Electron Microscopes

Dwi Marta Nurjaya

Electron Microscopy
Electron microscope is a type of microscope that uses
electrons to illuminate a specimen and create an
enlarged image.
electron microscope uses electrostatic and
electromagnetic lenses in forming the image by
controlling the electron beam to focus it at a specific
plane relative to the specimen in a manner similar to
how a light microscope uses glass lenses to focus light
on or through a specimen to form an image.
Types of Electron Microscopes:

TEM (Transmission EM)

SEM (Scanning EM)
REM (Reflection EM)
STEM (Scanning Transmission EM)
STM (Scanning Tunneling M)

Comparison of TEM with Slide Projector

Slide projector

Image

Specimen

TEM

Light source

Optical ray path with source,

condenser (illumination system),
specimen, and objective lens
Sample is transmitted.
(shadow projection)
Differences to SEM:
Transmission and true
optical ray path

(Source: FEI brochure All you want to know ...

Comparison of TEM with Light Microscope (LM)

Optical ray path with illumination,
specimen, objective lens, first
magnified intermediate image,
further lenses (projector), image
Real optical image
Difference:

Source

Condenser
Specimen
Objective lens

TEM
electrons (200 kV ~ 0.7c*)

glass lenses

= 0.0025 nm (200 kV)

= 450 650 nm

* c = speed of light

1st image

visible light

electro-magnetic lenses

resolution: 0.24 nm (d >> )

LM

final image

100 nm (d ~ )
(Source: Philips brochure Elektronenmikroskopie, p. 6)

TEM Specimens
Size
: Maximum 3 mm in diameter
Thickness : Typically 100 500 nm or even less
(HRTEM)
Biology
Tissue sections on thin carbon/formvar film
Particles (viruses) frozen in thin ice layer

Materials
Classical way: Thin disk thinned to perforation, e.g. by
mechanical grinding/polishing and ion thinning, or electropolishing; electron transparent at edge of hole
Nowadays: FIB techniques
5

What is SEM
Scanning electron microscope (SEM) is an EM that
designed for direct studying of the surfaces of solid
objects.

Advantages of Using SEM over OM

OM
SEM

Magnification

Depth of Field

4x 1000x
10x 3000000x

15.5mm 0.19mm
4mm 0.4mm

Resolution
~ 0.2mm
1-10nm

The SEM has a large DOF (depth of field)

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.

Advantages of Using SEM over OM

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.)

Characteristics of a SEM
MAGNIFICATION:
For most SEM: 10-300,000x (or more).
Useful Magnification: ~20,000x

RESOLUTION:
Theoretical Limit of Resolution: 50 (5 nm)
Practical Resolution: 200 (20 nm)

The resolving power of the SEM depends primarily on the effective

beam diameter of the probe (spot size). For two points having a
inter-point distance, d, to be resolved, spot size must be smaller
than d.

Inside SEM

Inside SEM
The electron beam produced by
an electron gun is focused to a
point on the sample surface by
two condenser lenses.
The second condenser lens
(sometimes also called as
objective lens) focuses the beam
to an extraordinarily small
diameter of only 10-20 nm.
Electrons, either SE or BSE,
from the sample surface are
detected by a detector and
amplified to form images on the
screen of a CRT.

Comparison of LM, TEM and SEM

Comparison of OM,TEM and SEM

Microscope

Resolution

Magnificatio
n

Light
source

Specimen

Bare eyes

10m

Visible light

Bulk Material

Optical
Microscope

0.1-1m

5-2000x

Visible light

Polished

Scanning
electron m.

0.01m

100-600,000x

electrons

Bulk material

Transmission
electron m.

0.1nm

1,0001,000,000x

electrons

Thin foils

Before SEM there was TEM

TEM takes electrons from a source and

through condenser and objective
electromagnetic lenses, focuses the beam on
an area of the sample. If the sample is thin
enough for the electrons to travel through, the
projector lens will project an interference
pattern, (or the image) onto a phosphorus
screen below.
ADV: extremely high resolution.
LIMITATIONS:
because the electron beam has to travel
through the sample, sample preparation is
usually required to make the sample thin
enough.
since the beam is traveling though the sample,
the sample bulk and not the surface is being
imaged.

