Hematoxylin Stain, Gill

Product Options:

Cytology...GILL I:
Part # 1180A 500 ml $24.70
Part # 1180C 1 gal $98.80
Cytology & Histology...GILL II:
Part # 1180D 500 ml $30.90
Part # 1180F 1 gal $124.50
Histology...GILL III:
Part # 1180G 500 ml $25.70
Part # 1180I 1 gal $153.60

  

STAIN SOLUTION:                                                                                      

500 ml 1 Gallon
Hematoxylin Stain, Gill I (single strength) Part 1180A Part 1180C
Hematoxylin Stain, Gill II (double strength) Part 1180D Part 1180F
Hematoxylin Stain Gill III (triple strength) Part 1180G Part 1180I

Additionally Needed For H&E Staining:

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Lithium Carbonate, Saturated Aqueous

                   or

Scott Tap Water Substitute

Part 12215

      or

Part 1380

Alcohol, Ethyl Denatured, 70% Part 10844
Eosin Y Working Solution

                 or

Eosin-Phloxine Stain Set

Part 1072

      or

Part 1082

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

 

APPLICATION:

Newcomer Supply Hematoxylin Stain, Gill, is a ready to use progressive nuclear stain available in three strengths for cytology and histology applications. Due to the progressive staining nature of Gill hematoxylin, over-staining is less likely and a differentiation step is not required.  Gill hematoxylin does not require filtering but if using any Gill hematoxylin for cytology specimen staining, filtering before each use is highly suggested to prevent solution contamination.

    • Gill I is termed “single strength”. ideal for both gynecological and non-gynecological cytology preparations.
    • Gill II, noted as “double strength”, used as a counterstain in immunohistochemistry (IHC) procedures, frozen section nuclear stain, routine hematoxylin and eosin (H&E) stain and more intense cytology staining.
    • Gill III is “triple strength”, used primarily for tissue sections when a darker nuclear stain is preferred with shorter staining time.
    • Goblet cells will stain with Gill III and moderately by Gill II but not by other hematoxylin solutions.

 

METHOD:

For Histology Specimens

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

Technique:  Paraffin sections cut at 4 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

H&E STAINING PROCEDURE WITH HEMATOXYLIN STAIN, GILL:

    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. Stain with Hematoxylin Stain, Gill of choice. Staining time may vary depending upon the solution strength and intended purpose.
        1. See Procedure Note #3.
    4. Wash well in three changes of tap water.
    5. Blue in Lithium Carbonate, Saturated Aqueous (Part 12215) or Scott Tap Water Substitute (Part 1380) for 10 dips.
    6. Wash in three changes of tap water; rinse in distilled water.
    7. Drain excess water; proceed to 70% alcohol for 10 dips.
    8. Counterstain in Eosin Y Working Solution (Part 1072) or prepared Eosin-Phloxine Working Solution (Part 1082) for 30 seconds to 3 minutes, depending on preference of intensity.
    9. Dehydrate in two changes of 95% ethyl alcohol for 1 minute each and two changes of 100% ethyl alcohol, 10 dips each.   Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Nuclei Blue
Mucin/Goblet cells Blue
Erythrocytes and eosinophilic granules Bright pink to red
Cytoplasm and other tissue elements Various shades of pink

 

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. Stain for a length of time for preferred nuclear stain intensity.
        1. Gill I recommended: Cytology – 5 minutes.
        2. Gill II recommended:
          • Frozen sections/Squash preps – 1 minute.
          • Paraffin H&E: 3-5 minutes.
          • IHC: 3-5 minutes.
        3. Gill III recommended:
          • Frozen sections/Squash preps – 1 minute.
          • IHC: 3-5 minutes.
    4. Store hematoxylin at room temperature in tightly capped container away from direct light to minimize oxidation and extend shelf life.
    5. 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. 112.
    2. Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 81-92.
    3. Modifications developed by Newcomer Supply Laboratory.