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Correct positioning of the condenser with relation to the cone of illumination and focus (a step in establishing Köhler illumination) is critical to quantitative microscopy and to ensure the best digital images.Ĭondensers are divided primarily into classifications of imaging modality (such as brightfield, darkfield, and phase contrast), but also according to their degree of optical correction. Condenser height is controlled by a rack and pinion gear system that allows the condenser focus to be adjusted for proper illumination of the specimen. The opening and closing of this iris diaphragm controls the angles of illuminating wavefronts (and thus the aperture size) that bathe the specimen.
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Specifically, appropriate use of the adjustable aperture iris diaphragm (incorporated into the condenser or just below it) is of significant importance in securing correct illumination, contrast, and depth of field.
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The second reason is that the numerical aperture increases from 0.12 to 0.65 or, expressed as aperture angles, from 15 degrees to 80 degrees.Īperture adjustment and proper focusing of the condenser (with regard to height in relation to the objective) are of critical importance in realizing the full potential of the objective. The viewable area then becomes as much as 100x smaller. If a switch is made to the 40x objective, the diameter of the viewfield of the specimen shrinks by the factor 10 (to only 0.5 millimeters). An objective with a low magnification (for example, 4x) provides a large field of observation (with a diameter of as high as 5 millimeters in this case, provided that the eyepiece permits an intermediate image of diameter 20 millimeters). This is, in part, because the size of the observed specimen field changes with every objective magnification. The condenser aperture diaphragm is responsible for controlling the angle of the illuminating light cone and, consequently, the numerical aperture of the condenser.Īnother reason for the existence of fixed and adjustable diaphragms, prisms, beamsplitters, and filters in the microscope is that the illumination often must be reset after each change of the objective. When properly adjusted, light from the condenser will fill the rear focal plane of the objective with image-forming light by projecting a cone of light to illuminate the field of view. Such a situation leaves little light for observation, and as a result, the images assume a dark character. Any lack of brightness is not a problem in simple brightfield microscopy, but if contrast-enhancing techniques, such as phase contrast, differential interference contrast, fluorescence, or polarization contrast are used, additional optical elements that consume a significant portion of the available light flow are inserted into the beam path. Illumination of the specimen is the most important controllable variable in achieving high-quality images in microscopy, critical photomicrography, and digital imaging. The design of an optical microscope must ensure that the light rays are organized and precisely guided through the instrument.