Hematoxylin and eosin stain (or haematoxylin and eosin stain or hematoxylin–eosin stain; often abbreviated as H&E stain or HE stain) is one of the principal tissue stains used in histology. It is the most widely used stain in medical diagnosis and is often the gold standard. For example, when a pathologist looks at a biopsy of a suspected cancer, the histological section is likely to be stained with H&E.

H&E is the combination of two histological stains: hematoxylin and eosin. The hematoxylin stains cell nuclei a purplish blue, and eosin stains the extracellular matrix and cytoplasm pink, with other structures taking on different shades, hues, and combinations of these colors. Hence a pathologist can easily differentiate between the nuclear and cytoplasmic parts of a cell, and additionally, the overall patterns of coloration from the stain show the general layout and distribution of cells and provides a general overview of a tissue sample's structure. Thus, pattern recognition, both by expert humans themselves and by software that aids those experts (in digital pathology), provides histologic information.

This stain combination was introduced in 1877 by chemist Nicolaus Wissozky at the Kazan Imperial University in Russia.

The H&E staining procedure is the principal stain in histology in part because it can be done quickly, is not expensive, and stains tissues in such a way that a considerable amount of microscopic anatomy is revealed, and can be used to diagnose a wide range of histopathologic conditions. The results from H&E staining are not overly dependent on the chemical used to fix the tissue or slight inconsistencies in laboratory protocol, and these factors contribute to its routine use in histology.

H&E staining does not always provide enough contrast to differentiate all tissues, cellular structures, or the distribution of chemical substances, and in these cases more specific stains and methods are used.

There are many ways to prepare the hematoxylin solutions (formulation) used in the H&E procedure, in addition, there are many laboratory protocols for producing H&E stained slides, some of which may be specific to a certain laboratory. Although there is no standard procedure, the results by convention are reasonably consistent in that cell nuclei are stained blue and the cytoplasm and extracellular matrix are stained pink. Histology laboratories may also adjust the amount or type of staining for a particular pathologist.

After tissues have been collected (often as biopsies) and fixed, they are typically dehydrated and embedded in melted paraffin wax, the resulting block is mounted on a microtome and cut into thin slices. The slices are affixed to microscope slides at which point the wax is removed with a solvent and the tissue slices attached to the slides are rehydrated and are ready for staining. Alternatively, H&E stain is the most used stain in Mohs surgery in which tissues are typically frozen, cut on a cryostat (a microtome that cuts frozen tissue), fixed in alcohol, and then stained.

The H&E staining method involves application of haematoxylin mixed with a metallic salt, or mordant, often followed by a rinse in a weak acid solution to remove excess staining (differentiation), followed by bluing in mildly alkaline water. After the application of haematoxylin, the tissue is counterstained with eosin (most commonly eosin Y).