TOPIC : TRANSMISSION ELCTRON MICROSCOPY
(TEM)
Submitted by:
Muhammad Awais (mtf23001900)
Zarnaqa Binte Furqan (mtf23001903)
Tayyaba Akhlaque (mtf23001904)
Submitted To:
Dr. Amna Asghar
Subject:
Bionanotechnology
DEPARTMENT OF CHEMISTRY
DIVISION OF SCIENCE AND TECHNOLOGY
UNIVERSITY OF EDUCATION , LAHORE
What is Transmission Electron Microscopy?
Transmission electron microscopy (TEM) is an analytical technique used to visualize the
smallest structures in matter. Unlike optical microscopes, which rely on light in the visible
spectrum, TEM can reveal stunning detail at the atomic scale by magnifying nanometer
structures up to 50 million times. This is because electrons can have a significantly shorter
wavelength (about 100,000 times smaller) than that of visible light when accelerated through a
strong electromagnetic field, thus increasing the microscope resolution by several orders of
magnitude
To form a TEM image, a high energy electron beam is accelerated through an extremely thin
“electron transparent” sample, typically thinner than 100 nm. A series of electromagnetic lenses
and apertures are placed throughout the microscope’s column to focus the beam on the sample,
minimize distortions, and magnify the resulting image onto a phosphor screen or a specialized
camera
Components of the Transmission Electron Microscope
TEMs are composed of five key components:
1. High voltage source
2. Vacuum system
3. Microscope column
4. Detectors (e.g., imaging cameras, spectrometers)
5. Control computers and software
Figure 1 Transmission electron microscopy
At the top of the column is the electron gun which couples to a high voltage source used to set
the kinetic energy of the electron beam. Typical accelerating voltages range from 80 kV up to
300 kV. The microscope column consists of a series of electromagnetic lenses and apertures to
focus the electron beam onto the sample and magnify the TEM image onto the viewing screen
(or detectors). A vacuum system is used to maintain the required vacuum levels throughout the
column. Due to the complexity of the instrumentation, most of the components
are automatically computer controlled with only a few key parameters controlled by the
microscopist. The main user-controlled settings include the sample stage position, magnification,
objective lens current, beam current (spot size), and the choice of which apertures and detectors
to use when acquiring data.
Modern TEMs are equipped with electronic detectors (such as charge coupled device, or CCD,
detectors), in addition to a retractable fluorescent viewing screen, to capture TEM images in a
digital format. Most TEMs will also include an X-ray detector that can be inserted between the
objective lens pole-pieces (near the sample) to analyze composition. Optional electron energy-
loss spectrometers located at the end of column are used for electron energy-loss spectroscopy
(EELS).
What is Transmission Electron Microscopy Used For?
Countless discoveries and innovations have been driven by applying TEM to the world around
us. The ability to “see” atoms enable scientists to understand materials and biological systems at
the most fundamental level. TEM images not only hold immense scientific value but are forms of
art on their own.
Electron-Sample Interactions
In order to identify the right imaging approach or correctly interpret image contrast or spectral
characteristics, it’s important to understand all of the relevant signals originating in the TEM. All the
signals generated in the TEM either originate from elastic scattering or inelastic scattering.
Elastic scattering:
Elastic scattering in TEM involves no energy transfer to incident electrons and occurs through
two main interactions:
1. Low-angle scattering due to interaction with the electron cloud.
2. High-angle (Rutherford) scattering caused by Coulombic attraction to the nucleus.
In crystalline materials, elastic scattering happens at characteristic angles (Bragg angles)
determined by the lattice spacings (d spacings). The resulting diffraction pattern reveals
structural information, including symmetry, strain, and defects.
Crystalline materials produce well-defined diffraction spots.
Polycrystalline/amorphous materials generate diffuse rings, where ring radii
correspond to lattice spacings, and increasing diffuseness indicates smaller crystallite sizes.
Inelastic scattering:
Inelastic scattering in TEM involves a measurable energy loss (meV to hundreds of eV) by
incident electrons. This process generates various signals, including energy-loss electrons, X-
rays, secondary electrons, and sometimes visible light (cathodoluminescence).
