What Is Immunohistochemistry

What Is Immunohistochemistry

Introduction of Immunohistochemistry

Immunohistochemistry (IHC) is a powerful technique widely utilized in the field of diagnostic medicine, particularly in pathology and research. This method plays a crucial role in the identification and localization of specific proteins within tissue sections, providing valuable insights into the molecular composition of cells and tissues. By employing the principles of immunology and microscopy, immunohistochemistry has become an indispensable tool for pathologists and researchers alike.

Principles of Immunohistochemistry

At its core, immunohistochemistry involves the use of antibodies to detect and visualize specific proteins in tissue samples. Antibodies are molecules produced by the immune system that bind to specific target proteins, known as antigens. In the context of IHC, these antibodies are labeled with a detectable marker, such as a fluorescent dye or an enzyme, allowing for the visualization of the target protein under a microscope.

The immunohistochemistry of Breast Cancer.Figure 1. The immunohistochemistry of Breast Cancer.

The process begins with the preparation of thin tissue sections, which are typically obtained through biopsy or surgical procedures. These sections are then treated to enhance their binding capacity and to preserve the structural integrity of the proteins within. Subsequently, specific antibodies are applied to the tissue sections, where they bind selectively to their corresponding antigens.

Visualization of Results

To visualize the results of immunohistochemistry, various methods can be employed depending on the type of marker used. Enzyme-linked antibodies can be visualized through the addition of a substrate that produces a colored reaction product, making it easily detectable under a microscope. Fluorescent markers, on the other hand, emit light of a specific wavelength when exposed to the appropriate light source, allowing for precise localization of the target proteins.

Clinical Applications

Immunohistochemistry finds extensive application in clinical settings, aiding in the diagnosis and classification of diseases. One of its primary uses is in the identification of cancerous cells. By targeting specific proteins associated with different types of cancer, IHC allows pathologists to distinguish between benign and malignant tissues, aiding in the determination of an accurate diagnosis and the formulation of appropriate treatment plans.

Additionally, immunohistochemistry is instrumental in predicting the prognosis of certain diseases. For example, the presence or absence of specific markers in tumor tissues can provide valuable information about the likelihood of disease recurrence and the patient's overall prognosis. This information is crucial in tailoring personalized treatment strategies for individuals based on the molecular characteristics of their disease.

Research Applications

In addition to its diagnostic utility, immunohistochemistry is widely employed in research settings to investigate the molecular mechanisms underlying various diseases. Researchers use IHC to study the expression patterns of specific proteins in normal and diseased tissues, elucidating the role of these proteins in cellular processes and disease progression.

Moreover, immunohistochemistry facilitates the discovery of new therapeutic targets. By identifying proteins that are overexpressed or mutated in disease states, researchers can develop targeted therapies aimed at modulating the activity of these proteins, thereby offering more effective and precise treatment options.

Challenges and Considerations

While immunohistochemistry is a valuable technique, it is not without challenges. Variability in tissue preparation, antigen retrieval methods, and antibody specificity can impact the accuracy and reproducibility of results. Standardization of protocols and quality control measures are essential to ensure reliable and consistent outcomes across different laboratories.


In conclusion, immunohistochemistry stands as a cornerstone in the fields of pathology and biomedical research. Its ability to detect and visualize specific proteins within tissue sections provides invaluable information for diagnostic purposes and contributes significantly to our understanding of the molecular basis of diseases. As technology advances and protocols are refined, immunohistochemistry continues to evolve, promising even greater insights into the intricate world of cellular and tissue biology.

  1. Tan WCC, et al.; Overview of multiplex immunohistochemistry/immunofluorescence techniques in the era of cancer immunotherapy. Cancer Commun (Lond). 2020, 40(4):135-153.

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