PHH114: LIFE SCIENCES

TOPIC PRINCIPLES
OF
1 MICROSCOPY

LEARNING OUTCOMES:
By end of the topic, you should be able:
1. To know the difference between Simple Microscope and Compound Microscope.
2. To identify the parts of microscope and its function.
3. To understand the characteristics of Scanning Electron (SEM) and Transmission
electron microscopy (TEM).

 INTRODUCTION

In this topic you will learn about microscope. A microscope is an instrument that makes an
enlarged image of a small object, thus revealing details too small to be seen by the unaided eye.
The most familiar kind of microscope is the optical microscope, which uses visible light focused
through lenses. The word “microscope” comes from the Latin “microscopium,” which is derived
from the Greek words “mikros,” meaning “small,” and “skopein,” meaning “to look at.”
It is not definitively known who invented the microscope. However, the earliest microscopes
seem to have been made by Dutch opticians Hans Janssen and his son Zacharias Janssen (Figure
1) and by Dutch instrument maker Hans Lippershey (who also invented the telescope) about
1590.

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Figure 1: Hans Janssen and his son Zacharias Janssen

The resolution of a microscope is a measure of the smallest detail of the object that can be
observed. Magnification is when the objects you see appear larger than they are whereas
resolution is you are able to see close together objects as distinct.

1.1 LIGHT MICROSCOPE

With the evolved field of Microbiology, the microscopes used to view specimens are both
simple and compound light microscopes, which all are using lenses. The difference between
simple and compound light microscope, Simple microscope has only one lens whereas
Compound microscope has two or more lenses that makes things look larger rather than using
one lens.

There are a few limitations of light microscope that you have to know. The light microscope
uses visible light to detect and magnify very small objects, and enlarging them and use lenses to
focus light on the specimen, magnifying it thus producing an image. The specimen is normally
placed close to the microscopic lens. Other limitations are the wavelengths of light are 400-750
nm, can enlarge cells up to 2,000 times and can resolve objects down to about 200 nm in size.

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Figure 2: 2 objects visualized with poor resolving power (left) and good resolving power (right).

Resolving power is the ability of an optical device to produce separate images of close objects
(Figure 2.2).
The modern types of Light Microscopes include:
i. Bright field Microscope
ii. Dark-Field Microscope
iii. Phase-Contrast Microscope

Bright field Microscope

The Bright Field microscope is the most basic optical Microscope used in microbiology
laboratories which produces a dark image against a bright background. It made up of two
lenses, and widely used to view plant and animal cell organelles including some parasites such
as Paramecium after staining with basic stains.

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Figure 3 below shows the parts of a bright-field microscope (Compound light microscope).

Figure 3: Parts of a microscope (Source: www.microbenotes.com)

Dark-Field Microscope
The dark field microscope creates a dark background to allow viewing of small unstained
objects. Which mean the specimen is bright with dark background. Light not scattered by the
specimen bypasses the objective, therefore making the “field” dark. This microscope is good for
unstained, live, and motile specimens.

Phase-Contrast Microscope
The Phase-Contrast microscope use refraction and interference that caused by structures in a
specimen to create high-contrast and high-resolution images without staining. It is the oldest
and simplest type of microscope that creates an image by altering the wavelengths of light rays
passing through the specimen. It provides high contrast, good resolution and often used to

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observe live specimens. This microscope is good for bacteria, flagella, cilia and certain
structures, such as organelles in eukaryotic cells and endospores in prokaryotic cells (Figure 4).

Figure 4: The image of Pseudomonas (left) and Desulfotomaculum (right) using phase-contrast
microscopy.

Total Magnification
Magnification is to make something larger in size than it really is by use of lenses. In
microscopes, this is usually indicated by the abbreviation “X”. Total magnification: In a
compound microscope the total magnification is the product of the objective and ocular lenses.
We know that the magnification of the ocular lenses on microscope is 10X. Hence, to get the
total magnification, we must multiply the magnification of objective lens with the magnification
of ocular lens. Figure 5 below are the examples of total magnification calculation based on the
difference magnification of objective lenses.

Figure 5: Total magnification calculation for different objective lenses.

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1.2 ELECTRON MICROSCOPE

Electron microscope uses a beam of accelerated electrons as a source of illumination. Electron

microscope produce a high resolution of images, which able to magnify objects in nanometers.
Electron microscope can resolve structures down to 0.5 nm. Specimens can be magnified more
highly with this microscope.
There are 2 types of EM:
1. Transmission Electron Microscope (TEM)
2. Scanning Electron Microscope (SEM)

Transmission Electron Microscope (TEM)

The transmission electron microscope (TEM) (Figure 6) can view image of specimens up to 1
micrometer in thickness. High-voltage electron microscopes are similar to TEMs but work at
much higher voltages. For TEM, electron form an image after they passed through a thin
specimen. This microscope is useful for studying the internal structures of cells and provide 2-D
image as well as have high resolution and high magnification.

Figure 6: Transmission Electron Microscope (TEM)

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Scanning Electron Microscope (SEM)
The scanning electron microscope (SEM) (Figure 7), in which a beam of electrons is scanned
over the surface of a solid object, is used to build up an image of the details of the surface
structure. For SEM, electron form image after they passed through a surface of a specimen. This
microscope is useful for studying surface structures of cells and provides a 3-D image. It also
has much higher resolution and magnification than possible in LM.

Figure 7: Scanning Electron Microscope (SEM)

Activity 1

Answer this set of Question online to determine your understanding of microscope.

https://www.biologycorner.com/quiz/qz_microscope.html

Watch this video to learn basic microscope setup and use.

https://www.youtube.com/watch?v=SUo2fHZaZCU&t=6s

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SELF-CHECK 1
Define microscope, resolving power and total magnification.
 Describe transmission and scanning electron microscopes.

PHH114: LIFE SCIENCES

SUMMARY

A microscope is an instrument that makes an enlarged image of a small object, thus revealing
details too small to be seen by the unaided eye. The most familiar kind of microscope is the
optical microscope, which uses visible light focused through lenses.

KEY TERMS
Microscope
Resolving power
Total Magnification
Light Microscope
Electron Microscope

REFERENCES

Campbell, N.A. (2021). Biology. (12th ed). Pearson Education, Inc.

https://www.britannica.com/technology/microscope/History-of-optical-microscopes

https://www.youtube.com/watch?v=SUo2fHZaZCU&t=6s

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