SEM vs TEM

G.Cambaz, G.Yushin, Y.Gogotsi, V.Lutsenko, Anisotropic Etching of SiC Whiskers, Nano Letters, 6, 3, p.548, 2006. (cover article)

Limitations of using electrons

electrons will not freely travel through air - there are
enough molecules in air to easily absorb an electron
beam. Therefore, the electron source, lenses, and
sample must all be under a vacuum
since electrons are electrically charged, the sample
needs to be conductive enough to dissipate this
charge.

Electron Gun

The electron beam comes from a filament,

made of various types of materials.
The most common is the Tungsten hairpin
gun. This filament is a loop of tungsten
which functions as the cathode.
A voltage is applied to the loop, causing it to
heat up.
The anode, which is positive with respect to
the filament, forms powerful attractive
forces for electrons. This causes electrons
to accelerate toward the anode.
Some accelerate right by the anode and on
down the column, to the sample.
Other examples of filaments are
Lanthanum Hexaboride filaments and field
emission guns.

Hairpin Electron Gun

The hairpin shape tungsten filament

functions as a cathode
a small voltage differential applied across
the terminals of the filament causes
heating of the wire
The flow of current through the wire is
called filament current
Electrons thermionically emitted from the
filament are accelerated rapidly towards
the anode

Material
Operation temp
Service life
Vacuum system

: Tungsten
: 2700 K to 3000 K
: 25 100 hours
: 10-5 Torr

Lanthanum Hexaboride - LaB6

A thermionic electron gun consists
essentially of a heated wire or
compound from which electrons are
given enough thermal energy to
overcome the work function of the
source, combined with an electric
potential to give the newly free
electrons a direction and velocity.
Material: Lanthanum Hexaboride (LaB6)
Operation temperature : 1700 - 1850 K
Service life
: 500 hours
Vacuum system
: 10-6 Torr

Field Emission Gun

Electron source is not heated to remove electrons and for this

reason are often referred to as being "cold" sources.
Electrons are then pulled from a very small area of the pointed
tip and proceed down the column. Often this is aided by a
second anode that lies beneath the first.
Acting like an electrostatic lens the two anodes serve to further
coalesce and demagnify the beam.
The lost electrons are replenished by an electron source
attached to the tungsten tip.
A primary electron beam generated by a field emission source
offers significant advantages over those produced by by
thermionic emitters.

Material
Operation temperature
Service life
Vacuum system

: Tungten (W)
: 300 K
: over 1000 h
: 10-9 Torr

Comparison of 3 common electron sources

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.
3. all electron source need a vacuum environment to operate
1 atmosphere = 760mm Hg = 760 torr = 1.013bar = 101.3 kPa

Vacuum Pump
There are four types of vacuum pumps that are at least somewhat
commonly employed in SEMs:
1.
2.
3.
4.

Roughing pump/mechanical pump

Diffusion pump
Turbo Molecular pump
Ion pump

Electron Sample Interaction

The interactions may be elastic or
inelastic
The elastic interactions:
incident electrons and nucleus
a large-angle deflection of incident
electrons.
little energy loss

The inelastic interactions:

incident electrons and orbital shell
electrons
a small-angle deflection of incident
electrons.
heavy energy loss

SEM Detector

Elastic Vs Inelastic

Elastic Vs Inelastic
Elastic
Back-scattered electrons

Inelastic

Secondary electrons
Bremsstrahlung X-rays
Characteristic X-rays
Auger electrons

Other
Cathodoluminescence
Specimen current

Electron Sample Interaction

SEM Electron - Photon

Secondary

Back scattered

Auger

Cathodoluminescence

Secondary Electron
Produced by inelastic interactions of high
energy electrons with valence (or
conduction) electrons of atoms in the
specimen, causing the ejection of the
electrons from the atoms. These ejected
electrons with energy less than 50eV are
termed "secondary electrons".
Each incident electron can produce
several secondary electrons.
Production of SE is very topography
related. Due to their low energy, only SE
that are very near the surface (<10nm)
can exit the sample and be examined
(small escape depth).

Secondary Electron Detector

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.

Secondary Electron Detector

A typical scintillator/PMT secondary-electron detector, often named an

Everhart-Thornley (ET) detector after its two inventors

A PMT works by converting the incoming photons into electrons which are
then drawn to dynodes kept at a positive bias. The dynodes are made of
material with a low work function and thus give up excess electrons for
every electron that strikes them. The result multiplies the signal contained
in each photon produced by the scintillator.

Backscattered electrons (BSE)

Formation
Caused when incident electrons collide with an atom in a specimen that
is nearly normal to the path of the incident beam.
Incident electron is scattered backward (reflected).

