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The correct immersion oil for an objective lens has to be used to ensure that the refractive indices match closely. Use of an oil immersion lens with the incorrect immersion oil, or without immersion oil altogether, will suffer from spherical aberration. The strength of this effect depends on the size of the refractive index mismatch.
Using aseptic technique, prepare and air dried heat fixed slide with the desired organism. ... view slide under the microscope under the oil immersion objective (100X).
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 ...
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 ...
Immersion oils are transparent oils that have specific optical and viscosity characteristics necessary for use in microscopy. Typical oils used have an index of refraction around 1.515. [2] An oil immersion objective is an objective lens specially designed to be used in this way.
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.
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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.