| The
microscope is an essential tool for studying microbes. The brightfield
(compound) microscope pictured here is the type of scope most frequently
employed. Its usefulness depends on the magnification and resolution capabilities
of the instrument.
Magnification is the enlargement of an object. Two
lenses are used to magnify in a compound scope: an objective lens and an
ocular (eyepiece) lens. The microscope shown here has lenses in four objectives:
scanning (4 X), low power(10 X), high dry (40 X), and oil immersion (100
X). The ocular lens also magnifies ten times (10 X). Thus the total magnification
equals the product of the objective and ocular magnification values. This means that
an organism viewed through the ocular with the low power (10 X) objective
locked in place would have a total magnification of 100 times (100 X).
The oil immersion objective with its total magnification of 1000X is needed to magnify bacteria sufficiently.
Resolution (resolving power) is the ability of
the lenses to distinguish between two points a specified distance apart.
The brightfield microscope has a resolution of 0.2 micrometers.
This means that the microscope can distinguish two points as separate objects
if they are at least 0.2 micrometers apart. If two objects are any closer
than 0.2 micrometers, they will be magnified as one object.
The various parts of the microscope are identified
in the images shown here. In addition to the objectives and the oculars,
microscope parts include the following:
arm - supports the
microscope and is used for carrying it
base - support
structure; used in carrying microscope
body tube - holds
the oculars and channels the light rays
coarse adjustment knob-
used to initially focus specimen
condenser - collects
the light rays into a single cone of light
diaphraghm - provides
contrast by regulating brightness of light
fine adjustment knob-
used for final focus of specimen
mechanical stage
- platform for holding slide; contains clips to hold
slide; movement of stage is controlled by mechanical
stage knobs
revolving nosepiece
- holds the objectives
Specimens are initially found on scanning or low power.
(Bacteria are so small that the scanning power is of little use in locating
them. Low power is much more effective.) The microscope shown here is parfocal.
This means that a specimen in focus on one objective will be approximately
in focus on other objectives. It is relatively simple to bring the
organism into focus with high power and oil immersion if it has been focused
on low power.
The oil immersion objective requires immersion
oil to focus properly. Because the oil has the same refractive index as
the glass, it keeps the light rays traveling through to the objective.
In contrast, oil that is on the slide or the objective interferes with
proper imaging of the scanning, low, and high dry objectives.
Click on each image to see an enlarged version
.
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