Electron Microscopy of Materials Review

Maria Paula Fernandez Jimenez (5801303)

Pedro Nel Martinez

Mechanical Engineer

Military University Nueva Granada

Faculty of Engineering

Industrial Engineering

Cajicá,Cundinamarca

2023
 Content
Introduction ............................................................................................................................. 4
1.Notions of optics .................................................................................................................. 5
Magnification ....................................................................................................................... 5
Resolution ........................................................................................................................... 5
Depth of field ....................................................................................................................... 6
Lens defects ........................................................................................................................ 6
Chromatic aberration....................................................................................................... 6
Spherical aberration ........................................................................................................ 7
Astigmatism ..................................................................................................................... 7
2.Scanning Electron Microscopy (SEM) ................................................................................ 7
3.Transmission electron microscopy(TEM)............................................................................ 9
4.Scanning Transmission Electron Microscopy (STEM) ..................................................... 15
5.Energy dispersive x-ray analysis (EDX); electron energy loss spectroscopy (EELS). .... 17
6.Processing and simulation ................................................................................................ 21
Image processing.............................................................................................................. 21
Image simulation ............................................................................................................... 22
7.Sample Preparation ........................................................................................................... 22
SEM................................................................................................................................... 23
TEM ................................................................................................................................... 24
Self-supported samples ................................................................................................ 24
Samples supported on grids ......................................................................................... 24
8.Preliminary design of the experiment (Which microscope do I need and why?) ............. 25
References ........................................................................................................................... 27
 Ilustration Content
Ilustration 1 Optical microscope with a two-lens system. ..........................................................5
Ilustration 2 Components of an optical reflection system ..........................................................7
Ilustration 3 Schematic representation of a scanning electron microscope ............................8
Ilustration 4 a) Scanning image of a silica sample supported on alumina. b) Compositional
map, where the clear contrast corresponds to the silica-rich composition
zones.composition, where the clear contrast corresponds to the zones of silica-rich
composition. .....................................................................................................................................9
Ilustration 5 Components of an optical transmission system ....................................................9
Ilustration 6. Contrast transfer function (CTF) for a 400 kV microscope and Cs = 1.0 mm at
the Scherzer focus (-405 A) .........................................................................................................13
Ilustration 7 Different contrast mechanisms depending on the lens aperture ......................14
Ilustration 8. Mode of operation in the transmission electron microscope as a function of
the condenser aperture. ...............................................................................................................15
Ilustration 9. Mode of operation in the transmission electron microscope as a function of
the condenser aperture. ...............................................................................................................15
Ilustration 10. Image Formation in STEM..................................................................................16
Ilustration 11 a) TEM BF image of the SBA15 catalyst with Co particles and b) STEM-
HAADF micrograph of the same area. .......................................................................................17
Ilustration 12. Schemes of electronic transitions that give rise to energy signals ................18
Ilustration 13. MoS EDX Line Analysis ......................................................................................19
Ilustration 14. EELS spectrum of an amorphous carbon aggregate showing the maxima
due to transitions at n* and a* levels ..........................................................................................20
Ilustration 15. scanning microscope ...........................................................................................23
Ilustration 16. Spectroscope........................................................................................................26
 Introduction

From the composition of the first optical microscope composed in 1590 by the Dutch

brothers Hans and Zacharias Janssen; and the first optical microscope designed in 1676 by

the also Dutch Antony van Leeuwenhoek, it was still more than three hundred years before

the German researchers Ernst Ruska and Max Knoll succeeded in constructing the first

transmission electron microscope in 1931. Even so, this first microscope did not allow to

obtain images as sharp as those already obtained in optical microscopy at that time.

Between the 1940s and 1970s, an exponential development took place, leading to the

construction of a microscope with an atomic resolution of 1,250 kV. It was not until 1965 that

the first scanning electron microscope was introduced.

