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TEM

Transmission ELectron
Microscopy
GROUP 6
1 MANSI 00101012021

2 SAKSHI 01801012021

3 MONALIKA PATNAIK 02301012021

4 SIMMI 03101012021

5 AYUSHI ARORA 18501012021


Before starting our topic TEM…..
What is Electron Microscopy?
ELECTRON
MICROSCOPY

The technique of electron microscopy (EM)


is used to obtain high-resolution images of
biological and non-biological specimens.
Some electron microscopy techniques have a
resolution of 0.2 nanometers, which is 1000 times
better than light microscopy.
Electron microscopes can magnify
specimens up to 1 million times their
original size.

In biomedical research, it is used to


investigate the detailed structure of tissues,
cells, macromolecular complexes etc.
Electron Microscopy

Scanning Electron Microscopy Transmission Electron Microscopy


(SEM) (TEM)
Transmission Electron Microscopy
(TEM)
What is TEM and how is it different
from SEM?
Transmission electron microscopy (TEM) is a microscopy
technique in which an image is formed by passing an
electron beam through a specimen.

Because electrons have a shorter de Broglie wavelength


(3.9*10-10 m), transmission electron microscopes can image
at much higher resolution than light microscopes.

This allows the instrument to capture fine detail, even down


to a single column of atoms, which is thousands of times
smaller than a resolvable object seen through a light
microscope.
TEM
SEM
SEM TEM
1. Transmitted electrons (electrons
1. Detects reflected electrons to
passing through the sample) are
generate an image.
used to create an image.

2. Provides information about the


2. Provides valuable information about
surface and composition of the
the sample's internal structure, such
sample.
as crystal structure, morphology, etc.

3. The resolution is restricted to 0.5 nm.


3. Images with spatial resolutions as
low as 50 pm (0.05 nm).
4. The maximum magnification is 1-2
million times.
4. The maximum magnification is over
50 million times.
PRINCIPLE OF TEM
● Electrons possess a wave like character.

● Electrons emitted into vacuum from a heated filament


with increased accelerating potential will have small
wavelength.

● Such higher-energy electrons can penetrate


distances of several microns into a solid.

● These transmitted electrons are used to focus images


with much better resolution.

● Focusing relies on the fact that, electrons also


behave as negatively charged particles and are
therefore deflected by electric or magnetic fields.
RAY DIAGRAM OF TEM
As electrons cannot pass through glass, the lenses that are used are
electromagnetic. They simply consist of coils of wires inside metal pole
pieces. When current passes through the coils, a magnetic field is
generated.

Condenser lens Objective lens Magnifying lens

This lens is used to form The Objective lens is Used to assist in


and control the used primarily to magnifying the
diameter of the focus and initially image and to
electron beam. magnify the image. project the
It limits the amount of The specimen stage magnified image
electrons in the beam. is inserted into the onto the screen.
objective lens for
imaging purposes.
CONSTRUCTION OF TEM
It consists of V shaped tungsten filament which
produces stable beam of electrons using
thermoionic emission.

The focusing and intensity of electron beam is


controlled by Electromagnetic lens placed
between electron gun and specimen. It consists
of copper coil. Current is made to pass through
coiled copper wire which creates magnetic field
thus responsible for focussing e- beam.

Specimen stage is used to hold sample under


investigation. It is in the form of mesh having
diameter 2.5mm thick. Specimen in TEM is
usually 100 nm thick.
Vacuum pumps are used to
evacuate the TEM to low
pressure upto 10-4 Pa.
Evacuation helps prevent
generation of arc.
TEM uses fluorescent screen at the base of
microscope column which produces light when
transmitted electrons strike it to form 2-D image
of sample surface.
SAMPLE PREPARATION
1. The specimen is fixed on a fixed copper grid.

2. The specimen must be an ultrathin section of less than 100 nm.

3. Fixation is done using chemical like glutaraldehyde to stabilises


the cell, preventing further change or damage to the cell.

4. Rinsing is done using sodium cacodylate to maintain the pH of


specimen after the process of fixation.

5. Staining is done using heavy metals like, osmium tetroxide to


increase the contrast of the minute structures inside the
specimen. Diamond Knife

6. Finally, the stained specimen must be dehydrated, embedded in


plastic resin and cut into ultra-thin slices with a diamond knife
attached to a device called ULTRAMICROTOME.
Working of TEM

https://youtu.be/a0G7iyz4McM
The given diagram
represents the working of
Transmission Electron
Microscope along with well
labelled diagram and the
arrows representing the
path of the electron beam.
● The electron gun consists of an electron source that produces electron beam travels via
anode in the column of the microscope.

● The Condensing lens are used to focus the electrons of the beam into a thin beam which is
allowed to fall on the specimen.

● On reaching the specimen, the specimen scatters the electrons. The beam of transmitted
electrons is directed to the magnetic objective lens and the aperture.

