Alcian Blue - H&E Metanil Yellow 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.
|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|
|Alcian Blue pH 2.5, Barrett’s Esophagus Control Slides
Alcian Blue pH 2.5, Goblet Cell Control Slides
|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.
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.
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.
- 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.
- Place in Acetic Acid 3%, Aqueous (Part 10017) for 3 minutes.
- Move directly into Alcian Blue Stain 1%, pH 2.5 Aqueous (Part 1003) for 15 minutes.
- Wash in running tap water for 5 minutes; rinse in distilled water.
- Stain with Hematoxylin Stain, Mayer Modified (Part 1202) for 2 to 4 minutes.
- Rinse in running tap water until water runs clear.
- Blue in Scott Tap Water Substitute (Part 1380); 2-3 dips.
- Wash well in running tap water; rinse in distilled water.
- Place in 70% ethyl alcohol for 1 minute.
- Stain with Eosin Y Working Solution (Part 1072) for 1 minute.
- Dehydrate in 95% ethyl alcohol for 30 seconds.
- Dehydrate in two changes of 100% ethyl alcohol; 30 seconds each.
- Place in Metanil Yellow Stain, Aqueous (Part 12235) for 1 minute.
- See Procedure Note #3.
- Rinse/dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
|Mucin/ Barrett’s esophagus goblet cells||Turquoise|
|Gastric mucin||Some stain faint blue|
- Drain slides after each step to prevent solution carry over.
- Do not allow sections to dry out at any point during procedure.
- If over-stained in Metanil Yellow Stain, Aqueous increased background staining will occur.
- If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.
- Edgett, Wendi. “Alcian Blue-H&E-Metanil Yellow Stain for Diagnosing Barrett’s Esophagus”. Histologic 37.2 (2004). 35-38
- Staples, Terri. “Alcian Blue-H&E-Saffron Stain for Gastrointestinal Biopsies”. Histologic 26.1 (1996). 2-3.
- Modifications developed by Newcomer Supply Laboratory.