Eyepieces, like objectives, are classified in terms of their ability to magnify the intermediate image.

Their
magnification factors vary between 5X and 30X with the most commonly used eyepieces having a value of 10X-
15X. Total visual magnification of the microscope is derived by multiplying the magnification values of the
objective and the eyepiece. For instance, using a 5X objective with a 10X eyepiece yields a total visual
magnification of 50X and likewise, at the top end of the scale, using a 100X objective with a 30X eyepiece gives a
visual magnification of 3000X.
Total magnification is also dependent upon the tube length of the microscope. Most standard fixed tube length
microscopes have a tube length of 160, 170, 200, or 210 millimeters with 160 millimeters being the most common
for transmitted light biomedical microscopes. Many industrial microscopes, designed for use in the semiconductor
industry, have a tube length of 210 millimeters. The objectives and eyepieces of these microscopes have optical
properties designed for a specific tube length, and using an objective or eyepiece in a microscope of different
tube length will lead to changes in the magnification factor (and may also lead to an increase in optical aberration
lens errors). Infinity-corrected microscopes also have eyepieces and objectives that are optically-tuned to the
design of the microscope, and these should not be interchanged between microscopes with different infinity tube
lengths.
Modern research microscopes are very complex and often have both episcopic and diascopic illuminators built
into the microscope housing. Design constrictions in these microscopes preclude limiting the tube length to the
physical dimension of 160 millimeters resulting the need to compensate for the added physical size of the
microscope body and mechanical tube. This is done by the addition of a set of parallelizing lenses to shorten the
apparent mechanical tube length of the microscope. These additional lenses will sometimes introduce an
additional magnification factor (usually around 1.25-1.5X) that must be taken into account when calculating both
the visual and photomicrographic magnification. This additional magnification factor is referred to as a tube
factor in the user manuals provided by most microscope manufacturers. Thus, if a 5X objective is being used
with a 15X set of eyepieces, then the total visual magnification becomes 93.75X (using a 1.25X tube factor) or
112.5X (using a 1.5X tube factor).

In addition to the parallelizing lenses used in some microscopes, manufacturers may also provide additional
lenses (sometimes called magnification changers) that can be rotated into the optical pathway to increase the
magnification factor. This is often done to provide ease in specimen framing for photomicrography. These lenses
usually have very small magnification factors ranging from 1.25X up to 2.5X, but use of these lenses may lead
to empty magnification, a situation where the image is enlarged, but no additional detail is resolved. This type of
error is illustrated in Figure 7 with photomicrographs of liquid crystalline DNA. The photomicrograph in Figure 7(a)
was taken with a 20X plan achromat objective under polarized light with a numerical aperture of 0.40 and
photographically enlarged by a factor of 10X. Detail is crisp and focus is sharp in this photomicrograph that
reveals many structural details about this hexagonally-packed liquid crystalline polymer. Conversely, the
photomicrograph on the right (Figure 7(b)) was taken with a 4X plan achromat objective, having a numerical
aperture of 0.10 and photographically enlarged by a factor of 50X. This photomicrograph lacks the detail and
clarity present in Figure 7(a) and demonstrates a significant lack of resolution caused by the empty magnification
factor introduced by the enormous degree of enlargement.
Care should be taken in choosing eyepiece/objective combinations to ensure the optimal magnification of
specimen detail without adding unnecessary artifacts. For instance, to achieve a magnification of 250X, the
microscopist could choose a 25X eyepiece coupled to a 10X objective. An alternative choice for the same
magnification would be a 10X eyepiece with a 25X objective. Because the 25X objective has a higher numerical
aperture (approximately 0.65) than does the 10X objective (approximately 0.25), and considering that numerical
aperture values define an objective's resolution, it is clear that the latter choice would be the best. If
photomicrographs of the same viewfield were made with each objective/eyepiece combination described above,
it would be obvious that the 10x eyepiece/25x objective duo would produce photomicrographs that excelled in
specimen detail and clarity when compared to the alternative combination.
The range of useful total magnification for an objective/eyepiece combination is defined by the numerical
aperture of the system. There is a minimum magnification necessary for the detail present in an image to be
resolved, and this value is usually rather arbitrarily set as 500 times the numerical aperture (500 × NA). At the
other end of the spectrum, the maximum useful magnification of an image is usually set at 1000 times the
numerical aperture (1000 × NA). Magnifications higher than this value will yield no further useful information or
finer resolution of image detail, and will usually lead to image degradation, as discussed above. Exceeding the
limit of useful magnification causes the image to suffer from the phenomenon of empty magnification (see
Figures 7 (a) and (b)), where increasing magnification through the eyepiece or intermediate tube lens only
causes the image to become more magnified with no corresponding increase in detail resolution. Table 1 lists the
common objective/eyepiece combinations that lie in the range of useful magnification.

Range of Useful Magnification

(500-1000 × NA of Objective)
OBJECTIVE EYEPIECES
(NA) 10x 12.5x 15x 20x 25x
2.5X
--- --- --- x x
(0.08)
4X
--- --- x x x
(0.12)
10X
x x x x x
(0.35)
25X
x x x x ---
(0.55)
40X
x x x --- ---
(0.70)
60X
x x x --- ---
(0.95)
100X
x x --- --- ---
(1.40)
x = good combination

Table 1
These basic principles underlie the operation and construction of the compound microscope which, unlike a
magnifying glass or simple microscope, employs a group of lenses aligned in series. The elaboration of these
principles has led to the development, over the past several hundred years, of today's sophisticated instruments.
Modern microscopes are often modular with interchangeable parts for different purposes; such microscopes are
capable of producing images from low to high magnification with remarkable clarity and contrast.