Immunofluorescence is one of the methods of immunosuppression, which first developed by Coons et al. In 1941. In this method, antibodies or fluorescent labeled antigens are used to determine the position of antigens and antibodies in the tissue sections. Fluorescence materials used as markers in this method include Fluorescein isothiocyanate (FITC), Ficvaririte (PE), Rhodamine B (RB), and Red (TX Red). These markers are affected by the impact of UV light, the surface orbitals of electrons being transmitted to the layer above their initial orbitals. When the light source is interrupted, the electrons that are stimulated return to their initial layer, which in turn returns their excess energy in the form of light. This light is visible and measured by special devices. Currently, in our country fluorescence microscopy is most widely used in fluorescence methods, and these methods are divided into direct and indirect groups.
Direct immunofluorescence assay
This method is often used to identify specific antigens, such as germs or cells and sedimentary complexes. Identification of each specific antigen is carried out by the antibodies labeled with flavors. Direct immunofluorescence can be used to search for immunoglobulin deposits or components of the complement system in tissue in a biopsy sample.
Indirect immunofluorescence assay
In this method, the antibody labeled with fluorescence material is used to identify the antibody desired in the serum. In this way, the specific antibody to be detected in the sample should be connected to a solid surface fixed by the specific antigen. The solid phase is then washed and adhered to the antibody labeled with fluorescence. After a wash step, the fluorescence amount is measured.
Indirect immunofluorescence is one of the most common methods for detecting antibodies to anti-angiogenic antibodies in the body of patients with autoimmune diseases. In this method, use of different tissue sections or human tumor cell line (HEp-2) as an antigen source
Gets In the indirect immunofluorescence assay, the antigens were identified by the patient’s autoantibodies and created special patterns. Each of these specific patterns can be safe in relation to one of their diseases.
- It is commonly used for connective tissue disorders.
- The immunofluorescence method has a high sensitivity and specificity and is easy and inexpensive to do.
- In this test, the presence of anti-nucleoside antibodies (ANA) is detected in the patient’s blood.
- In this test, the patient’s serum antibodies are attached to the substrate (cells containing anti-genes, such as HEp-2) that produce a distinct fluorescence pattern, each of which is associated with a specific autoimmune disease.