Electron Microscopy

• An electron microscope is a microscope that uses a beam of

accelerated electrons as a source of illumination.
• It is a special type of microscope having a high resolution of
images, able to magnify objects in nanometres, which are
formed by controlled use of electrons in a vacuum captured on
a phosphorescent screen
• The key difference between electron and optical microscopy is
right there in the name. SEMs use a beam of electrons rather
than a beam of light. An electron source located at the top of
the microscope emits a beam of highly concentrated electrons.
Working Principle of Electron microscope
• The electron gun generates electrons
• Two sets of condenser lenses focus the electron beam on the specimen and then
into a thin tight beam.
• To move electrons down the column, an accelerating voltage (mostly between 100 kV
-1000 kV) is applied between the tungsten filament and anode
• The specimen to be examined is made extremely thin, at least 200 times thinner than
those used in the optical microscope. Ultra-thin sections of 20-100 nm are cut which
is already placed on the specimen holder.
• The electronic beam passes through the specimen and electrons are scattered
depending upon the thickness or refractive index of different parts of the specimen
• The denser regions in the specimen scatter more electrons and therefore appear
darker in the image since fewer electrons strike that area of the screen. In contrast,
transparent regions are brighter.
• The electron beam coming out of the specimen passes to the objective lens, which
has high power and forms the intermediate magnified image.
• the ocular lenses then produce the final further magnified image.
Scanning Electron Microscpe
•morphology
•composition

• It is termed a scanning electron microscope because the image

is formed by scanning a focused electron beam onto the
surface of the specimen in a raster pattern.
• Topography
• Morphology
• Composition
Electron sources
• In SEMs, there are three different types of electron sources:
• Thermionic filament – A Tungsten filament inside the microscope is heated
until it emits electrons. A Tungsten filament operates at white-hot
temperatures which means it gradually evaporates with time and eventually
breaks which can contaminate the upper part of the electron column. The
average lifetime of a Tungsten source is about 100 hours, depending on the
vacuum.
• Field emission gun (FEG) – Generate a strong electrical field that pulls
electrons away from their atoms. This is typically the more popular choice in
SEMs as it generates high resolution images, however it requires a vacuum
design that often comes at a high price.
• Cerium Hexaboride cathode (CeB6) – provides ten times the brightness
compared to Tungsten which means a better signal-to-noise ratio and better
resolution. A CeB6 source typically provides more than fifteen times the
service life of Tungsten: 1500+ hours. A CeB6 source is used in all the
desktop Phenom SEM series of instruments.
Applications of SEM