Use

Imaging and diffraction analysis in the SEM.

Production varies with atomic number (Z).
Higher Z elements appear brighter than lower Z elements.
Differentiate parts of specimen having different atomic number

Backscattered electrons are not as numerous as others. However, they

generally carry higher energies than other types of electrons

EBSD (Electron Back Scattered Detector)

The most common detector used is
called a surface barrier detector. It sits
above the sample, below the objective
lens. BSE which strike it are detected.
1.

2.

3.

When a BSE electron strikes the

detector, electrons in the material move
from valence to conduction band.
The electrons are now free to move in
the conduction band or drop back into
the valence band.
If a potential is applied, the e- and e+
can be separated, collected, and the
current measured. The strength of the
current is proportional to the number of
BSE that hit the detector.

QBSD (Quadrant Back Scattered Detector)

The most common design is a four quadrant solid state

detector that is positioned directly above the specimen

Characteristic X-Ray
When the incident beam bounces
through the sample creating
secondary electrons, it leaves
thousands of the sample atoms with
holes in the electron shells where
the secondary electrons used to be.
If these "holes" are in inner shells,
the atoms are not in a stable state.
To stabilize the atoms, electrons
from outer shells will drop into the
inner shells, however, because the
outer shells are at a higher energy
state, to do this the atom must lose
some energy in the form of X-ray

How EDX works ?

X-rays emitted from the sample atoms are characteristic in energy (wavelength) not only
the element of the parent atom, but also which shells lost electrons and which shells
replaced them.
Example: Iron
If innermost shell (the K shell) electron of an iron atom is replaced by an L shell electron, a 6400
eV K alpha X-ray is emitted from the sample
Or, if the innermost shell (the K shell) electron of an iron atom is replaced by an M shell
electron, a 7057 eV K beta X-ray is emitted from the sample
Or, if the L shell electron of an iron atom is replaced by an M shell electron, a 704 eV L alpha Xray is emitted from the sample
EDX Spectrum of Iron would have three peaks; An L alpha at 704 eV, a K alpha at 6400 eV, and
a K Beta at 7057 eV.

How to Quantify elements by EDX

Element
Al
Si
Ti
Cr
Mn
Fe
Ni
Nb
Mo

Atomic %
2.588
4.247
0.365
21.793
0.229
3.931
8.337
.261
.249

Weight %
1.203
2.056
0.301
19.529
0.216
3.783
59.013
5.221
8.677

Mapping the elements

By placing dots on the screen when an X-ray count of the particular element is
received, an image is formed that mimics the SEM image, except the contrast is
formed by the elemental X-ray emission.

combination of the
nickel, lead, and tin
maps.

Magnification
An image is obtained by taking the signal from the sample and
transferring it to a CRT screen. By decreasing the size of the
scanned area (from which we get the signal), magnification is
produced.

Magnification is determined by taking the ratio of the lengths of

the scans:
Mag. = L / I

Magnification

Resolution
Resolution is the ability to resolve two closely spaced points. While you
may have to be at a high magnification to see small features, resolution is
NOT the same as magnification.
One way to improve resolution is by reducing the size of the
electron beam that strikes the sample:
dmin = 1.29Cs1/4 3/4[7.92 (iT/Jc)x109 + 1]3/8
at low current: dmin = 1.29Cs1/4 3/4
Jc = current density of the source,
= electron wavelength
Cs = spherical aberration,
i = current,
T = temperature.

Resolution

Improving Resolution
1.
2.
3.
4.
5.
6.

reducing spot size by using different types of filament.

using optimum accelerating voltages.
changing the angle of tilt (i.e. emissive area hence number of SEs).
using high beam current
short working distance
using secondary electron (SE) image, as the optimum resolutions
are:
7. 5 nm for SE; 25 nm for BSE and 2 m for X-ray mapping.
8. long exposure times, leading to more electrons being detected.
9. reducing lens aberrations to minimum

Effect of Spot Size

A) using smallest spot,

B) using a slightly larger size of spot
C) using a large size of spot

Effects of Accelerating Voltage

Higher voltage
generates shorter
wavelength of
electrons better
resolution.
Higher voltage
causes an increase
of the volume of
electrons/specimen
interactions
worse resolution

Effects of Accelerating Voltage

3.0 KeV

20.0 KeV

More signal (brighter)

Effects of Accelerating Voltage

3.0 KeV

20.0 KeV

But (sometimes) reduced resolution

Depth of Field
Depth field of the SEM is the greatest among microscopes:
20 mm at 10X
5 m at 10,000X

Large depth of field is a great advantage for keeping in focus all parts of a
rough topography, but a compromise must be made between field depth
and resolution
Means to improve depth of field:
1. using large spot size.
2. using long working distance.
3. using small final aperture size.

