THE MICROSCOPE

INTRODUCTION
The microscope is one of the basic tools in biology for studying structures that could not be
seen by the naked eye. The first microscopes were light microscopes. This used the principle of the
refraction of light rays in producing enlarged images. Simple lenses were among the first
microscopes. Earliest reports on its use was by Pliny and later by Leeuwenhoek who fabricated
one so well that he was able to discover bacteria with it.
Many changes and developments have been done to improve the light microscope. The first of
these was the introduction of the compound microscope which is the type that you will use and
study in this laboratory course. Its design includes two sets of lenses, the ocular, and the objectives
on opposite ends of a closed tube. This was done to overcome the problem of the early
microscope which was not about magnification but about focal distance which would decrease
as magnification increases. This means that if magnification is increased the viewer suffers the
inconvenience of thrusting his eye into the surface of the lens in order to view the object. More
improvements have been made to overcome other problems later encountered in light
microscopy. The phase contrast, the light interference, the dark field and the confocal
microscopes are a few of these.
The electron microscope is also an important breakthrough which uses electron beams instead
of refracted light in producing highly magnified images. The major problems however of all these
new improvements on the microscope are cost and size. For instance, an electron microscope
requires one big room which adds to its cost of acquisition and maintenance. However recent
developments in microfluidics and optics produced a new concept on design of microscopes. This
is known as Optofluidic microscopy (OFM). In this new concept, the lens is replaced with a metal
coated sensor called Complementary Metal Oxide Semiconductor (CMOS) etched with an array
of micrometer sized apertures.

PARTS OF A MICROSCOPE
Mechanical Parts
These are the parts which are made of metal and function to support and adjust the optical parts.
BASE - the structure that supports the whole weight of the microscope; may have horse-shoe
shape or may just be a rectangular flatbed.
PILLAR - a supporting piece which arises from the base and forms a short column to join the
arm.
ARM - short curved structure used in holding the instrument.
BODY TUBE - hollow cylinder which bears the two separate lens systems, the objectives at the
lower end and the eyepiece at the upper end.
DRAW TUBE - upper portion of the body tube which bears the eyepiece.
REVOLVING NOSEPIECE - structure at the lower end of the body tube which can be rotated
and bears the objectives.
DUST SHIELD - part above the revolving nosepiece which protects the objectives from the
dust.
STAGE - a square platform where the slide to be examined is placed. Note the presence of an
aperture at the center.
STAGE CLIPS - found on the stage to hold the slide.
DIAPHRAGM - found under the stage to regulate the amount of the light admitted to the
specimen. Your microscope may have either of these two types:
1. Iris - immediately beneath the stage aperture whose opening is regulated by a lever.
2. Disk - a plate with circular openings of varied diameters
ADJUSTMENT SCREW - maybe attached to the upper part of the arm or pillar. This will move
either the body tube or stage in order to get a clear view of the specimen.
1. Coarse Adjustment - produces greater movement. Be careful when manipulating this
screw.
2. Fine Adjustment - produces fine upward or downward movement.
Optical Parts
These consist chiefly of special type of glass and aligned on an optical axis.

WARNING:
Do not touch the lenses or glass with your hands nor allow these to
come in contact with stains, sharp, or dirty materials.

MIRROR - found below the stage and is usually double-faced, one is flat and the other is
concave. For electric microscopes, a built-in light system is instead present. Note: Electric
microscopes should be switched off when not in use to avoid overheating.
CONDENSER - if present, it is a secondary lens found immediately beneath the stage to
concentrate light rays and is provided with separate adjustment screw. This maybe absent in
the microscope.
OCULAR OR EYEPIECE - mounted in the draw tube where one peeps into when viewing the
microscope.
Objectives - attached to revolving nosepiece.
1. Very low power objective or scanner (may be absent) - if present this is the shortest tube
which enables the viewer to see a larger area of the object. It bears the number 3.5X or
4X
2. Low power objective- gives a bigger image than the scanner and more details of the
specimens. Its magnification number is 10X.
3. High power objective- longer than the LPO; shows more details of the specimen than LPO.
Its magnification number is 40X or 45X.
4. Oil-immersion objective (may be absent)- if present, this is the longest objective and gives
the highest magnification; immersion oil is placed in mounting the slide to enable the
viewer to see the object. It magnifies 100X

OCULAR OR EYEPIECE

BODY TUBE DRAW TUBE

DUST SHIELD
COARSE ADJUSTMENT
REVOLVING NOSEPIECE
OBJECTIVES
STAGE FINE ADJUSTMENT
CONDENSER
ARM
DIAPHRAGM
MIRROR
PILLAR
BASE

Figure 1. A Microscope and its Parts

MICROSCOPE CARE AND USE

Handling
When moving and setting up a microscope, proper care should be given to how the device is
held. Remove the microscope from the cabinet by gripping the arm firmly with one hand while
placing the other hand underneath the base of the device for support Do not remove the dust
cover until the microscope is placed and positioned on the table where it will be used
Lens Care
Never touch the lenses with your fingers. Oil from your hands can smudge the lens and in some
cases even leave scratches. If you must dean the lens, use only the appropriate lens tissues to
avoid damaging the lens
Storage
When your work with the microscope is finished, remember to rotate the nosepiece back to the
lowest power setting. Lower the nosepiece to its lowest position to the stage. Make sure the
power cord is neatly wrapped up and replace the dust cover before returning the microscope to
the cabinet.

