NANO MATERIAL CHARACTERISATION

TECHNIQUES

Mathews K. Manayani
Assistant Professor
Nirmala College, Muvattupuzha
 Characterization of materials

✓Important methods for the characterization of materials –

✓Optical Microscopy
✓Scanning Electron Microscopy (SEM)
✓Transmission Electron Microscopy (TEM)
✓Scanning Transmission Electron Microscopy (STEM)
✓Environmental Transmission Electron Microscopy (ETEM)
✓Scanning Probe Microscopy (SPM)
 Characterization of materials
Microscopy is any technique for producing visible images of
structures or details of too small objects using a microscope or
other magnification tool

Microscopic Technique
1. Optical microscopy
❖ Light Microscopy
❖ Fluorescence Microscopy/ Confocal Microscopy
2. Electron microscopy
❖ Transmission electron Microscopy (TEM)
❖ Scanning Electron Microscopy (SEM)

3. Scanning Probe Microscopy

❖ Atomic Force Microscopy (AFM)

❖ Scanning Tunnelling Microscopy (STM)
 Milestones in Microscopy
1000AD – The first vision aid was invented (inventor
unknown) called a reading stone. It was a glass sphere that
magnified when laid on top of reading materials.

1674 – Anton van Leeuwenhoek built a simple microscope with

only one lens to examine blood, yeast, insects and many
other tiny objects

1931 – Ernst Ruska co-invented the electron microscope for

which he won the Nobel Prize in Physics in 1986

1981 – Gerd Binnig and Heinrich Rohrer invented the

scanning tunnelling microscope

1986 -Quate and Gerber invented the AFM

 Introduction
Optical and electron microscopy involve the diffraction, reflection, or

refraction of electromagnetic radiation interacting with the subject of

study, and the subsequent collection of this scattered radiation in order to

build up an image.

Fundamentals of Optics
➢ Properties of Light

➢ Theory of Image Formation

➢ Resolution

➢ Aberrations in Lenses
 Fundamental Requirement

✓ In order to perceive or image a structural feature it is

necessary that the wavelength of the probing radiation
should be similar in size to that of the feature.

✓ In other words, resolution is a function of wavelength

Electromagnetic Spectrum
 Properties of Light

✓ Electromagnetic Radiation
✓ Dual nature- Particle and Wave properties
✓ Relation with energy, frequency and wavelength
✓ Photons (can be measured quantitatively using CCD/other devices)
✓ Propagation can be depicted graphically
 Properties of Light

Mono chromatic Non chromatic

Linearly Polarised Non Polarised

Coherent Non Coherent

Collimated Divergent
 Theory of Image Formation

Optical instruments like microscopes, telescopes and binoculars use

optical elements to produce an image of an object. The two most
common elements for imaging objects are the converging lens and
the concave mirror

Common Optical Instruments

Theory of Image Formation
 Theory of Image Formation

Rules of ray tracing from a simple lens

Theory of Image Formation
 Theory of Image Formation
Focal Point: The point on the axis of a lens or mirror to which parallel rays of
light converge or from which they appear to diverge after refraction or reflection

Focal Length: It is the distance over which initially collimated rays are brought to a
focus. The focal length of a lens is defined as the distance in mm from the optical
centre of the lens to the focal point.
 Theory of Image Formation

Radius of Curvature: The distance from the vertex to the center of curvature is
the radius of curvature of the surface. Smaller radius of the curvature results in
a shorter focal length

Numerical aperture: It is defined NA = n sin α, where n is the refractive index

of the medium filling the space between the object and the lens, and α is the
half-angle of the maximum cone of light that can enter the lens
 Image Formation through Lens

Case1: The object has an infinite distance to the lens

In this case parallel rays from the object to the lens are assumed. These
are redirected in the lens to meet in the plane of the rear focal point
and generate an image in the plane of the focal point.
 Image Formation through Lens
Case 2: The object is situated at a relatively large distance (e.g. 100
times the focal length)

This situation produces an image that is smaller than the object (approx.
one 100th of the size of the original object)
 Image Formation through Lens
Case3: The object is located at a distance twice the focal length in
front of the lens.

This position creates an image of the object which is the same size
as the object itself (reproduction scale 1:1). The image is found at a
position twice the focal length from the rear side of the lens.
 Image Formation through Lens
Case 4: The object is situated in front of the focal point but within
the range of twice the focal length.

Image is generated which is larger than the object.