It is noted here that the machine variables for larger depth of field would
cause a reduce in the resolution power of the SEM.

Depth of Field
Working distance and aperture size affect depth of field and resolution
Working distance is the distance between the surface of the specimen and the front
surface of the objective lens, which is around 5-25 mm for most SEMs.
Aperture size controls the semi-angle, .
The working distance between objective lens and specimen can also control the semiangle. Thus, smaller aperture and longer working distance larger depth of field

ds is the spot size.

can be expressed as aperture size A and working distance, WD

However,
The smaller aperture reduces the current of beam worse resolution.
The longer working distance increases lens spherical aberration worse resolution.

Depth of Field
The depth of field is greater at
long working distance than
that at short working

Depth of Field

Contrast
Contrast is the ratio of the change in signal between any two points on the
specimenand the average signal.
Topographic Contrast: -- mainly given by secondary electron signal,
which is sensitive to the surface structure of specimens. Secondary
electrons can escape from the surface of most materials at depths about 5
nm (50 ).
Compositional Contrast: -- given by the backscattered electron signal,
which change with the differences in atomic number, i.e., nuclear charge of
the atoms composing the specimen. The higher the atomic number, the
greater the backscattering. In smooth plane surfaces we are dealing
entirely with compositional contrast

Contrast
Major Influences on Contrast
the incidence angle of impinging electrons onto the
specimen.
the collecting angle from the specimen to the positively
charged collector.

Machine Variables for Contrast Improvement

Reducing working distance
Using smaller aperture
Using larger electron beam size

Topography Contras

Composition Contrast
Because of this dependence of on atomic number,
images produced using BSE show characteristic
atomic number contrast.
That is, features of high average Z appear brighter
than those of low average Z

Contrast Comparison

The topography of the

specimen will also affect the
amount of backscatter signal
and so backscatter imaging is
often carried out on flat
polished samples

More information about SEM Image

Go to :
http://www.emal.engin.umich.edu/courses/semlectures/iform1.html
http://micro.magnet.fsu.edu/primer/java/electronmicroscopy/magnify1/in
dex.html
http://education.denniskunkel.com

Specimen Preparation

Stub

Specimen Preparation
Sampling: Cutting or Replica formation
Caution: surface contamination with dust must be avoid during cutting.

Specimen mounting: using of a conductive cement, gluing the

specimen onto a aluminum stub.
Caution: contamination of sample surface or the SEM by out gassing of solvent in the adhesive.

Specimen surface treatment:

a. No surface treatment is needed if the surface is conductive or if the insulating layer of
the specimen is thin enough for some electrons to penetrate an underlying conductor.
For example: fracture surface of a metal or thin plastic coating on a metal surface.

Specimen Preparation
b.

c.
d.

A thin layer (~100 ) of conductive coating (gold or gold plus palladium or

carbon) is needed, if the interesting surface is an insulator. For example:
fracture surfaces of polymer and ceramics.
Surfaces of wet specimens are preserved by freeze-drying or use of a
cold stage. For example, fibers of wet paper or liquid sample.
Morphology of biological and polymeric materials may be observed with
specially prepared samples. For example: Generating a fast fracture
surfaces, using chemical or plasma etching, etc.

Preparation for biological samples

Fixation: to stabilize samples in life like condition and to protect the
sample from disruption, dimensional change and loss of materials during
subsequent processing.
Chemical fixation: primary fixing in aldehyde or mixture of aldehydes, the secondary
fixing in osmium tetroxide (OsO4).
Cryogenic fixation: ultra rapid freezing

Dehydration:
Solvent drying: in a graded series of solutions containing acetone and ethanol.
Critical-point drying: the sample is infiltrated with liquid CO2 in a chamber.
As the temperature increases, the pressure increases, finally driving all the liquid into the
gas phase, at which the point sample is dry and gas can be exhausted from chamber.

Freeze-substitution method: for ultra-rapid freezing samples.

Air drying
Freeze-drying

Sputtering Equipment

Sputtering a thin gold or carbon layer (~ 10 nm) on a specimen

surface can improve the conductivity of specimens. Also you
need ensure the conductive layer connected to the base of
specimen holder.