Department of Metallurgical and Materials Engineering

Subject: Characterization of Engineering Materials (MME-308)

Title: Transmission Electron Microscope (TEM)
Submitted to: Dr. Muhammad Asif Rafiq
Submitted by: Raza Ali (2018-MM-34)
Date: June 16, 2021

University of Engineering and Technology, Lahore

 Table of Contents
1.0. Introduction
2.0. Components of TEM
2.1. Electron gun and condenser system
2.2. Image producing system
2.3. Image recording system
3.0. Working principle
3.1. Difference with light microscope
3.2. Imaging
3.3. Diffraction
3.4. Specimen preparation
4.0. Advantages of TEM
5.0. Limitations of TEM
6.0. Applications of TEM
7.0. Images using TEM
8.0. References

List of Figures
Figure 1: TEM microscope
Figure 2: left image: Cotton stem; area in the circle is the phloem tissue. Light microscope
x250 Right image: Enlarged image of cotton phloem tissue showing a sieve element (top cell)
and a companion cell (bottom cell), TEM x8,000.
Figure 3: Ray diagram of image formation and diffraction mechanism
Figure 4: Ultra-microtome and microtome grid 
Figure 5: Transmission electron micrograph of SARS-CoV-2 virus particles, isolated from a
patient. 
 Transmission Electron Microscope
1.0. Introduction
 Transmission electron microscopes (TEM) are microscopes that use a particle beam of
electrons to visualize specimens and generate a highly-magnified image. TEMs can
magnify objects up to 2 million times.
 In order to get a better idea of just how small that is, think of how small a cell is. It is no
wonder TEMs have become so valuable within the biological and medical fields.