MAGNIFICATION
Magnification is an important information on biological specimens. Two methods for computing
will be done in this exercise.
1. Magnification of Microscopic Specimens. Theoretically, it is a product of the magnification
power of ocular and magnification power of objective used. This is called LINEAR OR
THEORETICAL MAGNIFICATION.
2. Magnification of Macroscopic Specimens. Make three proportional drawings (1. reduced, 2.
same size and 3. enlarged) of the object (butterfly) below in your data sheet. Compute the
magnification index (X) as follows:

L x W of drawing
X=
L x W of object

Where: L = length and

W = width

OBJECTIVE LENSES
Eyepiece
One of the most important parts of the microscope are the lenses used to actually view the
specimen, The microscope uses the eyepiece lens, or the lens you look through, to present the
image of the specimen Eyepiece lenses are typically 10X, but may also come in 5X, 15X or 20X.
The power is generally found labeled on the eyepiece itself.
Objective Lenses
The lens nearest the object being viewed is called the objective lens. These lenses can come in a
range of powers such as 4X (Scanning), 10X (Low Power), 4OX (High Power), or 100X (Oil
Immersion Objective). Some lenses, such as the 100X lens, are specifically designed to be used
with immersion oil.
Objective Markings
The power of each lens can be found labeled on the side of the tube housing. Additionally, each
lens has a specific, color-coded band that denotes its resolution, red for 4X, yellow for 1OX, blue
for 40X, and white for 100X.
Calculating Magnification
To calculate the total magnification of the image, multiply the power of the eyepiece by the
power of the objective lens. For example, an eyepiece of 10X and an objective lens of 40X yields
a 40OX total magnification.
Oil Lens
Most lenses are used "dry" or without oil. Some lenses use a special type of oil to increase their
resolution. Oil is typically used with higher powered lens such as 100X. These lenses will typically
have special markings on the side of the housing to indicate it is designed to be used with oil.
Immersion Oil
When used with oil, the lens is lowered until it just contacts a small drop of oil placed on the
specimen slide. The oil serves as a bridge between the lens and the glass plate containing the
specimen, helping to focus the light and increase the resolution. Once oil is added to a slide, the
objective lens should not be switched back to a non-oil lens.
Light Refraction
Normally, some of the light is lost due to refraction when using the lens without immersion oil. The
addition of immersion oil helps to refocus the light as it passes from the slide to the lens. This
provides an image with better resolution and contrast.

