WELCOME TO OUR
ANAPHYSIO LABORATORY
CLASS !
MICROBIOLOGY LABORATORY EXERCISE 2
MICROSCOPY
OBJECTIVES:
At the end of this exercise, each student must be
able to:
1. Identify each major part of a compound light
microscope and describe its function
2. Describe the correct use and care of the
microscope
3. Determine the total magnification at different
settings
4. Learn the basic procedures on how to use a
microscope to observe prepared specimens ( LPO,
HPO )
Microscope
-[Link] C. Santos
two microscopes differ in the source of light
they use. One uses a mirror to converge sunlight
while other uses directly an illuminator.
4
MICROSCOPE
microscope instrument too
small for the naked eye.
small objects using such an
instrument is called microscopy.
Microscopic
Parts and Functions of a
Compound Microscope
Light Microscope
SIMPLE COMPOUND
Uses set lenses or a
Uses single lens
lens system
Simple Light
Microscope
Compound Microscope
Compound Microscope
Mechanical Parts Magnifying Parts Illuminating Parts
Adjustments Enlarge the Provide the
and specimen light
Support
Mechanical Parts
o Base
– Bottommost portion supports
o Pillar
– Part above the base that supports the other
parts, connect the stage to the base
o Inclination Joint
– tilting
Inclination Joint
Pillar
Base
Mechanical Parts
o Arm/Neck
– Curved/slanted part which is held while
carrying the microscope
o Stage
– Platform where object to be examined is
placed
o Stage Clips
– Secures the specimen to the stage
Mechanical Parts
o Stage Opening
o Body Tube
– Attached to the arm and bears the lenses
– Light passes from objectives to the
eyepiece
o Draw Tube
– Cylindrical structure on top of the body tube
that holds the ocular lenses
Draw Tube
Body
Tube
Arm /
Neck Stage
Mechanical Parts
o Revolving Nosepiece
– Rotating disc where the objectives are
attached
o Dust Shield
– Lies atop the nosepiece and keeps dust from
settling on the objectives
Dust Shield
Revolving
Nosepiece
o Coarse Adjustment Knob
– Geared to the body tube which elevates or
lowers when rotated bringing the object into
approximate focus
o Fine Adjustment Knob
– A smaller knob for delicate focusing bringing
the object into perfect focus
Coarse
Adjustment
Knob
Fine
Adjustment
Knob
Mechanical Parts
• Condenser Adjustment Knob
– Elevates and lowers the condenser to
regulate the intensity of light
• Iris Diaphragm Lever
– Lever in front of the condenser and which is
moved horizontally to open/close the
diaphragm
Iris Diaphragm
Lever
Condenser
Adjustment Knob
Illuminating Parts
o Mirror
– Located beneath the stage and has concave and
plane surfaces
to gather and direct light in order to illuminate the
object
o Electric Lamp
– A built-in illuminator beneath the stage that may eb
used if sunlight is not preferred or is not available
Mirror /
Electric Lamp
MAGNIFYING
PARTS
• Ocular / Eyepiece
– functions to further magnify the image
produced by the objective lenses. It usually
ranges from 5x to 15x.
Ocular/ Eyepiece
Objectives
MAGNIFYING
• PARTS
Objectives
– Metal cylinders attached below the nosepiece and
contains especially ground and polished lenses
• LPO / Low Power Objective
– Gives the lowest magnification, usually 10x
– General outline /features
• HPO / High Power Objective
– Gives higher magnification usually 40x or
43x
– In detail
• OIO / Oil Immersion Objective
– Gives the highest magnification, usually 97x or
100x, and is used wet either with cedar wood oil or
synthetic oil
Total Magnification
Magnification = Objective lens X Eyepiece lens
e.g. What is the total
magnification if the objective lens
is twenty times (X20) and the
eyepiece lens five times (X5)?
Magnification = 20 X 5 = X100
As magnification increases, detail
increases but
Onion cell
40x
Onion cell 100x
Onion cell
400x
less of the cell is seen
Images From
A Compound Light
Microscope.
(Elodea canadensis)
40X 100X 400X
29
Caring for the
1.
Microscope
Do not let any liquids to come in contact with
the microscope.
2. Always store the microscope inside a box
after use.
3. Return the objective lens onto low power after
use.
4. Carry the microscope by the arm.
5. Use a soft clean tissue to wipe the lenses
Uses Of Compound Light Microscope.
to identify diseases.
identify presence of minerals or metals in
human cells so as to solve criminal cases.
find out the origin of a drug by viewing its
component particles under a Microscope.
31
for conducting their academic experiments.
help to see the bacteria and virus
Plant cells and also the microorganisms living
Compound Microscope beneficial to
biologists too.
32
Coverslip
s
Microscope
slides
Preparing a
slide as a
wet mount.
Use of
some parts of a plant cell can be clearly
seen whenstains
the cell is mounted in water
E.g. an Elodea leaf cell:
cell wall
several chloroplasts
other cell structures which are not so
obvious can often be shown up more clearly
by the addition of dyes called STAINS
Iodine Solution Methylene Blue
Used to stain plant Used to stain animal
cells cells
Blood cells
Human cell
Plant cell
37
3
1 2
One of the fleshy scale
An onion is cut leaves is removed.
into quarters.
Snapping leaf
backwards exposes
5 the epidermis.
4
6 Epidermis is placed on
slide & covered with 2-3
drops of distilled A thin inner layer of
water . Coverslip is epidermis is
lowered. peeled off.
7
A drop of stain is put Stain is drawn over
at one end of slide. specimen using a small
piece of filter paper.
End
[Link] [Link]
