Transmission Electron

Microscopy

DR. HARSH MOHAN

DEPARTMENT OF PHYSICS
M.L.N. COLLEGE
YAMUNA NAGAR
(HARYANA)
 Transmission Electron
Microscopy (TEM): Principle,
Instrumentation and
Applications
 Microscopy
• main branches: optical, electron and scanning
probe microscopy.
• Optical and electron microscopy involves the
diffraction, reflection, or refraction of radiation
incident upon the subject of study, and the
subsequent collection of this scattered
radiation in order to build up an image.
• Scanning probe microscopy involves the
interaction of a scanning probe with the
surface or object of interest.
 Optical microscopy - definition

• Optical or light microscopy involves passing

visible light transmitted through or reflected from
the sample through a single or multiple lenses to
allow a magnified view of the sample.
• The resulting image can be detected directly by
the eye, imaged on a photographic plate or
captured digitally.
• The single lens with its attachments, or the system
of lenses and imaging equipment, along with the
appropriate lighting equipment, sample stage and
support, makes up the basic light microscope.
Optical microscopy - scheme
Optical microscopy - magnification
Resolution (not magnification!) is the ability to
separate two objects optically

Unresolved

Partially resolved

Resolved
 Optical microscopy - limitations
OM can only image dark or strongly
refracting objects effectively
.
Out of focus light from points outside the
focal plane reduces image clarity.
Compound optical microscopes are limited
in their ability to resolve fine details by the
properties of light and the refractive
materials used to manufacture lenses. A
lens magnifies by bending light..
Optical microscopes are restricted in their ability to
resolve features by a phenomenon called diffraction
which, based on the numerical aperture AN of the
optical system and the wavelengths of light used (λ),
sets a definite limit (d) to the optical resolution.
Assuming that optical aberrations are negligible, the
resolution (d) is given by:

In case of λ = 550 nm (green light), with air as medium,

the highest practical AN is 0.95, with oil, up to 1.5.
Due to diffraction, even the best optical microscope is
limited to a resolution of around 0.2 micrometres
 What is a Microscope?

• A tool that magnifies and improves resolution

of the components of a structure

• Has three components:

• sources of illumination,
• a magnifying system,
• detectors.
What is scale all about?

Scanning
Electron
Microscope
Remember that there are 1000 micrometers (µm) in 1 mm and
1000 nanometers (nm) in 1 µm.

The human eye can separate 0.2 mm at a normal viewing

distance of 25 cm

The light microscope can separate 0.2 µm (0.002mm)

depending on wavelength of light used

Electrons have a smaller wavelength than light therefore

provide the highest resolving power – about 2 nm
(0.000002mm)
 Electron Microscope vs. Optical Microscope
(first one built in 1931 by Ruska and Knoll) (Leeuwenhoek in 17 th century)

• Electron vs. Photon

Electron: charged, has rest mass, not visible

Photon: neutral, has no rest mass, visible at the

wavelength ~ 400 nm-760 nm.

Because of these differences, the microscope construction will also be different

What is the common property?

 The Compound Microscope
• Magnification
– Multiply magnifying power of the
objective lens X magnifying power of the
eyepiece lens
• Mechanical system
– Supports the microscope
• Optical system
– Illuminates object
– Passes light through a series of lenses
16
• Optical instrument
– Lens or combination of lenses
– Magnify, resolve fine details
• Earliest methods for examining physical
evidence
• Magnified image = virtual image
• Image viewed directly = real image.

17
Light Microscopes
 Sources of Illumination
• Light microscopes use a beam of light for
illumination and include fluorescence and
confocal microscopes

• Electron microscopes use electrons as a source

of illumination and include transmission and
scanning electron microscopes.
A short history
 A short history
• TEM constructed in 1931
• Von Ardenne first STEM in 1938 by rastering
the electron beam in a TEM
• Zworykin et al. 1942, first SEM for bulk
samples
• 1965 first commercial SEM by Cambridge
Scientific Instruments
Resolution at that time ~ 50 nm <-> Today < 1 nm