TERMINOLOGY
ABBE CONDENSER
This specially-designed lens, with an iris-type aperture, and mounted under the stage, moves
vertically to adjust the beam of light entering the lens system. Changing the size of the iris and
adjusting its position in relation to the stage controls the diameter and focal point of the cone of
light as it passes through the specimen.
ACHROMATIC LENSES
Achromatic lenses are used to correct the bending or refraction of light as it passes through the
prisms or lenses in the microscope. Each achromatic lens has different indexes of refraction to
realign certain colors of light at the focal point, resulting in a much clearer image.
ARM
The arm is the area of the microscope between the tube and the base. To carry a microscope,
grip the arm with one hand and place the other hand undermeath the base.
ARTICULATED ARM
This stand holds the microscope body, clamps to a table, and provides movement in three
dimensions.
BASE
The base is the bottom support structure of the microscope. To carry a microscope, grip the arm
with one hand and place the other hand underneath the base.
BINOCULAR HEAD
A binocular microscope features a head with two eyepiece lenses, one eyepiece for each eye.
BODY
The body refers to the main section of the microscope, minus the stand (base) or any illuminators.
The body components generally include the eyepiece and objective lenses, but not the focusing
block.
C-MOUNT
A c-mount is an adapter for use with a video camera. In general, the lens from the camera is
removed and replaced with an adapter which then connects to the trinocular port of the
microscope.
COARSE FOCUS
The larger of two adjustment knobs that moves the objective lenses closer to or farther away from
the specimen in large steps. (See also Fine Focus).
COAXIAL FOCUS
This focusing system features the coarse and fine focus knobs on a single rotation axis. With this
setup, the coarse focus knob is generally larger and on the outside while the fine focus knob is
smaller and on the inside.
CONDENSER LENS
Mounted in or below the stage, this lens focuses or "condenses" the light onto a specimen.
Microscopes with higher-powered objective lenses with smaller diameters require concentrated
light to work properly. The use of a condenser lens helps increase illumination and resolution.
CONTRAST PLATE
Used on tow -power microscopes, this circular opaque plate is placed on the stage, and can be
flipped between a white or black side depending on the coloration of the specimen.
COVERSLIP
A cover slip is a very thin square of plastic or glass used to cover the specimen on a slide With a
liquid sample, the cover slip assists with single-plane focusing by flattening out the liquid.
DIAPHRAGM
Most useful at higher powers, the diaphragm is typically a five-hole disc housed under the stage
that adjusts the amount of light passing through the stage opening. Each opening is a different
diameter controlling how much light passes through to help properly illuminate the specimen
while also increasing contrast and resolution.
DIN OPTICS
This German standard for the manufacture of microscope lenses features a uniform thread and
works with a 160mm tube length. While not notably better than non-DIN, they are
interchangeable between DIN microscopes.
DLOPTER ADJUSTMENT
Used on microscopes with a binocular head, this adjustment allows each eyepiece to be focused
independently to compensate for the difference in vision between your two eyes.
DUAL HEAD
Generally found on high-power microscopes, this unit features a single eyepiece on one side and
an additional eyepiece on the top or opposite side. Dual heads are useful for the teacher to
verify a student's view and they can also be used for video or camera work.
EYEPIECE LENS
These lenses, also known as ocular lenses, are typically 10X, but also come in 5X, 15X and 20X
varieties. The eyepiece lens is what you actually look through to see your specimen. The
interocular distance is adjustable so that you can keep both eyes open when looking into the
microscope.
FINE FOCUS
The smaller of two adjustment knobs moves the objective lens closer to or farther away from the
specimen in very small steps. It can be used to fine-tune the focus on various parts of a specimen
after first using the Coarse Focus to get dose (See also Coarse Focus).
FIELD OF VIEW
The field of view, or "FOV", is the diameter of light visible when looking into the eyepiece lens.
Increasing lens power equals a smaller FOV. The FOV is measured by placing a clear metric ruler
on the stage and counting the millimeters across from side to side. For example, a 40X objective
averages about 4.5mm, whereas 1000X averages about 0.18mm. (See also Micrometer).
FIXED ARM
A type of stand used for low-power microscopes where the are and body are integral parts and
fixed firmly to the base.
FOCUS
The method by which the specimen's distance to the objective lens is adjusted to provide a sharp
image. Depending on the microscope, this adjustment will either be made by moving the stage or
moving the tubes.
HEAD
The head, or upper portion of the microscope, contains the eyepiece tube or tubes and prisms.
Various heads are available, including a Monocular Head (one eyepiece), Binocular Head (two
eyepieces, one for each eye), Dual Head (two singular eyepieces) and Trinocular Head (two
eyepieces with a third typically used for a camera).
ILLUMINATOR
This light source mounted underneath the stage commonly uses one of three types of lights:
tungsten is the most common and cheapest, fluorescent runs cool and provides a bright white
light, and halogen is very bright and white but gives off heat like tungsten.
IMMERSION OIL
A special oil is used exclusively with a 100X or higher objective lens, typically at 1000X total
power. A drop of oil is placed on the cover slip with the objective lens lowered until it touches the
drop. Acting as a bridge between the glass lens and the glass slide, the oil concentrates the path
of light to increase the resolution of the image.
INCLINATION JOINT
Some microscopes feature a rotating arm pivot point that can be adjusted to allow for more
comfortable viewing. The downside of adjusting the microscope is that wet samples may run off
the slide.
INTERPUPILLARY ADJUSTMENT
This adjustment allows the distance between the eyepiece lenses, found on stereo or binocular
