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Critical illumination acts to form an image of the light source on the specimen to illuminate it. [2] This image is formed by the condenser or collector lens. This illumination is bright but not always even, as any structure in the light source (for example the filament of a light bulb) will be visible in the
Fluorescence and confocal microscopes operating principle. Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser scanning confocal microscopy (LSCM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation. [1]
In both cases the numerical aperture of the objective is 1.49 and the refractive index of the medium 1.52. The wavelength of the emitted light is assumed to be 600 nm and, in case of the confocal microscope, that of the excitation light 500 nm with circular polarization. A section is cut to visualize the internal intensity distribution.
Köhler illumination is a method of specimen illumination used for transmitted and reflected light (trans- and epi-illuminated) optical microscopy.Köhler illumination acts to generate an even illumination of the sample and ensures that an image of the illumination source (for example a halogen lamp filament) is not visible in the resulting image.
A condenser is an optical lens that renders a divergent light beam from a point light source into a parallel or converging beam to illuminate an object to be imaged. Condensers are an essential part of any imaging device, such as microscopes , enlargers , slide projectors, and telescopes.
Typical components of a fluorescence microscope are a light source (xenon arc lamp or mercury-vapor lamp are common; more advanced forms are high-power LEDs and lasers), the excitation filter, the dichroic mirror (or dichroic beamsplitter), and the emission filter (see figure below).
Monochromatic aberrations are caused by the geometry of the lens or mirror and occur both when light is reflected and when it is refracted. They appear even when using monochromatic light, hence the name. Chromatic aberrations are caused by dispersion, the variation of a lens's refractive index with wavelength. Because of dispersion, different ...
This means the lens's F-number value is fixed to the overall designed focal ratio of the optical system (the diameter of the primary mirror divided into the focal length). The inability to stop down the lens results in the catadioptric lens having a short depth of field.