H&E STAININGLab Lec # 4

What is H&E ?• The hematoxylin and eosin stain (H&E)• Use

– in histology and histopathology laboratories.

• ability to demonstrate a wide range of – normal and – abnormal cell and tissue components

• Benefit!• it is a relatively simple stain to carry out on paraffin or frozen sections.• The first method of choice to determine whether a biopsy tissue is

cancerous or not.

How it works?• Hematoxylin

– has a deep blue-purple color and – stains nucleic acids by a complex, incompletely understood reaction.– nuclei are stained blue,

• Eosin– is pink and stains proteins nonspecifically. – cytoplasm and extracellular matrix have varying degrees of pink staining.

– if abundant polyribosomes are present, the cytoplasm will have a distinct blue cast.

• Well-fixed cells show considerable intranuclear detail. Nuclei show varying cell-type- and cancer-type-specific patterns of condensation of heterochromatin (hematoxylin staining) that are diagnostically very important.

Procedure

• Dewax – (paraffin wax –hyrdophobic/impervious to aqueous

reagents)• Hydrate – (aqueous reagents can penetrate)

• Add Hematoxylin– Add mordant (for binding)– Add alkaline solution (for more blueing)

• Apply Eosin• Dehydrate & Mount