Electron Microscope
Dr. K. C. Barick
E-mail: [email protected]
Scale and microscopic techniques
Structure determines properties
Microscope
❖ Microscope came from Greek word (Mikron: small and
scopeos: to look)
❖ It is an instrument for viewing objects that are too small and
the science of investigating small objects using such an
instrument called microscopy.
Microscope
Simple microscope Compound microscope
Light microscope Electron microscope
Phase Fluorescent Dark field Bright Scanning Transmission Confocal
contrast field
Terms used in Microscope
❖ Magnification: Degree of enlargement, depends on focal length of
objective, magnifying power of eyepiece
❖ Resolution: Ability to reveal closely adjacent structural details as
separate and distinct.
❖ Limit of resolution (LR): Minimum distance between two visible
objects at which they can be separate and not contact with each other
❖ Numerical aperture: Ratio of diameter of lens to its focal length
❖ Working distance: Distance
between the front surface of the
lens and surface of object
Conventional (lens) microscopy
f f
u v
1 1 1
+ =
u v f
v
Magnification =
u
Image formation in conventional microscope
Optical microscopes: Instrumentation
❖ Illumination system: Radiation source and
condenser lens to focus beam on the
specimen.
❖ Sample stage: Situated
between the illumination
and imaging systems.
❖ Imaging system:
➢ Objective lens to focus beam after it passes
through sample and forms an intermediate
image and
➢ Projector lens which magnifies a portion of the
intermediate image to form final image.
❖ Image recording system:
➢ Converts the radiation into a permanent image.
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Optical miscroscope: Sample Preparation
❖ Polishing:
❖ To make flat sample finish as optical microscopes have
low depth of field
❖ Etching:
❖ To reveal the details
Chemical etching removes
Material from surfaces at
different rate, depending
on the orientations and
compositions of the phase
Brass stainless steel deformed Nb
Optical microscopy images
Bronze revealing a cast
dendritic structure
Beo grown on a substrate Al –Si alloy
Why electron microscopy?
❖ Electrons have much a shorter wavelength than visible light,
and this allows electron microscopes to produce higher-
resolution images than standard light microscopes.
❖ Electrons have the strongest
interaction with matter of any
radiation so many types of
information are available
❖ High intensity, high energy,
beams of electrons are relatively
easy to generate
❖ It is used in conjunction with a
variety of ancillary techniques
(e.g. thin sectioning, immuno-
labeling, negative staining) to
answer specific questions.
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for electrons is 0.002 - 0.1 nm depending on energy
Electron microscope: Types of electron
❖ When an electron beam strikes a sample, a large number of
electron and photon signals are generated:
High energy
electrons
Low energy
electrons
(~10-50 eV)
Scanning Electron Microscope
(SEM)
Image Source: JEOL USA, Inc.
Scanning electron microscope (SEM)
❖ A scanning electron microscope (SEM) is a
type of electron microscope that produces
images of a sample by scanning (raster
scan pattern) the surface with a focused
beam of electrons.
❖ Electrons interact with atoms in the sample,
producing various signals that contain
information about its surface .
❖ Historical background:
➢ First SEM built by Knoll in Germany and by
Zworykin in the USA (1940s)
➢ It became a practical reality in Prof. Sir Charles
Oatley’s lab at Cambridge in the 1950s
➢ The concept is now mature – but progress
continues unabated
www-g.eng.cam.ac.uk
Information from SEM
❖ Topography
➢ Surface feature of an object or how
its look, its texture; relation between
these features and materials
characteristics
❖ Morphology SEM image of BaTiO3 sample
➢ Shape and size of the particles in an object, relation between these
features and material characteristics
❖ Composition
➢ Elements and compounds that the object is made up and its
relative amount, relationship between composition and materials
characteristics
❖ Crystallographic information
➢ How the atoms are arranges in the object, relationship between
these arrangements and properties
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SEM: Important features
❖ Magnification range 15x to 200,000x
10 kV ——> 0.12 Å
❖ Resolution of 50 Å
❖ Excellent depth of focus 100 kV ——> 0.039 Å
❖ Relatively easy sample preparation
Specimen:
Conducting -
➢ little or no preparation
➢ attach to mounting stub for insertion into instrument
➢ may need to provide conductive path with Ag paint
Non-conducting -
➢ usually coat with very thin layer of conducting (Au, C, Cr)
❖ Sample stage can be tilted, Specimen size limited by size of sample
chamber
Optical microscope (OM) vs. SEM
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SEM: Schematic representation
1. Vacuum system: It is required when using an
electron beam as electrons quickly disperse or
scatter due to collisions with other molecules.
2. Electron beam generation system: It is found at
the top of the microscope column. This system
generates the "illuminating" beam of electrons
(primary electron beam).