Electron microscopy is a science in full expansion due to its relative novelty,

compared to optical microscopy. Thus, concepts such as magnification, depth of field,

resolution and aberrations used in electron microscopy have the same meaning as in optical

microscopy. It is the illumination system of the object that suggests an obvious division

between scanning and transmission electron microscopy. The scanning microscope

provides images of external morphology similar to those formed by the eye, while a

transmission microscope reports on the internal structure of solids. The electron microscope

can be considered as a communication channel between an object (problem) and the

observer. The information obtained is a function of the various physical processes of

interaction of the high-energy electron beam (20-1500 kV) with the object.

Microscopy can also have disadvantages such as:

1. Low representativeness of the data obtained, especially in the case of transmission

microscopy, because the

1. Low representativeness of the data obtained, especially in the case of transmission

microscopy, since the amount of sample observed is always a minuscule amount of the

original sample.

2.Difficulty in quantifying electron diffraction and HREM results. The electron interaction is

so intense that it is dominated by dynamic effects, so any quantification is affected by the

thickness of the diffracted zone of the crystal.

3.Difficulty in interpreting HREM images.

1.Notions of optics

Magnification

Magnification is the magnification of the final object, it is proportional to the number

of lenses and focal lengths. The closer the focal length and the object distance, the greater

the magnification.

Ilustration 1 Optical microscope with a two-lens system.

Resolution

The resolution is the smallest distance between two points that can be viewed

separately as independent. According to Rayleigh's Criterion (equation [1]), to obtain a

higher resolution, smaller (r), we can either decrease the wavelength of the radiation or

increase the angle of incidence:

𝑟 = 0,61 𝜆/𝑛 𝑠𝑒𝑛 𝛼 (1)

Where:

𝑟 is the resolution

𝜆 is the wavelength of the radiation

𝑛 is the refractive index of the medium

𝛼 is the angle between the light source and the object (depends on the size of the lens

apertura

Depth of field

The depth of field limits the thickness of the object to be studied. A greater depth of

field (h) means obtaining sharp images of objects of appreciable thickness, for which the

wavelength of the illumination source is increased or the angle of convergence (a) is

decreased, in both cases sacrificing resolution.

0,61𝜆
ℎ= 𝑠𝑒𝑛 𝛼 tan 𝛼 (2)
𝑛

Where:

ℎ is the depth of field

𝜆 is the wavelength of the radiation

𝑛 is the refractive index of the medium

𝛼 is the angle between the light source and the object (depends on the size of the

objective aperture)

Lens defects

Chromatic aberration

It occurs because light is not monochromatic, it depends on the refractive index of

the lens which in turn varies with the wavelength. As many images as colors are

produced.

Spherical aberration

This is related to rays entering the lens at greater angles to the optical axis. The

effect is that the image is not sharp.

Astigmatism

It is produced by the lack of axial symmetry of the lens. It is manifested by the

deformation of the image in a certain direction.

2.Scanning Electron Microscopy (SEM)

According to the basic principles of optics, the system of image formation in

scanning electron microscopy is based on the optical system of reflection. The object is

illuminated frontally and the reflected beams are responsible for the final information.

Ilustration 2 Components of an optical reflection system

In a scanning electron microscope, the beam passes through condenser and

objective lenses, and is scanned across the sample by scanning coils, while a detector

counts the number of low-energy secondary electrons emitted by each spot on the

surface.
 A scanning electron microscope consists of a fine electron beam with energy up to

40 kV that is focused and scanned over the surface of a sample.

Ilustration 3 Schematic representation of a scanning electron microscope

The two major advantages of the scanning microscope are the range of

magnification and the depth of field of the image. Depth of field is the property by which

SEM images can focus on surfaces at different heights at the same time.

The accelerating voltage is between 20 and 40 kV, its resolution between 50 and

20 A. The higher magnification is a function of the scanning system rather than the lenses,

the smaller the scanned area the higher the magnification, a surface at a focus can be

magnified between 3x and 150,000x.

Among the phenomena that take place in the sample under electron impact, the

most important in SEM is the emission of secondary electrons with energies of a few tens

of eV, followed by the emission of backscattered electrons with higher energies. There are

suitable detectors that discriminate electrons according to their energy, thus allowing

images to be formed with both secondary and backscattered electrons.

Because of this the images formed by secondary electrons represent the surface

characteristics of the sample.