Morphology only at that time <-> Today analytical instrument

 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Electron Microscopy
• beams of electrons
are used to
produce images
• wavelength of
electron beam is
much shorter than
light, resulting in
much higher
resolution
29
 Types of Electron Microscope
• Transmission Electron Microscope (TEM) uses a
wide beam of electrons passing through a thin
sliced specimen to form an image. This
microscope is analogous to a standard upright or
inverted light microscope

• Scanning Electron Microscope (SEM) uses

focused beam of electrons scanning over the
surface of thick or thin specimens. Images are
produced one spot at a time in a grid-like raster
pattern.
 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

The Transmission Electron

Microscope
• electrons scatter when they pass
through thin sections of a specimen
• transmitted electrons (those that do
not scatter) are used to produce
image
• denser regions in specimen, scatter
more electrons and appear darker
31
 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

32
 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

EM

33
Transmission Electron Microscopy (TEM)

 Ultrathin sections of
specimens
 Light passes through
specimen, then an
electromagnetic lens,
to a screen or film
 Specimens may be
stained with heavy
metal salts

Copyright © 2010 Pearson Education, Inc. Figure 3.10a

Transmission Electron Microscopy (TEM)

 10,000–100,000×; resolution 2.5 nm

Copyright © 2010 Pearson Education, Inc. Figure 3.10a

 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Specimen Preparation
• analogous to procedures used for
light microscopy
• for transmission electron
microscopy, specimens must be cut
very thin
• specimens are chemically fixed and
stained with electron dense material

36
 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

Other preparation methods

• shadowing
– coating specimen with a thin film of a
heavy metal
• freeze-etching
– freeze specimen then fracture along
lines of greatest weakness (e.g.,
membranes)

37
 Electron Microscopy - definition and types
• developed in the 1930s that use electron beams instead of light.
• because of the much lower wavelength of the electron beam
than of light, resolution is far higher.

TYPES
• Transmission electron microscopy (TEM) is principally
quite similar to the compound light microscope, by sending an
electron beam through a very thin slice of the specimen. The
resolution limit (in 2005) is around 0.05 nanometer.

• Scanning electron microscopy (SEM) visualizes details on

the surfaces of cells and particles and gives a very nice 3D
view. The magnification is in the lower range than that of the
transmission electron microscope.
 Transmission Electron Microscopy (TEM)
• beam of electrons is transmitted through a specimen, then an
image is formed, magnified and directed to appear either on a
fluorescent screen or layer of photographic film or to be
detected by a sensor (e.g. charge-coupled device, CCD camera.
• involves a high voltage electron beam emitted by a cathode,
usually a tungsten filament and focused by electrostatic and
electromagnetic lenses.
• electron beam that has been transmitted through a specimen
that is in part transparent to electrons carries information
about the inner structure of the specimen in the electron beam
that reaches the imaging system of the microscope.
• spatial variation in this information (the "image") is then
magnified by a series of electromagnetic lenses until it is
recorded by hitting a fluorescent screen, photographic plate, or
CCD camera. The image detected by the CCD may be
displayed in real time on a monitor or computer.
 The resolution is proportional to the
wave length!

Electron equivalent wavelength and accelerating voltage

The dualism wave/particle is quantified by the

De Broglie equation:
λ = h/p = h/mv
λ : wavelength; h: Planck constant; p:
momentum
The energy of accelerate electrons is equal to their kinetic
energy:
E = eV = m0v2/2
V: acceleration voltage
e / m0 / v: charge / rest mass / velocity of the electron
These equations can be combined to calculate the wave
length of an electron with a certain energy:
p = m0v = (2m0eV)1/2
λ = h / (2m0eV)1/2 (≈ 1.22 / V1/2 nm)

At the acceleration voltages used in TEM, relativistic effects

have to be taken into account (e.g. E>100 keV)
λ = h / [2m0eV (1 + eV/2m0/c2)]1/2
Resolution
 Resolution
Can increase the resolution by:
 Light and Electron Microscopes
• Lenses are
used to
control a
beam of
illumination,
magnify, and
direct an
image to a
detector
 Comparison of OM,TEM and SEM
Source of
Light source electrons
Condenser
Magnetic
lenses
Specimen
Objective

Projector Specimen
Eyepiece CRT
Cathode Ray
Tube

detector

OM TEM SEM

Principal features of an optical microscope, a transmission electron microscope

and a scanning electron microscope, drawn to emphasize the similarities of
overall design.
TEM

