COURSE:

Characterization Techniques
2024

Dr. Rabeay Hassan

[email protected]
Scanning Electron Microscope
(SEM)
 Discovery of Atomic Structure

This scanning tunneling microscope (STM) Sketch of a scanning probe microscope

image of a graphene sheet reveals that a
‘Wigner crystal’ a honeycomb arrangement
of electrons has formed inside a layered
structure underneath. Credit: H. Li et
al./Nature
https://www.nature.com/articles/d41586-
021-02657-6
AFM and SEM images of the perovskite surface and after treatment with NiO x . (a,b)
AFM characteristics of the perovskite layer and NiO x layer coated on the perovskite film.
The scan size is 3 μm × 3 μm. (c,d) Top-view SEM images of the perovskite layer and NiO
x layer coated on the perovskite film.
Source: ACS Energy Lett. 2020, 5, 1, 79–86
 How to decide which technique is most appropriate for characterizing
common material properties including surface quality, grain size, and local
chemistry……….???

What are the practical ways to: a) increase the depth of field; b) improve the
image quality; c) increase signals; d) enhance resolution on images of bulk
samples; e) enhance resolution on images of nanomaterials?

Materials Characterization Using Non-destructive Evaluation (NDE) Methods

http://dx.doi.org/10.1016/B978-0-08-100040-3.00002-X
Of the following, __________ radiation has the shortest wavelength.
A) X-ray
B) radio
C) microwave
D) ultraviolet
E) infrared
 Electromagnetic Radiation
Electromagnetic radiation (EMR) refers to the waves of the electromagnetic field,
propagating (radiating) through space carrying electromagnetic radiant energy. It
includes radio waves, microwaves, infrared, (visible) light, ultraviolet, X-rays, and
gamma rays.
All electromagnetic radiation travels at the
same velocity: the speed of light (c), 3.00 
108 m/s.
Therefore,
c = 
So, they differ in only wavelength and
frequency

Electromagnetic radiation consists of two

perpendicular waves, one electric and one
magnetic, propagating at the speed of light (c).
Discovery of Atomic Structure

Or
 The Electromagnetic Spectrum

c = 

Waves with short wavelengths have a high frequency

Ultraviolet light, with wavelengths of ≤ 400 nm, has enough energy to cause
severe damage to our skin in the form of sunburn. Because the ozone layer absorbs
sunlight with wavelengths less than 350 nm, it protects us from the damaging
effects of highly energetic ultraviolet radiation.
 Wavelength, Frequency, Energy
• Electromagnetic wave characteristics:
– short wavelengths have a high frequency
– long wavelengths have a low frequency
c = 

E = h
• Electromagnetic waves & Energy:
– high frequency waves have high energy
– low frequency waves have low energy

smaller wavelengths increasing frequency increasing energy

Electromagnetic energy comes in the form of particles (known as photons). Each photon
carries a certain amount of energy.

c = 
 QUANTIZED ENERGY

In 1900, Max Planck proposed that light and other electromagnetic waves
were emitted in discrete packets of energy, which he called "quanta”.

Planck's constant, h, relates the

energy in one quantum (photon)
of electromagnetic radiation to the
frequency of that radiation.
6.62 x 10-34 joule-seconds.

If we know the wavelength of light, the

energy in one photon, or packet, of that
light could be calculated: c = , E = h

Max Planck postulated that electromagnetic energy is absorbed or

emitted in discrete packets, or quanta.
A photon is a quantum of electromagnetic energy.
 Bohr Theory
1. Electrons exist only in certain discrete
energy levels, which are described by
quantum numbers.
2. Energy is involved in the transition of
an electron from one level to another.

Electrons exist only in orbits with specific

amounts of energy called energy levels.

 Electrons can only gain or lose certain

amounts of energy.