 A fraction of the incident beam electrons may leave the material as backscattered

electrons with their energy reduced by inelastic interactions with the specimen.

Today it is possible to make combined use of both types of electrons thanks to the

great advances that have been made in backscattered detectors that allow good images to

be obtained with large numbers of beads.

Another important emission that occurs when the electron beam interacts

inelastically with the sample, both in SEM and TEM, is that of X-ray photons with energy

and wavelength characteristic of the elements that make up the sample.

Ilustration 4 a) Scanning image of a silica sample supported on alumina. b) Compositional

map, where the clear contrast corresponds to the silica-rich composition

zones.composition, where the clear contrast corresponds to the zones of silica-rich

composition.

3.Transmission electron microscopy(TEM)

Ilustration 5 Components of an optical transmission system

In a transmission electron microscope the sample is illuminated by a beam of

electrons produced in the barrel at the top of the microscope.

 When the electrons are emitted from the filament they pass through a large

potential difference (accelerating voltage) and acquire a kinetic energy. The wavelength

corresponding to them is then given by the De Broglie equation:

1⁄
𝜆 = ℎ/[2𝑚0 𝑒𝑉(1 + 𝑒𝑉/2𝑚0 𝑐 2 )] 2 (3)

𝜆 = wavelength

𝑚0 = electron residual mass

𝑉= potential difference

ℎ = Planck's constant

𝑒 = electron load

𝑐= speed of light

In the table 1 lists the different electron wavelengths for the most common

accelerating voltages in microscopy.

Table 1. Wavenlenghts of the electron when subjected to different accelerating voltages

In the table 2 lists the different properties of the different electron sources

commonly used to form the electron gun.

 Table 2. Electron emitting sources in comercial electron microscopes

The fact that the electrons are charged means that after a diffraction phenomenon

equivalent to that occurring with X-rays, the scattered electrons can be focused to form an

image. Spherical aberration consists in the reduction of the focal length of the electrons

passing through the outer regions of the objective lens with respect to the electrons

passing through the center of the lens. The spherical aberration coefficient (Cs) of the

objective lens is, together with the accelerating voltage and beam coherence, one of the

limiting factors of the resolving power of an electron microscope, so that the smallest

distance (d) reflected in a TEM image is given by the expression:

1⁄ (4)
𝑑 = 0,66(𝐶𝑠𝜆3 ) 4

Where λ is the wavelength of the incident electrons.

When the beam passes through the crystal, the electrons are scattered or reflected

by the crystal planes by the planes of the crystal with Miller hkl indices. According to

Bragg's law, constructive interference between the constructive interference between the

scattered beams takes place when:

2𝑑ℎ𝑘𝑙 𝑠𝑖𝑛 𝜃 = 𝑛𝜆 (5)

Where:

𝑑ℎ𝑘𝑙 is the interplanar distance

𝜃 is the incident angle

𝑛 is an integer

The image is the inverse Fourier Transform of the diffraction which, in turn, is the

Fourier Transform of the object. Due to the effect of the objective lens the electron beams

have their amplitudes and in some cases their phases altered. The degree of the alteration

will be determined by the Contrast Transfer Function (CTF) of the lens which varies with

the focus conditions. The CTF indicates the contrast transferred by the object to the image

for a given reciprocal vector q and is given by an expression of the type:

𝐶𝑇𝐹 = 𝐴(𝑞). 𝑒𝑥𝑝[𝑖𝜒(𝑞)] (6)

Where A(q) is an aperture function that defines the truncation of the diffraction pattern by

the objective aperture and where the phase function exp[ix(q)] describes the phase

distortion imposed on the output wavefunction1 by the objective lens. If the CTF is

negative then the atoms will show a black contrast on a white background and if positive

they will give a white contrast on a black background. If the CTF is large for a given value

of q it means that the periodicity corresponding to that value of q in the object will be

strongly transmitted to the image. Thus, to preserve the relative amplitudes and phases of

the scattering process this function has to be close to unity and of the same sign. It has

been shown by Scherzer that the largest frequency range with sin x(q) = 1 is obtained

when:

1⁄ (7)
≜ 𝑓 = 2.5(𝐶 𝜆/2𝜋) 2

The blur value given by the expression [] is the best blur value given to obtain a

high resolution image.

