Hematoxylin and Eosin (H&E)
Validation Stain: H&E
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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:
With Hematoxylin and Eosin (H&E) Stain: | Individual Stain Solution |
Hematoxylin Stain, Harris Modified OR Hematoxylin Stain, Harris |
Part 1201 OR Part 12013 |
Acid Alcohol 1% | Part 10011 |
Lithium Carbonate, Saturated Aqueous OR Scott Tap Water Substitute |
Part 12215 OR Part 1380 |
Eosin Y Working Solution | Part 1072 |
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:
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- Heat dry sections in oven according to your laboratory protocol.
- 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.
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- See Procedure Notes #1 and #2.
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- Stain with Hematoxylin Stain, Harris Modified or Hematoxylin Stain, Harris for 1 to 5 minutes, depending on nuclear stain preference.
- Wash well in three changes of tap water.
- Differentiate quickly in Acid Alcohol 1%.
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- See Procedure Note #3.
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- Rinse immediately in three changes of tap water.
- Blue slides in Lithium Carbonate, Saturated Aqueous or Scott Tap Water Substitute for 10 dips.
- Wash in three changes of tap water; rinse in distilled water.
- Drain excess water from slides; proceed to 70% alcohol for 10 dips.
- Counterstain in Eosin Y Working Solution for 30 seconds to 3 minutes, depending on counterstain preference.
- 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 |
Cytoplasm of epithelium, connective tissue, muscle | Shades of pink |
PROCEDURE NOTES:
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- Drain slides after each step to prevent solution carry over.
- Do not allow sections to dry out at any point during procedure.
- Differentiate to suit preference of nuclear stain intensity.
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- Check microscopically for satisfactory nuclear staining.
- Nuclei should be distinct and background light to colorless.
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- If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.
REFERENCES:
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- Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 118-120.
- Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 143-144, 153-154.
- Modifications developed by Newcomer Supply Laboratory.