Figure 1: TEM
microscope
 The transmission electron microscope is a very powerful tool for material science. A high
energy beam of electrons is shone through a very thin sample, and the interactions
between the electrons and the atoms can be used to observe features such as the crystal
structure and features in the structure like dislocations and grain boundaries [1].
 Chemical analysis can also be performed. TEM can be used to study the growth of layers,
their composition and defects in semiconductors.
 High resolution can be used to analyze the quality, shape, size and density of quantum
wells, wires and dots.
 Among all microscopes both light and electron microscopes, TEM are the most powerful
microscopes used in laboratories. It can magnify a mall particle of about 2nm, and
therefore they have a resolution limit of 0.2um.
2.0. Components of TEM
Transmission electron microscope (TEM), type of electron microscope has three essential
systems [2]:
 An electron gun, which produces the electron beam, and the condenser system, which
focuses the beam onto the object.
 The image-producing system, consisting of the objective lens, movable specimen stage,
and intermediate and projector lenses, which focus the electrons passing through the
specimen to form a real, highly magnified image.
 The image-recording system, which converts the electron image into some form
perceptible to the human eye. The image-recording system usually consists of
a fluorescent screen for viewing and focusing the image and a digital camera for
permanent records. In addition, a vacuum system, consisting of pumps and their
associated gauges and valves, and power supplies are required.
2.1. Electron gun and condenser system
 The source of electrons, the cathode, is a heated V-shaped tungsten filament or, in high-
performance instruments, a sharply pointed rod of a material such
as lanthanum hexaboride.
 The filament is surrounded by a control grid, sometimes called a Wehnelt cylinder, with a
central aperture arranged on the axis of the column; the apex of the cathode is arranged to
lie at or just above or below this aperture.
 The cathode and control grid are at a negative potential equal to the desired accelerating
voltage and are insulated from the rest of the instrument.
 The final electrode of the electron gun is the anode, which takes the form of a disk with
an axial hole. Electrons leave the cathode and shield, accelerate toward the anode, and, if
the stabilization of the high voltage is adequate, pass through the central aperture at a
constant energy. The control and alignment of the electron gun are critical in ensuring
satisfactory operation.
 The intensity and angular aperture of the beam are controlled by the
condenser lens system between the gun and the specimen. A single lens may be used to
converge the beam onto the object, but, more commonly, a double condenser is
employed.
 In this the first lens is strong and produces a reduced image of the source, which is then
imaged by the second lens onto the object.
 Such an arrangement is economical of space between the electron gun and the object
stage and is more flexible, because the reduction in size of the image of the source (and
hence the final size of illuminated area on the specimen) may be varied widely by
controlling the first lens.
 The use of a small spot size minimizes disturbances in the specimen due to heating and
irradiation.
2.2. Image producing system
 The specimen grid is carried in a small holder in a movable specimen stage. The
objective lens is usually of short focal length (1–5 mm [0.04–0.2 inch]) and produces a
real intermediate image that is further magnified by the projector lens or lenses.
 A single projector lens may provide a range of magnification of 5:1, and by the use of
interchangeable pole pieces in the projector a wider range of magnifications may be
obtained.
 Modern instruments employ two projector lenses (one called the intermediate lens) to
permit a greater range of magnification and to provide a greater overall magnification
without a commensurate increase in the physical length of the column of the microscope.
 For practical reasons of image stability and brightness, the microscope is often operated
to give a final magnification of 1,000–250,000× on the screen.
 If a higher final magnification is required, it may be obtained by photographic or digital
enlargement.
 The quality of the final image in the electron microscope depends largely upon the
accuracy of the various mechanical and electrical adjustments with which the various
lenses are aligned to one another and to the illuminating system.
 The lenses require power supplies of a high degree of stability; for the highest standard
of resolution, electronic stabilization to better than one part in a million is necessary.
 The control of a modern electron microscope is carried out by a computer, and dedicated
software is readily available.
2.3. Image recording system
 The electron image is monochromatic and must be made visible to the eye either by
allowing the electrons to fall on a fluorescent screen fitted at the base of the microscope
column or by capturing the image digitally for display on a computer monitor.
 Computerized images are stored in a format such as TIFF or JPEG and can be analyzed
or image-processed prior to publication.
 The identification of specific areas of an image, or pixels with specified characteristics,
allows spurious colors to be added to a monochrome image. This can be an aid to visual
interpretation and teaching and can create a visually attractive picture from the raw
image.
3.0. Working principle
 TEMs employ a high voltage electron beam in order to create an image. An electron gun
at the top of a TEM emits electrons that travel through the microscope’s vacuum tube.
 Rather than having a glass lens focusing the light (as in the case of light microscopes), the
TEM employs an electromagnetic lens which focuses the electrons into a very fine beam.
 This beam then passes through the specimen, which is very thin, and the electrons either
scatter or hit a fluorescent screen at the bottom of the microscope.
 An image of the specimen with its assorted parts shown in different shades according to
its density appears on the screen.
 This image can be then studied directly within the TEM or photographed.
3.1. Difference with light microscope
Although TEMs and light microscopes operate on the same basic principles, there are several
differences between the two. The main difference is that TEMs use electrons rather than light
in order to magnify images. The power of the light microscope is limited by the wavelength
of light and can magnify something up to 2,000 times. Electron microscopes, on the other
hand, can produce much more highly magnified images because the beam of electrons has a
smaller wavelength which creates images of higher resolution [3]. 

Figure 2: left image: Cotton stem; area in the circle is the phloem tissue. Light
microscope x250 Right image: Enlarged image of cotton phloem tissue showing a
sieve element (top cell) and a companion cell (bottom cell), TEM x8,000.
 3.2. Imaging
 The beam of electrons from the electron gun is focused into a small, thin, coherent beam
by the use of the condenser lens.
 This beam is restricted by the condenser aperture, which excludes high angle electrons.
The beam then strikes the specimen and parts of it are transmitted depending upon the
thickness and electron transparency of the specimen.
 This transmitted portion is focused by the objective lens into an image on phosphor
screen or charge coupled device (CCD) camera. Optional objective apertures can be used
to enhance the contrast by blocking out high-angle diffracted electrons. The image then
passed down the column through the intermediate and projector lenses, is enlarged all the
way.
 The image strikes the phosphor screen and light is generated, allowing the user to see the
image. The darker areas of the image represent those areas of the sample that fewer
electrons are transmitted through while the lighter areas of the image represent those
areas of the sample that more electrons were transmitted through.
3.3. Diffraction
 Figure 3 right image shows a simple sketch of the path of a beam of electrons in a TEM
from just above the specimen and down the column to the phosphor screen.
 As the electrons pass through the sample, they are scattered by the electrostatic potential
set up by the constituent elements in the specimen.
 After passing through the specimen they pass through the electromagnetic objective lens
which focuses all the electrons scattered from one point of the specimen into one point in
the image plane.
 Also, shown in figure 3 right image is a dotted line where the electrons scattered in the
same direction by the sample are collected into a single point. This is the back focal plane
of the objective lens and is where the diffraction pattern is formed.

