H-E Stain (H-E)

H-E Stain (H-E)

Assay-kits

Product code: ACMBS-02
Specifications: 4x250mL (Box)
Intended use: Used for staining of cell tissue
Store temperature: 10℃～30℃
Expiry date: 15 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationPrecautions

Principle

H-E Stain (H-E) is mainly used to display the general morphological structure of normal tissues and lesions of various tissues, and comprehensive observation. H-E Stain is an essential basic staining method in biology, histology, pathology and cytology. It is widely used in pathological diagnosis, teaching and scientific research and has important value.

The nucleus in the cell is composed of an acidic substance, which has a stronger affinity with the basic dye (Hematoxylin), while the cytoplasm has the opposite effect. It contains a higher affinity for the alkaline substance and the acid dye (Eosin). Therefore, after the cells or tissue sections are stained with H-E Stain, nuclei are stained with hematoxylin by a bright blue-violet color, and the cytoplasm, muscle fibers, collagen fibers and the like are red in different degrees, and the red blood cells are orange-red.

Main components

Reagent composition	Main ingredient
1. Hematoxylin stain (Harris)	Hematoxylin
2. Eosin stain (Aqueous)	Eosin
3. 1% hydrochloric acid ethanol solution	Hydrochloric acid, ethanol
4. Dilute lithium carbonate solution	Lithium carbonate

Sample requirement

The smears and slices must be adequately fixed.

Test procedure

1. Paraffin sections are deparaffinized in xylene I and II for 5 minutes respectively, and then rinsed with absolute ethanol, 95% ethanol, and 80% ethanol for 1 minute each, and rinsed with tap water for 1 minute.
2. Stain for 5～10 minutes with Hematoxylin stain (Harris). Rinse with tap water for 1～2 minutes.
3. Place tissue in 1% hydrochloric acid ethanol for a few seconds, Rinse with tap water for 1～2 minutes.
4. With Dilute lithium carbonate solution for 30 seconds, and then water rinse for 1～2 minutes.
5. Place tissue in Eosin stain (Aqueous) for 30～60 seconds. Rinse with water.
6. About 10 seconds each in 95% ethanol I and II, the time should not be too long.
7. Dehydrate in absolute ethanol I and II for 1～2 minutes, and transparent in xylene I and II for 1～2 minutes.
8. Sealed with neutral resin, and examine microscopically.

Result interpretation

Nuclei appear indigo blue while cytoplasm, stroma and fibers appear varying shades of red.

Precautions

1. Avoid Hematoxylin stain (Harris) cold storage and freezing! Hematoxylin stain (Harris) may cause precipitation and redness due to reduced solubility after long-term storage in an environment of<10℃. Before using hematoxylin stain, pay attention to observe if there is a large amount of precipitation or hematoxylin stain turns red. Hematoxylin stain was reconstituted in a 37℃water bath or oven for 2 hours before filtering.
2. Because Hematoxylin stain (Harris) is a dynamic oxidation process, its coloring strength may vary slightlydepending on the date of use. Therefore, users are kindly requested to perform a staining test on each bottle of hematoxylin after unsealing it. Adjust the hematoxylin staining time to ensure better staining effect.
3. Excessive storage temperature of hematoxylin staining solution will cause excessive oxidation. Care should be taken to store it at 10℃～30℃ in the dark.
4. It is normal to produce a layer of oxide film on the surface of hematoxylin stain (Harris) and a little precipitation of aluminum sulfate crystals on the bottom. Before use, the oxide film should be removed and filtered regularly. Dilute lithium carbonate solution (a package of lithium carbonate powder is dissolved in 1000ml distilled water) is used for blueing.
5. Hematoxylin stain (Harris) is not easy to color when the temperature is low, and the dyeing time can be appropriately extended.
6. The color separation is to use 1% hydrochloric acid alcohol solution to wash away the excessive hematoxylin absorbed by the cells, so that the nuclear and cytoplasmic contrast is distinct. The differentiation time should not be too long, otherwise the nucleus will be light stained.
7. After the eosin stain is passed through ethanol, it should not be immersed for too long, especially the low concentration of ethanol. so, it should be avoided to decolorize eosin.
8. Smears and prints of H-E stain method is the same as paraffin section staining from the third step after washing and fixing.
9. The production of good H-E stained sections has a lot to do with whether they are fixed in time and fully fixed.
10. When storing reagents, try to avoid high, low temperature and bright environment, so as not to affect the quality and effect.
11. 80% ethanol, 95% ethanol and absolute ethanol should be replaced regularly to ensure their purity.
12. This reagent should be used by professionals and interpretation of the results. Read the instruction manual carefully before use, use it within the validity period, and do personal hygiene protection.
13. Dispose of waste according to the requirements of the hospital or environmental protection department after use.
14. The production date, production batch number and expiration date are shown in the outer packaging.

References

1. Gao Mingdu, Huang Yilu. Pathological tissue sectioning technique [M]. Nanshantang Publishing House.

2. Liu Zenghui. Pathological staining technique [M]. People's Health Publishing House.

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