3. Electron beam manipulation system: It
consists of electromagnetic lenses and coils
located in microscope column and control the size,
shape, and position of the electron beam on
specimen surface.
4. Beam specimen interaction system: It involves the interaction of the electron beam
with the specimen and the types of signals that can be detected.
5. Detection system: It consists of several different detectors, each sensitive to
different energy/particle emissions that occur on sample.
6. Signal processing system: It is an electronic system that processes the signal
generated by detection system and allows additional electronic manipulation of the
image.
7. Display and recording system: It allows visualization of an electronic signal and
permits recording of the results using photographic or magnetic media
SEM: Schematic representation
Specimen High energy
compositional contrast
Backscattered electrons (BSE)
Secondary electrons (SE)
Fluorescent X-rays
Low energy
topographic
Composition - EDS
contrast
❖ Brightness of regions in image
increases as atomic number
increases
❖ Less penetration gives more
backscattered electrons
Bone Research 7 (2019) 15
SEM: Image design
❖ Beam scans the specimen in a raster
manner
❖ Signal produced as beam spot is Amplify the signal and
placed at each position in turn is control the display beam
collected and used to control the
brightness of the display beam which Scan the
writes the image on screen object Scan the
❖ Pixel by pixel scanning and then
reconstruction of image with one Collector image
pixel at a time
Object
Image
Synchronous
rasters
3rd century Mosaic
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Electron beam source
❖ Electron gun generates electron beam – thermionic or field emission
❖ Thermionic sources rely on heat to generate electrons (filament kept in
Wehnelt which controls the number of electrons leaving the gun).
❖ Field emission sources (field emission gun or FEG) use a strong electrostatic
field to induce electron emission
❖ SEM requires high current density (electrons per time unit per area) and as
small electron spot as possible
❖ Electrons are primarily accelerated toward an anode that is adjustable from
200V to 30 kV.
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Electron gun (thermionic)
Tungsten wire LaB6
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780323461412000018
o Tip of a tungsten needle is made
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copy
Electron gun (FEG)
❖ In FEG, a strong electric field is used to
extract electrons)
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/search_result.html?keyword=Schottky%
20emission
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s/TEM.pdf
❖ A smaller spot and higher current densities compared to
thermionic guns
Electron gun (FEG)
❖ Thermionic emission is the process of releasing electrons from a metal that
has been heated to a high temperature (2700 K for W filament). The emitted
electrons are called thermions.
❖ Schottky-type electron gun emits sufficient electrons with the aid of a strong
electric field at a lower temperature (～1800 K) than the temperature that can
effectively emit thermoelectrons.
➢ Schottky FEGs works with the principle of Schottky effect, a
phenomenon where the potential barrier decreases when a strong
electric field is applied to a substance.
➢ This is done by coating the tungsten tip in zirconium oxide, which
facilitates the thermal emission of electrons when the source is heated
❖ Cold FEG is a type of electron source that emits electrons from a sharp
metal tip by applying a strong electric field, with the key feature being that
the emitter is kept at room temperature (not heated to facilitate electron
emission)
➢ Cold FEG is superior to Schottky FEG with respect to brightness, source
size, and energy spread, whereas the Schottky FEG can emit a higher
and stable current, which is highly advantageous for X-ray chemical
analysis and electron backscatter diffraction (EBSD).
Applied voltage and vacuum
❖ Effect of applied voltage:
➢ The accelerating voltage in
SEM varies from 200V-30 kV.
➢ Choosing right accelerating
voltage is critical
❖ Role of vacuum:
➢ Chemical (corrosion) and
thermal stability is necessary
for a well-functioning filament
➢ FEG :~ 10-10 Torr
➢ W/LaB6: ~ 10-6 Torr
➢ The signal electrons must travel from the sample to the detector
(chamber pressure)
➢ Vacuum requirements is dependant of the type of detector
SEM: Optics
Schematic cross-section of a magnetic lens and light optical analogue with a convex glass lens. Copper
wire (green) is coiled inside an iron cylinder (black). The rotationally symmetric but radially
inhomogeneous magnetic field (red) focusses the electron beam (blue).