• Materials science
• Nanowires for gas sensing
• Semiconductor inspection
• Biological sciences
This image shows 50 µm of solar
panel surface and highlights the
pyramidal structures that help trap
light and reduce reflection.
Photovoltaic researchers are
seeking ways to optimize the texture
of these surfaces, since the shape,
size and uniformity of the pyramids
affect optical reflectance and energy
capture.
This is what 20 µm of
foundation from the make-up
industry looks like. A typical
product like this contains 15 to
50 ingredients. The most
common minerals used as a
base for foundation are mica,
bismuth oxychloride, titanium
dioxide and zinc oxide. The
spheres here are probably
silica, which acts as an
anticaking agent, bulking
agent, opacifying agent and
suspending agent. Silica also
typically absorbs sweat and oil,
reduces light reflection and
improves spreadability.
A hydrogen-absorbing alloy
magnified 30,000 times.
Hydrogen-storage alloys are
metallic materials that can
reversibly absorb and release
hydrogen from the gas phase
or electrochemically. These
alloys are already being used in
electrodes, particularly in
electric vehicles, to improve
performance and to avoid
using common alternative
materials that contain toxic
lead or cadmium, as these can
leach into landfill
SEM SAMPLE
PREPARATION
SAMPLE PREPARATION
 Sample preparation is crucial if you require a good SEM image. Unfortunately,
not all samples can be easily imaged, but this guide will help you with tips and
tricks to obtain good results from the most common samples
 Feel free to combine different techniques to boost the beneficial effects and
never underestimate your creativity. Scanning electron microscopes (SEMs)
are versatile instruments and they can do much more than you would expect.
 This sample preparation guide is meant for those who are approaching
scanning electron microscopy for the first time, or are relatively new to it
 The content is valid for macro categories of samples . For more detailed
information on specific kinds of samples, please contact your SEM
manufacturer
BASIC SAMPLE PREPARATION
Every SEM is equipped with a sample holder or a loading chamber where the
sample can be inserted . To load a sample in a SEM, the use of aluminium
stubs is recommended . These come in different, standard sizes and are
readily available on a commercial basis.
It is also recommended that you remove all loose particles from your sample.
To do this, you can:
• Hold the aluminium stub with tweezers, tilt it by 90° and gently tap it on its
side
• Spray dry air on the sample . Remember to take the following precautions: •
Be careful while handling your sample to prevent damage .
• Always use tweezers to prevent contamination .
• Make sure that the mounting procedure is solid, so that you do not introduce
mechanical vibrations due to incorrect mounting .
• DO NOT spray dry air in the direction of any electronics or scanning
electron microscope, because it might be flammable .
• Make sure there is no condensed liquid in your spray air straw by
f ir st spraying away from your sample . These precautions will
dramatically reduce the risk of contamination of your system and
sample holder and will guarantee better performance over time . For
any of the samples on the following list, please refer to the dedicated
section of the guide for suggestions on how to get the best images
To stick the sample to the pin stub, you can use:
• Double-sided carbon sticker
• Conductive paint
• Conductive tape
• Special clamps
• A combination of the above
 NON-CONDUCTIVE SAMPLES
When a non-conductive material is imaged, the electrons shot onto the
sample surface don’t have a path to the ground potential, causing them
to accumulate on the surface . The image will become increasingly
bright or entirely white until details are no longer visible . Mild
movement can also be detected, caused by the mutual interaction of
the electrons . This will cause blurriness in the collected image
Several solutions are widely used:
Conductive tapes or paints
By covering part of the sample with a piece of conductive tape (e .g .
copper tape) or some conductive paint, a bridge to the surface of the
aluminum stub is created This will allow the sample to partially
discharge and is enough to image mildly non-conductive samples
when imaging areas close to the tape edge .
Low vacuum:
Introducing an atmosphere in the sample chamber allows beam
interaction with air molecules . Positive ions are generated and
attracted by the large number of electrons on the sample surface .
The ions will further interact with the electrons, discharging the
sample . While this technique adds some noise to the final image,
you can analyze the sample faster and inexepensively without
further processing
Sputter coating:
By using a sputter coater, it is possible to create a thin layer of a
conductive material on the sample surface . This creates a
connection between the surface of the aluminum pin and the ground
potential . The choice of coating material is strongly dependent on
the kind of analysis to be performed on the sample . Gold and
platinum are ideal materials for high-resolution images because both
have extremely high conductivity . Lighter elements, like carbon, can
be used when Energy Dispersive Spectroscopy (EDS) analysis on
non-organic samples is required .
An alloy of indium oxide and titanium oxide (ITO) can create
transparent, conductive layers, to be used on optical glasses to
m a k e t h e m s u i t a b l e f o r S E M . H o w e v e r, t h e r e a r e
d i sad v antage s to usi ng a sputte r c o ate r: A d d i ti o nal
instrumentation is required, the analysis becomes more time
consuming, and the samples undergo more pumping cycles .
Also, any advantage of using a backscatter electron detector
(BSD) to image the sample is lost, as the contrast becomes
very homogeneous and there is no difference in gray intensity
for different elements
MAGNETIC SAMPLES
Samples that generate a magnetic field can interfere with the accuracy of the