 Transmission Electron
Microscope

 Illumination source is
beam of electrons from
tungsten wire
 Electromagnetic lenses
perform same function
as glass lenses in LM
 Higher resolution and
higher magnification of
thin specimens
 FEI Tecnai 20
For TEM, since the electrons
need to penetrate the specimen, it
must be very thin (< 100 nm)
 Comparison of TEM and LM

a. Similarities (Arrangement and function of components are similar)

1) Illumination system: produces required radiation and directs it onto the
specimen. Consists of a source, which emits the radiation, and a
condenser lens, which focuses the illuminating beam (allowing variations
of intensity to be made) on the specimen.
2) Specimen stage: situated between the illumination and imaging
systems.
3) Imaging system: Lenses which together produce the final magnified
image of the specimen. Consists of i) an objective lens which focuses the
beam after it passes through the specimen and forms an intermediate
image of the specimen and ii) the projector lens(es) which magnifies a
portion of the intermediate image to form the final image.
4) Image recording system: Converts the radiation into a permanent
image (typically on a photographic emulsion) that can be viewed.
 Comparison of TEM and LM
b. Differences
1) Optical lenses are generally made of glass with fixed focal lengths whereas magnetic lenses are
constructed with ferromagnetic materials and windings of copper wire producing a focal length which
can be changed by varying the current through the coil.
2) Magnification in the LM is generally changed by switching between different power objective lenses
mounted on a rotating turret above the specimen. It can also be changed if oculars (eyepieces) of
different power are used. In the TEM the magnification (focal length) of the objective remains fixed
while the focal length of the projector lens is changed to vary magnification.
3) The LM has a small depth of field, thus different focal levels can be seen in the specimen. The large
(relative) depth of field in the TEM means that the entire (thin) specimen is in focus simultaneously.
4) Mechanisms of image formation vary (phase and amplitude contrast).
5) TEMs are generally constructed with the radiation source at the top of the instrument: the source is
generally situated at the bottom of LMs.
6) TEM is operated at high vacuum (since the mean free path of electrons in air is very small) so most
specimens (biological) must be dehydrated (i.e. dead !!).
7) TEM specimens (biological) are rapidly damaged by the electron beam.
8) TEMs can achieve higher magnification and better resolution than LMs.
9) Price tag!!! (100x more than LM)
 Comparing light and electron microscopes
Light microscope Electron microscope

Magnification x1500 X500,000

Resolution 250nm 0.25nm

Type of radiation used Light Electrons

Focussed by Glass lenses Electromagnets

Type of material that can Living/moving/dead/abiotic Dead/abiotic

be viewed

Size Small and portable Large and static

Preparation and cost of Cheap and easy Difficult and expensive

material

© OCR 2016
•All rays from a point in the object are gathered by the lens and
converge to a point in the image.
•All parallel rays are focused in the focal plane.
•The back focal plane of the objective lens contains groupings of
rays that have left the object at the same angle.
•The back focal plane contains the diffraction pattern of the sample.
•Diffraction pattern and image are both formed in the imaging
process
•The intermediate lens is then focused on either the image plane
(for the image), or the back focal plane (for the diffraction pattern).
 Dark Field – Advantages and Disadvantages
•Advantages and disadvantages of Dark Field Imaging :-
•Advantages
•Provides high contrast for examining molecules with
very low contrast such as DNA.
•For crystalline objects, specific diffraction spots can be
selected in the back focal plane of the objective lens in
order to form a dark field image only from the electrons
scattered by a chosen set of crystal planes.
•Disadvantages
•More difficult to focus and correct for astigmatism since
phase contrast is not present.
•Image brightness is low, since the objective aperture
transmits only a small fraction of the scattered beam,,
•Longer exposure times needed to get good
photographic images.
•Consequently, specimens are subjected to greater
levels of radiation damage.
 Imaging Modes

•Two principle modes of TEM operation, A – Projecting the diffraction pattern,

B – Projecting the image.
• The intermediate lens selects either the Back Focal Plane or the image plane of
the objective lens.
 TEM imaging modes

Bright field/dark field depends on the aperture position. Modern way for
diffraction tilts the beam instead of moving the aperture
 Magnification in TEM

