Products

Alcian Blue - H&E Metanil Yellow Stain

Alcian Blue H&E Metanil Yellow Histology Stain

Reagents for this procedure are sold as individual stain solutions and are available for purchase under separate part numbers with storage requirements and expiration date designated per bottle.

Product Options:

Part # Non-Kit

SOLUTIONS:

250 ml 500 ml  1 Liter 1 Gallon
Acetic Acid 3%, Aqueous Part 10017A Part 10017B
Alcian Blue Stain 1%, pH 2.5 Aqueous  Part 1003A Part 1003B
Hematoxylin Stain, Mayer Modified Part 1202A Part 1202B Part 1202C
Scott Tap Water Substitute Part 1380A  Part 1380B Part 1380C
Eosin Y Working Solution Part 1072A Part 1072B Part 1072C
Metanil Yellow Stain, Aqueous Part 12235B Part 12235C

 

Additionally Needed:

Alcian Blue pH 2.5, Barrett’s Esophagus Control Slides
                              OR
Alcian Blue pH 2.5, Goblet Cell Control Slides		 Part 4023
     OR
Part 4051
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Alcohol, Ethyl Denatured, 70% Part 10844

 

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

 

APPLICATION:

Newcomer Supply Alcian Blue-H&E-Metanil Yellow Stain is a useful diagnostic screening method for Barrett’s esophagus and distinguishing from other gastrointestinal conditions/disorders.  The use of three separate stains and an eosin counterstain, provides a colorful combination of the gastrointestinal tracts various components.

 

METHOD:

Fixation:  Formalin 10%, Phosphate Buffered (Part 1090)
Technique:  Paraffin Sections cut at 4 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 provided staining procedure.

 

STAINING PROCEDURE:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. 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.
  3. Place in Acetic Acid 3%, Aqueous (Part 10017) for 3 minutes.
  4. Move directly into Alcian Blue Stain 1%, pH 2.5 Aqueous (Part 1003) for 15 minutes.
  5. Wash in running tap water for 5 minutes; rinse in distilled water.
  6. Stain with Hematoxylin Stain, Mayer Modified (Part 1202) for 2 to 4 minutes.
  7. Rinse in running tap water until water runs clear.
  8. Blue in Scott Tap Water Substitute (Part 1380); 2-3 dips.
  9. Wash well in running tap water; rinse in distilled water.
  10. Place in 70% ethyl alcohol for 1 minute.
  11. Stain with Eosin Y Working Solution (Part 1072) for 1 minute.
  12. Dehydrate in 95% ethyl alcohol for 30 seconds.
  13. Dehydrate in two changes of 100% ethyl alcohol; 30 seconds each.
  14. Place in Metanil Yellow Stain, Aqueous (Part 12235) for 1 minute.
    1. See Procedure Note #3.
  15. Rinse/dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Nuclei Blue
Cytoplasm Pink-Red
Mucin/ Barrett’s esophagus goblet cells Turquoise
Gastric mucin Some stain faint blue
Collagen Yellow
Smooth Muscle Salmon

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. If over-stained in Metanil Yellow Stain, Aqueous increased background staining will occur.
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Edgett, Wendi. “Alcian Blue-H&E-Metanil Yellow Stain for Diagnosing Barrett’s Esophagus”. Histologic 37.2 (2004). 35-38
  2. Staples, Terri. “Alcian Blue-H&E-Saffron Stain for Gastrointestinal Biopsies”. Histologic 26.1 (1996).  2-3.
  3. Modifications developed by Newcomer Supply Laboratory.