Haematoxylin Eosin (HE) Staining Kit

Product Information

DescriptionHE staining, the combined staining of Hematoxylin and Eosin, is the most basic and widely used technical method in the teaching and research of histology, embryology and pathology. Hematoxylin stain is alkaline and mainly causes chromatin in the nucleus and nucleic acids in the cytoplasm to color blue to blue-purple; eosin is an acidic dye that mainly causes components in the cytoplasm and extracellular matrix to color red. HE staining can be used to observe and compare the morphology and structure of normal and diseased tissues, and can be used to initially determine or identify abnormal substances appearing in diseased tissues and cells.

In this HE stain set, the concentration of hematoxylin in Hematoxylin Staining Solution is 0.5%, and the concentration of eosin in Eosin Y Solution (Alcoholic) is 0.05%. Tissue or cell stained by this product, the nucleus is blue to blue-purple, the cytoplasm is red, and the contrast between cytoplasm and nucleus is sharp.

Storage and Handling Conditions

Store and transport at room temperature. Valid for 18 months.

Product Content

Pre-experiment preparation

Prepare your own hematoxylin differentiation solution (G1039 recommended), hematoxylin bluing solution (G1040 recommended), xylene, gradient ethanol, and neutral gum.

Directions for Use

1. Sample pre-processing

(1) For paraffin sections: sections were sequentially deparaffinized by xylene for 10 min, replaced by fresh xylene for 10 min, anhydrous ethanol for 5 min, fresh anhydrous ethanol for 5 min, 90% ethanol for 5 min, 75% ethanol for 5 min, and washed in tap water.

(2) For frozen sections: frozen sections stored at -20°C should be allowed to stand for 5-10 min to return to room temperature. If fixation is required, the sections should be washed in tap water at room temperature with the required fixative.

2. HE staining

(1) Hematoxylin staining: the above-treated sections were stained into Hematoxylin staining solution for 3-5 min and washed with tap water;

(2) Differentiation: sections were rinsed well by hematoxylin differentiation solution for 2-5 s under running tap water;

(3) Back to blue: Hematoxylin back to blue solution for 2-5 s, tap water running water to rinse well.

(4) Eosin staining: the sections were dehydrated by 85% and 95% gradient ethanol for 5 min each. Then it was stained in eosin (alcoholic) for 5 min, and dehydrated twice with absolute ethanol for 5 min each.

(5) Transparent: The slices are then dehydrated with fresh absolute ethanol for 5 min, the xylene is transparent for 5 min, and the fresh xylene is replaced with transparent for 5 min.

(6) Mounting: Add neutral gum dropwise for mounting.

Note

1. After staining, a specific solution is used to remove the excess bound stain from the tissue, a process known as differentiation, and the solution used is called a differentiation solution. A low concentration of hydrochloric acid is commonly used as a differentiation solution in HE staining because the acid destroys the quinone-type structural structure of hematoxylin, causing the pigment to separate from the tissue and fade. After the tissue has been stained with hematoxylin, it must be differentiated to remove excess bound and nonspecifically adsorbed dye from the tissue before eosin staining can be performed to ensure that the nuclei and cytoplasm are distinctly colored. G1039 Hematoxylin Differentiation Solution can be used. During the differentiation process, the differentiation time should be adjusted appropriately according to the thickness of the tissue section, tissue type, etc. in combination with microscopic observation.

2. The return of blue after hematoxylin staining in HE staining is important. Hematoxylin is ionic and red under acidic condition; it is blue under alkaline condition. Tissue sections are red or pink after differentiation by acidic differentiation solution, and need to be washed immediately to terminate the differentiation, and then use weak alkaline solution to make hematoxylin, this process is the return to the blue effect. If there is no special bluing solution, with warm water immersion can also make the cell nucleus return to blue, just take a little longer. We recommend G1040 Hematoxylin Bluing Solution.

3. The degree of hematoxylin and eosin staining can be adjusted according to the need for staining time.

4. If the storage temperature is lower than 15, alum crystals may precipitate out of the hematoxylin solution, which will not affect the product quality. It should be sealed and stored after each use to prevent the active ingredients from evaporating.

5. The purity of anhydrous ethanol used for dehydration after dyeing needs to be greater than 99.9%. If the ethanol contains water, the color of eosin will fade and lead to uneven dyeing.

6. Both Hematoxylin and Eosin Stain (alcohol soluble) are reusable, and each 100 mL of Stain can be used to stain approximately 150 sections. Replace the stain with a new one when the tissue or cell coloring is significantly lighter or abnormal.