electron beam, reshaping it and producing deformed, blurry images, usually
elongated along one axis . This problem is known as stigmation alteration and
consists of an increase in the eccentricity of the beam cross section .
Stigmation correction
All SEMs offer the chance to tune the stigmation . Certain instruments require
the user to fix stigmation values every time, while others can store standard
values that are valid for most samples . The procedure alters the magnetic
field of the lenses, which reshapes the beam . When the shape is circular
again, the best image can be produced . When changing the stigmation, it
might be necessary to finetune the focus again .
BEAM-SENSITIVE SAMPLES
Delicate samples, like thin polymeric foils or biological samples, can be damaged by the
electron beam due to the heat generated in the interaction area or the rupture of
chemical bonds . This will result in either a hole in the surface or a progressive
deformation of the scanned area .
Beam settings
The easiest way to reduce this effect is to use lower values for voltage and current . In
these cases, the smallest possible values are recommended .
Sputter coating
In the worst cases, a thin coating layer can be applied to the sample in order to shield
the sensitive surface . Increased conduction will also improve image resolution .
Cooling
Thermal effects can be reduced by using a temperature controlled device . Removing
the heat generated by the beam will protect the sample from thermal-induced surface
modifications .
Time
Spending a long time on a specific spot will cause damage to the sample, over time .
Being quick during the analysis will prevent excessive alterations, but might not produce
the best results in terms of image quality
Magnification
Zooming in implies having the same number of electrons shot on a smaller area . The
thermal drift is increased and the deformation effects will become more evident . When
possible, low magnification is recommended
POWDERS AND PARTICLES
When imaging particles, information like particle size or shape are
crucial in the design of the process flow . The easiest way to prepare
a powder or particles sample is to collect a small amount of sample
with a spoon and let it fall on a carbon double-sided sticker, then
using spray air to remove the excess particles . Unfortunately, this
method will cause many particles to overlap, hiding important
features, or to be blown off, inducing errors in particle counting
routines
Particles disperser:
The best way to prepare a powder sample is by using a particle
disperser unit . This will allow an even distribution of the sample on
the sticker, reducing the incidence of overlapping particles and
generating a pattern that can be used to study granulometry .
Operational parameters, such us the vacuum level and the amount of
sample needed, depend largely on the nature of the powder .
Factors to consider:
• Fine powders require a smaller amount of sample .
• Delicate samples might break due to strong pressure outburst .
• Hydrophilic samples might need a higher vacuum burst to be
separated
SAMPLES CONTAINING MOIST
OR OUTGASSING ELEMENTS
When electron microscopes operate in high vacuum levels, every wet
sample that is loaded in the imaging chamber will immediately start
to outgas . Certain samples have microstructures that will resist the
phase change, providing excellent results without major concerns . A
typical example is a fresh leaf . A sample without a rigid structure can
be imaged if force drying or critical point drying is used to prepare it.
Force drying
To verify whether the sample will resist the vacuum, the use of
another instrument, such as a desiccator or a sputter coater, is
recommended . Eventual changes in the sample should be
immediately noticeable.
Critical point drying
Also known as supercritical drying, this technique forces the liquids
in the sample to evaporate, maintaining a low temperature . The
evaporation is driven by the pressure level, which is brought below
the vapor tension of the liquid in the sample . During this process, the
liquids will create fractures in the sample, causing modifications in
the structure
Cooling
This is an alternative to drying techniques that will preserve the
structure of the sample completely intact by freezing the sample . If
the phase change is quick enough, the liquids in the sample will not
form crystals and the structure will be perfectly preserved . It is
important to consider that the phase change is not permanent and a
prolonged exposure to a high vacuum will increase the evaporation
rate .
Low vacuum
If the sample does not have a particularly high moisture content,
using a small amount of sample at a reduced vacuum level can SEM
images of a cucumber’s intact structure. SEM images of (left)
spreadable hazelnuts and (right) cacao cream. SEM image of a
tomato’s peel and interior structure. be enough to collect images .
The overall image quality will be lower, but the sample can be
imaged in its original state .
Small amount
of sample Using a small quantity of sample is sometimes enough to
contain the effects of vacuum and evaporation . The sample can be
collected with a toothpick and a veil of it can be deposited on the
stub.
This technique is particularly effective with gels and emulsions
Transmission Electron
Microscopy (TEM)
TEM
• 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. 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.
• The TEM operates on the same basic principles as the light microscope
but uses electrons instead of light. Because the wavelength of electrons is
much smaller than that of light, the optimal resolution attainable for TEM
images is many orders of magnitude better than that from a light
microscope. Thus, TEMs can reveal the finest details of internal structure -
INSTRUMENTATION
• The TEM consists of an Electron gun made from a Tungsten
filament or a Lanthanum hexaboride crystal
• The e-gun is charged with about 100-300 kV before any
electrons of reasonable energy will be released from the source.
• Then a series of electromagnetic and electrostatic lenses direct
