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  Fluorescence Microscope

Fluorescence Microscope:

Fluorescence microscope is therefore designed that the specimen can be illuminated at one wavelength of light and noticed by a light emitted at various wavelengths (figure is as shown below).

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                                    Figure: Fluorescence Microscopy-Example of Principle of Light pathway

Certain organisms exhibit fluorescence naturally under UV light. When the specimen does not have natural fluorescence and it has to be tainted with a fluorescent dye termed as fluorochromes. Whenever microorganisms are stained with fluoro-chrome and noticed under fluorescence microscope with an UV or close to UV source. They emerge as luminescent, bright objects against a dark background.

The Fluorescence Microscope exposes a specimen to UV violet or blue light and forms an image of the object with resultant fluorescent light. A mercury vapor arc lamp generates an intense beam and heat transfer is emitted by a particular infrared filter. The light passes via an exited filter that transmits only the necessary wavelength. A dark field condenser gives a black background against which the fluorescent object glows.

The microscope creates an image of the fluorochrome stained microorganism from the light emitted whenever they fluoresce. The barrier filter situated after the objective lens eliminates the remaining UV light which may damage the eyes or blue and violet light that will decrease the image contrast.

The fluorescent microscope is employed in medical microbiology to observe pathogens like Mycobacterium tuberculosis. In microbial biology the fluorescence microscope is employed to observe microorganisms after staining the sample with fluorochrome acridine orange dye. The stained organisms having fluorescent orange or green can be noticed among another particulate matter.

 

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