Immunofluorescence is one of the widely used techniques in modern biology and medicine, and it is developed by Coons et al. (1950), and it is a combination of immunofluorescence technique and morphological technology to develop immune fluorescent cells (or tissue).
Immunofluorescence may be used to analyze the distribution of proteins, glycans, and small biological and non-biological molecules.
Immunofluorescence has been widely used in biological research and medical research yield and becomes one most important and effective method. There are two different immunofluorescence assay which include indirect immunofluorescence assay and direct
There are several microscope designs can be used for analysis of immunofluorescence samples. The simplest is the epifluorescence microscope, and now the confocal microscope is also widely used. With the improvement of technology, various super-resolution microscope designs with much higher resolution can also be used.
There are also some limiting factors for Immunofluorescence / IF/ ICC. One of that is fixing cells, molecules on the outside of the cell membrane or in the supernatant can be bound by the antibodies which allow for living cells to be stained, and molecules in cell membrane must be stained after fixation which may make protein cross-linked and false negative or false positive signals due to non-specific binding.
1. Clear positioning. This assay can give research the clear subcellular localization of molecules.
The expression of molecules can be observed directly.
2. High specificity. The preparation of sample can effectively protect the natural structure of the antigen
3. High sensitivity. The target with low expression can be detected with immunofluorescence / IF/ ICC.
4. Easy to operate. The test result can be get quickly. This assay can be used vivo experiments.
5. Multiple staining. The multiple targets can be detected in the same sample.
6. Beautiful result. It is more advantageous to publish the article through the beautiful image display result.