T. Clark Brelje, Ph.D.
University of Minnesota
Department of Genetics, Cell Biology and Development
6-160 Jackson Hall
321 Church St SE
Minneapolis, MN 55455
Robert L. Sorenson, Ph.D.
University of Minnesota
Department of Genetics, Cell Biology and Development
6-160 Jackson Hall
321 Church St SE
Minneapolis, MN 55455
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Hematoxylin and eosin (or H&E) are the most widely used dyes in histology. They stain many components found in the cytoplasm, nucleus, and extracellular matrix.
Hematoxylin
Click the thumbnail to show this specimen stained with hematoxylin.
Hematoxylin is a positively charged, blue dye complex that stains components of cells and tissues that are negatively charged.
Phosphate groups of nucleic acids (DNA and RNA)
Sulfate groups of some extracellular polysaccharides (glycosaminoglycans) and proteins (mucus)
Tissue components that stain with basic dyes are referred to as basophilic structures.
The overall blue color shows a general staining of this specimen with hematoxylin.
At higher magnification (click this link or the above image), structures with a range of staining intensities become evident.
Nuclei are intensely stained
Cytoplasm of cells are lightly stained
Eosin
Click the thumbnail to show this specimen stained with eosin.
Eosin is a negatively charged, pink dye that stains components of cells and tissues that are positively charged.
Amino groups in proteins (mostly side chains of lysine and arginine)
Tissue components that stain with acid dyes are referred to as acidophilic (or eosinophilic) structures.
The overall pink color shows a general staining of this specimen with eosin.
At higher magnification (click this link or the above image), the range of staining intensities of structures is much less than with hematoxylin. Granules are seen in the cytoplasm of most cells.
Nuclei are lightly stained
Cytoplasm of most cells is intensely stained
Hematoxylin & Eosin
Click the thumbnail to show this specimen stained with hematoxylin and eosin.
Staining with the combination of hematoxylin and eosin (H&E) reveals even more cellular structures than either dye used alone.
At higher magnification (click this link or the above image), the nuclei and cytoplasm of individual cells are easily identified. This reflects the complementary binding affinities of these dyes for many cellular compotents.
These structures are described as:
Basophilic - structures stained blue by hematoxylin
Acidophilic (or Eosinophilic) - structures stained pink by eosin
Hydrophobic structures (such as those rich in fats) do not stain well with H&E.
