Eosin Y - Phloxine B Working Solution

STAIN SOLUTION:                                                                                           

Additionally Needed For H&E Staining:

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

APPLICATION:

Newcomer Supply Eosin Y- Phloxine B Working Solution provides an enhanced counterstain touch to hematoxylin and eosin stains, which can be used in either manual or automated staining platforms.  With the addition of the phloxine B component, differentiation of muscle, connective tissue and epithelial elements tend to be sharper and better demonstrated than with the traditional eosin y solution.

Hematoxylin and eosin (H&E)  staining is used for screening specimens in anatomic pathology, for research, smears, touch preps and other applications.  Its two primary coloring agents stain all cellular material: nuclei (blue), and cytoplasmic elements (pink-red).  Popularity of this stain is due to its simplicity, ability to clearly demonstrate a variety of tissue components, dependability, repeatability, and speed of use.

Quality Control: Since hematoxylin and eosin staining is the foundation of the diagnostic process, maintaining quality is of critical importance. Procedures will vary between laboratories depending upon volume of slides, automation vs manual staining, chemical hygiene and solution integrity.  The longevity of eosin depends upon these factors and stain quality should be regularly screened with an H&E control slide.

METHOD:

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 EOSIN Y- PHLOXINE B:

1. 1. 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. 1. See Procedure Notes #1 and #2.
   2. Stain with Hematoxylin Stain, Harris Modified (Part 1201) or Hematoxylin Stain, Harris (Part 12013) 1-5 minutes, depending on preference of nuclear stain intensity.
   3. Wash well in three changes of tap water.
   4. Differentiate quickly in Acid Alcohol 1%.
      1. 1. Nuclei should be distinct and background very light to colorless.
   5. Rinse well in three changes of tap water.
   6. Blue slides in Lithium Carbonate, Saturated Aqueous (Part 12215) or Scott Tap Water Substitute (Part 1380) for 60-90 seconds.
   7. Wash in three changes of tap water; rinse in distilled water.
   8. Drain excess water; proceed to 70% ethyl alcohol for 10 dips.
   9. Counterstain in Eosin Y- Phloxine B Working Solution for 1 minute.
   10. 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:

PROCEDURE NOTES:

1. 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 using a xylene substitute, follow manufacturer recommendations for deparaffinization and clearing steps.

REFERENCES:

1. 1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 126-127.
   2. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 116-117.
   3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 143-144, 153-154.
   4. Modifications developed by Newcomer Supply Laboratory.