What is a fluorescence microscope?

A fluorescence microscope is a conventional light microscope that is fitted with a complementary lighting fixture, called “fluorescence.” This makes it a very useful tool in the field of scientific research, since it allows reaching high levels of sensitivity and microscopic resolution, allowing a different appreciation of the information that can be obtained from specimens and that generally goes unnoticed, with conventional light.

Fluorescence is part of the luminescence processes. In which susceptible molecules emit light from excited states. That they are created electronically by a physical (for example, light absorption), mechanical (friction), or chemical mechanism. The generation of luminescence through the excitation of a molecule by photons of ultraviolet or visible light is a phenomenon called photoluminescence. This is formally divided into two categories, fluorescence and phosphorescence. Depending on the electronic configuration of the excited state and the emission pathway.

Fluorescence is the property of some atoms and molecules to absorb light at a particular wavelength. And subsequently emitting longer wavelength light after a short interval, called the fluorescence lifetime. The phosphorescence process occurs in a similar way to fluorescence, but with a much longer excited lifetime.

How is the fluorescence process?

The fluorescence process is governed by three important events. They all occur on timescales that are several orders of magnitude apart. Excitation of a susceptible molecule by an incoming photon occurs in femtoseconds (1015 sec). While the vibratory relaxation of electrons in the excited state at the lower energy level is much slower; and can be measured in picoseconds (1012 sec). The final process is the emission of a photon of longer wavelength and the return of the molecule to the ground state. This occurs in the relatively long time period of nanoseconds (109 sec).

The entire lifetime of molecular fluorescence, from excitation to emission, is measured in just one billionths of a second. The phenomenon is a surprising manifestation of the interaction between light and matter. This interaction forms the basis for expansive steady-state fields and time-resolved fluorescence spectroscopy and microscopy. Due to the highly sensitive emission profiles, spatial resolution, and high specificity of fluorescence investigations, the technique has become an important tool in genetics and cell biology.

Fluorescence microscope applications

There are many applications of fluorescence microscopy, especially in biology and medicine:

  • Labeling of molecules in cells and tissues for their characterization and identification.
  • Study of normal and pathological cells.
  • Immunological studies.
  • Mineralogy.

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