CLINICAL PARASITOLOGY

Microscope

magnify objects up to two million

Introduction times.

● The microscope is a fundamental tool Transmission Electron Microscope

for biologists. This instrument has
been perfected over the past 300 ● also uses electrons, but instead of
years. It has, within limits, allowed the scanning the surface (as with SEM's)
invisible to become the visible. The electrons are passed through very
extension of the eye or vision makes thin specimens. You will first learn to
much information available to the properly use the Compound Light
curious student. For instance: shape, Microscope.
size, position, connections, colours,
number, texture and even chemical
Terms
composition are some of the facts that
may be recorded by the eye and the Magnification
microscope. - How much larger an object appears
under a scope than it actually is;

Types of Microscope oculars are engraved with their

magnifying powers; oculars magnify
10X (magnification = 10 multiplied by,
Light Microscope
for instance 4 for scanning power = 40
● The models found in most schools times normal size).
use compound lenses and light to
magnify objects. The lenses bend or Resolution
refract the light, which makes the - The rendering of detail; high
object beneath them appear closer. magnification without good resolution
is worthless; light is the limiting factor
Scanning Electron Microscope of resolution; the blue light of your
scope allows resolution of 0.2 microns
● allow scientists to view a universe too
small to be seen with a light
microscope. SEMs don't use light
waves; they use electrons (negatively Depth of Field
charged electrical particles) to
 - The distance through which you can cleaning solution. Never use your
move the specimen and still have it in finger, handkerchief, paper towels or
focus; the higher the magnification, spit to clean the lenses. Do not
the closer the objective is to the slide remove any parts for cleaning; it only
and so the depth of field decreases, allows dust to enter the microscope.
this requires a more delicate
technique. ✓ PUTTING AWAY:
● Turn off light & center mechanical
In order to see such small structures, the stage.
subject must be made larger or magnified. ● Position the nosepiece so that the
Thus in microscopy, we talk a lot about lowest scanning (4X) objective is in
magnification: However, the most important place.
function of your microscope is not ● Remove the slide from the stage, put
magnification, but the rendering of detail or it in the proper place.
resolution. If a mouse was magnified to the ● Clean the stage and lenses with
size of an elephant, but you could not gauze and lens cleaner, wipe off any
distinguish it from an elephant, would this oil.
extend your knowledge of the mouse? ● Wrap the cord around the arm.
Accordingly, resolution is the costly feature of ● CAREFULLY carry with two hands
a microscope and GENTLY place the microscope in
the proper cabinet
Here are some important precautions for
handling our microscopes that will keep Parts of the Microscope
them in great condition for many years.

1. OCULAR LENS or EYEPIECE - On a

✓ CARRYING:
binocular scope there are two ocular
● Always carry your microscope with
lenses,one for each eye. These
two hands, one grasping the arm or
lenses magnify the image at 10X
back slot and the other supporting the
power. The power of the ocular
base.
The lens multiplied by the objective
✓ TABLE PLACEMENT:
lens gives the total magnification of
● Set the microscope on a flat, solid
the microscope.
support and in a position where it will
2. ARM - A support for the upper portion
not easily be knocked off. Coil the
of the scope. It also serves as a
cord to avoid tripping over it.
convenient carrying handle.
✓ CLEANING:
● Lenses must be clean for resolution.
Use only lens paper or gauze and
3. MECHANICAL STAGE CONTROLS -
Objective Power Objective Name
A geared device to move the slide
(placed in the slide clamp) precisely. 4× Scanning
4. COARSE ADJUSTMENT KNOB - A
10× Low power
rapid control which allows for quick
focusing by moving the objective lens 40× High power
or stage up and down. It is used for
100× Oil Immersion
initial focusing.
5. FINE ADJUSTMENT KNOB - A slow
11. SLIDE CLAMP - A clamp to hold the
but precise control used to fine focus
slide on the stage.
the image when viewing at higher
12. STAGE - A platform for placement of
magnifications.
the microscope slide.
6. BASE - The part of your microscope
13. CONDENSER - A lens that
that sits on a level, stable support
concentrates or directs the light onto
7. OCULAR ADJUSTMENT - An
the slide.
adjustment for differences in the
14. IRIS DIAPHRAGM CONTROL - A
focusing abilities of your eyes.
lever (or rotating disk) that adjusts the
8. DIOPTIC ADJUSTMENT - A
amount of light illuminating the slide.
horizontal adjustment of the oculars.
Use just enough light to illuminate the
Adjust for your eyes so you see only
object on the slide and give good
one field of view with both eyes open.
contrast.
9. NOSEPIECE - A circular plate with 4
15. FILTER HOLDER - A blue filter rests
objective lenses that can be rotated
in this holder below the substage
into position for different
condenser.
magnifications.
16. CONDENSER HEIGHT CONTROL -
10. OBJECTIVE LENS - Four separate
A knob that controls the height of the
lenses that magnify the image (4X,
condenser.
10X, 40X and 100X) depending on
17. LAMP - The light source.
the objective in use. The lens is
18. LAMP SWITCH - Turns the lamp "on"
positioned just above the object being
and "off
viewed
 Introduction
❖ A microscope is a tool that allows us
to view things that are too small to
see with the naked eye. The most
common microscope is the compound
light microscope.
❖ It uses a system of two or more
lenses to collect and focus the
transmitted visible light through the
specimen to the eye. Often this is
referred to as bright field microscopy.
Animal cells, plant cells, bacteria and
protozoa can be easily seen using this
type of microscope
❖ It uses a system of two or more lens
to collect and focus the transmitted
visible light through the specimen to
the eye. Often this is referred to as
bright field microscopy. Animal cells,
plant cells, bacteria and protozoa can
be easily seen using this type of
Set up and use
microscope