Mob × Mint × Mproj = Total Mag

Depends on the magnification, some lens may not be used

Electron-specimen interaction
Electron Beam and Specimen Interactions
Sources of Image Information
Electron/Specimen Interactions

(1-50KeV)

Electron Beam Induced Current (EBIC)

Distribution
Distributionand
andsize
sizeof
of
micropores
micropores
lattice
latticedistortion
distortion

Polymer
Polymercrystalline
crystalline
structure
structureand
and
morphology
morphology
Application

Polymer
Polymercomposition
composition Distribution
Distributionof
ofdispersed
dispersed
phase
phase
 Examples

Graphene
Carbon
nanotube

PE/SWNT NHSK
Transmission Electron Microscopy
(TEM)
Neuron growing on astroglia Black Ant

House Fly
House Fly

Human stem
cells

Human red blood cells

Neurons
CNS
Transmission Electron Microscopy
(TEM)
View inside cell via sections
magnification 120,000 X

50,000X
Conventional TEM Micrographs
Bacteria in cell
Apoptosis

Skin

Collagen Virus in cell

Chloroplast
Scanning Electron Microscopy (SEM)
 Scanning Electron Microscopy (SEM)

Scanning electron microscopy is used for inspecting topographies of specimens at

very high magnifications using a piece of equipment called the scanning electron
microscope. SEM magnifications can go to more than 300,000 X but most
semiconductor manufacturing applications require magnifications of less than 3,000
X only. SEM inspection is often used in the analysis of die/package cracks and
fracture surfaces, bond failures, and physical defects on the die or package surface.
During SEM inspection, a beam of electrons is focused on a spot volume of the
specimen, resulting in the transfer of energy to the spot. These bombarding
electrons, also referred to as primary electrons, dislodge electrons from the
specimen itself. The dislodged electrons, also known as secondary electrons, are
attracted and collected by a positively biased grid or detector, and then translated
into a signal.

To produce the SEM image, the electron beam is swept across the area being
inspected, producing many such signals. These signals are then amplified, analyzed,
and translated into images of the topography being inspected. Finally, the image is
shown on a CRT.
 Scanning Electron Microscopy (SEM)

• The energy of the primary electrons determines the quantity of

secondary electrons collected during inspection. The emission of
secondary electrons from the specimen increases as the energy of
the primary electron beam increases, until a certain limit is reached.
Beyond this limit, the collected secondary electrons diminish as the
energy of the primary beam is increased, because the primary beam
is already activating electrons deep below the surface of the
specimen. Electrons coming from such depths usually recombine
before reaching the surface for emission.
•
• Aside from secondary electrons, the primary electron beam results
in the emission of backscattered (or reflected) electrons from the
specimen. Backscattered electrons possess more energy than
secondary electrons, and have a definite direction. As such, they
can not be collected by a secondary electron detector, unless the
detector is directly in their path of travel. All emissions above 50 eV
are considered to be backscattered electrons.
 Scanning Electron Microscopy (SEM)

• Backscattered electron imaging is useful in distinguishing one

material from another, since the yield of the collected backscattered
electrons increases monotonically with the specimen's atomic
number. Backscatter imaging can distinguish elements with atomic
number differences of at least 3, i.e., materials with atomic number
differences of at least 3 would appear with good contrast on the
image. For example, inspecting the remaining Au on an Al bond pad
after its Au ball bond has lifted off would be easier using backscatter
imaging, since the Au islets would stand out from the Al background.
•
• A SEM may be equipped with an EDX analysis system to enable it
to perform compositional analysis on specimens. EDX analysis is
useful in identifying materials and contaminants, as well as
estimating their relative concentrations on the surface of the
specimen.
 Scanning Electron Microscopy (SEM)

• type of electron microscope capable of producing high-

resolution images of a sample surface.
• due to the manner in which the image is created, SEM
images have a characteristic 3D appearance and are
useful for judging the surface structure of the sample.

