Oil immersion
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 aperture of the objective lens.
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. An oil immersion objective is an objective lens specially designed to be used in this way. Many condensers also give optimal resolution when the condenser lens is immersed in oil.
Lenses reconstruct the light scattered by an object. To successfully achieve this end, ideally, all the diffraction orders have to be collected. This is related to the opening angle of the lens and its refractive index. The resolution of a microscope is defined as the minimum separation needed between two objects under examination in order for the microscope to discern them as separate objects. This minimum distance is labelled δ. If two objects are separated by a distance shorter than δ, they will appear as a single object in the microscope.
A measure of the resolving power, R.P., of a lens is given by its numerical aperture, NA:
where λ is the wavelength of light. From this it is clear that a good resolution (small δ) is connected with a high numerical aperture.
The numerical aperture of a lens is defined as
where α0 is half the angle spanned by the objective lens seen from the sample, and n is the refractive index of the medium between the lens and specimen (≈1 for air).
State of the art objectives can have a numerical aperture of up to 0.95. Because sin α0 is always less than or equal to unity (the number "1"), the numerical aperture can never be greater than unity for an objective lens in air. If the space between the objective lens and the specimen is filled with oil however, the numerical aperture can obtain values greater than unity. This is because oil has a refractive index greater than 1.
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