Histology
The histological analysis of brain tissue is performed to understand the microscopic structure of these tissues and how they change under different conditions. There are many commercial markers available that can give us insights and answers.
The principle
Firstly, the tissue is collected and preserved using a fixative to maintain its structure. Hereafter the tissue is quickly frozen using liquid nitrogen. This method prevents the degradation and preserves the proteins and enzymes. The frozen tissue is then sliced in sections of 30-40 µm. Next, a free-floating immunohistochemical (IHC) staining is performed that targets the protein, enzyme, or cell of interest.
IHC staining is a technique that uses antibodies to detect specific proteins within the tissue. A primary antibody is used to bind to the specific protein of interest. After washing away the unbound antibodies, a secondary antibody is added. This antibody binds to the primary; it also contains a detectable marker, eg. fluorescent dye. This dye is then visualized using a fluorescent microscope. The fluorescent signal corresponds with the targeted protein. Multiple primary and secondary antibodies and fluorescent dyes can be used, which gives the researcher the opportunity to visualize multiple targets.