Niels Bohr adopted Planck’s assumption and explained these phenomena in this way:
Electrons in an atom can only occupy certain orbits (corresponding to certain
energies).
Bohr Theory and the Ionization Energy of Hydrogen

The energy absorbed or emitted from the process

of electron promotion or demotion can be
calculated by the equation:

1 1
E = −RH ( nf2 - ni2 )
where RH is the Rydberg constant, 2.18  10−18 J,
and ni and nf are the initial and final energy levels of
the electron.
 -RH -RH
ΔE = Ef – Ei = –
nf2 ni2

1 1
= RH ( – ) = h = hc/λ
ni2 nf 2

Energy-level diagram
Rydberg constant, 2.179  10−18 J

E = h
 The Photoelectric Effect and Photons
Hot objects and the Quantization of
Energy…..
how electromagnetic radiation and Photo-electron
atoms interact?

Electrons can gain energy by interacting

with photons.

The photoelectric effect refers to the

ejection of electrons by a metal when struck
with light of sufficient energy.
Einstein concluded that energy is
proportional to frequency:
E = h
where h is Planck’s constant, 6.63
 10−34 J-s
Shorter wavelengths of radiation interact more strongly with
nanoscale materials, and can produce higher resolution
images

wave-particle duality
Scanning Electron Microscope
(SEM)
 Handbooks
Materials Characterization: Introduction to Microscopic and Spectroscopic
Methods, Second Edition. Yang Leng. 2013 Wiley-VCH Verlag GmbH & Co.
KGaA. Published 2013 by Wiley-VCH Verlag GmbH & Co. KGaA.

A Beginners' Guide to Scanning Electron Microscopy

A Beginners' Guide to Scanning Electron Microscopy | SpringerLink

978-3-319-98482-7.pdf (springer.com)
a machine for looking at small things
What material features can we analyze
using electron microscopy?

The basic and key

component of a microscope
is its illumination source.
 Resolution vs Magnification
Magnification range: Electron microscopes offer a very high magnification range,
typically in the range 10 to 500,000 times for SEM, and 2000 to 1 million times for TEM.
This enables characterization of microstructures at many different length scales, from
micro- to nanoscale, within an imaging session. However, it is important to understand the
difference between magnification and resolution.

The magnification of an image is the size of a feature in the image relative to the
actual size of the same feature on the sample.

Magnification is making the image bigger, whereas resolution is the ability to

distinguish the features of the object in the image. High magnification is useless if the
microscope operating conditions have not been optimized to achieve high resolution.

Imaging with electrons

Electrons with wavelengths of 40 pm down to 1 pm can be readily
achieved using accelerating voltages of 1-300 kV (Reimer and Kohl,
2008; Williams and Carter, 2009).
imaging with electrons
Electron Beam Interaction with Matter
 How do we get an image?

Electrons in
Electrons out

or: x-rays out

We shoot high-energiezed electrons and analyze the

out-coming electrons/x-rays
imaging with electrons
 Signals from the sample
Incoming electrons (Primary electron)
Secondary electrons (SE)
Auger electrons (AE)
Backscattered
electrons (BSE) Cathodo-
luminescence (light)
X-rays

Sample
 when electrons hit matter ..
Primary electrons (incident):
The electrons which are targeted
at the specimen.

Secondary electrons (SEs):

that originate from inside the
specimen:
Secondary electrons: Electrons
with kinetic energy produced by
primary electron ematter
interactions. SE emission occurs
when they escape from the
sample.

The spectrum of electrons-

specimen interactions can be
divided into elastic interactions
and inelastic interactions.
when electrons hit matter ..
 when electrons hit matter …....
(1) they may collide with an inner shell electron, ejecting
electrons
 the ejected electron is a low-energy, secondary electron (Detected & used to form
SEM images)……
the original high-energy electron is scattered (known as a ‘back-scattered’ electron)
an outer-shell electron drops into the position formerly occupied by the ejected electron,
this is a quantum process, so a X-ray photon of precise wavelength is emitted - basis for
X-ray microanalysis.

