Microscope Resolution

Microscope Magnification and Optical Resolution

A microscope's total magnification is a combination of the eyepieces and the objective lens. For example, a biological microscope with 10x eyepieces and a 40x objective has 400x magnification. There are however, a few limits to the amount of total magnification that can be reached before empty magnification comes into play. Empty magnification occurs when the image continues to be enlarged, but no additional detail is resolved. This is often the case when higher magnification eyepieces are used. In order to avoid empty magnification, there are a few simple steps that are helpful to follow.

Eyepiece and Objective Combinations for Optimal Magnification

When selecting a combination of eyepieces and objective lenses for the optimal magnification, without ending up with "empty magnification" it is important to consider the numerical aperature (NA) of the objective. The numerical aperture of a microscope objective defines the objective's resolution. Each microscope objective has a minimum and maximum magnification necessary for the details in an image to be resolved. A simple formula for the minimum value is (500 x NA). And for the maximum magnification (1000 x NA). Magnifications higher than this value will result in empty magnification, or an image that has a poor resolution. The table below shows some typical NA values with their corresponding objective and provides a range of useful magnification combinations. The blank boxes in the table would provide empty magnification and should be avoided. For example, pairing 20x eyepieces with a 100x objective would not provide good resolution and would result in empty magnification. To determine this, we took 1.25NA x 1000 = 1250 magnification maximum. However, the combination of the 100x objective x 20x eyepieces = 2000, which is above the maximum magnification.

Range of Useful Magnification based on NA of Objectives

Objective
(Numerical Aperture)
10x
12.5x
15x
20x
2.5x
(0.08 NA)
x
4x
(0.10 NA)
x
x
10x
(0.25 NA)
x
x
x
x
25x
(0.40 NA)
x
x
x
x
40x
(0.65 NA)
x
x
x
60x
(0.85 NA)
x
x
100x
(1.25 NA)
x
x
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