the electrons into a beam
• The electron optical system of a TEM consists of an electron source
and several electron lenses stacked vertically to form a lens column.
The TEM can be conveniently divided into three sections:
1) The illumination system consists of the electron source, electron
accelerator, together with two or more condenser lenses which,
together with a condenser aperture, determines the diameter of the
electron beam at the specimen and the intensity level in the TEM
image. Typically there will be gun and condenser alignment coils to
allow the optical center of the gun and the condenser system to be
aligned on the optical axis of the objective lens and also a condenser
stigmator to correct for the imperfections in the condenser lenses.
2) The objective lens and specimen stage
are the heart of the instrument. The specimen (which is typically 3mm
in diameter and less than 100nm thick in the region of interest) is
mounted in the specimen stage within the strong magnetic field of the
objective lens (~2T). The electron optical properties of the objective
lens will define the ultimate resolution of the microscope. The
specimen holder and goniometer allows specimens to be held
stationary while imaging at atomic resolution while also allowing
movement in up to 5 axes (X, Y, Z and tilt X, tilt Y), depending on
specimen holder, and easy transfer into and out of the microscope
vacuum system. There is an objective stigmator in the lower bore of
the objective lens which corrects for the axial asymmetry of the pole
piece and an objective aperture, in the back focal plane, which can
increase contrast by defining which electrons form the image.
3) The imaging system
consists of at least three lenses that together form a magnif ied image (or
diffraction pattern) of the specimen on the f lu orescent screen or CCD
camera. Small changes to the intermediate lenses focal lengths allow the
magnif ication to be changed in discrete steps over a large range (x2,000
– x1,500,000). Larger changes to the excitation of the first of these lenses
(intermediate lens 1) are used to switch between imaging and diffraction
on the viewing screen. In conjunction with the selected area aperture, an
area of the specimen can be def in ed in imaging mode from which a
diffraction pattern can be obtained in diffraction mode (selected area
diffraction). The intermediate lenses are relatively weak with focal
lengths of a few centimeters. Alignment coils in the imaging system
allow f in e movement of the image (image alignment) on the viewing
screen and alignment of the imaging system with the center of the
various cameras and detectors (projector alignment).
• Most TEM images are collected with an objective aperture
inserted around the optic axis of the microscope. If a small
aperture is used, selecting only the direct beam, then any
scattered electrons will fall outside this aperture and as a result
the image will show contrast variations. If the specimen is
amorphous this contrast will depend on the specimen thickness
and density and a mass-thickness contrast image will be
obtained. If the specimen is crystalline then any scattering
contrast will be dominated by diffraction contrast caused by
Bragg diffraction of electrons from suitably aligned lattice
planes.
• In addition to scattering contrast, features seen in some TEM
images depend on the phase of the electron wave at the exit plane
of the specimen. This cannot be measured directly in the TEM but
does give rise to interference between electron beams that have
passed through different parts of the specimen which can be
bought together by defocussing the TEM image. A large diameter
(or no) objective aperture is needed to enable many beams to
contribute to the phase contrast image. In the special case of a
crystalline specimen oriented to be on a zone axis this can give
rise to atomic resolution images, however it must be remembered
that this is not a direct image of the structure. This can be seen if
the defocus is changed from one side of focus to the other - the
contrast will reverse with bright becoming dark and vice versa.
Solid state chemistry
• Solid-state in Chemistry is the study of the
structure, properties and the synthesis of
solid materials. It is also sometimes called
as materials chemistry. More significantly,
in solid-state chemistry, we study the
concept of a compound in a more deeper
level. It basically helps us understand the
compound from a molecular level to the
crystal structure level.
 Properties of solid
• Solids have definite mass, volume and shape
• Solids are rigid. This is due to lack of space between
the constituent particles which make it rigid or fixed
• The constituents particles can only oscillate about
their mean positions
• The intermolecular distance between molecules is
short
• Solids are incompressible, meaning the constituent
particle is arranged close to each other and because
of that, there is negligible space between the
constituent particle
 Types of solid state
• Crystalline Solids
Have a typical geometry. In such type of
solids, there are definite arrangements of
particles (atoms, molecules or ions )
throughout the 3-dimensional network of a
crystal in a long-range order. Examples
include Sodium Chloride, Quartz, Diamond,
etc.
• Crystalline solids have a sharp melting point and start
melting when it reaches a particular temperature.
• The shape of crystalline solids are definite and having
typical arrangements of particles.
• They show cleavage property i.e. when they are cut with
the edge of a sharp tool they split into two pieces and the
newly generated surfaces are smooth and plain.
• They have definite heat of fusion (amount of energy
needed to melt a given mass of solid at its melting point).
• Crystalline solids are anisotropic which means their
physical properties like electrical resistance or refractive
index show different values when they are measured along
with different directions in the same crystal.
• Crystalline solids are true solid
 Types of Crystalline Solids

