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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.
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]
Additional flexibility can be added by using digital light-sheet microscopy to generate the illumination patterns. In digital LSM, the light sheet is created by rapidly scanning a laser beam through the sample. [3] [2] [14] This allows for fine control over the specific illumination pattern by modulating the intensity of the laser as it scans ...
Also the good optical sectioning capability reduces the background signal and thus creates images with higher contrast, comparable to confocal microscopy. Because light sheet fluorescence microscopy scans samples by using a plane of light instead of a point (as in confocal microscopy), it can acquire images at speeds 100 to 1,000 times faster ...
A total internal reflection fluorescence microscope (TIRFM) is a type of microscope with which a thin region of a specimen, usually less than 200 nanometers can be observed. TIRFM is an imaging modality which uses the excitation of fluorescent cells in a thin optical specimen section that is supported on a glass slide.
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
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