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In confocal laser-scanned microscopes, the full-width half-maximum (FWHM) of the point spread function is often used to avoid the difficulty of measuring the Airy disc. [1] This, combined with the rastered illumination pattern, results in better resolution, but it is still proportional to the Rayleigh-based formula given above.
An optical instrument is said to be diffraction-limited if it has reached this limit of resolution performance. Other factors may affect an optical system's performance, such as lens imperfections or aberrations, but these are caused by errors in the manufacture or calculation of a lens, whereas the diffraction limit is the maximum resolution ...
Optical microscope. 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.
Super-resolution microscopy is a series of techniques in optical microscopy that allow such images to have resolutions higher than those imposed by the diffraction limit, [1][2] which is due to the diffraction of light. [3] Super-resolution imaging techniques rely on the near-field (photon-tunneling microscopy [4] as well as those that use the ...
In a dry objective or condenser, this gives a maximum NA of 0.95. In a high-resolution oil immersion lens, the maximum NA is typically 1.45, when using immersion oil with a refractive index of 1.52. Due to these limitations, the resolution limit of a light microscope using visible light is about 200 nm.
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]
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 ...
The resolution of a good optical microscope is mainly determined by the numerical aperture (A Num) of its objective lens. The numerical aperture in turn is determined by the refractive index n of the medium filling the space between the sample and the lens and the half collection angle of light θ according to Carlsson (2007): [ 46 ] : 6 A N u ...