● The final image is produced due to transmitted beam when it is passed through the
Projective lens which helps in magnifying the image of the specimen.

● The magnified image is obtained on the fluorescent screen.


Applications of TEM

A TEM is ideal for a number of diverse fields such as life sciences,


nanotechnology, medical, biological and material research, forensic
analysis, as well as industry and education.

TEMs provide topographical, morphological, compositional and


crystalline information. The images allows the researchers to view
samples on molecular level, making it possible to analyze structure
and texture.
But what was the objective behind
using TEM….?
Resolution is the quality or definition that we can see something.
The resolving power of a microscope is directly related to the
wavelength of the irradiation used to form an image

Light Microscopes high wavelength of visible light


results in
low resolution power of microscope

Electron Microscopes low wavelength of electrons


results in
high resolution power of microscope
High resolution results in clear and
magnified image generation and allows
study on molecular level.
Early Applications of TEM in Material
Sciences (1940)
Early progress in material sciences in 1940s was mostly
limited to studies of small particles, such as the carbon
black that gave strength to automobile tires and the
pigments that were used in color paints and cosmetics.

TEM made it possible to analyze the outlines of small size


particles, determine their sizes, shapes and atomic
properties.
Figure shows dislocation loops in aluminium observed by
TEM.
TEM in Nanotechnology

TEM has an unparalleled ability to provide structural and


chemical information over a range of length scales down
to the level of atomic dimensions.

It has developed into an indispensable tool for scientists


who are interested in understanding the properties
of nanostructured materials and manipulating their
behaviour.
TEM in Nano Industry
A - Schematic representation of
SARS-CoV-2 as a core shell
nanoparticle.
B - TEM image of SARS-CoV-2 virions.
TEM in Semiconductor Industry

Semiconductor designers need analytical tools that offer


excellent flexibility and scalability, as well as high resolution
imaging, in order to properly analyze and optimize device
performance.

TEM characterisation allows manufacturers to calibrate


toolsets, diagnose failure mechanisms, and optimize overall
process yields.
TEM in Semiconductors

TEM is used for process


development and failure analysis
in semiconductor.
Forensic Application of TEM

TEM can be instrumental in forensic trace evidence


analysis due to its ability to analyze the morphology
of small particles, gather elemental information on very
small particles and determine the internal structures of
small particles.
Gunshot residue analysis, examination of paint and
fibres, trace comparisons, etc.
Forensic analysis
results from TEM

TEM image of paint pigments


found in an architectural paint
during a forensic analysis.
Drawbacks of TEM
1. Sampling Abilities

Any high-resolution imaging technique has its own in-built limitation: at any given
time one can examine just a small part of a specimen; the higher the resolution,
the lower will be its sampling

2. Projection Limitation

TEM works in such a way that we see 2D images of 3D


specimens, viewed as part of transmission. This specific
drawback in TEM is termed as projection limitation. One
particular aspect of this limitation is that the images,
diffraction patterns, or spectra information obtained by
TEM is averaged through the thickness of the specimen.
This means that there is no depth sensitivity in a single
TEM image.
3. Damages due to radiation
Ionizing radiation can always damage the specimens used in TEM. Polymers,
organic materials, certain minerals, and ceramics are examples of materials that
can get damaged by ionizing radiation. Such damages become worse at voltages
as high as 400 kV, which are possible to achieve in many commercial
instruments. There is also the possibility of exposing oneself to such damaging
radiation.

4. Specimen Preparation
Specimens need to be thin - the materials have to be electron transparent.
This in turn means that electrons passing through the material and falling on
the screen or photographic plate must have sufficient intensity for generating
an image within a reasonable timeframe. This is often a function of the
electron energy and the average atomic number (Z) of the specimen under
observation.
Extending Capabilities of TEM to
overcome drawbacks
1. Cryogenic TEM
Cryogenic transmission electron microscopy (Cryo-TEM) is widely used in nanoparticle
research. The specimens are kept in vitreous ice so that they are close to their native
environment. Then they are imaged whilst maintained at the temperature of liquid helium or
liquid nitrogen. This process of specimen preparation decreases sample damage from
radiation by almost 6-fold. 3D images of large biological structures at nanometer resolutions
can also be obtained using this method of sample preparation. A great advantage of
Cryo-TEM is that the sample environment can be controlled and hence the image retains
the native structural features without any distortions.

2. Low voltage electron microscope


The low voltage electron microscope (LVEM) may soon become the mode of choice for
electron imaging because of its ability to give high contrast images. Use of low accelerating
voltages of between 5 and 25 Kv leads to image contrast with a resolution more than
twenty times that of conventional TEMs using accelerating voltages of 10–1000 kv. The
increased contrast is due to the profuse electron scattering in LVEM.
THANK YOU
ANY QUESTIONS?

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