Ilustration 6. Contrast transfer function (CTF) for a 400 kV microscope and Cs = 1.0 mm at

the Scherzer focus (-405 A)

The resolution given for this focus value is determined by the first zero of the CTF,

which is given by the expression:

1
𝑟𝑠𝑐ℎ = 0,66 𝐶𝑠 4𝜆3⁄4 (8)

This contrast transfer function changes with focus and as the transfer function

changes, so does the transfer of reflections.

In TEM images there are three types of contrast mechanisms: i) intensity contrast

from absorption, i.e. from the loss of incident electrons by multiple inelastic scattering; ii)

amplitude contrast is determined by the objective aperture which eliminates scattered

electrons with a divergence angle greater than the diameter of this aperture and which, as

a consequence, will not contribute to the image. This aperture is therefore often referred to

as the contrast aperture; iii) the phase contrast which is determined by the focus.

The fundamental principle, when obtaining TEM images, is to look at the electron

diffraction diagram, since it is this that gives us information about how a sample scatters.

Given the relationship between the image and the fraction, the diffraction pattern can be

used to choose the contrasted mechanism to be used depending on the sample to be

studied. So far we have focused on the usual technique, called brightfield, in which the
diaphragm (aperture) of the objective lens is focused on two or more diffracted beams in

addition to the central beam to form a brightfield image, an image that will contain the

spacings of the lattice planes that gave rise to those beams.

Ilustration 7 Different contrast mechanisms depending on the lens aperture

A diffracted beam with a certain spacing can be chosen with the objective aperture

instead of the transmitted beam and a dark field image is formed. By choosing electrons

scattered in a certain direction, a dark field image is obtained which contains interpretable

information about the amplitude of the sample. In general, the areas from which the

selected diffracted beam comes will appear bright in the image while the rest of the

unselected beams will form the dark background of the so-called dark field image.

In electron diffraction, there is a choice between the area selection method (SAED),

where the area to be diffracted is determined by an aperture (area section aperture), and

the microdiffraction and convergent beam electron diffraction (CBED) methods, where no

aperture is used but the beam is converged to a point on the sample.

 Ilustration 8. Mode of operation in the transmission electron microscope as a function of

the condenser aperture.

4.Scanning Transmission Electron Microscopy (STEM)

When operating a modern transmission microscope, we can do it in two ways; in

TEM mode, which has been discussed at length in this chapter.

in two ways; in TEM mode, which has been discussed extensively in this chapter,

and also, if the equipment allows it, in STEM (Scanning Transmission Electron

Microscopy) mode.

and, in addition, if the equipment allows it, in STEM mode (Scanning Transmission

Electron Microscopy). A scanning transmission electron microscope (STEM) is essentially

a scanning transmission electron microscope (TEM).

A scanning transmission electron microscope (STEM) is essentially a transmission

electron microscope to which is attached a system of deflector coils, which allow the

electron beam to be swept over the surface of the sample. The coils control that the

electron probe sweeps parallel to the optical axis at all times, i.e., that it does not change

direction as it sweeps.

Ilustration 9. Mode of operation in the transmission electron microscope as a function of

the condenser aperture.

 A TEM/STEM has the objective lens and the under-sample projector lenses, but in

STEM mode these are only used to change the camera length between the sample and

the detector plane and also to form the electron probe image. Therefore, most of the optics

act on the beam before it passes through the sample. Now no objective lens is needed to

focus the image, but the signal from the interaction of electrons with matter is collected by

a detector that transforms these pulses into an image, and therefore certain defects of this

lens, such as chromatic aberration, are no longer important. This will be an advantage

when analyzing samples with considerable thickness. In the same way as in SEM, it is

now the detectors that will capture scattered electrons to form the images. The difference

is that in STEM mode the beam passes through the sample.

In STEM mode we can obtain three types of images depending on the detector

used (Figure 13.13): Bright Field (BF) images, Dark Field images and HAADF or Z-

contrast images. BF images are formed by detectors that collect electrons diffracting close

to the optical axis, i.e. at low angles. On the other hand, DF images are formed by

electrons diffracted at high angles, which are collected by a ring-shaped detector (ADF).

