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where n is the index of refraction of the medium in which the lens is working (1.00 for air, 1.33 for pure water, and typically 1.52 for immersion oil; [1] see also list of refractive indices), and θ is the half-angle of the maximum cone of light that can enter or exit the lens.
The fluorescence light emitted from the lightsheet is then collected perpendicularly with a standard microscope objective and projected onto an imaging sensor (usually a CCD, electron-multiplying CCD or CMOS camera). In order to let enough space for the excitation optics/lightsheet an observation objective with high working distance is used.
Using a small-angle approximation, the angular resolution may be converted into a spatial resolution, Δℓ, by multiplication of the angle (in radians) with the distance to the object. For a microscope, that distance is close to the focal length f of the objective. For this case, the Rayleigh criterion reads:
Barlow lenses for microscopes can be found with magnifications ranging from 0.3× to 2×. Some standard lenses are 2×, which decreases the working distance by half and doubles the magnification, 0.75× (3/4×), which increases the working distance by 4/3× (1.33×) and decreases the magnification by 0.75×, and a 0.5× Barlow doubles the ...
A Scanning SQUID Microscope is a sensitive ... The details of this mathematical calculation ... This allows one to scan currents of 500 nA at a working distance of ...
The near-field optical (NFO) microscope involved a sub-wavelength aperture at the apex of a metal coated sharply pointed transparent tip, and a feedback mechanism to maintain a constant distance of a few nanometers between the sample and the probe. Lewis et al. were also aware of the potential of an NFO microscope at this time. [14]
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 observation of sub-wavelength structures with microscopes is difficult because of the Abbe diffraction limit.Ernst Abbe found in 1873, [2] and expressed as a formula in 1882, [3] that light with wavelength , traveling in a medium with refractive index and converging to a spot with half-angle will have a minimum resolvable distance of
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