COMPOUND

MICROSCOPE
PARA311 LAB : Lecture 1
Rich King T. Valdez, RMT
Illustration by Smart-Servier Medical Art

Illustration by Smart-Servier Medical Art

 LEARNING OBJECTIVES
PARA311 LAB: Lecture 1

I. Identify the major components of the microscope and their function

II. Describe the process to correctly focus on the appropriate field of view
III. Discuss the ocular micrometer to
IV. Demonstrate the ability to troubleshoot encountered problems with the
microscope
V. Identify how to maintain a microscope
PARA311 LAB: Lecture 1

What is a Microscope?
- Used to magnify organism, it helps professional
- Greek word: mikros - small, skopein - to see or look out
-through this we can look out the organismthat we can't
see with our naked eye
 Compound Microscope
1. Ocular: provide 10 x magnification power
2. Objectives: it has different objectives, starting from the scanner, low power objectives
(10x magnification),high power (40x magnification),and oil immersion objectives
○ with two lenses, the compound microscope offers better
PARA311 LAB: Lecture 1

magnification than a simple microscope; the second lens magnifies the

image of the first.
○ Bright field microscopes (the specimen is lit from underneath, and
they can be binocular or monocular).
○ provide a magnification of 1,000 times, which is considered to be high,
although the resolution is low. This high magnification, however,
allows users to take a close look at objects too small to be seen with
the naked eye, including individual cells.
Capable on one activity:
• far focal - ability of the equipment to remain at focus,
even if change objectives and move side-side or up and
down, backward or for ward
 Slide holder

Slide holder
PARA311 LAB: Lecture 1

Revolving Nosepiece
 MAJOR PART OF COMPOUND MICROSCOPE
PARA311 LAB: Lecture 1

● Mechanical Parts: used to support and adjust the parts

● Magnifying Parts: used to enlarge the specimen

● Illuminating Parts: used to provide light

 MECHANICAL PARTS
● Slide Holder: The slide holder secures the microscope slide in place on
PARA311 LAB: Lecture 1

the stage, allowing for smooth movement of the slide so that different
areas of the specimen can be viewed.
● Revolving Nosepiece: This is the rotating part of the microscope that
holds the objective lenses. It allows you to easily switch between
different magnifications by rotating the desired objective lens into
position.
● Stage: The stage is the flat platform where the slide is placed. It can be
moved vertically (for focusing) and horizontally (to scan the specimen)
using the adjustment controls.
 MECHANICAL PARTS
The base support the whole weight of the microscope.
You can destroy a very important or component of
the microscope which is the bulb or the mirror,
because inside the base there is structure of bulb,
that provides the illumination of the microscope.
Make sure to handle them very carefully
● Centering Screws: These screws allow for the precise alignment of the
condenser with the center of the field of view, ensuring optimal
PARA311 LAB: Lecture 1

illumination of the specimen.

● Base: bottom post portion that supports the entire microscope,
provides stability and support for the microscope when it is upright.
The base also typically holds the illuminator, or light source.
● Pillar: part above the base that supports the other parts.
● Arm: connects the eyepiece tube to the base ,curve or slanted part
which is held while carrying the microscope.
● Stage and stage clip: is a platform for the slides, which hold the
specimen. The stage typically has a stage clip on either side to hold the
slide firmly in place. Some microscopes have a mechanical stage, with
adjustment knobs that allow for more precise positioning of slides.
 MECHANICAL PARTS
PARA311 LAB: Lecture 1

● Body tube: attached to the arm and bears the lenses.

● Draw tube: cylindrical structure on the top of the body tube that holds the
ocular lenses.
● Dust shield: lies atop the revolving nose piece and keeps dust from settling
on the objectives.
Mechanical parts:
- slide holder
-revolving nose piece
-stage and stage clip
- centering screw
- base
-Arm
- pillar
-body tube
- draw tube
- dust shield
 MECHANICAL PARTS
Use only in scanner and LPO
● Coarse Adjustment Knob: moves stage (or body tube) up and down, used
PARA311 LAB: Lecture 1

to focus the image on the microscope under low or medium power.

Used in HPO and OIO
● Fine Adjustment Knob: small knob, used to fine-tune the focus of your
specimen after using the coarse adjustment knob.
 MAGNIFYING PARTS
PARA311 LAB: Lecture 1

● Ocular/ Eyepiece contains the ocular lens, which the user looks through to
see the magnified specimen, further magnify the image produced by the
objective lenses by 5x to 15x.

● Objective lenses: combine with the eyepiece lens to increase magnification

levels. Microscopes generally feature three or four objective lenses, with
magnification levels ranging 4x to 100x.
Magnifying parts
- ocular / eyepiece
-objective lenses
 ILLUMINATING PARTS
● Illuminator or Electric Lamp: microscopes require a light source for
viewing. This can come in the form of a built-in, low-voltage
PARA311 LAB: Lecture 1

illuminator light, or a mirror that reflects an external light source

like sunlight.
● Condenser Diaphragm: focus the light onto the specimen,
concentrating light rays on the specimen.
● Field Diaphragm: usually located near the light source, controls
the diameter of the light beam that enters the condenser,
affecting the illumination and resolution of the specimen.
● Iris Diaphragm: controls the amount of light reaching the specimen
and regulates the light necessary to obtain a clear view of the object
Illuminating parts:
-Illuminator
-Condenser diaphragm
-Field diaphragm
-Iris diaphragm
 How to focus the Microscope?
1. Plug in the microscope into a power outlet and turn on the microscope
using the main switch.
PARA311 LAB: Lecture 1

