immunohistokjemi

Immunohistochemistry (IHC) is a laboratory technique used to detect the presence of specific proteins in cells and tissues. It is widely used in medical research, pathology, and clinical diagnostics to identify and localize antigens within tissue sections. The process involves several key steps: tissue fixation, sectioning, antigen retrieval, blocking, primary antibody incubation, secondary antibody incubation, and visualization.

Tissue fixation is the first step, where the tissue is preserved to maintain its structure and antigenicity. This is typically achieved using formaldehyde or other fixatives. The tissue is then embedded in a suitable medium, such as paraffin wax, and cut into thin sections using a microtome.

Antigen retrieval is performed to expose the antigens of interest, which may be masked by the tissue matrix. This can be done using various methods, such as heat-induced epitope retrieval (HIER) or enzymatic digestion.

Blocking is the next step, where non-specific binding sites on the tissue section are saturated with a blocking agent, such as normal serum or bovine serum albumin (BSA), to reduce background staining.

The primary antibody, specific to the antigen of interest, is then incubated with the tissue section. This antibody binds to the antigen, forming an antigen-antibody complex.

A secondary antibody, which is conjugated to an enzyme or fluorescent dye, is then incubated with the tissue section. This antibody binds to the primary antibody, allowing for the visualization of the antigen-antibody complex.

Finally, the tissue section is developed using a chromogenic substrate or fluorescent dye, resulting in a visible stain that indicates the presence and location of the antigen within the tissue.

Immunohistochemistry is a powerful tool for the study of cellular and molecular biology, as well as for the diagnosis and prognosis of various diseases. It allows for the visualization of proteins at the cellular level, providing valuable insights into the biology of cells and tissues.