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For practical purposes, in objective based TIRF, medium 1 is typically a high refractive index glass coverslip, and medium 2 is the sample in solution with a lower refractive index. There may be immersion oil between the lens and the glass coverslip to prevent significant refraction through air.
Oil immersion objective lenses look superficially identical to non-oil immersion lenses. In light microscopy , oil immersion is a technique used to increase the resolving power of a microscope . This is achieved by immersing both the objective lens and the specimen in a transparent oil of high refractive index , thereby increasing the numerical ...
Electrostatic immersion objective lens is used to form a real image of the sample by way of a 2/3-magnification virtual image behind the sample. The uniformity of the electrostatic field between the objective lens and specimen, limited by spherical and chromatic aberrations larger than those of any other lenses, ultimately determines the ...
In optical microscopes a darkfield condenser lens must be used, which directs a cone of light away from the objective lens. To maximize the scattered light-gathering power of the objective lens, oil immersion is used and the numerical aperture (NA) of the objective lens must be less than 1.0. Objective lenses with a higher NA can be used but ...
Some microscopes make use of oil-immersion objectives or water-immersion objectives for greater resolution at high magnification. These are used with index-matching material such as immersion oil or water and a matched cover slip between the objective lens and the sample. The refractive index of the index-matching material is higher than air ...
The maximum NA is limited by the refractive index of the medium between the lens and the sample. As with objective lenses, a condenser lens with a maximum numerical aperture of greater than 0.95 is designed to be used under oil immersion (or, more rarely, under water immersion), with a layer of immersion oil placed in contact with both the ...
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Two Leica oil immersion microscope objective lenses; left 100×, right 40×. The objective lens of a microscope is the one at the bottom near the sample. At its simplest, it is a very high-powered magnifying glass, with very short focal length. This is brought very close to the specimen being examined so that the light from the specimen comes ...