• Molecular Solids
• Non-Polar Molecular Solids
• These solids are formed from molecules
or atoms that share a nonpolar bond e.g.
Cl , H , I , the atoms or molecules are held
2 2 2

by weak dispersion force or by London

forces.
• Polar Molecular Solids
• These solids are held together by polar
bonds e.g. HCl, SO 2
 Ionic Solids

• Constituent particles are ions. These are

formed by the arrangement of cations and
anions by strong attraction forces.
• These are hard and brittle in nature.
• Ionic solids act as an insulator in a solid-
state but are conductors in a molten and
aqueous state.
• They have a high melting point.
• Example: NaCl, MgO, ZnS, CaF etc.
2
 Metallic Solids

• Positive metal ions in a sea of delocalized

electrons. These electrons are evenly spread out
throughout the crystal.
• Due to the presence of free and mobile electrons,
they are responsible for high electrical and
thermal conductivity.
• They are conductors in both solid and molten
state.
• The physical nature of these solids are hard but
they are malleable and ductile.
• They have high melting point than ionic solids.
• Examples: Fe, Cu, Ag, Mg, etc.
 Covalent or Network Solids

• Awide range of crystalline solids of non-

metal form covalent bond between
adjacent atoms throughout the crystal and
form a giant molecule or large molecules.
• These solids are hard like diamond and
soft like graphite which are isotopes of
carbon.
• They are insulators as in the case of a
diamond but in case of graphite due to
free electrons, they conduct electricity and
act as a conductor.
 Amorphous Solid State
• Amorphous solid-state comprises of those solids which have the property of
rigidity and incompressibility but to a certain extent. They do not have a
definite geometrical form or long range of order. Examples include glass,
rubber, plastic, etc.
• Properties of Amorphous Solids
• Amorphous solids are gradually softened over a range of temperature and
they can be moulded into different shapes on heating.
• Amorphous solids are pseudo solids or super cooled liquids which means
they have a tendency to flow very slowly. If you observe that the glass pans
which is fixed to windows of old buildings they are found to be slightly thicker
from the bottom than at the top.
• Amorphous solids have irregular shape i.e. their constituent particles do not
have definite geometry of arrangements.
• When amorphous solids are cut with a sharp edge tool they form pieces with
irregular surfaces.
• Amorphous solids do not have definite heat of fusion due to its irregular
arrangement of the particles.
• Amorphous solids are isotropic in nature which means the value of any
physical property would be same along any direction because of the
irregular arrangement of particles.
 Crystal Structures
• All substances, except helium, if cooled
sufficiently form a solid phase; the vast majority
form one or more crystalline phases, where the
atoms, molecules, or ions pack together to form a
regular repeating array
• Crystal Structures are usually determined by the
technique of X-ray crystallography
• This technique relies on the fact that the
distances between atoms in crystals are of the
same order of magnitude as the wavelength of X-
rays (of the order of 1 Å or 100 pm): a crystal thus
acts as a three-dimensional diffraction grating to
a beam of X-rays.
 Close pack crystal structure
• Efficient space saving lattice
• Close-packing represents the most
efficient use of space when packing
identical spheres
• Close-packing represents the most
efficient use of space when packing
identical spheres
 Hexagonal close packed
Repeated stacking sequence build up the
layers ABABABA
hollows marked with a dot are never
occupied by spheres, leaving very small
channels through the layers

hexagonal close-packing (hcp)

 cubic close-packing (ccp)

the third layer could be positioned over

those hollows marked with a dot

when repeated would be ABC ABC

 BODY-CENTRED AND PRIMITIVE
STRUCTURES
• slightly less efficient packing method: this is
the body-centred cubic structure (bcc)
• this structure an atom in the middle of a cube
is surrounded by eight identical and
equidistant atoms at the corners of the cube
• the coordination number has dropped from
twelve to eight
• the packing efficiency is now 68%, compared
with 74% for close-packing
• The simplest of the cubic structures is the
primitive cubic structure
majority of metals have one of the three
basic structures: hcp, ccp, or bcc
 Symmetry
symmetry operation

symmetry elements

The line about which the

molecule rotates is called an
axis of symmetry