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SEM: Optics
Detectors
❖ Backscattered electron detector - Solid state detector
➢ Electron energy up to 30-50 keV (high energy)
➢ Annular around incident beam to repel
secondary electrons with Biased mesh
➢ Detector sites above the sample, below the
objective lens. BSE which strikes it are
detected
Sample
➢ Images are more sensitive to chemical
composition (electron yield depends on
atomic number)
❖ Secondary electron detector – PMT detector
Detector
or Scintillator detector
➢ Bias mesh (100 – 300 V) needed in front of
detector to attract low energy electrons
Sample
Magnification in SEM
❖ Magnification is accomplished by scanning a progressively
smaller portion of the specimen and displaying the image on
the CRT. Thus total magnification is square area of CRT
divided by area scanned:
➢ Magnification is controlled by change
in the strength of scanning coil
Change in magnification at Change in working distance
constant working distance at constant magnification
(No image rotation) (image rotation)
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microscope-sem
SEM: Role of voltage
Images were taken at
different accelerating voltage
- resolution is improved with
increased voltage
Images were taken with
different accelerating voltages -
Surface structures appear
clearer at lower voltages
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SEM: Depth of field
❖ Depth of field (DOF) is distance along
the optical axis either side of the focus
point that remains in acceptable focus
❖ This is the height over which sample
can be clearly focused
❖ SEM has large DOF which produce the
images that appear 3d in nature
❖ DOF is improved by longer working
distance, smaller objective apertures
and lower magnification
Focal
plane
SEM image of pollen (Top: 8 mm WD
100µm aperture, Below: 37 mm WD,
Effect of working distance 100 µm aperture
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SEM: Spot size
❖ Spot size mainly restricts
the beam current and will
thereby cause for
brightness and contrast
compensations.
❖ Smaller spot sizes will
require higher brightness
and contrast levels, thus
there can be a limits when
using a small spot size.
❖ Typically, smaller spot
sizes allow for higher
resolution and a greater
depth of field.
Effects of spot size on image quality.
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SEM imaging
❖ An SEM image focused at high
magnification will still be in
focus at low magnification
❖ Charging effect has to
nullify by coating with C
or Au
10X
110X
200X
400X
4K
16K 45K
Courtesy: Dr. D. Srivastava, NFC
SEM images: SE vs. BSE vs. X-rays
Cracked structure on the surface of SEM image of a bacteria
the insect hair
BSE + SE BSE
SE BSE (Topography images)
high energy (compositional contrast)
Backscattered electrons Brightness of regions in image increases as
atomic number increases
Secondary electrons low energy (topographic contrast), high
resolution)
Fluorescent X-rays composition - EDS
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SEM imaging: EDS
❖ During the process of generation of SE,
vacancy is created in electronic state
❖ Outer shell electron fills the inner shell
vacancy producing an X-ray photon (X-ray
fluorescence)
❖ Each element has a fingerprint X-ray signal
❖ Energy dispersive spectroscope based on EDS spectrum of mineral crust of
energy of X-rays vent shrimp Rimicaris exoculata
(https://en.wikipedia.org).
❖ Determine amount of element present (~ 0.1 at %) by measuring X-ray
line intensity
❖ Full, point, line and area scan analysis, and elemental mapping
Mn doped ZnO Mn K Co doped ZnO Co K
SEM vs. ESEM
❖ Environmental SEM (ESEM) used to image challenging
samples such as:
➢ Insulating samples
➢ Vacuum-sensitive samples (i.e., biological samples)
➢ Irradiation-sensitive samples (i.e., thin organic films)
➢ “Wet” samples (oily, greasy)
❖ It is also used to image chemical and physical processes in-
situ such as:
➢ Mechanical stress-testing
➢ Oxidation of metals
➢ Hydration/dehydration (i.e., Watching paint dry)
Traditional SEM chamber pressure: ~ 10-6 Torr
ESEM: 0.08 – 30 Torr
Various gases can be used
Sample preparation
❖ All water must be removed from the sample
❖ All samples must also be trimmed to an appropriate size
❖ No preparation required for metallic samples
❖ Non-metallics cover the sample with a thin layer of a conductive
material
➢ The sputter coater is used to coat non-metallic
samples (bugs, plants, human hair, etc.) with a
thin layer of gold.
➢ This makes them conductive, and ready to be
viewed by the SEM.
❖ Powder samples should properly dispersed in
acetone/ethanol medium and then drop casted
on stub, and dried at room temperature.
Advantages and disadvantages
❖ Advantages:
➢ High resolution of images
➢ A superior level of magnification
➢ A large dept of field
➢ Uses electromagnets as an alternative to
lenses
➢ 3D data of sample can be measured
➢ Preparation of sample is relatively easy
❖ Disadvantages:
➢ Costly to purchase and maintain
➢ Sample have to be viewed in vacuum
➢ Non-conductive materials need coating
➢ Sensitive to vibration and external magnetic fields
➢ Sensitive to electrical supply
➢ They should be maintained with access to cooling systems
Take home message: SEM
❖ The scanning electron microscope is a versatile instrument
that can be used for many purposes and can be equipped with
various accessories
❖ An electron probe is scanned across the surface of the sample
and detectors interpret the signal as a function of time
❖ A resolution of 1–2 nm can be obtained when operated in a
high resolution setup
❖ The introduction of ESEM and the field emission gun have
simplified the imaging of challenging samples
❑ Topography
❑ Morphology
❑ Composition SEM
❑ Crystallographic information
❑ ESEM – Biological samples