microscopes, to be set either closer or farther apart for more comfortable viewing.
MECHANICAL STAGE
A slide holder featuring two knobs that allow the slide position to be adjusted forwards or
backwards and from side to side. This is very useful when dealing with moving specimens like
protozoans. Microscopes can have either a bolt-on mechanical stage added or come with a
built-in integral mechanical stage.
MICROMETER
Micrometer, or micron, is the metric linear measurement used for microscopy. At 1000 microns per
millimeter, a 1.8mm long specimertcan be expressed as 1,800 microns (or micrometers) long.
MIRROR
This reflective surface directs light through the opening in the stage to illuminate a specimen.
MONOCULAR HEAD
A microscope head featuring a single eyepiece lens.
NOSEPIECE
Also called a revolving nosepiece or turret, this structure holds the objective lenses. Rotating it
allows the objective lens to be switched.
NUMERICAL APERTURE (N.A.)
Important on high-power microscopes, this number is an expression of the lens' ability to resolve
fine detail in the observed object. Calculated using a complex mathematical formula, it is
derived from the angular aperture of the lens and the refraction index of the medium between
the lens and specimen. For best results, the condenser system, an optical lens responsible for
illuminating the object, should match or exceed the rating of the highest power objective lens on
the microscope.
OBJECTIVE LENS
The lens closest to the object being viewed is known as the objective lens. Each objective lens has
a different magnification, including the scanning lens (4X), low-power lens (10X), mid-power lens
(40X), and high-power lens (100X).
OIL IMMERSION LENS
An objective lens, generally 100X or more, designed to work with a drop of immersion oil placed
on the slide to increase the resolution of the lens.
ON/OFF SWITCH
Controls the power to the microscope.
PARCENTERED
A microscope that is calibrated this way will maintain the specimen in the center of the view
when the objective lens is rotated.
POINTER
Some microscopes feature a pointer that is visible when looking through the eyepiece lens. This
pointer can be rotated by turning the eyepieces around.
POST STAND
Typically used with low-power microscopes, this type of stand features a single post attached to
the base allowing the body of the microscope to be rotated on adjusted vertically.
RESOLUTION
A lens system's ability to resolve fine details of the observed object.
RETICLE
This special grid pattern is inserted into the eyepiece lens for taking measurements of objects
seen through the microscope.
REVOLVING NOSEPIECE
Also called a nosepiece or turret, this structure holds the objective lenses. Rotating it allows the
objective lens to be switched.
RING LIGHT
A separate light connected to the microscope body that produces a ring of light. An independent
light that usually connects to the microscope body and gives off a ring of light.
SEMI-PLAN LENSES
This type of lens improves image quality by presenting a sharper image with less distortion in the
perimeter of the field of view, or FOV. It provides better image quality than standard achromatic
lenses but is also more expensive.
SLIDE
This rectangular plate, made of either plastic or glass, is used to hold the specimen. In some
cases, it may also contain a depression or well to hold several drops of liquid.
SLIP CLUTCH
This mechanical device prevents gear damage to the microscope when the focus adjustment
reaches its maximum travel.
STAGE
This area is the main, flat surface that holds the slides for observation.
STAGE ADJUSTMENT KNOB
The mechanical stage allows for adjustment of the slide position forward, backward, and left or
right.
STAGE CLIPS
The stage has several of these clips that are used to hold a slide in place.
STAGE PLATE
Found on low-power microscopes, this frosted circular glass plate fits in over the lower illuminator.
(See also Contrast Plate).
STAND
For Low-power microscopes, there are typically three types of connections between the
microscope body and the base including Post Stand, Fixed Arm, and Universal Stand.
STEREO
When using two objectives slightly offset, the image is presented in 3D. (See also Binocular Head).
SUB-STAGE
This area is located directly below the stage itself.
T-MOUNT
This type of adapter is useful for attaching still cameras to the microscope.
TENSION ADJUSTMENT
A factory adjustment of the focusing mechanism that makes the microscope easy to focus, but
tight enough to prevent the stage from drifting due to the weight of the stage or tube.
TRINOCULAR HEAD
This type of microscope features two eyepieces for viewing and a third for attaching a camera.
This setup is available on both high- and low-powered microscopes, with some models providing
the option to adjust the ratio of light being sent between the third port and the two viewing
eyepieces. Dual power trinoculars transmit images through a set of separate lenses not used by
the eyepieces.
TURRET
Also called a revolving nosepiece, or simply, nosepiece, this structure holds the objective lenses.
Rotating it allows the objective lens to be switched.
UNIVERSAL STAND
A long, boom-type arm used to support a (low-power) microscope body. It has many adjustments,
allowing the microscope to be aligned in a wide variety of configurations. Generally, one uses an
external (like a fiber optic) light source with a universal stand. Some low-power microscopes use
a boom-style arm to support the body, allowing alignment in a wide variety of positions. Typically,
an external light source is used with this setup.
WIDEFIELD EYEPIECE LENSES
These wide-diameter eyepiece lenses offer a broader FOV when viewing a specimen.
X
This represents the magnification factor of a lens or view. For example, a lens with a 200X means
the object is viewed at 200 times its actual size. To calculate the total magnification of a
microscope, multiply the power of the eyepiece lens by the power of the selected objective lens
(eyepiece lens power * objective lens power).
XR
In this notation, the X represents the magnification of the lens (see X), while the R stands for
“retractable”. XR lenses contain a spring-loaded mechanism that allows them to telescope
inward to prevent damage to the lens and slide in case of slide contact.