Finally, the Z contrast images are formed by the electrons that are also collected by this

ring detector but with the electrons that are scattered at very high angle. The STEM BF

and STEM DF images are thus formed by the contribution of elastic and inelastic

scattering, but in the case of the Z-contrast images only incoherent, inelastic scattering is

used.

Ilustration 10. Image Formation in STEM

 HAADF images are formed with electrons that are scattered at very high angles.

This type of interaction is known as Rutherford scattering. The incident electron interacts

with the nuclei present in the sample and the result is scattered electrons with an atomic

number squared (Z2) dependence at very high angles. The detector that collects these

electrons is ring-shaped (ADF) and this same detector will be able to collect scattered

electrons at a wide variety of angles by simply varying the length of the chamber.

Ilustration 11 a) TEM BF image of the SBA15 catalyst with Co particles and b) STEM-

HAADF micrograph of the same area.

The disadvantages of STEM mode lie in its dynamic character. To form an image,

about 2,048 lines have to be scanned. The detectors analyze the signal they pick up and

convert it into images synchronously on a low-scan TV screen or PC, which means that

the process is slow and it takes a few seconds to capture a good image, where both

microscope and sample instabilities play a decisive role.

5.Energy dispersive x-ray analysis (EDX); electron energy loss spectroscopy

(EELS).

The electron microscope has the ability to concentrate a beam of high-energy

electrons onto a certain area of the sample in a controlled manner. These electrons can

yield part of their energy to the specimen, giving rise to a whole series of phenomena of

energy transitions in the material studied, which have given rise to a large number of

spectroscopic techniques.
 When the electron beam is focused on the sample it causes a series of electron

transitions between different energy levels. The excited ion relaxes to its initial state by the

transfer of an electron from an outer orbital to an inner shell, which results in the emission

of X-rays.

Ilustration 12. Schemes of electronic transitions that give rise to energy signals

To carry out the compositional analysis of the material it is necessary to adequately

separate the different maxima in order to carry out their quantification. This is done by the

detection system. This method is the most common and it is rare to find an electron

microscope that does not have an EDX detector attached, unless this microscope is

exclusively dedicated to high resolution. This method uses a semiconductor detector that

characterizes X-ray photons according to their incident energies by recording the entire

spectrum simultaneously. In TEM the thin film method is used in which it is assumed that

the area of the crystal where the analysis is performed is thin enough to be able to neglect

absorption and fluorescence.

In STEM mode we will distinguish between two types of EDX analysis: standard

analysis (point, line or area integral) and elemental maps. A standard analysis is

considered to be the spectrum obtained from the selected area, whether it is a point, a line

or a certain area. The point analysis consists of analyzing the area where the electron
probe is placed. The resolution depends on the size of the probe and allows analysis at

atomic resolution. On the other hand, line analysis consists of drawing a line on the image

in such a way that the probe sweeps the line and performs the analysis along it.

Ilustration 13. MoS EDX Line Analysis

These experimental conditions must satisfy an optimal compromise between high

X-ray excitation and high spatial resolution. This requires limiting parameters such as

image resolution, number of dots and counts per second, electron probe size, mapping

magnification, etc., to avoid contamination and displacement effects. Very high acquisition

times are therefore required to obtain a good signal-to-noise ratio. The problem of low

signal levels becomes more apparent with low atomic number elements.

As for EELS, it consists in the detection, thanks to a magnetic prism usually placed

after the phosphorescent screen, of the energy loss suffered by the electrons after

interacting with the sample. One of the major differences of EELS spectroscopy with

respect to EDX is the higher energy resolution that can be achieved, being around 1 eV or

even less, while the energy resolution of an EDX detector is around 135 eV. This high

energy resolution implies that by EELS spectroscopy one can see more subtle effects

caused on the different electron levels by the type of bond, the ionization state of the atom,

its environment, etc., which makes it difficult to quantify the analysis.