 Image Formation through Lens
Case 5: The object is located in the focal point of the lens.

In this case a virtual image, not a real one, is generated. The rays
will leave the lens in a parallel manner. No image can be found unless
we use another optical system e.g. our eye,
 Defects in Lenses
Spherical Aberration – Peripheral rays and axial rays have different
focal points
➢ This causes the image to appear hazy or blurred and slightly out of
focus.
➢ This is very important in terms of the resolution of the lens because
it affects the coincident imaging of points along the optical axis and
degrades the performance of the lens.
 Defects in Lenses
Chromatic aberration is a type of distortion in which there is a failure of a
lens to focus all colours to the same convergence point.

Chromatic aberration is caused by lens dispersion, with different colours of

light travelling at different speeds while passing through a lens. As a
result, the image can look blurred or noticeable coloured edges (red,
green, blue, yellow, purple, magenta) can appear around objects, especially
in high-contrast situations.
 Resolution
Resolution refers to the minimum distance between two points at which
they can be visibly distinguished as two points. The resolution of a
microscope is theoretically controlled by the diffraction of light.

Maximum Resolution = 0.61 * λ / N.A

λ= wavelength of illumination, N.A. = Numerical Aperture

The N.A. is a measure of the light gathering capabilities of an objective lens.

N.A. = n sin α where: n = index of refraction of medium, α= Angle subtended by

the lens
 Optical Microscopy

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
Imaging Modes
 Bright Field Optical Microscopy

Bright field microscopy is the most elementary form of microscope

illumination techniques and is generally used with compound microscopes

Name "bright field" is derived from the fact that the specimen is
dark and contrasted by the surrounding bright viewing field.
 Bright Field Optical Microscopy

Uses
➢ Because it is a simple method, this is the first type of microscopy
students learn in schools.
➢ The life sciences, particularly microbiology and bacteriology, have
always relied on the bright field technique.

Advantages
➢ Very Simple operation technique
➢ Specimens can be viewed without staining

Disadvantages
➢ Bright field microscopy has very low contrast
 Dark Field Optical Microscopy

Dark field microscopy is a technique for improving the contrast of

unstained, transparent specimens.

In a dark field microscope the light source is blocked off, causing

light to scatter as it hits the specimen.

The specimen appears lit up against a dark background

Bright Field x Dark Field

Bright-field& dark-field
images of carbon steel.
 Optical Microscopy

✓ 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 technique can only image dark or strongly refracting objects

effectively

❖ Diffraction limits resolution to approximately 0.2 micrometre

❖ Out of focus light from points outside the focal plane reduces image
clarity
 Electron Microscopy
✓ The electron microscope uses a beam of electrons to create an
image of the specimen. It is capable of much higher magnifications
and has a greater resolving power than a light microscope, allowing
it to see much smaller objects in finer detail

✓ The high resolution of electron microscopes results from short

wavelengths of the electrons used for microscope illumination

✓ The wavelength of electrons in electron microscopes is 10,000

times shorter than that of visible light

✓ The resolution of electron microscopes reaches the order of 0.1 nm

✓ Two type of electron microscopes:

✓ Transmission electron microscopes (TEM)
✓ Scanning electron microscopes (SEM)
 Scanning Electron Microscopy (SEM)

✓ SEM is a type of electron microscope that produces images of a sample

by scanning it with a focused beam of electrons. The electrons interact
with atoms in the sample, producing various signals that contain
information about the sample's surface topography and composition

What can we study in a SEM?

• Topography and morphology
• Chemistry
• Crystallography
• Orientation of grains
• In-situ experiments:
– Reactions with atmosphere
– Effects of temperature
 Various SEM images

AFM Cantilever Tip Ant Head Blood Cells

Diamond Thin Film Microstructure of a plain carbon Calcium Phosphate

(Numerous Multifaceted Micro- steel that contains 0.44 wt% of Crystal
crystals) carbon
 Components of Scanning Electron Microscopy
➢ Electron gun (filament)
➢ Electromagnetic optics
➢Sample stage
➢ Detectors
➢ Vacuum system
➢ Computer hardware and software
 Components of Scanning Electron Microscopy
✓ Electron Gun- It is a thermionic emission gun which produces –e. The thermo
electrons are emitted from a filament made of a thin tungsten wire by heating
the filament at high temperature.

✓ Condenser lens/Electromagnetic lens- placing a lens below the e- gun enables

to adjust the diameter of the e- beam.