7. Please wear lab coat and disposable gloves during operation.

Product Information

DescriptionHE staining, the combined staining of Hematoxylin and Eosin, is the most basic and widely used technical method in the teaching and research of histology, embryology and pathology. Hematoxylin stain is alkaline and mainly causes chromatin in the nucleus and nucleic acids in the cytoplasm to color blue to blue-purple; eosin is an acidic dye that mainly causes components in the cytoplasm and extracellular matrix to color red. HE staining can be used to observe and compare the morphology and structure of normal and diseased tissues, and can be used to initially determine or identify abnormal substances appearing in diseased tissues and cells.

In this HE stain set, the concentration of hematoxylin in Hematoxylin Staining Solution is 0.5%, and the concentration of eosin in Eosin Y Solution (Alcoholic) is 0.05%. Tissue or cell stained by this product, the nucleus is blue to blue-purple, the cytoplasm is red, and the contrast between cytoplasm and nucleus is sharp.

Storage and Handling Conditions

Store and transport at room temperature. Valid for 18 months.

Product Content

Pre-experiment preparation

Prepare your own hematoxylin differentiation solution (G1039 recommended), hematoxylin bluing solution (G1040 recommended), xylene, gradient ethanol, and neutral gum.

Directions for Use

1. Sample pre-processing

(1) For paraffin sections: sections were sequentially deparaffinized by xylene for 10 min, replaced by fresh xylene for 10 min, anhydrous ethanol for 5 min, fresh anhydrous ethanol for 5 min, 90% ethanol for 5 min, 75% ethanol for 5 min, and washed in tap water.

(2) For frozen sections: frozen sections stored at -20°C should be allowed to stand for 5-10 min to return to room temperature. If fixation is required, the sections should be washed in tap water at room temperature with the required fixative.

2. HE staining

(1) Hematoxylin staining: the above-treated sections were stained into Hematoxylin staining solution for 3-5 min and washed with tap water;

(2) Differentiation: sections were rinsed well by hematoxylin differentiation solution for 2-5 s under running tap water;

(3) Back to blue: Hematoxylin back to blue solution for 2-5 s, tap water running water to rinse well.

(4) Eosin staining: the sections were dehydrated by 85% and 95% gradient ethanol for 5 min each. Then it was stained in eosin (alcoholic) for 5 min, and dehydrated twice with absolute ethanol for 5 min each.

(5) Transparent: The slices are then dehydrated with fresh absolute ethanol for 5 min, the xylene is transparent for 5 min, and the fresh xylene is replaced with transparent for 5 min.

(6) Mounting: Add neutral gum dropwise for mounting.

Note

1. After staining, a specific solution is used to remove the excess bound stain from the tissue, a process known as differentiation, and the solution used is called a differentiation solution. A low concentration of hydrochloric acid is commonly used as a differentiation solution in HE staining because the acid destroys the quinone-type structural structure of hematoxylin, causing the pigment to separate from the tissue and fade. After the tissue has been stained with hematoxylin, it must be differentiated to remove excess bound and nonspecifically adsorbed dye from the tissue before eosin staining can be performed to ensure that the nuclei and cytoplasm are distinctly colored. G1039 Hematoxylin Differentiation Solution can be used. During the differentiation process, the differentiation time should be adjusted appropriately according to the thickness of the tissue section, tissue type, etc. in combination with microscopic observation.

2. The return of blue after hematoxylin staining in HE staining is important. Hematoxylin is ionic and red under acidic condition; it is blue under alkaline condition. Tissue sections are red or pink after differentiation by acidic differentiation solution, and need to be washed immediately to terminate the differentiation, and then use weak alkaline solution to make hematoxylin, this process is the return to the blue effect. If there is no special bluing solution, with warm water immersion can also make the cell nucleus return to blue, just take a little longer. We recommend G1040 Hematoxylin Bluing Solution.

3. The degree of hematoxylin and eosin staining can be adjusted according to the need for staining time.

4. If the storage temperature is lower than 15, alum crystals may precipitate out of the hematoxylin solution, which will not affect the product quality. It should be sealed and stored after each use to prevent the active ingredients from evaporating.

5. The purity of anhydrous ethanol used for dehydration after dyeing needs to be greater than 99.9%. If the ethanol contains water, the color of eosin will fade and lead to uneven dyeing.

6. Both Hematoxylin and Eosin Stain (alcohol soluble) are reusable, and each 100 mL of Stain can be used to stain approximately 150 sections. Replace the stain with a new one when the tissue or cell coloring is significantly lighter or abnormal.

7. Please wear lab coat and disposable gloves during operation.

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