Hematoxylin and Eosin (H&E)

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining small intestine.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  H&E quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity. 
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

CONTROL SLIDE VALIDATION:

APPLICATION:

Newcomer Supply Hematoxylin & Eosin (H&E) Control Slides are for the clear demonstration of nuclei, epithelium, connective tissue and muscle, for documentation of high quality nuclear and cytoplasmic staining.

For quality control measures, use H&E Control Slides each day that H&E staining is performed and additionally after any H&E solution changes.

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 1. Heat dry sections in oven according to your laboratory protocol.
   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. 1. See Procedure Notes #1 and #2.
   3. Stain with Hematoxylin Stain, Harris Modified or Hematoxylin Stain, Harris for 1 to 5 minutes, depending on nuclear stain preference.
   4. Wash well in three changes of tap water.
   5. Differentiate quickly in Acid Alcohol 1%.
      1. 1. See Procedure Note #3.
   6. Rinse immediately in three changes of tap water.
   7. Blue slides in Lithium Carbonate, Saturated Aqueous or Scott Tap Water Substitute for 10 dips.
   8. Wash in three changes of tap water; rinse in distilled water.
   9. Drain excess water from slides; proceed to 70% alcohol for 10 dips.
   10. Counterstain in Eosin Y Working Solution for 30 seconds to 3 minutes, depending on counterstain preference.
   11. 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. Differentiate to suit preference of nuclear stain intensity.
      1. 1. Check microscopically for satisfactory nuclear staining.
         2. Nuclei should be distinct and background light to colorless.
   4. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

REFERENCES:

1. 1. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 118-120.
   2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 143-144, 153-154.
   3. Modifications developed by Newcomer Supply Laboratory.