 Ilustration 14. EELS spectrum of an amorphous carbon aggregate showing the maxima

due to transitions at n* and a* levels

To perform this type of spectra, either a magnetic prism spectrometer or an omega

filter (so called because of its shape) is used, which is mainly used for EFTEM energy

filtering maps, which is another way of obtaining elemental maps similar to those obtained

with EDX mapping. The way in which the spectrum originates is due to electrons passing

through the spectrometer and being deflected by the surrounding magnetic field with an

angle greater than or equal to 90°. Electrons with higher energy are deflected farther away

than those with zero loss. A spectrum is thus formed in the scattering plane consisting of a

distribution of electron counts versus energy loss. Three regions can be considered in an

EELS spectrum:

a) Peak zero loss: consists of elastically backscattered electrons, but also contains

electrons that have undergone energy loss.

also contains electrons that have undergone very small energy losses. It is essential to

calibrate the spectrum.

b) Small loss region: collects electrons that suffer a loss of up to 50 eV.

up to 50 eV. It contains electrons that have interacted with the electrons of the outermost

layer and that are weakened. outermost layer and are weakly bonded. It provides

information on the electronic properties.

c) High-loss region: contains information on electrons that have interacted with electrons in

the innermost shells. They provide characteristic information about the atoms in the

sample.

6.Processing and simulation

Digital image processing is the experimental image manipulation aimed at

obtaining more information about the sample.

Image simulation is the artificial generation of images (simulated images) from ideal

structural models and computer calculations.

Image processing

It is necessary to filter the experimental images and eliminate possible noise to

obtain an improved image, with sharper contrasts and it is also essential to quantify the

information contained in an image. For all this it is necessary to have the digitized image

and a suitable program to process it, in other words, to convert it into numbers.

Digitized images are stored as formations of intensity values, usually in two

dimensions. The size (number of pixels) and depth (number of grays) of the image

depends on the way it is digitized. The usual way today is directly on the microscope

through a CCD (Slow-scanCharged Coupled Device) camera, which usually consists of

1024 x 1024 pixels and 16,000 gray levels. They can also be recorded on negatives, which

are either digitized in a densitometer or printed on positives and then recorded with a

scanner.

To convert the image into numbers, the Fourier transform is calculated using the

so-called Fast Fourier Transform (FFT). Thus, the operations that can be performed are

several and go through the obtaining of a Fourier Transform from the image that is the real

space. From this FFT the image can be "cleaned" by simply applying masks to the
reflections obtained and then reconstructing the image. It is also possible to vary the size

of the apertures (imitated in the computer) obtaining contrasts different from those

obtained experimentally in the microscope, being even possible to choose the beams of

transmitted electrons that form a reconstructed and real image.

Image simulation

The idea of simulating high resolution images comes from the fact that information

about the phases is lost when forming an experimental intensity map except under certain

conditions for very thin crystals at the Scherzer focus. In general, the image does not have

a direct correspondence between the dark spots in the image and the atomic positions, so

one assumes a structure, simulates the image and observes how it fits the experimental

image, so that by modifying the original proposed structure and repeating the process one

can arrive at a fine structure fit from the comparison of simulated and experimental

images. The main problem lies in the sensitivity of the image to various experimental

factors, such as the correct alignment of the beam with respect to the sample and the

optical axis, the thickness of the sample, the focus, the coherence of the beam, the

chromatic and spherical aberrations, etc. The best known image simulation programs are

SHRLI (by M. A. O'Keeffe, a pioneer in this field), NCEMSS (by R. Kilaas), Mactempas

(also by R. Kilaas), EMS (by P. Stadelman) and Cerius. The high-resolution image

simulations shown in this paper have been performed with the NCEMSS program.

7.Sample Preparation

This section is intended to summarize and emphasize the most commonly used

techniques in materials science. In any case, samples should be prepared according to the

information to be obtained from them, both in scanning and transmission.

SEM

One of the great advantages of scanning electron microscopy is the fact that many

species can be examined with little or no preparation. The thickness of the specimen is not

a consideration in this case, so the size is determined only by the size of the specimen

holder. To examine the topographic contrast it is sufficient to cut and polish the surface to

avoid contaminating the microscope, with as little damage as possible and using only the

minimum of cleaning necessary. Solvents such as acetone, ultrasound or mechanical

polishing are used for this purpose. The sample is then supported on a metal specimen

holder, usually with glue or conductive paint, and dried.

