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Basics of Microscopes

What is a fluorescence microscope?

A fluorescence microscope enables cells and proteins to be observed by using a fluorescent protein or antibody as a label. This type of microscope is indispensable for modern cell biology.

Principle Fluorescent materials absorb a specific wavelength of light (excitation light) and emit light of a longer wavelength (fluorescence), which is based on Stokes’ law. For example, the fluorescence emitted by a target molecule can be observed by adding a specific fluorescent reagent to cells and then applying excitation light. A fluorescent microscope has all of the components to induce this fluorescence and capture the resulting image.
Features - Allows observation of fluorescence images, in addition to observation with transmitted light.
- It is possible to only observe specific areas by using different fluorescent labels.
- Fluorescent dyes enable users to view localization of particular proteins in cells.
- The use of fluorescent proteins such as GFP allows for observation of living cells.
Structure Generally, a fluorescence microscope is a combination of a biological microscope and fluorescent incident illumination equipment. The structure includes a focusing knob, an XY stage handle for positioning specimens, and a revolver for switching objective lenses. For lighting, it is also equipped with a cube turret that adjusts the wavelength of the excitation light, a shutter that prevents photobleaching of samples, and an ND (neutral density) filter that adjusts the strength of the excitation light.

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