for demonstration of DNA

Schiff Reagent, McManus is a stable and reliable product that is conveniently stored at room temperature for ready-to-use staining.   Although initially colorless, Schiff Reagent, McManus when in direct contact with skin, clothing, countertops, floors and other surfaces, will react and leave a bright magenta stain that is difficult to remove.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 5 microns

Solutions:  All solutions manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below. 

 

PAS STAINING PROCEDURE:

  1. 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.
    1. See Procedure Notes #1 and #2.
  2. Place in Periodic Acid 0.5%, Aqueous (Part 13308) for 10 minutes.
  3. Wash in three changes of tap water; rinse in distilled water.          
  4. Place in Schiff Reagent, McManus for 20 minutes.
    1. See Procedure Notes #3, #4 and #5.
  5. Wash slides in lukewarm tap water for 5-10 minutes.
  6. Stain with Hematoxylin Stain, Harris Modified (Part 1201), 1-5 minutes, depending on preference of nuclear stain intensity.
  7. Wash in tap water for 2-3 minutes.
  8. Differentiate in Acid Alcohol 1% (Part 10011); 1-2 quick dips.
  9. Wash in tap water for 1 minute.
  10. Blue sections in Lithium Carbonate, Saturated Aqueous (Part 12215); 3-4 dips.
  11. Wash in several changes of tap water; rinse in distilled water.
  12. 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. 

RESULTS:

Glycogen Magenta
Acid & neutral epithelial mucin Magenta
Fungal cell walls Red to purple
Basement membranes Red to purple
Nuclei Blue

 

PROCEDURE NOTES:

  1. Drain staining rack/slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during staining procedure.
  3. Once used, do not return Schiff Reagent, McManus back to the original bottle and mix with fresh solution.
  4. To test quality of Schiff Reagent, McManus reactivity;
    1. Add a few drops of Schiff Reagent, McManus to 10 ml of Formaldehyde 37-40%, ACS (Part 1089). Formalin 10%, Phosphate Buffered can also be used.
    2. A good Schiff reagent will rapidly turn reddish-purple.
    3. A deteriorating Schiff reagent will have a delayed reaction and turn deep blue-purple.
  5. All glassware/plasticware that contained Schiff Reagent, McManus in the staining process should be thoroughly rinsed with running tap water to remove residual stain prior to standard glassware cleaning procedure.
  6. Digestion steps can be employed in the PAS procedure for further identification of mucosubstances.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 168-174, 180.
  2. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009.137-141.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 164-168, 245.
  4. Modifications developed by Newcomer Supply Laboratory.

 

Technical Memo 2: 

Schiff Reagent, McManus for Fungus Stain, PAS/Light Green 

 

SOLUTION:                                                                                                                                           

  125 ml 500 ml  1 Liter 4 Liters
Schiff Reagent, McManus Part 1371A Part 1371B Part 1371C Part 1371D

                                               

Additionally Needed For Fungus Stain, PAS/Light Green:                                                    

Fungus, PAS, Aspergillus sp., Artificial Control Slides

                         OR

Fungus, PAS, Candida sp., Artificial Control Slides

Part 4232

 

Part 4233

Periodic Acid 0.5%, Aqueous Part 13308
Light Green SF Yellowish Stain 0.1%, Aqueous Part 12203
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

For storage requirements and expiration date refer to individual product labels.


APPLICATION:

Newcomer Supply Schiff Reagent, McManus, a crucial element in the Fungus Stain, PAS/Light Green procedure, is used for staining fungus and glycogen in tissue sections.

Schiff Reagent, McManus is a stable and reliable product that is conveniently stored at room temperature for ready-to-use staining.   Although initially colorless, Schiff Reagent, McManus when in direct contact with skin, clothing, countertops, floors and other surfaces, will react and leave a bright magenta stain that is difficult to remove.

 

METHOD:

Fixation:  Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin section cut at 5 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below. 

 

STAINING PROCEDURE:

  1. 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.
    1. See Procedure Notes #1 and #2.
  2. Place slides in Periodic Acid 0.5%, Aqueous (Part 13308) for 5 minutes.
  3. Wash in three changes of tap water; rinse in distilled water.
  4. Drain slides of excess water and stain in Schiff Reagent, McManus for 20 minutes.
    1. See Procedure Notes #3, #4 and #5.
  5. Wash gently in lukewarm tap water for 10 minutes to allow pink color to develop.
  6. Counterstain in Light Green SF Yellowish Stain 0.1%, Aqueous (Part 12203) for 5 seconds.
    1. See Procedure Note #6.
  7. 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. 

 

RESULTS:

Fungal cell walls and glycogen Red to magenta
Background Pale green
 

PROCEDURE NOTES:

  1. Drain staining rack/slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during staining procedure.
  3. Once used, do not return Schiff Reagent, McManus back to the original bottle and mix with fresh solution.
  4. To test quality of Schiff Reagent, McManus reactivity;
    1. Add a few drops of Schiff Reagent, McManus to 10 ml of Formaldehyde 37-40%, ACS (Part 1089). Formalin 10%, Phosphate Buffered can also be used.
    2. A good Schiff reagent will rapidly turn reddish-purple.
    3. A deteriorating Schiff reagent will have a delayed reaction and turn deep blue-purple.
  5. All glassware/plasticware that contained Schiff Reagent, McManus in the staining process should be thoroughly rinsed with water to remove residual stain prior to standard glassware cleaning procedure.
  6. Increase or decrease staining time in Light Green SF Yellowish Stain 0.1%, Aqueous for a length of time to suit preference of counterstain intensity.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 321-323.
  2. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245.
  3. Modifications developed by Newcomer Supply Laboratory.