TRANSMISSON

ELECTRON MICROSCOPY
(TEM)
Presented by :-
Ankitasoreng
Smriti Rita Beck
Biva Mahata
Asma kerketta
Maxima Kandulna
Amisha Hembrom
CONTENTS
1. Introduction
2. principles
3. parts
4. how does it works?
5. Sample preparation
6. Application
7. Advantages
8. Disadvantages
 INTRODUCTION
It is a type of electron microscopy.
Electron microscopy involves the study of different specimen by using an
electron microscope.
Electron microscopes are scientific instrument that uses a beam of energetic
electrons to examine objects on a very fine scale.
Through the use of on electron microscope we can see things that we would
not normally be able to see with our naked eyes and has a grater
magnification than light microscope.
Transmission electron microscopy (TEM) is a technique use to observe the
features of very small specimen. the technology uses an accelerated beam
of electrons, which passes through a very thin specimen to enable a scientist
to observe features such as structure and morphology.
Transmission electron microscopy (TEM) was first developed by German
scientist Max Knoll and Ernst Ruska in 1931 and has evolved over the
years to become a common technique that is used globally in science
and engineering to look at micro and nano particles.
PRINCIPLES
The working principle of TEM is similar to the light microscopes.
The major difference is that light microscopes uses light rays to focus and
produce an image while the TEM uses a beam of electron to focus on the
specimen, to produce an image.
Electrons have a shorter wavelength in comparison to light which has a
long wavelength.
The wavelength of the electron is about 0.005nm which is 10,000X shorter
than that of light, hence TEM has a better resolution than that of the light
microscope, of about 1000times.
This can accurately be stated that the TEM can be used to detail the
internal structures of the smallest particles like a viron particle.
 PARTS OF TEM
It has three main parts:
1. Electron gun
2. Image producing system
3. Image recording system
 z

1. Electron gun-

To produce the electrons.

Electrons are produced by a cathode that is a tungsten filament that is
V-shaped and it is normally heated.The cathode is negatively charged
with an end of the anode which is disk-shaped .
It also has the condenser lens system which works to focus the
electron beam on the specimen by controlling the energy intensity.
The TEM uses two condenser lenses to converge the beam of electrons
to the specimen. The two condenser lens each function to produce an
image i.e the first lens which has strong magnification, produces a
smaller image of the specimen, to the second condenser lens, directing
the image to the objectives.
2. Image producing system-
It is made up of the objective lens, a movable stage for holding the
specimen, intermediate and projector lenses. They function by
focusing the passing electrons through the specimen forming a highly
magnified image.
The objective lens has a short focal length of about 1-5mm and it
produces an intermediate image from the condenser which are
transmitted to the projector lenses for magnification.
The projector lenses are of two types, i.e the intermediate lens which
allows great magnification of the image and the projector lens which
gives a generally greater magnification over the intermediate lens.
To produce efficient high standard images, the objectives and the
projector lenses need high power supplies with high stability for the
highest standard of resolution
3. Image recording system-
Its made up of the fluorescent screen used to view and to focus on the
image. They also have a digital camera that permanently records the
images captured after viewing.

The image that is formed is called a monochromatic image, which is

greyish or black and white. The image must be visible to the human eye,
and therefore, the electrons are allowed to pass through a fluorescent
screen fixed at the base of the microscope.
 HOW DOES IT WORKS?
A heated tungsten filament in the electron gun produces electrons that get focus
on the specimen by the condenser lenses.
Magnetic lenses are used to focus the beam of electrons of the specimen. By the
assistance offered by the column tube of the condenser lens into the vacuum
creating a clear image, the vacuum allows electrons to produce a clear image
without collision with any air molecules which may deflect them.
On reaching the specimen, the specimen scatters the electrons focusing them on
the magnetic lenses forming a large clear image, and if it passes through a
fluorescent screen it forms a polychromatic image.
The denser the specimen, the more the electrons are scattered forming a darker
image because fewer electron reaches the screen for visualization while thinner,
more transparent specimens appear brighter.
 PRPARATION OF SAMPLE
The specimen to be viewed under the TEM must undergo a special preparation technique
to enable visualization and creation of a clear image.
The specimen should be thin, because electrons are easily absorbed and easily scattered
on solid elements, showing poor visualization for thick samples.
To get thin slice samples following steps are need to be followed:-
Fixation - the specimen is first fixed on a plastic material with glutaraldehyde or
osmium tetraoxide.
Dehydration - Washing with increasing ethanol concentration, followed by the final
wash in another polar substance like propylene oxide .
Embedding - Epoxy resin is used to penetrate the cell, which will then occupy the
space and make the sample hard enough to bear the pressure of sectioning or cutting.
Sectioning & Staining - The specimen is then cut into thin slices or sections (less
than 100nm) with the help of ultramicrotome.The specimen is then stained
appropriately (with the appropriate stain) for the uniform scattering of electrons. They
are treated with heavy metal compounds
 APPLICATIONS OF TEM
TEM is used in a wide variety of fields From Biology, Microbiology,
Nanotechnology, forensic studies, etc. Some of these applications include:
1. To visualize and study cell structures of bacteria, viruses, and fungi
2. To view bacteria flagella and plasmids
3. To view the shapes and sizes of microbial cell organelles
4. To study and differentiate between plant and animal cells.
5. Its also used in nanotechnology to study nanoparticles such as ZnO
nanoparticles
6. It is used to detect and identify fractures, damaged microparticles which
further enable repair mechanisms of the particles.
 ADVANTAGES OF TEM
1. It has a very powerful magnification of about 2 million times that
of the Light microscope.
2. It can be used for a variety of applications ranging from basic
Biology to Nanotechnology, to education and industrial uses.
3. It can be used to acquire vast information on compounds and
their structures.
4. It produces very efficient, high-quality images with high clarity.
5. It can produce permanent images.
6. It is easy to train and use the Transmission Electron Microscope
 DISADVANTAGES OF TEM
1. Generally, the TEMs are very expensive to purchase
2. They are very big to handle.
3. The preparation of specimens to be viewed under the TEM is very tedious.
4. The use of chemical fixations, dehydrators, and embedments can cause the dangers of
artifacts.
5. They are laborious to maintain.
6. It requires a constant inflow of voltage to operate.
7. They are extremely sensitive to vibrations and electro-magnetic movements hence
they are used in isolated areas, where they are not exposed.
8. It produces monochromatic images, unless they use a fluorescent screen at the end of
visualization.
Thank
you!