✓ Objective lens- the objective lens is used for focusing.

✓ Specimen stage- It supports the specimen. It can perform horizontal (x,y

axis) & vertical movements(z axis), tilting of specimen, and rotation.

✓ Secondary e- detector- It is used for detecting the secondary e- emitted from

the specimen.

✓ Image display unit- The out put signals from the secondary electron detector
are amplified and then transferred to the display unit.

✓ Vacuum system- The electron optical system and the specimen chamber must
be kept at a high vacuum of 10-3 to 10-4 Pa
 Working principle of SEM
➢A beam of electrons is produced by the gun

Anode
➢ The electron beam pass through the column

➢Electromagnetic lenses which focus and direct the

beam down towards the sample.

➢ Hits the sample produce backscattered or

secondary electrons from the sample.

➢Detectors collect the secondary or backscattered

electrons, and convert them to signal
 Working principle of SEM
An electron gun, located at the top of the device, shoots out a beam of
highly concentrated electrons. The electron guns used by SEMs are Thermionic
guns. In the case of Thermionic guns, the thermo electrons are emitted from a
filament made of a thin tungsten wire by heating the filament at high
temperature.

The microscope is composed of a series of lenses within a vacuum

chamber. These lenses direct the electrons towards the specimen in order to
maximize efficiency. The more electrons that are used, the more powerful the
magnification. The SEM usually requires a vacuum chamber to function, as the
electron beam must not be obstructed as it passes through the body of the
microscope. Small particles could deflect the electrons onto the specimen itself,
obscuring the results.

When a specimen is hit with a beam of the electrons known as the

incident beam, it emits X-rays and mainly three kinds of electrons: backscattered
electrons, secondary electrons and Auger electrons. The SEM uses backscatter
electrons and secondary electrons. An electron recorder picks up the rebounding
electrons and records their imprint. This information is translated onto a screen
which allows three-dimensional images to be represented clearly. One of the
SEM's greatest advantages is its ability to reproduce textual information in a
consistent and coherent manner.
Scanning Electron Microscopy
Thermionic Emission Gun

Electron energy, E = eVo, electron energy determines

the wavelength of electrons and wavelength determines
resolution of the microscope
Field Emission Gun
 Detector: Everhart–Thornley (E–T) detector

Backscattered electrons- Elastic Scattering (BSEs: incident electrons

scattered by atoms in the specimen)

Secondary electrons- Inelastic Scattering (SEs: electrons ejected from

atoms in the specimen)

SEs travel with large deflection angle, while BSEs travel directly toward
the detector.

SEs energy is about 3–5 eV, SEs are used for topographic contrast

BSEs energy 60-80% of incident electron, used for elemental composition

contrast.
 Detector: Everhart–Thornley (E–T) detector

✓ The Faraday cage is positively or negatively charged having 250 or -50 V

✓ Detector attracts SEs when +ve, and screen out SEs with energy –ve

✓ Scintillator converts signal electrons into photons by accelerating the electrons

✓ The photons are enhanced by the PMT and display on a display screen
Electron–Specimen Interactions

✓ Pear Shaped interaction zone, and its size

increases with the energy of incident electrons

✓ SEs from a surface with a depth of 5–50 nm

✓ BSEs from depths of about 50–300 nm

✓ Spatial resolution of an SEM image is affected

by the size of the volume from where the signal
electrons escapes.

✓ SEs should have a better spatial resolution than

that formed by BSEs
✓ For imaging in the SEM, specimens must be
Electrically conductive
Electrically grounded

1. Cleaning the surface of the specimen

2. Stabilizing the specimen
3. Rinsing the specimen
4. Dehydrating the specimen
5. Drying the specimen
6. Mounting the specimen
7. Coating the specimen
Cleaning the surface of the specimen
Very important: Surface contains many unwanted deposits,
such as dust, mud, soil etc

Stabilizing the specimen

Hard, dry materials such as wood, bone, feathers, dried

insects, or shells can be examined with little further
treatment, but living cells and tissues and whole, soft-bodied
organisms usually require chemical fixation to preserve and
stabilize their structure.
Stabilization is typically done with fixatives

Fixation performed by incubation in a solution of a buffered

chemical fixative, such as glutaraldehyde, sometimes in combination
with formaldehyde and other fixatives.

Fixatives that can be used are:-

1.Aldehydes.
2.Osmium tetroxide.
3.Tanic acid.
4.Thiocarbohydrazides.

