Physics of Medical Devices

Ten lecture

Microscope
Msc. Eman Ahmed

Third Stage
Department of medical physics
Al-Mustaqbal University-College
2021- 2022

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 Introduction
The microscope is a valuable instrument. There are many small objects or

details of objects which cannot be seen by the unaided human eye. The

microscope magnifies the image of such objects thus making them visible

to the human eye. Microscopes are used to observe the shape of bacteria,

fungi, parasites and host cells in various stained and unstained

preparations.

History of Microscope
 In the 1st Century AD, the Romans invented the glass and used them to

magnify objects.

 In the early 14th Century AD, eyeglasses were made by Italian spectacle

makers.

 In 1590, two Dutch spectacle makers, Hans, and Zacharias Jansen

created the first microscope. It was a simple tube with 2 lenses system

and had 9X magnification.

 In 1670, Robert Hooke, an English Chemist, Mathematician, Physicist,

and Inventor, improvise the microscope of that time and developed the

compound microscope. He first developed the 3 lenses microscope.

 In 1675, Anton Van Leeuwenhoek ground a glass ball into a convex lens

and used it to make a single-lens microscope with 270X magnification.

Using this microscope, he first observed the bacterial cells.

in the microscope.

 In 1830, Joseph Jackson Lister suggested the use of multiple low-power

lenses to achieve clear magnification.

 In 1878, Ernst Abbe, a German Physicist and Optical Scientist,

developed a mathematical theory relating wavelength with image

resolution. He was the first to develop and use water and oil immersion

lenses.

 In 1903, Richard Zsigmondy invented the ultramicroscope. This could

view objects smaller than the wavelength of light.

 In 1932, Frits Zernike invented the phase-contrast microscope.

 In 1938, Max Knoll and Ernst Ruska invented the first electron

microscope. It was a transmission electron microscope (TEM). It used a

beam of an electron instead of light to make an enlarged image.

 In early 1940, Russian physicist Sergey Y. Sokolov developed concept

of ultrasound microscope. But, only in 1970, its working model was

developed in America.

 In 1942, Ernst Ruska improved TEM into a scanning electron

microscope (SEM). In this type, electron beams are passed across the

specimen instead of passing through it as in TEM.

 In 1951, British physicists William Nixon and Ellis Coslett invented X-

ray microscope.

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 In 1981, Gerd Binnig and Heinrich Rohrer invented the scanning

tunneling microscope. This allowed us to get the 3-D image of an object.

Types of Microscopy

Microscopes used in clinical practice are light microscopes. They are

called light microscopes because they use a beam of light to view

specimens. A compound light microscope is the most common microscope

used in microbiology. It consists of two lens systems (combination of

lenses) to magnify the image. Each lens has a different magnifying power.

A compound light microscope with a single eye-piece is called monocular;

one with two eye-pieces is said to be binocular. Microscopes that use a

beam of electrons (instead of a beam of light) and electromagnets (instead

of glass lenses) for focusing are called electron microscopes. These

microscopes provide a higher magnification and are used for observing

extremely small microorganisms such as viruses.

Parts of the Microscope

The main parts of the microscope are the eye-pieces, microscope tube,

nosepiece, objective, mechanical stage, condenser, coarse and fine

focusing knobs, and light source

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What is Refractive Index?
Refractive Index can be defined as velocity of light in a vacuum to velocity

of light in a medium (substance). Simply it is the measure of bending of a

light ray when passing from one medium to another.

Mathematically it can be defined as;

n = c/v

where,

n = refractive index

c = speed of light in vacuum

v = velocity of light in a medium

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Resolution

in microscopy, this refers to the ability of the lens system to separate the

image into small parts OR the ability to separate two elements in

thviewing field from each other. As the light increases, resolution should

increase or as the light decreases, resolution in general decreases. Since

resolution is a function of diffraction the very best optical microscopes are

limited to a resolution of 0.2 micrometers. NOTE – in modern usage, the

resolution of a projector or a video monitor or any other display unit, has a

different meaning, as noted below.

What is Resolution?
Resolution can be defined as the shortest distance between two points on a

specimen that can be distinguished by a microscope in its image. It is the

ability of a microscope to distinguish details on a specimen.

Mathematically it is given as;

r = ⋋/2NA

where,

r = resolution

⋋ = imaging wavelength

NA = numerical aperture

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Monocular

A single eyepiece, is set at 45o to the head for viewing by one observer.

Inexperienced microscopists will tend to close the eye not looking through

the eyepiece, which may result in headaches and eye strain to the intense

light in one eye and darkness in the other.

Binocular

Two eyepieces, side by side, one for each of the Viewer’s eyes. The two

eyepieces are generally set at a comfortable 45° angle. The inter-ocular or

inter-pupillary distance may be adjusted to match the distance between an

individual’s eyes.

Polarizing Microscope

Polarizing Microscope is a special type of light microscope that uses

polarized light to illuminate a specimen and develop its magnified image.

It is similar to a regular optical microscope but uses polarized light instead

of normal natural light. It enhances image quality and image contrast.

They are also called petrographic microscopes.

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 Polarizing Microscope Principle

Normal light produced from illuminator is passed through polarizer which

converts the normal light into plane-polarized light. The polarizing

microscope focuses the plane-polarized light on anisotropic (substance

having multiple refractive indexes) specimen. When the polarized light

waves strike such anisotropic specimen, birefringence (double reflection)

occurs generating two waves, ordinary and extraordinary waves, which are

perpendicular to each other. These two waves get transmitted in different

phases. An analyzer combines these two waves and passes through the ocular

lens to develop an enlarged image.

Uses of Polarizing Microscope

 Used in geological studies to study rocks, minerals and soil components.

 Used is studying internal structures of transparent planktons, diatoms,

protozoans, etc.