GENERAL ZOOLOGY LABORATORY

S.Y. ‘23 - ‘24 | SMAE WEEK / CHAPTER 1: MICROSCOPY

— MODULE BASED — - One of the lens systems

formed an enlarged image of the object and the
1.0 MICROSCOPE second lens system magnified the image formed
by the first. The modern compound microscope
● optical instrument that uses a lens or consists of two lens systems, the objective and
combination of lenses to produce the ocular or eyepiece. The first magnified
magnified images that are too small to image obtained with an objective lens, is again
be seen by an unaided eye magnified by the eyepiece to give a virtual
● provides the enlarged view that helps in inverted image. The total magnification is the
examining and analyzing the image product of the magnifications of two lens
a. theory microscopes (Light systems.
microscope)
- In this type of microscope, there are
b. electron microscopes (eg.TEM,
SEM) ocular lenses in the binocular eyepieces and
c. scanning probe microscopes. objective lenses in a rotating nosepiece closer to
(eg.AFM, PSTM) the specimen. Because It contains its own light
source in its base, a compound light microscope
is also considered a bright field microscope.
1.1 PARTS OF MICROSCOPE Bright field microscopy simply means that the
● Mechanical part - base, c-shaped arm specimen is lit from below and viewed from
and stage. above.
● Magnifying part - objective lens and
ocular lens. NOTE
● Illuminating part - substage condenser,
iris diaphragm, light source.

1.2 LIGHT MICROSCOPY ● Microscopes have come a long way

● uses the properties of light to produce since then—today's strongest
an enlarged image. compound microscopes have
magnifying powers of 1,000to 2,000X.
● is the simplest type of microscope.
a. Simple microscope
- double convex or plano convex
lens.
- uses only a single lens, e.g.:
hand lens.
- The developments of
advanced techniques for grinding and
shaping lenses allowed professionals
such as Hans Janssen and Anton van
Leeuwenhoek to develop simple
microscopes which advanced the study
of biology significantly. 1.3 MAGNIFICATION
b. Compound microscope
● the ability to view an object as larger
- used two lenses or lens
● A good image is obtained when the
systems.
amount of specimen detail is also

BS PSYCHOLOGY / FIRST YEAR | SEMESTER 1 | QUARTER 1 | NATSCIE ELEC-1 | SMAE PAGE 1

GENERAL ZOOLOGY LABORATORY
S.Y. ‘23 - ‘24 | SMAE WEEK / CHAPTER 1: MICROSCOPY

increased. Magnification alone will not

eyepiece usually
achieve this. contains a 10X or 15X
● In order to ascertain the total power lens.
magnification when viewing an image
with a compound light microscope, take Diopter Adjustment Useful as a means to
the power of the objective lens which is change focus on one
eyepiece so as to
at 4x, 10x or 40x and multiply it by the
correct for any
power of the eyepiece which is typically difference in vision
10x. between your two
● Therefore, a 10x eyepiece used with a eyes.
40X objective lens, will produce a
magnification of 400X. The naked eye Body tube (Head) The body tube
connects the eyepiece
can now view the specimen at a
to the objective
magnification 400 times greater and so
lenses.
microscopic details are revealed.
Arm The arm connects the
body tube to the base
1.4 RESOLUTION of the microscope.
● the ability to measure the separation of
Coarse adjustment Brings the specimen
images that are close together.
into general focus.
● Good resolution or the resolving power
of the microscope is necessary to see Fine adjustment Fine tunes the focus
the valuable details in an image. and increases the
● Be especially careful with oil immersion detail of the
lenses. This objective has the smallest specimen.
working distance and your careful
Nosepiece A rotating turret that
handling is important. houses the objective
lenses. The viewer
spins the nosepiece to
1.5 WORKING DISTANCE select different
● At low magnification your working objective lenses.
distance is longer and so vice versa
Objective lenses A standard
when increasing magnification.
microscope has three,
● Damage to your specimen is inevitable if four, or five objective
you are not cautious of the shorter lenses that range in
working distance when increasing your power from 4X
magnification. to100X.
Parts and Function of a Microscope
Stage The flat platform
Parts of a Compound Function where the slide is
Microscope placed.

