Can be used for confocal microscopy without fluorescence

Fluorescence microscopy

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Fluorescence is one of the most frequently used physical phenomena in biological and analytical microscopy, mainly due to its high sensitivity and high specificity. Fluorescence is a form of luminescence. Fluorescence microscopy even enables users to determine the distribution of a single type of molecule, its amount and its location in a cell. Colocalization and interaction studies can be performed and ion concentrations as well as intra- and intercellular processes such as endocytosis and exocytosis can be observed. With the help of high-resolution fluorescence microscopy, it is even possible to image structures that are below the resolution limit.

Fluorescence light microscopes

Fluorescence microscopes used in research applications are based on a set of optical filters:

  • an excitation filter
  • a dichroic beam splitter and
  • an emission filter

The filters are often put together in a filter cube (compound microscope) or in a flat plate (mainly stereo microscope).

During the Excitation filter lets through the wavelengths for the excitation of a certain color in the sample, serves the Emission filter as a kind of quality control in that it only lets through the wavelengths of interest emitted by the fluorophore. The dichroic mirrors has the task of reflecting light in the excitation area and transmitting light in the emission area, which enables classic incident light illumination with epifluorescence.

Fluorescence stereomicroscopes

The fluorescence stereomicroscopes from Leica Microsystems use TripleBeam technology, a separate (third) beam path for fluorescence illumination of the sample without dichroic mirrors.

An excitation filter for the beam path and two emission filters, one for each observation beam path, are therefore required.