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| Transmitted light – polarization contrast |
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In this method, polarized light is used; it consists of light waves which all feature the same direction of vibration, i.e. which are linearly polarized. This very “ordered” light is generated by polarizers which filter out a privileged plane from the statistical confusion of vibration directions prevailing in natural light.
It is an important fact that two filters of this type do not let any light pass when they are arranged one behind the other in the beam path at an angle of 90° to each other. The first filter sorts out the vibration directions in such a way that the second filter cannot let pass this very selection. The second filter is called “analyzer”, since it allows the privileged direction of the first filter – called “polarizer” – to be checked.
The appropriate arrangement is relatively easy to implement in the microscope. The polarizer (1) on the condenser – near the aperture diaphragm – ensures that the specimen (3) is illuminated with linearly polarized light via the condenser. The analyzer (5), arranged at an angle of 90° to the polarizer (1), is located behind the objective. The tube lens (6) forms the intermediate image (7).
If no specimen is on the microscope stage – or only an empty, clean microscope slide – the image will remain completely dark. When illuminated, many specimens turn the vibration direction of the polarized light out of the plane produced by the polarizer.
Such specimens are mainly birefringent materials, in which the refractive index depends on the vibration direction of the incident light. This is mainly the case with crystals, such as starch or minerals, but also with polymers. If such materials are viewed under the polarization microscope between the crossed polarizer and the analyzer, bright areas can be seen in the image because light is partially transmitted by the analyzer.
The drawing of the beam path also includes a so-called auxiliary object (5a), also termed lambda plate. In polarized light, this lambda plate converts contrast to colors. As in phase contrast, path differences are used for this purpose, although this time with polarized light and birefringent material in the auxiliary object. The path differences generated lead to an extinction of certain wavelengths in the light, i.e. only certain colors remain from the white light and create beautiful, colored pictures.
Mechanical stress in the glass results in so-called stress-induced birefringence, which – in turn – influences the polarized light. Therefore, Pol examinations in the microscope require condensers and objectives which are free of such internal stress. Such objectives can be recognized by the “Pol” marking inscribed in red.
The unstained polymer fiber is only indistinctly visible in brightfield (left) but can be seen in detail in polarized light (center). The lambda plate (right) converts contrast to colors.
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