What is Hematoxylin-Eosin Staining

What is Hematoxylin-Eosin Staining

In the realm of biology and medicine, delving into the intricate details of tissues and cells is a crucial aspect of research and diagnosis. One indispensable tool that aids in this exploration is the Hematoxylin-Eosin (H&E) staining technique. This method has become a cornerstone in histology, allowing scientists and medical professionals to peer into the microscopic world with enhanced clarity and distinction.


Before we delve into the specifics of Hematoxylin-Eosin staining, it is essential to appreciate the complexity of biological tissues. Tissues are composed of various types of cells, each contributing to the overall structure and function of organs. These cells are immersed in an intricate matrix of proteins, fluids, and other substances. To understand the architecture and composition of tissues, scientists employ staining techniques, of which Hematoxylin-Eosin is one of the most widely used.

Hematoxylin and Eosin

At the heart of the Hematoxylin-Eosin staining method are two dyes: hematoxylin and eosin. These dyes have distinct affinities for different cellular components, providing a dual-color contrast that enhances the visibility of microscopic structures.

Comparison of iHE and traditional H&E staining. Figure 1. Comparison of iHE and traditional H&E staining.(Li Y, et al.; 2018)

  • Hematoxylin

Hematoxylin, derived from the heartwood of certain trees, has an affinity for acidic structures in cells. It tends to bind with nucleic acids, particularly DNA, and imparts a blue-purple hue to the stained regions. In essence, hematoxylin acts as a blueprint stainer, highlighting the foundational elements of cellular structures.

  • Eosin

Eosin, on the other hand, is a dye that stains basic structures in cells, such as proteins found in the cytoplasm. This dye imparts a pink to red color to the stained areas. Eosin provides the contrasting color to hematoxylin, allowing for the differentiation of various cellular components and their spatial relationships.

The Staining Process

The Hematoxylin-Eosin staining process involves a series of meticulously orchestrated steps, each contributing to the final masterpiece under the microscope.

  • Fixation

The first step involves fixing the tissue sample. This process prevents decay and maintains the integrity of cellular structures. Formaldehyde is commonly used for this purpose.

  • Dehydration and Clearing

The fixed tissue is dehydrated using a series of alcohol solutions. Subsequently, the tissue is cleared using a substance such as xylene. These steps remove water from the tissue, making it ready for the staining process.

  • Hematoxylin Staining

The tissue is immersed in a hematoxylin solution. Hematoxylin selectively binds to nucleic acids in the cell, particularly DNA, creating a blue-purple color in the nucleus. This step is crucial for highlighting the cellular blueprint and delineating individual cells.

  • Differentiation

Excess hematoxylin is removed by washing the tissue with acid alcohol. This step is critical for achieving the desired contrast and preventing over-staining, which could obscure details.

  • Eosin Staining

The tissue is then immersed in an eosin solution. Eosin binds to basic cellular structures like proteins, imparting a pink to red color to the cytoplasm. This step completes the dual-color contrast, allowing for the visualization of cellular components in their entirety.

  • Dehydration and Mounting

The stained tissue is dehydrated once again and mounted on a microscope slide using a mounting medium. This step ensures that the stained tissue is preserved for microscopic examination.

  • Microscopic Examination

Once the staining process is complete, the stained tissue is ready for microscopic examination. The Hematoxylin-Eosin stain reveals a spectrum of colors, each representing specific cellular structures. Nuclei, stained blue-purple with hematoxylin, stand out prominently, allowing for the identification of individual cells. The contrast provided by eosin highlights the cytoplasmic details, aiding in the visualization of cell boundaries and tissue architecture.


The Hematoxylin-Eosin staining technique is not merely a tool for aesthetic appreciation under the microscope; it serves as a diagnostic powerhouse in pathology. Pathologists routinely use H&E-stained slides to examine tissues and identify abnormalities associated with diseases.

  • Tumor Identification

In cancer diagnosis, H&E staining plays a pivotal role in identifying tumor cells. The distinctive nuclear and cytoplasmic features revealed by the stain help pathologists differentiate between normal and cancerous cells, allowing for accurate diagnosis and classification.

  • Inflammatory Conditions

Inflammatory conditions, such as infections or autoimmune diseases, often alter the appearance of tissues. H&E staining aids in identifying the specific types of inflammatory cells present, providing valuable insights into the nature and severity of the condition.

  • Organ Assessment

H&E staining is instrumental in assessing the structural integrity of various organs. It enables pathologists to identify changes in tissue architecture, helping diagnose conditions ranging from liver diseases to cardiovascular disorders.


In the realm of microscopic exploration, the Hematoxylin-Eosin staining technique stands as a testament to the union of science and art. Through the careful application of these dyes, scientists and medical professionals unlock a world of cellular details, paving the way for a deeper understanding of tissues and their pathologies. As we peer through the lens of a microscope at an H&E-stained slide, we are not merely observing colors; we are deciphering the intricate stories written in the microscopic tapestry of life.

Related Products/Services

Hematoxylin-Eosin Staining
Hematoxylin Counterstainer
Pink Hematoxylin Counterstainer

  1. Li Y, et al.; Hematoxylin and eosin staining of intact tissues via delipidation and ultrasound. Sci Rep. 2018, 8(1):12259.

*If your organization requires the signing of a confidentiality agreement, please contact us by email.

Please note: Our services can only be used for research purposes. Do not use in diagnostic or therapeutic procedures!

Online Inquiry