Resolution
• depends on the size of the electron spot, which in turn
depends on the magnetic electron-optical system which
produces the scanning beam.
• is not high enough to image individual atoms, as is
possible in the TEM … so that, it is 1-20 nm
 Advantages of Using SEM over OM

Magnification Depth of Field Resolution

OM 4x – 1000x 15.5mm – 0.19mm ~ 0.2mm
SEM 10x – 3000000x 4mm – 0.4mm 1-10nm

The SEM has a large depth of field, which allows a large amount of the
sample to be in focus at one time and produces an image that is a good
representation of the three-dimensional sample. The SEM also produces
images of high resolution, which means that closely features can be
examined at a high magnification.

The combination of higher magnification, larger depth of field, greater

resolution and compositional and crystallographic information makes the
SEM one of the most heavily used instruments in research areas and
industries, especially in semiconductor industry.
 Scanning Electron Microscope
– a Totally Different Imaging Concept

• Instead of using the full-field image, a point-to-

point measurement strategy is used.
• High energy electron beam is used to excite the
specimen and the signals are collected and analyzed
so that an image can be constructed.
• The signals carry topological, chemical and
crystallographic information, respectively, of the
samples surface.
Principles of SEM

Magnification?
Resolution?
 Image Formation in SEM
M= C/x

-
A 10cm
e
beam
Detector

10cm
Amplifier

Beam is scanned over specimen in a raster pattern in synchronization with

beam in CRT.
Intensity at A on CRT is proportional to signal detected from A on specimen
and signal is modulated by amplifier.
 The Scanning Electron Microscope
• (SEM) bombards a specimen with a beam of
electrons instead of light
• Produces a highly magnified image from 100x to
100,0000
• Depth of focus 300X better than optical systems at
similar magnification
• Bombardment of the specimen’s surface with
electrons
– Produces x-ray emissions
– Characterize elements present in the material under
investigation

92
 Scanning Electron Microscopy (SEM)
• An electron gun produces a beam of electrons that
scans the surface of a whole specimen.
• Secondary electrons emitted from the specimen
produce the image.

Figure 3.9b
 Electron beam
produced here
Beam passes down the
microscope column
Cross section of
electromagnetic Electron beam now tends to
lenses diverge

But is converged by
electromagnetic lenses
Sample

Diagram of Scanning Electron Microscope or SEM

in cross section - the electrons are in green
 Scanning Electron Microscope

96
SEM components
 What is SEM
Column
TV Screens
Sample
Chamber
The SEM is designed
for direct studying of
the surfaces of solid
objects

Cost: $0.8-2.4M

Scanning electron microscope (SEM) is a microscope that uses electrons

rather than light to form an image. There are many advantages to using the
SEM instead of a OM.
A Look Inside the Column
Column
 Electron Gun

A more e- beam

detailed
look α
inside

Source: L. Reimer,
“Scanning Electron
Microscope”, 2nd Ed.,
Springer-Verlag, 1998, p.2
Cathode Ray Tube (CRT) accelerates electrons towards the
phosphor coated screen where they produce flashes of light
upon hitting the phosphor. Deflection coils create a scan
pattern forming an image in a point by point manner
Color CRT?
 Image Magnification

Example of a series of increasing magnification (spherical lead particles imaged

in SE mode)
How an Electron Beam is Produced?

• Electron guns are used to produce a

fine, controlled beam of electrons
which are then focused at the
specimen surface.
• The electron guns may either be
thermionic gun or field-emission gun
Electron beam Source

W or LaB6 Filament
Thermionic or Field Emission Gun
Thermionic Emission Gun
• A tungsten filament
heated by DC to
approximately 2700K or
LaB6 rod heated to around
2000K
• A vacuum of 10-3 Pa (10-4 -
Pa for LaB6) is needed to
prevent oxidation of the
filament +
• Electrons “boil off” from
the tip of the filament
• Electrons are accelerated
by an acceleration voltage
of 1-50kV
 Source of Electrons
Thermionic Gun E: >10MV/cm
T: ~1500oC
W

Filament
(5-50µm)

(5nm)

W and LaB6 Cold- and thermal FEG

Electron Gun Properties

Source Brightness Stability(%) Size Energy spread Vacuum
W 3X105 ~1 50µm 3.0(eV) 10-5 (τ )
LaB6 3x106 ~2 5µm 1.5 10-6
C-FEG 109 ~5 5nm 0.3 10-10
T-FEG 109 <1 20nm 0.7 10-9
Brightness – beam current density per unit solid angle
 Electron Gun