Lens the viewer looks Stage clips Metal clips that hold
through to see the the slide in place
specimen. The

BS PSYCHOLOGY / FIRST YEAR | SEMESTER 1 | QUARTER 1 | NATSCIE ELEC-1 | SMAE PAGE 2

GENERAL ZOOLOGY LABORATORY
S.Y. ‘23 - ‘24 | SMAE WEEK / CHAPTER 1: MICROSCOPY

the diffracted light causes destructive

Stage height These knobs move
adjustment (Stage the stage left and interference.
Control) right or up and down. ● One of the consequences is the
reduction in light intensity resulting
Aperture The hole in the middle
in greater or lesser dark areas. The
of the stage that
allows light from the patterns of light and dark that are
illuminator to reach recognized as an image of the
the specimen. specimen. Because our eyes are very

Illumination The light source for a sensitive to variations in brightness,

microscope. Older and then the image becomes a more
microscopes used or less faithful reconstitution of the
mirrors to reflect light
original specimen.
from an external
source up through the ● The objective lens at first formed a
bottom of the stage; real and inverted magnified image. And
however, most then the eye piece further magnifies
microscopes now use
the same image to a virtual magnified
a low-voltage bulb.
image.
Iris diaphragm Adjusts the amount of
light that reaches the
specimen.
2.0 FOCUSING ON MICROSCOPIC
Condenser Gathers and focuses
light from the OBJECTS
illuminator onto the
specimen being
viewed. 1 Start with clean lenses

Base supports the

microscope and it’s ● It is important that microscope
where the illuminator lenses be very clean. Before
is located. viewing through a microscope,
use lens paper to gently clean the
lenses.
1.5 IMAGE FORMATION 2 Begin at Low Power
Magnification

● direct or undeviated light from a

specimen is projected by the objective ● Always begin by viewing the object
and it spreads evenly across the through a low power lens. Depending
entire image plane at the diaphragm on how small the object is, start with
of the eyepiece. the scanning or low-power objective.
● Light diffracted by the specimen has ● Using a low-power objective lens, get
come to focus at different localized the target object centered in the
sites on the same image plane, and field-of-view and focus as much as

BS PSYCHOLOGY / FIRST YEAR | SEMESTER 1 | QUARTER 1 | NATSCIE ELEC-1 | SMAE PAGE 3

GENERAL ZOOLOGY LABORATORY
S.Y. ‘23 - ‘24 | SMAE WEEK / CHAPTER 1: MICROSCOPY

possible, first by using the coarse immersion. Manipulating the coarse

focus and then fine-tuning the clarity focus at high power can smash the
of the image with the fine focus. lens into the slide, potentially
● Once the object is in focus, switch to damaging the scope and the
the next higher objective power. Do specimen.
not change the focus or manipulate
5 Key points Magnification
the focus knobs in any way while
changing objectives.
● Magnification is defined as the degree of
3 Adjustment for oil immersion enlargement of an object provided by the
objective
microscope. Magnification of a microscope
is the product of the individual magnifying
● Without changing the adjustment ability of an ocular lens and objective lens.
of high power, turn to oil
Magnification Magnification Total
immersion objectives. One drop of ocular lens of objective magnification
of oil is added onto the slide. The lens
nose piece is turned such that
10x 4x 40x
the oil immersion objective
touches on the drop of oil. Open 10x 10x 100x
the iris diaphragm completely.
10x 40x 400x
Use only fine adjustments for
focusing. 10x 100x 1000x

4 Importance of Par Focal

6 Resolving power:

● A set of objectives on a microscope

are said to be par focal if the ● It is defined as an ability to
viewer can change from one to distinguish between two particles
another and still have the situated very close.
specimen nearly in focus. This is a
7 Numerical aperture
very convenient feature, because
as the magnification increases,
● It is defined as the property of the lens
even small manipulations of the
that decides the quantity of light that
focus knob can take a specimen
can enter. The angle of the cone of
far out of focus.
light entering an objective is known as
● After changing to a higher objective
theta. NA=nsinθ ; NA=numerical
(such as high-dry or oil-immersion)
aperture
the viewer needs only manipulate
● n=refractive index of the imaging
the fine focus knob. Never
medium between the front lens of
manipulate the coarse focus at oil
the objective and the specimen cover

BS PSYCHOLOGY / FIRST YEAR | SEMESTER 1 | QUARTER 1 | NATSCIE ELEC-1 | SMAE PAGE 4

GENERAL ZOOLOGY LABORATORY
S.Y. ‘23 - ‘24 | SMAE WEEK / CHAPTER 1: MICROSCOPY

glass, a value that ranges from 1.00

for air to 1.51 for specialized
immersion oils. θ= one-half of the
angular aperture (A)
● Resolution is a subjective value
in microscopy because at high
magnification, an image may
appear not very sharp but still it
can be resolved to the maximum
ability of the objective.
Numerical aperture is defined as
the resolving power of lens
objective, but the entire
resolution of a microscope
system also depends on the
numerical aperture of the
substrate condenser. For getting
a better resolution, higher the
numerical aperture of the total
system.

BS PSYCHOLOGY / FIRST YEAR | SEMESTER 1 | QUARTER 1 | NATSCIE ELEC-1 | SMAE PAGE 5