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.

Product Options:

Part # Non-Kit

SOLUTIONS:                                                                                                                  

  250 ml 500 ml 1 Liter
Fouchet Reagent Part 1095A Part 1095B  
Van Gieson Stain Solution   Part 1404A Part 1404B

 

Additionally Needed:

Bile Control Slides Part 4060
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 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.

 

METHOD:

Fixation: 10% Phosphate Buffered Formalin (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. Filter Fouchet Reagent prior to use.
  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.
  1. Place slides in freshly filtered Fouchet Reagent for 5 minutes.
  2. Wash in three changes of tap water; rinse in distilled water.
  3. Stain sections in Van Gieson Stain Solution for 5 minutes.
  4. Rinse slides quickly in 95% alcohol.
  5. 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:

Bile Emerald green to olive drab
Connective tissue Pink to red
Background Yellow

               

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. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 268-269.
  2. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 219.
  3. Modifications developed by Newcomer Supply Laboratory.