Hematoxylin and eosin stain (H and E stain or HE stain)
Hematoxylin and eosin stain (H&E stain or HE stain) is one of the most commonly used staining techniques in histology and pathology. It is used to reveal the general structure and organization of tissues and cells in various specimens, such as biopsies, surgical resections, autopsies, and cytology smears. H&E stain can help identify normal and abnormal features of tissues and cells, such as inflammation, necrosis, neoplasia, differentiation, and malignancy. H&E stain can also provide clues to the etiology, pathogenesis, and prognosis of various diseases and conditions.
Hematoxylin and eosin (H&E) stain is a common technique to reveal the morphologies of tissues and cells by using two different dyes: hematoxylin and eosin. Hematoxylin is a basic dye that stains acidic structures of the cell, such as nuclei and ribosomes, in a purplish-blue color. Eosin is an acidic dye that stains basic structures of the cell, such as cytoplasm and extracellular matrix, in a pink or red color.
The reagents used in the H&E staining process are:
- Distilled water: Used to clean the sections and rinse off the excess dyes.
- Alum hematoxylin: A basic dye that stains the nuclei and other acidic structures of the tissues purple.
- Acid alcohol: A differentiator that removes the excess hematoxylin from the sections.
- Scott`s tap water: A bluing agent that restores the blue color of the hematoxylin after differentiation.
- Eosin dye: An acidic dye that stains the cytoplasm and other basic structures of the tissues pink.
- Ethanol: A dehydrating agent that removes the water from the sections and prepares them for clearing.
- Xylene: A clearing agent that makes the sections transparent and ready for mounting.
- Mounting medium: A substance that covers and protects the sections and adheres them to the glass slides.
The procedure of H&E staining can be divided into several steps: dewaxing, dehydration, staining, differentiation, bluing, and cover-slipping. The following is a detailed description of each step:
- Dewaxing: This step is necessary if the tissue sections are embedded in paraffin wax.
- Dehydration: This step is required to remove any water from the tissue sections before staining with hematoxylin.
- Staining: This step involves applying the hematoxylin and eosin dyes to the tissue sections.
- Differentiation: This step is optional but recommended for improving the contrast and clarity of the staining.
- Bluing: This step is necessary to restore the blue color of the hematoxylin after differentiation.
- Cover-slipping: This step is the final step of the procedure and involves mounting the stained tissue sections on glass slides.
Hematoxylin and eosin staining produces a contrast between the nuclei and the cytoplasm of cells, as well as between different types of tissues and extracellular components. The nuclei are stained blue or purple by hematoxylin, which binds to the acidic components of chromatin, such as DNA and RNA. The cytoplasm and other structures are stained pink or red by eosin, which binds to the basic components of proteins, such as amino acids and collagen.
H&E staining has many applications in the field of pathology and histology, including:
- Diagnosis of diseases
- Monitoring the effectiveness of treatment
One of the main advantages of H&E staining is that it is quick to execute, cheap, and can be altered.
Some limitations of the H&E staining technique include:
- Not specific for certain cellular components or molecules
- May be affected by various factors
- May not be compatible with some other techniques or methods
- May not be suitable for some types of tissues or cells
These are some of the limitations of the H&E staining technique that should be taken into account when using it for histological analysis.
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