Clinical Parasitology

LABORATORY

THE COMPOUND MICROSCOPE to the ocular lenses. Don't forget to

tighten the clamp
Microscope  Check the lenses to make sure they are
 produce magnified visual or photographic clean
images of small objects. Putting the microscope away
 Gk. mikrós = “small”  Rotate the lowest-power objective into
 Gk. skopeîn = “to look or see” position.
 Light compound microscope  Remove the slide from the stage.
 uses visible light to illuminate the  Clean the microscope surfaces free of
specimen, and passes that light dust or debris. If immersion oil has been
through two separate lens to magnify used, wipe it off the lens and stage with
the image. lens tissue.
 Coil the power cord around the base of
the microscope.
 Replace the dust cover and/or return the
microscope to its correct place in the
cabinet.

Parts of the Microscope

Support System
A F
B G
C H
D I
E J

Care of the Microscope

Transport and setting-up a microscope 1 Base or the overall support

foot
 Hold both sides
2 Arm or supports the observation
around the hole of
Limb tube
the arm when
carrying the 3 Revolving objective changer
microscope nosepiece
 do not carry 4 Stage holds the slide specimen
two microscopes at one time in place
 do not hold the microscope by the
stage (1) or observation tube (2) Magnification System
 Don’t let the electric cord of the  consist of a system of lenses
microscope dangle in such a way as to  mounted in two groups
hazard foot entanglement
 Place the microscope in a stable flat
surface. Keep the workstation uncluttered
 Microscope w/ rotatable observation tube
may be used in 2 ways:
 w/ microscope arm near the
observer or 1 Objective
 w/ microscope arm away from the first group of lenses at the bottom of the
observer. tube, just above the object
To change from one position to another,
hold the observation tube firmly with one
hand while loosening the observation tube
adjustment clamp and rotating the
observation tube with the other hand to
prevent it from accidentally falling off
from the microscope and causing damage
4x Scanning magnifies 4
Angelika Marie B. Reyes
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 Clinical Parasitology
LABORATORY

times source the microscope beneath

10x Low-power magnifies 10 / the stage
objective times Illuminato -Microscopes that do not
40x High-power magnifies 40 r have built-in light bulb
objective times have a mirror that reflects
100 Oil-immersion magnifies 100 rays from the sunlight or
x times external lamp onto the
object.
2 Condenser brings the rays of light
4x air is the imaging (illumination) to a
Dry common focus on the
10x medium between the
objectives object to be examined.
40x cover glass
100 Oil- the tip of the OIO 3 Iris -a series of thin plates
x immersion lens is immersed in diaphragm found inside the
objective oil condenser
-has a central aperture
which control the amount
2 Eyepiece or Ocular of light that passes into
the condenser.
second group of lenses at the top of the tube 4 Filter -allows the passage of
light of desired
wavelength only
-Daylight blue filter is
used to balance the light
created by tungsten or
halogen microscope
10x magnifies the image produced by lights.
the objective 10 times
20x magnifies the image 20 times Adjustment System

A microscope may be:

 monocular – has only 1 eyepiece
 binocular – has 2 eyepieces; used
most clinical laboratories because
both eyes are used view an object,
thus eyestrain

Magnification
 refers to the ability of an optical system to
enlarge an image of a specimen
 expressed as magnifying power - the
number of times the image of an object is
enlarged

Illumination System

1 Mechanical used to move the

stage object slide on the
control stage
knobs
(1a) – x-axis knob
 horizontal
(1b) – y-axis knob
1 Light -an electric bulb built into  vertical

Angelika Marie B. Reyes

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 Clinical Parasitology
LABORATORY

2 Observation enables the observation  Magnification is a function of the

tube tube of the microscope interaction between the visible light rays
adjustment to rotate and the curvature of bi-convex lens, one
clamp that is thicker at the center than at the
3 Coarse -largest screw periphery
adjustment -used first to achieve
knob an approximate focus
-rotated to bring the
specimen as close as
possible to the
objective
4 Fine -moves the objective
adjustment more slowly  When parallel rays of light pass through a
knob used to bring the object bi-convex lens, light rays are refracted
into perfect/precise (bent) and converge at one point called
focus the focal point.
5 Condenser used to raise the  The vertical plane in which the focal
adjustment condenser for greater point lies is the focal plane.
screw illumination or to
 The distance from the center of the bi-
lower it to reduce the
convex lens to the focal plane is known as
illumination
the focal distance (focal length).
6 Iris can be moved to close
 The distance between the front principal
diaphragm or open the diaphragm,
plane of the lens and the object is known
ring thus reducing or
as the object distance.
increasing both the
 In a similar manner, the distance from the
angle and the intensity
rear principal plane to the image is termed
of the light.
the image distance.
7 Light -adjusts the light
intensity
adjustment
knob
8 Interpupilla regulates the eyepieces
ry distance according to the
adjustment distance between your
eyes so that you
observe a single image
through the eyepieces.
9 Diopter -compensates for the  When object is situated two focal lengths
adjustment difference in the in front of the lens, the image is also two
ring eyesight between the 2 focal lengths behind the lens.
eyes  The lens is performing 1:1
-focuses first with the magnification, i.e., the image size is the
right eye same as the object size.
10 Pre- controls the  For a magnified image to be observed, the
focusing mechanism for object distance must be shorter than the
knob preventing collision focal length of the lens
between the specimen
and the objective

Principles of Light Microscopy

 The microscope must accomplish three
tasks: produce a magnified image of the
specimen, separate the details in the
image, and render the details visible to the
human eye or camera.

Magnification  A real image is an image which is

located in the plane of convergence for

Angelika Marie B. Reyes

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 Clinical Parasitology
LABORATORY

the light rays (back focal plane) that  also expressed as resolving power, i.e.,
originate from a given object. If a screen the shortest between two objects so that
is placed in the plane of a real image, the they can be and still appear separate; the
image will generally become visible on smaller the value, the greater the
the screen and appears inverted. resolution.
 A virtual image is produced on the same  The limit of resolution of unaided human
side of the lens as the object, such as the eye is 0.1 mm
image produced by a simple magnifying  The maximum resolving power of a light
lens. It is formed on the retina of the eye, microscope with the highest
therefore cannot be projected on a screen. magnification possible is 0.2 µm. (0.0002
These images always appear upright. mm). It, therefore, cannot resolve
 Parasitologic work uses brightfield structures smaller than 0.2 µm.
microscope which is best for observing not resolvable
stained specimens and living organisms.

intermedi
ary image
resolvable
 RP is expressed by the following
mathematical equation:

final
where:
image
 λ (lambda) = wavelength of light
 Light from illuminator passes the
 NA = numerical aperture
substage condenser, which forms a well-
 Wavelength of light.
defined light cone that is concentrated
onto the object.  Numerical aperture (NA).
 Light is transmitted through the specimen
and into the objective which then projects
a primary enlarged image, called
intermediate image, to a fixed plane
within the body tube.
 The intermediate image becomes the
“object” for the eyepiece to produce a
secondarily enlarged image, called the
final image.
 When the human eye is placed above the
eyepiece, the lens and cornea of the eye
“look” at the final image as if it were 10
inches from the eye, near the base of the
microscope
 Total magnification of the microscope is
derived by multiplying the magnification
values of the objective and the eyepiece.
For instance, using a 10X objective with a
10X eyepiece yields a total magnification
of 100X and likewise.

Resolution
 ability of the optical system to separate
two closely adjacent objects into 2
distinct entities.
 determines microscopic image clarity and
richness of detail

Angelika Marie B. Reyes

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