If it is an insulating material, it must be coated with a conductive agent. If

observations are to be carried out at very low voltage or in an environmental SEM, this

coating is not necessary; they can be coated with carbon or metals, the most commonly

used being gold. The metal gives a better and more uniform coating and prevents the

sample from loading more effectively than carbon does. When the time comes to perform

X-ray analysis, it is necessary that the signal of the conductive material is known and

above all, does not mask any of the signals of our sample, in this case, we will have to

resort to carbon coating.

Ilustration 15. scanning microscope

TEM

There are many ways to prepare samples for TEM, the method of choice depends

on the type of material and the information required.

The sample to be studied by TEM has to be mainly thin, thin enough to be

transparent to the electron beam, and representative of the material to be studied.

Generally the samples are separated into two groups: self-supported and

supported on a grid which is usually made of copper but can also be aluminum, nickel,

tungsten, etc. The type of sample to be prepared will depend mainly on the type of

material, thus, a uniform sample composed of a single material is prepared as self-

supported, while other types of species can be supported on grids that are normally 3 mm

in diameter and from 100 to 300 meshes.

Self-supported samples

For its preparation it is necessary a first stage of initial thinning and cutting and a

later thinning until reaching the transparency to the electron beam. The first stage of initial

thinning supposes the obtaining of sheets of between 100 and 200 ^m of thickness and of

these a disc of 3 mm of diameter is cut. It is the central region of this disk that is finally

thinned on one or both sides, to a thickness of only a few microns. final thinning of the disk

is usually carried out by chemical polishing, electrochemical, ion bombardment or

ultramicrotomy.

Samples supported on grids

The alternative to self-supported samples is to make small portions of sample or

particles transparent to electrons, and deposit them on a thin film supported on a metal

grid. The classic example is a copper grid covered with a film of amorphous carbon with

holes in it, so that, hopefully, some particles will be partially localized in the holes so that
they can be studied without interference. The usual way to deposit the particles on the

copper grid with the carbon film with holes in it is by suspension. The material is first

fractured in an agate mortar, then dispersed in a suitable inert solvent such as aceto na,

ethanol, butanol or n-hexane, with the aid of ultrasound if the agglomerated nature of the

sample requires it. After deposition of the larger particles, invisible particles will be found in

suspension, perfect for observation under the transmission electron microscope. One or

two drops of the suspension are deposited on the grid, the solvent is allowed to evaporate

and the grid is introduced into the microscope. This is the general method of preparation of

supported samples in materials science, however, there are other common methods such

as ultramicrotomy. This is most suitable for biological materials and also when dealing with

particles or fibers, which are too small to be individually thinned, and too large to be

transparent to the electron beam.

The procedure to be followed involves a first step of embedding the material or

particles in an epoxy resin or similar, followed by cutting them into thin flakes that fall onto

water or an inert medium from which they are collected on the grids.

8.Preliminary design of the experiment (Which microscope do I need and why?)

The electron microscope can provide us with chemical information through the

different spectroscopies, real space information from the images and reciprocal space

information through the electron diffraction diagrams.

In general, when studying an unknown sample, it is convenient to begin by studying

it by X-ray diffraction, which will give us global information of the sample. Subsequently, its

examination by SEM microscopy will allow us to discern the crystallinity of the sample, the

presence of various phases and to carry out a preliminary study of its composition by EDX

spectroscopy. With these data we would continue its study in an analytical microscope of

100-200 kV where we would proceed to study by SAED its reciprocal network, by EDX the
composition of each microcrystal and by medium resolution microscopy, both in brightfield

or darkfield mode, the presence of extensive defects, intergrowths, twinning, etc.

Ilustration 16. Spectroscope

Finally, when the problem requires it, the sample is taken to a high resolution

electron microscope already knowing perfectly the necessary orientation of the crystal and

having simulated the high resolution images for different thicknesses and defocus values.
 References

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