Modern S/TEM enables a holistic approach to materials characterization, combining high-
resolution imaging with analytical techniques for composition, chemistry, and bonding analysis
—crucial for nanotechnology and materials science.
The two most widely used inelastic signals are:
1. Electron energy-loss spectroscopy (EELS) – measures energy-loss electrons.
2. Energy-dispersive X-ray spectroscopy (EDS/EDX) – detects characteristic X-rays.
X-ray Spectroscopy:
X-ray spectroscopy is one of the most popular chemical analyses that can be performed in the
TEM or STEM. An EDX spectra provides a snapshot of the elements present in the sample.
There are two types of X-rays produced via inelastic scattering:
Characteristic X-rays: used for compositional analysis in nano-sized regions like defects,
surfaces, precipitates, and interfaces.
Bremsstrahlung X-rays: translates from German to “braking radiation” is a type of
continuum X-ray emission involving inelastic interaction with atomic nuclei. It is commonly
used in biological applications.
interaction of the incident electron beam with the sample, which can result in characteristic X-
ray emission.
Electron energy-loss spectroscopy:
EELS measures the energy distribution of all electrons that have interacted inelastically within
the sample. Unlike X-ray spectroscopy, which detects de-excitation events, the EEL spectrum
represents excitations or ionization within the specimen and therefore can probe a wider variety
of phenomena than EDX spectroscopy. These include core-shell transitions, plasmon resonance,
valence-to-conduction band transitions in semiconductors, and even vibrational modes
(phonons).
Figure 2Here is a simulated EELS spectrum related to Transmission Electron Microscopy (TEM). It includes characteristic
energy loss peaks, such as the Carbon K-edge (~284 eV), Oxygen K-edge (~532 eV), and an additional peak around 710 eV.
Here is a simulated EELS spectrum for biomolecules composed of carbon (C) and oxygen (O). It
highlights.
Carbon K-edge (~284 eV) – Represents C–C, C=C, and C=O bonds in organic molecules.
Oxygen K-edge (~532 eV) – Associated with oxygen-containing functional groups like C=O and
C–O–C.
Minor peak around 400 eV – Could indicate nitrogen (N) if proteins are present.
Figure 3Here is a simulated EELS spectrum for biomolecules composed of carbon (C) and oxygen (O)
TEM for Materials Science and Nanotechnology
Zooming into the atomic scale allows scientists to view the fundamental building blocks of
functional materials like catalyst nanoparticles, batteries, and semiconductor devices. Focused
electron beams can also be used to manipulate materials in situ, allowing “nanofabrication” and
novel phenomena to be studied and discovered. The level of detail on this scale is nothing short
of stunning and provides a way for understanding the connections between structure, property,
and performance, allowing engineers to design nanomaterials from the bottom up.
Figure 4TEM showing atomic resolution
TEM for Life Science
The world of biology is full of fascinating and dynamic yet invisible phenomena. Cryogenic
TEM allows structural biologists to visualize the architecture of macromolecular assemblies like
proteins, viruses, and intracellular structures at near atomic resolution. Recent technological
advancements such as the use of direct electron detectors, automation, and data processing have
propelled the technique into the mainstream. Using a method known as single particle analysis,
scientists have been able to determine the structure of the novel coronavirus spike protein, which
allows the virus to bind to host cells, allowing targeted vaccines to be developed.
Figure 5TEM micrograph of a poferritin particles and 3D reconstruction
REFRENCES:
Williams, D. B., & Carter, C. B. (2009). Transmission Electron Microscopy: A Textbook
for Materials Science. Springer.
Egerton, R. F. (2011). Electron Energy-Loss Spectroscopy in the TEM. Reports on
Progress in Physics, 72(1), 016502.
Koster, A. J., & Klumperman, J. (2003). Electron Microscopy in Cell Biology: The
Basics. Molecular Biotechnology, 23, 33–61.
Reimer, L., & Kohl, H. (2008). Transmission Electron Microscopy: Physics of Image
Formation. Springer.
Buseck, P. R., Cowley, J. M., & Eyring, L. (2018). High-Resolution Transmission
Electron Microscopy. Oxford University Press.
University of California, Berkeley: Electron Microscopy Lab – [Link]
[Link]