(2) they may collide or nearly collide with an atomic nucleus

 undergo varying degree of deflection (inelastic scattering)
undergo loss of energy (EELS) - again varying lost energy appears as X-rays of
varying wavelength this X-ray continuum is identical to that originating from an X-ray
source/generator .
 when electrons hit matter ..
(3)they may collide with outer shell electrons
 either removing or inserting an electron results in free radical formation, this
species is extremely chemically active. Reactions with neighbouring atoms induce
massive change in the specimen, especially in the light atoms. This radiation
damage severely limits possibilities of EM. So, examination of cells in the live state
NOT POSSIBLE.
all examinations need to be as brief (low dose) as possible

(4) they may pass through unchanged

these transmitted electrons can be used to form an image, this is called imaging by
subtractive contrast and can be recorded by either:
(a) TV-type camera (CCD)
(b) photographic film - direct impact of electrons
Photographic film: silver halide grains detect virtually every electron
Elastic versus inelastic electron
scattering
•Elastic scattering occurs when there is no loss of energy of the incident
(primary) electron.
•Elastically scattered electrons can change direction but do not change their
wavelength. Coherent elastic scattering produces the effect of electron
diffraction, which is used to analyze crystal structure.

•Inelastic scattering occurs when there is an interaction that causes loss of

energy of the incident primary electron.
•Inelastically scattered electrons have a longer wavelength. Inelastic scattering
occurs by many mechanisms, and the energy is transferred to the specimen
generating a range of useful signals that are exploited to characterize the material.

the ratio of elastic to inelastic scattered electrons decreases with

increasing specimen thickness.

SEM detects scattered electrons emitted from the surface of the

sample, while TEM detects transmitted electrons.
The interaction of incoming primary electrons with a sample.
(a) Signals generated by electron-ematter interactions in a thin sample.
(b) Absorption of SE, BSE, and X-rays in thick samples, by inelastic scattering within the
interaction volume, limits the sample depth from which they can escape.

For a given specimen thickness, materials containing heavier elements (higher atomic number Z)
have stronger electron interactions and absorb electrons faster. To be electron transparent, a
sample has to be thinner than the electron range in the specimen.
 Electron-induced X-ray emission

Characteristic X-ray generation.

An X-ray is generated when an Schematic of an energy dispersive
electron moves to fill the empty X-ray (EDX) spectrum exhibiting
electron energy level vacated by large numbers of characteristic X-
the secondary electron. rays at specific energies.
Scanning electron microscope
Scanning electron microscope (SEM) images of
various scaffold structures used in bone tissue
engineering.
https://tel.archives-ouvertes.fr/tel-
01139717/document
Schematic diagram of the core components of an SEM
microscope
BSE vs SE

Comparison of SEM techniques: Top:

backscattered electron analysis
(composition)
Bottom: secondary electron analysis
(topography)
Electron source:
The electron gun in an SEM typically accelerates electrons through ~20 keV accelerating
voltage. Low voltage SEM (LVSEM) operating mode can be used to reduce electron
penetration and achieve higher resolution SE imaging.

Vacuum:
The pressure inside the SEM chamber is usually low vacuum (10-5 Torr). Variable Pressure
SEM (VPSEM) and Environmental SEMs (ESEM) can operate in reduced vacuum to inhibit
evaporation of volatile components of the specimen.

Why Need a Vacuum?

When a SEM is used, the electron-optical column and sample chamber must always be at a
vacuum.

1. If the column is in a gas filled environment, electrons will be scattered by gas molecules
which would lead to reduction of the beam intensity and stability.

