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Preparation

The Fluorescence Microscope
- Preparation of Specimen

Studying the dynamics in the cell is essential for understanding cell function. Fluorescence microscopy is one of the most used approaches in studying the location and movement of molecules and subcellular components in the cell. Usually, cellular components do not fluoresce themselves. Fluorescent markers are therefore introduced.

Fluorescent Dyes
Fluorescent dyes are directly taken up by the cells. They are incorporated and concentrated in specific subcellular compartments. The living cells are then mounted on a microscope slide and examined in a fluorescence microscope.

Immunofluorescence
Immunofluorescence is another common technique. It involves the use of antibodies to which a fluorescent marker has been attached. Antibodies are molecules that recognize and bind selectively to specific target molecules in the cell. The fluorescent signal can be amplified by using an unlabelled primary antibody and detecting it with labelled secondary antibodies.

Tagging of Proteins
It is possible to modify cells so that they create their own fluorescing molecules. These protein molecules are tagged with a fluorescing marker. When a specific protein is modified in this way, the location of that protein can be studied. It is also possible to watch the movements of the proteins and its interactions with other cellular components inside the cell.

 

 
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