W hairpin FEG
LaB6 crystal
 Thermionic Sources

Increasing the filament current will increase the beam current but
only to the point of saturation at which point an increase in the
filament current will only shorten the life of the emitter
Beam spot image at different stage of heating
 Magnetic Lenses
• Condenser lens – focusing
determines the beam current
which impinges on the sample.
• Objective lens – final probe
forming
determines the final spot size of
the electron beam, i.e., the
resolution of a SEM.
 Electromagnetic Lenses

An electromagnetic lens is essentially soft iron core wrapped in

wire

As we increase the current in the wire we increase the strength

of the magnetic field

Recall the right hand rule electron will move in a helical path
spiralling towards the centre of the magnetic field
Electromagnetic lens
 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.
 The Condenser Lens
• For a thermionic gun, the diameter of
the first cross-over point ~20-50µm
• If we want to focus the beam to a size
< 10 nm on the specimen surface, the
magnification should be ~1/5000, which
is not easily attained with one lens (say,
the objective lens) only.
• Therefore, condenser lenses are added
to demagnify the cross-over points.
The objective lens
 The objective lens aperture

Aperture in SEM: either to limit the amount of electrons or enhance contrast

 How Is Electron Beam Focused?
A magnetic lens is a solenoid designed to produce
a specific magnetic flux distribution.
Magnetic lens
(Beam diameter) (solenoid)

F = -e(v x B) p

q
Lens formula: 1/f = 1/p + 1/q

Demagnification: M = q/p

f ∝ Bo 2

f can be adjusted by changing Bo, i.e., changing the current through

coil.
The Condenser
Lens

Demagnification:
M = f/L
 Condenser-lens system

C1 controls the spot size

The condenser aperture must be centered
C2 changes the convergence of the beam
 The Objective Lens
• The objective lens
controls the final
focus of the electron
beam by changing the
magnetic field strength
• The cross-over image is
finally demagnified to
an ~10nm beam spot
which carries a beam
current of
approximately 10-9-10-
10-12 A.
 The Objective Lens - Focusing
• By changing the Objective
current in the lens

objective lens, the

magnetic field
strength changes
and therefore the
focal length of
the objective lens
is changed.
Out of focus in focus out of focus
lens current lens current lens current
too strong optimized too weak
Depth of Field
Detector and sample stage
Electron Detectors and Sample Stage

Objective
lens
Sample stage
 Topographical Contrast
Everhart-Thornley
SE Detector

Bright lens polepiece

e-
SE Scintillator
light pipe PMT
Dark
sample Quartz
Faraday window
cage +200V +10kV
Photomultiplier
tube

Topographic contrast arises because SE generation depend on the

angle of incidence between the beam and sample.
Electron beam – Specimen Interaction. Note the two types
of electrons produced.
Electrons from the focused beam interact with the sample
to produce a spray of electrons up from the sample. These
come in two types – either secondary electrons or
backscattered electrons.

As the beam travels across (scans across) the sample the

spray of electrons is then collected little by little and forms
the image of our sample on a computer screen.

We can look more closely at these two types of electrons

because we use them for different purposes.
A new electron is knocked An incoming electron rebounds
out (as a secondary back out (as a backscattered
electron) - electron) -

+ +

Energy of electron from beam is

lost to atom

Inelastic scattering Elastic scattering

• Secondary Electrons:
Source
Caused by an incident electron passing "near" an atom in the specimen, near
enough to impart some of its energy to a lower energy electron (usually in the K-
shell). This causes a slight energy loss and path change in the incident electron and
the ionization of the electron in the specimen atom. This ionized electron then
leaves the atom with a very small kinetic energy (5eV) and is then termed a
"secondary electron". Each incident electron can produce several secondary
electrons.
Utilization
Production of secondary electrons is very topography related. Due to their low
energy, 5eV, only secondaries that are very near the surface (< 10 nm) can exit the
sample and be examined. Any changes in topography in the sample that are larger
than this sampling depth will change the yield of secondaries due to collection
efficiencies. Collection of these electrons is aided by using a "collector" in
conjunction with the secondary electron detector. The collector is a grid or mesh
with a +100V potential applied to it which is placed in front of the detector,
attracting the negatively charged secondary electrons to it which then pass
through the grid-holes and into the detector to be counted.
A conventional secondary electron detector is positioned off to the
side of the specimen. A faraday cage (kept at a positive bias) draws
in the low energy secondary electrons. The electrons are then
accelerated towards a scintillator which is kept at a very high bias
in order to accelerate them into the phosphor.
The position of the secondary electron detector also affects
signal collection and shadow. An in-lens detector within the
column is more efficient at collecting secondary electrons that
are generated close to the final lens (i.e. short working distance).
 Secondary Electron Detector