2. Other gas molecules, which could come from the sample or the microscope itself, could
form compounds and condense on the sample. This would lower the contrast and
obscure detail in the image.
 Electron guns
•Traditional electron-guns: thermionic electron gun (electrons are emitted when a solid is
heated)
–Tungsten (W) -wire, LaB6-crystal
•Modern electron-guns: Field emission guns (FEG) (cold guns, a strong electric field is
used to extract electrons)
–Single crystal of Tungsten, etched to a thin tip
•With field emission guns we get a smaller spot and higher current densities compared to
thermionic guns
•Vacuum requirements are tougher for a field emission guns

Single crystal of LaB6 Tungsten wire Field emission tip

 Electron Detectors
Backscattered electron
detector: (Solid-State
Detector)

Secondary electron detector:

(Everhart-Thornley)
 Electron Detectors
Electron Detectors:

Various detectors (Secondary Electron (SE),

Back Scattered Electron (BSE), characteristic x
rays, Auger Electron (AE) etc.) pick up the
signals. SEs are emitted from the surface top
<20 nm.

Everhart-Thornley (ET) Detector is a

secondary electron and back-scattered electron
detector used in SEM imaging.
ET which comprises a biased grid to attract the The Everhart-Thornley Detector (E-T
electrons, a scintillator to convert the SE signal to detector or ET detector) is a secondary
light, and a photomultiplier tube to amplify the electron and back-scattered electron
signal. detector used in scanning electron
ET detectors are usually fitted on one side of the microscopes (SEMs). ... A low positive
vacuum chamber. SE signal intensity can often be voltage is applied to the Faraday cage to
enhanced by tilting the specimen surface toward attract the relatively low energy (less
than 50 eV by definition) secondary
the detector.
electrons.
 Electron Detectors

Secondary electrons: Everhart-Thornley Detector

Backscattered electrons: Solid State Detector

X-rays: Energy dispersive spectrometer (EDS)

 Energy Dispersive X-ray (EDX) detectors

X-ray spectrometers:
X-rays are abundant and emitted all directions from the specimen. Since X-rays are
hazardous to human health, the specimen chamber must absorb X-rays, usually by having
a thick lead lining. The X-ray detector is located in line-of-sight of the specimen inside the
chamber.

X-rays are counted according to their energy in energy dispersive X-ray analysis
(EDX)

EDX detectors are based on semiconductor chips that convert individual X-rays to
electron-hole pairs, which then form an electronic current. The electronic signal generated
is proportional to the energy of the incoming X-ray. In this way during a collection time of
a few minutes, the detector can count the number of X-rays detected of given energy at a
chosen location on the specimen. This forms an energy dispersive X-ray spectrum
Sample Preparation
Sample chamber SEM of calcified particle in
cardiac tissue
 Coating needed prior to SEM imaging

Sputter coating for SEM is the process of applying an ultra-thin coating of electrically-
conducting metal – such as gold (Au), gold/palladium (Au/Pd), platinum (Pt), silver (Ag),
chromium (Cr) or iridium (Ir) onto a non-conducting or poorly conducting specimen.
Sputter coating prevents charging of the specimen, which would otherwise occur because of
the accumulation of static electric fields. It also increases the amount of secondary electrons
that can be detected from the surface of the specimen in the SEM and therefore increases the
signal to noise ratio.
Sputtered films for SEM typically have a thickness range of 2–20 nm.
Benefits for SEM samples sputtered with metal:
Reducing microscope beam damage
Increasing thermal conduction
Reducing sample charging (increasing conduction)
Improving secondary electron emission
Reducing beam penetration with improved edge resolution
Protecting beam sensitive specimens
Specimen composition
EDX Elemental Mapping Directly from SEM Image
 Specimen composition
A modern SEM can be equipped with various accessories:

Chemical composition of materials and the distribution of different phases can be key to their
structural and functional behavior. Quantification of chemical composition can be used to
understand and control key features such as oxidation, surface reactivity, mechanical strength,
and conductivity. Chemical imaging in SEM can be carried out by both BSEs and X-ray
analysis.

BSE imaging: Primary electrons are backscattered by electrostatic interaction between the
positive atomic nuclei and negative primary electrons (Rutherford backscattering).
The repulsive power of atomic nuclei is proportional to their positive charge, determined by
the number of protons (element Z-number). Heavier elements (high Z) generate more BSEs
than light elements (low Z). The BSE signal will be proportional to the average Z-number in
the sampled part of the interaction volume (the BSE escape depth being dependent on
incident electron energy). Therefore in a BSE image, the contrast will vary both due to
topographic contrast and chemical contrast.

