MLS 108 (LAB) – Parasitology (PRELIM)
L1: THE MICROSCOPE
❖ Virtually all organisms studied in microbiology
cannot be seen with the naked eye, but require the
use of optical systems for magnification.
❖ Either Zacharias Janssen of the Netherlands or
Galileo Galilei of Italy invented the microscope
shortly before 1600. History has been unable to
determine which scientist should be given credit.
The microscope was not used to examine
microorganisms until the 1680’s, when a clerk in a
dry goods store, Anton van Leeuwenhoek,
examined scrapings of his teeth and any other
substances he could find.
❖ The early microscopes, called the simple
microscopes, consisted of biconvex lenses and
were essentially magnifying glasses. To see
microbes, a compound microscope, which has two
lenses between the eye and the object, is required.
Parts and Function . ➢ found below the stage and attached to the
According to their functions, the parts of the compound
microscope are generally divided into:
mechanical and illuminating & magnifying parts.
MECHANICAL PARTS .
➢ The mechanical parts comprise all those structures
that support the whole device and which enable
the operator to use the illuminating and magnifying
parts.
❖ Base
➢ may be a U or Y shaped stand that supports
the microscope.
❖ Pillar
➢ a short piece of metal attached to one end of
the base. It also supports the microscope.
❖ Handle/Arm
➢ curved metallic part arising from the pillar. It is
used in holding the microscope.
❖ Inclination Screw
➢ found at the junction of the pillar and the
handle. It is used to tilt the microscope.
❖ Body Tube
➢ cylindrical structure arising vertically from the
handle. It holds the dust shield and the
nosepiece with the objectives at the lower end.
❖ Draw Tube
➢ upper and smaller end of the body tube
bearing the eyepiece.
❖ Revolving Nosepiece
➢ circular structure at the lower end of the body
tube to which the objectives are attached.
❖ Dust Shield
➢ a thin circular structure at the lower end of the
body tube. It is used to protect the objectives
and the specimen from dust.
❖ Adjustment Screws
➢ two pairs of wheels attachéd to either side of
the body tube.
➔ Coarse Adjustment Knob
➢ bigger wheels used to adjust the low
power objective in focusing.
➔ Fine Adjustment Knob
➢ smaller wheels used for delicate focusing
in connection with the high power and oil
immersion objectives. It is also used to
make the specimen more clear and vivid.
❖ Stage
➢ square platform with an opening at the center.
It is where the slide is placed during focusing.
❖ Stage Clips
➢ paired structures on either side of the stage. It
is used to hold the slide in place.
❖ Mirror Rack
pillar. It holds the mirror.
❖ Rack Stop
➢ This is an adjustment that determines how
close the objective lens can get to the slide.
MLS 108 (LAB) – Parasitology (PRELIM)
ILLUMINATING AND MAGNIFYING PARTS
➢ The illuminating and magnifying parts consist
mainly of lenses and glass parts.
❖ Ocular or Eyepiece
➢ found on the draw tube through which the
operator peeps during actual focusing.
❖ Mirror
➢ found below the stage near the base provided
with concave and plane surfaces. It is used to
collect and direct light to the specimen.
❖ Iris Diaphragm
➢ found below the stage consisting of regularly
arranged circular blades used to regulate the
central opening to decrease the increase of
the light reflected on the object.
❖ Objectives
➢ tube-like structures attached to the revolving
nosepiece.
❖ Low Power Objective (LPO)
➢ often marked with 10X or 6X. It is usually
shorter than the two other objectives and it
usually forms the general outline or wider
portion of the object.
❖ High Power Objective (HPO)
➢ often marked with 45X or 43X. It is longer than
the LPO. It forms a bigger image of the object
in focus and is used to examine living
microorganisms suspended in drops of water.
In most cases, it is used to enlarge specimens
that are so small under LPO.
❖ Oil Immersion Objective (OIO)
➢ often marked with 100X or 90X. It has the
highest degree of magnification and is used to
examine stained preparations of
microorganisms using immersion oil as their
medium.
❖ Aperture
➢ is the hole in the stage through which the base
(transmitted) light reaches the stage.
All of the objectives (with the possible exception of the
scanner objective) are PARFOCAL; that is, when a
subject is in focus with one lens, it will be in focus with
all the lenses.
➔ The magnification of a microscope depends on
the type of objective lens used with the ocular; it is
stamped on the barrel
➔ Compound microscopes have three or four
objective lenses mounted on a nosepiece:
scanning, low power, high dry, and oil
immersion.
➔ The total magnification (mag) of the object =
mag of ocular (usually 10X) x mag of objective lens
. ➔ Resolution or resolving power
◆ ability of lenses to reveal fine detail or two points
distinctly separated; two objects are seen as distinct
even though they are very close together.
➔ It is a function of the wavelength of light used and a
characteristic of the lens system called
numerical aperture.
➔ Resolving power is expressed in units of length;
the smaller the distance, the better the resolving
power
Wavelength of light used
Resolving = —----------------------------------
2 X numerical aperture
The microscope is a very important tool in microbiology,
and it must be used carefully and correctly. Follow
these GUIDELINES every time you use a
microscope:
1. Carry the microscope with one hand beneath the
base and one hand on the arm.
2. Do not tilt the microscope, instead, adjust your stool
so you can comfortably use the instrument.Observe
the slide with both eyes open, to avoid eyestrain.
3. Always focus by moving the lens away from the
slide.
4. Always focus slowly and carefully.
5. When using the low power lens, the iris diaphragm
should be barely open so that good contrast is
achieved. More light is needed with higher
magnification.
6. Before using the oil immersion lens, have your slide
in focus under high power. Always focus with your
low power first.
7. Keep the stage clean and free of oil. Keep all
lenses except the oil immersion lens free of oil.
8. Keep all lenses clean. Use only lens paper to clean
them. Wipe oil off before putting your microscope
away. Do not touch the lenses with your hands.
9. Clean the ocular lens carefully with lens paper. If
dust is present, it will rotate as you turn the lens.
10. After use, remove the slide, wipe off oil, put the dust
cover on, and return the microscope to the
designated area.
11. When a problem does arise with the microscope,
obtain help from the instructor. Do not use another
microscope unless yours is declared “out of action”.
MLS 108 (LAB) – Parasitology (PRELIM)

How to Focus Your Microscope .

1. The proper way to focus a microscope is to start

with the lowest power objective lens first and while
looking from the side, crank the lens down as close
to the specimen as possible without touching it.

2. Now, look through the eyepiece lens and

focus upward only until the image is sharp.

3. If you can't get it in focus, repeat the process again.

4. Once the image is sharp with the low power lens,

you should be able to simply click in the next power
lens and do minor adjustments with the focus knob.

5. If your microscope has a fine focus adjustment,

turning it a bit should be all that's necessary.

6. Continue with subsequent objective lenses and fine

focus each time.