2. Turn on the light source by rotating the light intensity adjustment knob.
3. Place the specimen on the mechanical stage by placing it inside a bow-
shaped lever. The specimen should be centered in the aperture of the
stage.
4. Use the scanner objective for initial observation of the specimen.
5. Hold and rotate the revolving nosepiece until the 10x objective aligned to
the specimen. A lock-in feature will be obtained once the objective is
positioned.
 How to focus the Microscope?
5. Rotate the condenser knob to bring the condenser down and partially
open the iris diaphragm.
PARA311 LAB: Lecture 1

6. Turn the coarse focus adjustment knob so that the stage moves upward
towards the objective. Move it as far as it can without touching the slide.
7. Now, look into the eyepiece and adjust the coarse focus adjustment knob
so that the stage moves away from the slide. Continue moving until a
clear image of the slide comes into focus.
8. Turn the fine adjustment knob to precise the focus of the image.
9. Move the microscope slide until the object of focus is at the center of the
field of view. Then readjust the illuminator or diaphragm in order to
attain the maximum, or comfortable light intensity.
 HPO High power objective
PARA311 LAB: Lecture 1

a. Hold and rotate the revolving nosepiece until the 40x is obtained.
b. Rotate the condenser knob to bring the condenser half-way up and open the
iris diaphragm to attain a clear image.
c. Look in the eyepiece, rotate the fine adjustment knob to focus the specimen.
(DO NOT USE THE COARSE FOCUS ADJUSTMENT KNOB! Minimal use of
focusing adjustment is necessary in a high-power objective once a clear
image is obtained under the low-power objective.)
 OIO Oil immersion
PARA311 LAB: Lecture 1

1. When changing the objective lens of the microscope into oil-immersion

objective, a small drop of immersion oil is added into the slide under the
low-power or scanning objectives. (If the HPO is still used, change the
objective into LPO or scanning objective using the revolving nosepiece
before dropping the immersion oil.)
2. Hold and rotate the revolving nosepiece to change the objective into 100x.
Wait for a “click” until it is positioned. The 40x objective must never come
into contact with the immersion oil in the slide. The lens of OIO should be
slightly dipped into the immersion oil.
 OIO
PARA311 LAB: Lecture 1

3. Rotate the condenser focus knob to bring the condenser up and adjust the
iris diaphragm to get a clearest image. The light intensity may also be
adjusted to maximum if necessary.
4. Look into the eyepiece, rotate the fine adjustment knob to bring the image
into precise focus. (DO NOT USE THE COARSE FOCUS ADJUSTMENT KNOB!)
5. After use, remove the oil from the objective by wiping the front lens with
lens paper or gauze with 70% alcohol or mixture of 70% ether – 30%
alcohol.
 OCULAR MICROMETER
PARA311 LAB: Lecture 1

No. Of stage micrometer units

Formula: calibrated multiplier x No. Of microns = X 100
No. Of ocular micrometer units

OD (ocular division) - each line or spaces

bet ween is called OD
-they used to determine the size of organisms
This spaces bet ween those lines
will give you the information
on how long or thick is the
parasitic ova
OCULAR MICROMETER
PARA311 LAB: Lecture 1
OCULAR MICROMETER
PARA311 LAB: Lecture 1
 TROUBLE SHOOTING
PARA311 LAB: Lecture 1

- unplug
-power is not available Black field in the Oculars
- no lock in or the objective are not lock into
After focusing:
-dirty objective Blurred Image
place -dirt on the coverslip
- bulb is burnt out or inserted incorrectly -microscope slide is dirty
-bulb is damaged - forgot to clean the stage, there is the
- condenser is too low material present (can be oil)
- the diaphragm is completely close
 TROUBLE SHOOTING
PARA311 LAB: Lecture 1

Problem Focusing Partial Illumination

-Far focal - not fully open the condenser, or too low, or not
-too much oil centered correctly.
-microscope is not aligned properly -not fully open the diaphragm (it contains the
illuminator)
- e.g. Only half area has the light:possible that Hindi
nag lock in yung objectives
 TROUBLE SHOOTING
PARA311 LAB: Lecture 1

Light Flickers
 Things to take note
PARA311 LAB: Lecture 1

● Always carry the microscope with two hands.

● Never touch the lenses with your fingers.
● Only use lens paper for cleaning.
● Keep desk clear of objects and cords.
● When you are finished with your "scope", rotate the nosepiece so that it's
on the low power objective, roll the stage down to lowest level, rubber
band the cord.
PARA311 LAB: Lecture 1
 Evaluation
PARA311 LAB: Lecture 1

I. Identify the major components of the microscope and their function

II. Describe the process to correctly focus on the appropriate field of view
III. Discuss the ocular micrometer to
IV. Demonstrate the ability to troubleshoot encountered problems with the
microscope
V. Identify how to maintain a microscope
 References
PARA311 LAB: Lecture 1

Mescher, A. (2015). Junquiera’s Basic Histology: Text and Atlas (15th ed.). New
York: McGraw-Hill

Dr. Jose Jurel M. Nuevo. Diagnostic Parasitology Laboratory Manual. Our Lady of
Fatima University. Valenzuela City.

Mikhail A. Valdescona, RMT, MPH. PAR313 Lecture. Our Lady of Fatima

University. Valenzuela City.

Sherlyn Joy P. Isip, RMT, MSMT. PARA311 Laboratory. Our Lady of Fatima
University. Valenzuela City.
 THANK
Illustration by Smart-Servier Medical Art

Illustration by Smart-Servier Medical Art

YOU