For a flat specimen (minimized topographic contrast), regions of higher average Z will emit
more BSEs (be brighter) than regions of lower average Z. This is very rapid method to look
for distributions of two or more phases which have sufficiently different average Z-number to
cause chemical contrast modulation in the BSE imaged.
 EDX element distributions (maps)

Scanning electron microscopy image (A) of a superficial triple junction of a grain boundary
in a control TiO2 coin with the corresponding elemental maps (B and C). For better visibility
oxygen distribution (green) is pictured separately (C). EDX spectra (D) show quantitatively
the composition of the grain boundary and of one of the surrounding grains.
 Elemental compositions
Ce

Fe Sr
Surface crystallography
 Surface crystallography
Electron backscatter diffraction (EBSD): is a technique that can determine the crystal
structure and crystal orientation. The methodology collects elastically scattered BSEs which
have undergone coherent Bragg scattering. A dedicated EBSD detector collects the scattered
BSEs over a large solid angle, which forms electron backscatter diffraction patterns. The
EBSD patterns can be analysed to give the crystalline structure and orientation of the crystal
that the BSEs were scattered by.

Bragg scattering

EBSD analysis in SEM is now very automated, and has found widespread application in the analysis of
crystallography of metallic alloys.

EBSD can be used to quantify which crystal structures are present and their orientation, sizes and
morphology of individual grains, the collective texture of alloys, and crystallographic relationships
between phases.
Electron backscatter diffraction (EBSD)

Experimentally, EBSD is conducted in a scanning electron microscope (SEM)

equipped with an EBSD detector.
 Summary
 The scanning electron microscope is a versatile instrument that can be used for
many purposes and can be equipped with various accessories.
 An electron probe is scanned across the surface of the sample and detectors
interpret the signal as a function of time.
 A resolution of 1 – 2 nm can be obtained when operated in a high resolution
setup
 The introduction of ESEM and the field emission gun have simplified the
imaging of challenging samples

–Secondary electrons (SE): mainly topography

•Low energy electrons, high resolution
•Surface signal dependent on curvature
–Backscattered electrons (BSE): mainly chemistry
•High energy electrons
•“Bulk” signal dependent on atomic number
–X-rays: chemistry
•Longer recording times are needed
Disadvantages of imaging with
electrons
 Disadvantages of imaging with
electrons

High electron energy leads to the materials destruction.

Samples with a high vapour pressure such as biological materials and

liquids will change by loss of water and other volatiles in vacuum.
SEM Applications
 In-situ imaging: oxidation of steel at high
temperatures
•800 °C
•Formation of Cr2O3

2 min 10 min 90 min

https://www.nature.com/articles/s41563-
019-0387-3

https://www.nature.com/articles/s41563-
019-0387-3.pdf
 SEM vs TEM

SEM detects scattered electrons emitted from the surface of the sample, while TEM
detects transmitted electrons. In the case of SEM, the electron beam scans over the
surface of the sample, while in the case of TEM, the electron beam passes through the
sample. The accelerated voltage ranges from 10 to 40 kV for the SEM, while for TEM, it is
>100 kV.

SEM provides information about surface morphology and composition of materials while
TEM gives details about internal composition of materials. Thus, TEM can illustrate several
characters of a material (morphology, crystallization, stress or even magnetic domains).
SEM is a better tool for surface characterization as compared to TEM which is better for
internal structure analysis.

Both need electrically conductive materials to be tested. Non-conductive materials should

be coated with a conductive layer of metal or carbon.

SEM requires a very easy preparation technique, while TEM needs skill to prepare a very
thin sample.
The thickness of the specimen is not important in SEM, while specimen thickness is very
important in TEM.
The thickness of the specimens to be examined under TEM should be less than 150 nm.