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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]
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.
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.
The stereo, stereoscopic or dissecting microscope is an optical microscope variant designed for low magnification observation of a sample, typically using light reflected from the surface of an object rather than transmitted through it. The instrument uses two separate optical paths with two objectives and eyepieces to provide slightly ...
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.
The optical microscope, also referred to as a light microscope, is a type of microscope that commonly uses visible light and a system of lenses to generate magnified images of small objects. Optical microscopes are the oldest design of microscope and were possibly invented in their present compound form in the 17th century.
Historically, microscopes were nearly universally designed with a finite mechanical tube length, which is the distance the light traveled in the microscope from the objective to the eyepiece. The Royal Microscopical Society standard is 160 millimeters, whereas Leitz often used 170 millimeters. 180 millimeter tube length objectives are also ...
By 1978, the first theoretical ideas had been developed to break the Abbe limit, which called for using a 4Pi microscope as a confocal laser-scanning fluorescence microscope where the light is focused from all sides to a common focus that is used to scan the object by 'point-by-point' excitation combined with 'point-by-point' detection. [14]
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