Alcian Blue Stain 1%, pH 2.5 Aqueous

Alcian Blue Stain 1%, pH 2.5 Aqueous

Product Options:

Part # 1003A250 ml $69.80
Part # 1003B500 ml $138.70

 

 

Tech Memo 1:

Alcian Blue Stain 1%, pH 2.5

 

SOLUTION:                                                                                                                                    

  250 ml 500 ml
Alcian Blue Stain 1%, pH 2.5 Aqueous Part 1003A Part 1003B

 

 Additionally Needed:

Alcian Blue pH 2.5, Umbilical Cord Control Slides

                                    OR

Alcian Blue pH 2.5, Multi-Tissue Control Slides

Part 4020

     OR

Part 4021

Acetic Acid 3%, Aqueous Part 10017
Nuclear Fast Red Stain, Kernechtrot  Part 1255
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 Alcian Blue Stain 1%, pH 2.5 Aqueous is designed to stain acid epithelial mucin (sialomucin, sulfomucin) as well as stromal (mesenchymal) mucin in the Alcian Blue Stain 1%, pH 2.5 procedure.

 

METHOD:

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

Technique:  Paraffin sections 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.
  1. Place slides in Acetic Acid 3%, Aqueous (Part 10017) for 3 minutes.
  2. Put slides directly into Alcian Blue Stain 1%, pH 2.5 Aqueous and stain for 30 minutes at room temperature or for 15 minutes in a 37°C water bath.
  3. Wash in running tap water for 10 minutes; rinse in distilled water.
  1. See Procedure Note #3.
  1. Counterstain in Nuclear Fast Red Stain, Kernechtrot (Part 1255) for 5 minutes.
  1. Shake solution well before use; do not filter.
  1. Rinse well in distilled water.
  1. See Procedure Note #4
  1. Dehydrate quickly through two changes of 95% ethyl alcohol and two changes of 100% ethyl alcohol. Clear in three xylene changes, 10 dips each; coverslip with compatible mounting medium. 

     

RESULTS:

Acid epithelial mucin Blue
Stromal (mesenchymal) mucin Blue
Nuclei Pink-red
Cytoplasm Pale pink

 

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. A brief dip in Acetic Acid 3%, Aqueous from Step #2 can be added before rinsing to remove excess Alcian Blue Stain 1%, pH 2.5 Aqueous if needed.
  4. Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in dehydration steps.
  5. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida. Histotechnology: A Self-Instructional Text. 2nd ed. Chicago: ASCP Press, 1997. 118-120.
  2. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 172-173.
  3. Modifications developed by Newcomer Supply Laboratory.

 

Tech Memo 2:

Alcian Blue/PAS Stain

 

SOLUTION: 

  250 ml 500 ml
Alcian Blue Stain 1%, pH 2.5 Aqueous Part 1003A Part 1003B

 

Additionally Needed:                           

Alcian Blue/PAS Control Slides Part 4022
Acetic Acid 3%, Aqueous Part 10017
Periodic Acid 0.5%, Aqueous Part 13308
Schiff Reagent, McManus Part 1371
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 Alcian Blue Stain 1%, pH 2.5 Aqueous, a crucial element in the Alcian Blue/PAS Stain procedure is used to differentiate between acidic epithelial mucins (sialomucin, sulfomucin) and neutral epithelial mucin and is a means of detecting the overall presence of mucins.  Acidic mucins are stained with the Alcian Blue technique while neutral mucins and glycogen are stained by the PAS reaction.

 

METHOD:

Fixation:  Formalin 10%, Phosphate Buffered (Part 1090

Technique:  Paraffin sections 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.
  1. Place slides in Acetic Acid 3%, Aqueous (Part 10017) for 3 minutes.
  2. Place slides directly into Alcian Blue Stain 1%, pH 2.5 Aqueous for 30 minutes.
  3. Wash slides in gently running tap water for 10 minutes; rinse in distilled water.
  4. Place slides in Periodic Acid 0.5%, Aqueous (Part 13308) for 10 minutes.
  5. Wash slides in running tap water for 5 minutes; rinse in distilled water.
  6. Place slides in Schiff Reagent, McManus (Part 1371) for 20 minutes.
  7. Wash in lukewarm tap water for 5-10 minutes.
  8. 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:

Acid epithelial mucin Violet
Neutral epithelial mucin Magenta
Glycogen Magenta
Stromal (mesenchymal) mucin 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. Newcomer Supply Schiff Reagent, McManus is stored at room temperature.  There is no benefit to store this product at 4°C.
  4. 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. 173-174.
  2. Carson, Freida, Histotechnology: A Self-Instructional Text. 2nd ed. Chicago: ASCP Press, 1997. 91-102, 121-123.
  3. Modifications developed by Newcomer Supply Laboratory.