Shorter wavelengths are capable of resolving details to a greater degree than are the longer wavelengths.
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There are several equations that have been derived to express the relationship between numerical aperture, wavelength, and resolution: Notice that equation 1 and 2 differ by the multiplication factor, which is 0. These equations are based upon a number of factors including a variety of theoretical calculations made by optical physicists to account for the behavior of objectives and condensers, and should not be considered an absolute value of any one general physical law. In some instances, such as confocal and fluorescence microscopy, the resolution may actually exceed the limits placed by any one of these three equations.
Other factors, such as low specimen contrast and improper illumination may serve to lower resolution and, more often than not, the real-world maximum value of r about 0. The following table Table 1 provides a list resolution r and numerical aperture NA values by objective magnification and correction.
When the microscope is in perfect alignment and has the objectives appropriately matched with the substage condenser, then we can substitute the numerical aperture of the objective into equations 1 and 2 , with the added result that equation 3 reduces to equation 2. An important fact to note is that magnification does not appear as a factor in any of these equations, because only numerical aperture and wavelength of the illuminating light determine specimen resolution.
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As we have mentioned and can be seen in the equations the wavelength of light is an important factor in the resolution of a microscope. Shorter wavelengths yield higher resolution lower values for r and visa versa. The greatest resolving power in optical microscopy is realized with near-ultraviolet light, the shortest effective imaging wavelength. Despite the objections of his wife, Michael decides to make one last attempt to save his friend from drug addiction.
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