1. Remove the dust cover. Carry your

microscope with two hands, one
under the base and the other gripping
the arm or frame to the appropriate
bench
❖ The ocular lens also magnifies at
2. Gently place the microscope on a flat,
generally a 10x magnification. The
level surface and plug the microscope
power of magnification will be marked
into a power outlet
on the lens. It may be monocular or
3. Turn on the light source The on button
binocular (one lens for one eye or two
may appear on the back or side of the
for both eyes). The majority of our
microscope
scopes are binocular
4. Adjust the brightness of the light
source. A low setting is generally best
for scanning a slide. This is done by
one of two ways depending on the
microscope that you are using. One
 way to adjust brightness is to adjust it madeWith the scanning or lowest
using the brightness adjustment objective in use- you may scan across
located on the side of the microscope. the slide by moving the stage from
This model generally also has an right to left and left to right.
adjustable condenser. It is generally This is done using the stage knobs
marked for the individual objectives. found just below the stage of the
Set the condenser accordingly. On microscope.
older microscopes the brightness is As you scan across the slide you may
adjusted by turning the Illumination need to use the fine focus (inner
Ring on top of the light source. By knob) to sharpen the image. Once
turning the ring you can decrease the you decide on an area of the slide you
amount of light or increase the may want to look at a higher
amount of light being shown through magnification of the area. This is done
the sample by rotating the nose of the microscope
5. Adjust the stage as low as possible to the next objective. On our
with the coarse focus knob. It is the microscopes that is a 10X objective.
outer or larger knob Once the object has been rotated and
6. Rotate the nose of the microscope so locked into place and you have made
that the lowest objective is in line with your light adjustments (generally
the light source. brighter); look through the eyepiece
7. Place a slide on the stage holding it and using only the fine adjustment
with the slide clip. If a slide has a knob sharpen the image. If the
coverslip –it should always face up magnification still does not give the
8. Once your slide is correctly seated, detail that you need the nose of the
look through the ocular lens, adjust microscope may be rotated again to
your brightness and using the coarse lock in the third objective which in our
adjustment knob (outer knob) bring case is a 40X. Again brightness may
the stage up towards the objective, need adjusted and the fine focus is
stopping when the slide image comes used to focus and sharpen the image.
into view. The slide should never be To view significantly small structures
pushed all the way up to the objective the oil immersion objective (100x)
9. Once the image is seen use the fine may need to be used. This is used by
focus adjustment knob (inner knob) to rotating the nose of the microscope to
sharpen in on the details of the between the third objective and the oil
structure. After this point you will only immersion objective. At this point a
use the fine focus on the slide. All small drop of oil is placed on the slide
coarse adjustments have been and then the oil immersion objective is
 rotated in passing through the oil on
the slide. Immersion oil should only be
used with the oil immersion objective.
Once oil is used the slide must be
properly cleaned prior to viewing it
with any of the other objectives on the
microscope. An increase in light will
be needed and only the fine focus
adjustment should be used to focus
the image. DO NOT under any
circumstance use the coarse
adjustment as you will crush the slide
into the objective and potentially
damage the objective and the slide
If you get oil on the other objectives,
notify the laboratory manager as soon
as possible so that the objectives can
be removed and properly cleaned. If
the oil is allowed to dry on the
objective it will not come off and all
images viewed thereafter will have a
haze and not focus correctly.
10. When finished viewing your image-
use the coarse adjustment knob to
lower (go away from) the objectives of
the microscope. Rotate the nose of
the microscope to the scanning or 4x
objective. If oil has been used, use
lens tissue and cleaner to remove the
oil from the slide, objective, and
stage.
11. Turn off the light and unplug the
microscope. Cover the microscope
with a dust cover.
12. Return the microscope to the shelf.
13. If you encounter any issues tell or ask
your lab assistant, instructor or
laboratory manager