Bile (Bilirubin), Hall
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|
|Fouchet Reagent||Part 1095A||Part 1095B|
|Bile Control Slides||Part 4060|
|Van Gieson Stain||Part 1404|
|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.
Newcomer Supply Bile Stain, Hall’s Method is for the demonstration of bile (bilirubin) substances in tissue sections and to distinguish bile pigments from other tissue pigments. The acidity of Fouchet Reagent works to oxidize bilirubin to biliverdin, resulting in a green color development with the Van Gieson Stain serving as a complimentary counterstain.
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.
- Filter Fouchet Reagent with Grade 1 filter paper prior to use.
- 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 slides in freshly filtered Fouchet Reagent for 5 minutes.
- Wash in three changes of tap water; rinse in distilled water.
- Stain sections in Van Gieson Stain (Part 1404) for 5 minutes.
- Rinse quickly in 95% ethyl alcohol.
- 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.
|Bile/bilirubin||Emerald green to olive drab|
|Connective tissue||Pink to red|
- Drain slides after each step to prevent solution carry over.
- Do not allow sections to dry out at any point during procedure.
- If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.
- Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 268-269.
- Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 219.
- Modifications developed by Newcomer Supply Laboratory.