Figure 3: Ray diagram of image formation and diffraction

mechanism

3.4. Specimen preparation

 Specimens must be very thin so that electrons are able to pass through the tissue.
 This may be done by cutting very thin slices of a specimen’s tissue using an ultra-
microtome.  The tissue must first be put in a chemical solution to preserve the cell
structure.  The tissue must also be completely dehydrated (all water removed). 
 Once preserved and dehydrated, tissue samples are placed in hard, clean plastic.  The
plastic supports the tissue while it is being thinly cut with the ultra-microtome.
 After sections are cut and mounted on grids, (tiny circular disks with openings,) a solution
of lead is used to stain the tissue (Figure 4).  The lead provides contrast to the tissue by
staining certain cell parts. 
 When placed in the electron microscope, the electrons are scattered by the lead.  They do
not penetrate the tissue or hit the fluorescent screen, leaving those areas dark. 

Figure 4: Ultra-microtome and microtome

grid 
The advantages, limitations and applications of TEM are as follows [4]:
4.0. Advantages of TEM
 It has a very powerful magnification of about 2 million times that of the light microscope.
 It can be used for a variety of applications ranging from basic biology to nanotechnology,
to education and industrial uses.
 It can be used to acquire vast information on compounds and their structures.
 It produces very efficient, high-quality images with high clarity.
 It can produce permanent images.
 It is easy to train and use the transmission electron microscope.
5.0. Limitations of TEM
 Generally, the TEMs are very expensive to purchase.
 They are very big to handle.
 The preparation of specimens to be viewed under the TEM is very tedious.
 The use of chemical fixations, dehydrators, and embedments can cause the dangers of
artifacts.
 They are laborious to maintain.
 It requires a constant inflow of voltage to operate.
 They are extremely sensitive to vibrations and electro-magnetic movements hence they
are used in isolated areas, where they are not exposed.
 It produces monochromatic images, unless they use a fluorescent screen at the end of
visualization.
6.0. Applications of TEM
TEM is used in a wide variety of fields from biology, microbiology, nanotechnology, forensic
studies, etc. Some of these applications include:
 To visualize and study cell structures of bacteria, viruses, and fungi.
 To view bacteria flagella and plasmids.
 To view the shapes and sizes of microbial cell organelles.
 To study and differentiate between plant and animal cells.
 It is also used in nanotechnology to study nanoparticles such as ZnO nanoparticles.
 It is used to detect and identify fractures, damaged micro particles which further enable
repair mechanisms of the particles.
7.0. Images using TEM

Figure 5: Transmission electron micrograph of

SARS-CoV-2 virus particles, isolated from a patient. 

8.0. References
[1] https://warwick.ac.uk/fac/sci/physics/current/postgraduate/regs/mpagswarwick/ex5/
techniques/structural/tem/ last accessed 15 June, 2021 (10:00am)
[2] https://www.britannica.com/technology/transmission-electron-microscope last accessed
15 June, 2021 (10:30am)
[3] https://www.ccber.ucsb.edu/ucsb-natural-history-collections-botanical-plant-anatomy/
transmission-electron-microscope last accessed 15 June, 2021 (11:15am)
[4] https://microbenotes.com/transmission-electron-microscope-tem/ last accessed 15 June,
2021 (12:00pm)