Side Mounted In-Lens

What are the differences between these two images?
• Backscattered Electrons:
Formation
Caused by an incident electron colliding with an atom in the
specimen which is nearly normal to the incident's path. The
incident electron is then scattered "backward" 180 degrees.
Utilization
The production of backscattered electrons varies directly with
the specimen's atomic number. This differing production
rates causes higher atomic number elements to appear
brighter than lower atomic number elements. This interaction
is utilized to differentiate parts of the specimen that have
different average atomic number.
 Backscatter Detector

The most common design is a four quadrant solid state detector that is positioned
directly above the specimen
Example of an image using a scanning electron microscope and
secondary electrons

Here the contrast of these grains is all quite similar.

We get a three-dimensional image of the surfaces.
 Example of an image using a scanning electron microscope and
backscattered electrons

Grain containing
of silica so it is
darker

Grain containing
Here the differing contrast of the
titanium so it is
grains tells us about composition
whiter
So how does this work – telling composition from
backscattered electrons?

The higher the atomic number of the atoms the more

backscattered electrons are ‘bounced back’ out

This makes the image brighter for the larger atoms

Titanium – Atomic Silica – Atomic Number

Number 22 14
Understanding compositional analysis using X-rays and the
scanning electron microscope
-
If the yellow electron falls
back again to the inner
ring, that is to a lower
energy state or valence,
+ then a burst of X-ray
energy is given off that
equals this loss.

This is a characteristic
packet of energy and can
tell us what element we
Inelastic scattering are dealing with
 Backscattered Electrons (BSE)
Primary

BSE image from flat surface of an Al

(Z=13) and Cu (Z=29) alloy
BSE are produced by elastic interactions of beam electrons with nuclei of
atoms in the specimen and they have high energy and large escape depth.
BSE yield: η=nBS/nB ~ function of atomic number, Z
BSE images show characteristics of atomic number contrast, i.e., high
average Z appear brighter than those of low average Z. η increases with tilt.
Effect of Atomic Number, Z, on
BSE and SE Yield
 Interaction Volume: I
e-

Monte Carlo simulations of 100 electron trajectories

The incident electrons do not go along a

straight line in the specimen, but a zig-zag
path instead.
 Interaction Volume: II

The penetration or,

more precisely, the
interaction volume
depends on the
acceleration voltage
(energy of electron)
and the atomic
number of the
specimen.
Escape Volume of Various Signals
• The incident electrons interact with specimen
atoms along their path in the specimen and
generate various signals.
• Owing to the difference in energy of these
signals, their ‘penetration depths’ are
different
• Therefore different signal observable on the
specimen surface comes from different parts
of the interaction volume
• The volume responsible for the respective
signal is called the escape volume of that
signal.
 Escape Volumes of Various Signals

If the diameter of primary

electron beam is ~5nm
- Dimensions of escape
zone of
•Secondary electron:
diameter~10nm; depth~10nm
•Backscattered electron:
diameter~1µm; depth~1µm
•X-ray: from the whole
interaction volume, i.e., ~5µm
in diameter and depth
 Electron Interaction Volume

Pear shape

5µm

a b

a.Schematic illustration of electron beam interaction in Ni

b.Electron interaction volume in polymethylmethacrylate
(plastic-a low Z matrix) is indirectly revealed by etching
 Magnification
e -

x Low M High M
Large x small x
40µm 7µm

1.2µm 15000x
2500x

The magnification is simply the ratio of the length of the scan C on the
Cathode Ray Tube (CRT) to the length of the scan x on the specimen. For a
CRT screen that is 10 cm square:
M= C/x = 10cm/x
Increasing M is achieved by decreasing x.
M x M x
100 1 mm 10000 10 µm
1000 100 µm 100000 1 µm
 Resolution Limitations
Ultimate resolution obtainable in an SEM image can be
limited by:

1. Electron Optical limitations

Diffraction: dd=1.22λ/α
for a 20-keV beam, λ =0.0087nm and α=5x10-3 dd=2.1nm
Chromatic and spherical aberrations: dmin=1.29λ3/4 Cs1/4
A SEM fitted with an FEG has an achievable resolution of ~1.0nm at 30 kV
due to smaller Cs (~20mm) and λ.
2. Specimen Contrast Limitations
Contrast dmin
1.0 2.3nm
0.5 4.6nm
0.1 23nm
0.01 230nm
3. Sampling Volume Limitations (Escape volume)
 How Fine Can We See with SEM?

• If we can scan an area with width 10 nm

(10,000,000×) we may actually see
atoms!! But, can we?
• Image on the CRT consists of spots called
pixels (e.g. your PC screen displays
1024×768 pixels of ~0.25mm pitch)
which are the basic units in the image.
• You cannot have details finer than
one pixel!
 Resolution of Images: I
• Assume that there the screen can display 1000
pixels/(raster line), then you can imagine that
there are 1000 pixels on each raster line on the
specimen.
• The resolution is the pixel diameter on
specimen surface.

P=D/Mag = 100um/Mag
Mag P(µm) Mag P(nm)
P-pixel diameter on specimen surface 10x 10 10kx 10
D-pixel diameter on CRT, Mag-magnification 1kx 0.1 100kx 1
 Resolution of Images: II
• The optimum condition for imaging is when
the escape volume of the signal concerned
equals to the pixel size.
 Resolution of Images: III
• Signal will be weak if escape volume,
which depends on beam size, is smaller
than pixel size, but the resolution is still
achieved. (Image is ‘noisy’)
 Resolution of Images: IV
• Signal from different pixel will overlap
if escape volume is larger than the
pixel size. The image will appeared
out of focus (Resolution decreased)
 Resolution of Images: V
In extremely good SEM, resolution can be a few nm. The
limit is set by the electron probe size, which in turn depends
on the quality of the objective lens and electron gun.
Pixel diameter on Specimen
Magnification µm nm
10 10 10000
100 1 1000
1000 0.1 100
10000 0.01 10
100000 0.001 1
 Depth of Field

Depth of Field

4x105W
D= (µm)
AM

To increase D
Decrease aperture size, A
Decrease magnification, M
Increase working distance, W (mm)
 SE Images
Image Contrast
Image contrast, C
is defined by

SA-SB ∆S

C= ________ = ____
SA SA

SA, SB Represent signals

generated from two
points, e.g., A and B, in
the scanned area.
In order to detect objects of small size and low contrast in an SEM it is
necessary to use a high beam current and a slow scan speed (i.e., improve
signal to noise ratio).
SE-topographic and BSE-atomic number contrast
 Scanning Electron Microscope

162
The Scanning Electron Microscope is analogous to the
stereo binocular light microscope because it looks at
surfaces rather than through the specimen.
 Main Applications
• Topography
The surface features of an object and its texture
(hardness, reflectivity… etc.)
• Morphology
The shape and size of the particles making up the
object (strength, defects in IC and chips...etc.)
• Composition
The elements and compounds that the object is
composed of and the relative amounts of them
(melting point, reactivity, hardness...etc.)
• Crystallographic Information
How the grains are arranged in the object
(conductivity, electrical properties, strength...etc.)
 SE Images - Topographic Contrast

1µm

Defect in a semiconductor device Molybdenum

The debris shown here is an oxide fiber trioxide crystals
got stuck at a semiconductor device
detected by SEM
BSE Image – Atomic Number Contrast

2µm

BSE atomic number contrast image showing a niobium-rich

intermetallic phase (bright contrast) dispersed in an alumina matrix
(dark contrast).

Z (Nb) = 41, Z (Al) = 13 and Z(O) = 8

Alumina-Al2O3
 Can you see the
difference?

TEM LIGH SEM

T
We learn more from mistakes than successes…