Rinsing the specimen

Sample must be rinsed - remove excessive fixatives.

Dehydrating the specimen: Water must be removed
✓ Air-drying causes collapse and shrinkage, this is commonly achieved by
replacement of water in the cells with organic solvents such as ethanol
or acetone.
✓ Dehydration - performed with a graded series of ethanol or acetone.

Drying the specimen

✓ Specimen should be completely dry - Otherwise the sample will be
destroyed

Mounting the specimen

✓ Specimen has to be mounted on the holder
✓ Mounted rigidly on a specimen holder called a specimen stub
✓ Dry specimen mounted on a specimen stub using an adhesive such as
epoxy resin or electrically conductive double-sided adhesive tape.
Coating the specimen
✓ To increase the conductivity of the specimen and to prevent the high
voltage charge on the specimen
✓ Coated with thin layer i.e., 20nm-30nm of conductive metal
✓ All metals are conductive and require no preparation before being used
✓ Non-metals need to be made conductive
✓ Done by using a device called a "sputter coater. ”
✓ Conductive materials:
Gold, Gold-palladium Alloy, Platinum,
Osmium, Iridium, Tungsten, Chromium, Graphite
Charge-up phenomenon can be prevented by coating the non-conductor
sample with metal (conductor)

Sample coating is intended to prevent charge-up phenomenon by allowing

the charge on the specimen surface go to ground through the coated
conductive film
EDS can be used to find the chemical composition of materials
down to a spot size of a few microns, and to create element
composition maps over a much broader raster area
Working of EDS

➢ The detector generates a charge pulse proportional to the X-ray

energy

➢ This pulse is first converted to a voltage, then the signal is amplified

through a field effect transistor (FET), isolated from other pulses,
further amplified, then identified electronically as resulting from an X-
ray of specific energy

➢ Finally, a digitized signal is stored in a channel assigned to that energy

in the MCA
 Principal of EDS

➢ The incident beam may excite an electron in an inner shell, ejecting it

from the shell while creating an electron hole where the electron was.

➢ An electron from an outer, higher-energy shell then fills the hole, and
the difference in energy between the higher-energy shell and the lower
energy shell may be released in the form of an X-ray

➢ The number and energy of the X-rays emitted from a specimen can be
measured by an energy-dispersive spectrometer.

➢ The energy of the X-rays are characteristic of the difference in energy

between the two shells, and of the atomic structure of the element from
which they were emitted, this allows the elemental composition of the
specimen to be measured.
 Advantages and Disadvantages
▪ Advantages of a Scanning Electron Microscope include its wide-array of
applications, the detailed three-dimensional and topographical imaging and the
versatile information garnered from different detectors.

▪ SEMs are easy to operate and advances in computer technology make operation
user-friendly.

▪ This instrument works fast and allows the generation of data in digital form.

▪ Most of the SEM samples require minimal preparation actions.

▪ SEMs are expensive, large and must be housed in an area free of any possible
electric, magnetic or vibration interference

▪ Maintenance involves keeping a steady voltage, currents to electromagnetic coils

and circulation of cool water

▪ SEMs are limited to solid, inorganic samples small enough to fit inside the vacuum
chamber that can handle moderate vacuum pressure.
 Applications

➢ SEMs have a variety of applications in a number of scientific and industry-

related fields, especially where characterizations of solid materials is
beneficial.

➢ In addition to topographical, morphological and compositional information,

a Scanning Electron Microscope can detect and analyze surface fractures,
provide information in microstructures, examine surface contaminations,
reveal spatial variations in chemical compositions, provide qualitative
chemical analyses and identify crystalline structures.

➢ SEMs can be as essential research tool in fields such as life science, biology,
gemology, medical and forensic science, metallurgy.
Characterization of Nanomaterials

Transmission Electron Microscopy (TEM)

High Resolution Transmission Electron Microscopy (HRTEM)
Environmental Transmission Electron Microscopy (ETEM)
 Transmission Electron Microscope
Transmission electron microscopy (TEM) is a microscopy technique
in which a beam of electrons is transmitted through an ultra-thin specimen,
interacting with the specimen as it passes through. An image is formed from
the interaction of the electrons transmitted through the specimen; the image
is magnified and focused onto an imaging device, such as a fluorescent
screen, on a layer of photographic film, or to be detected by a sensor such as
a CCD camera.
 Transmission Electron Microscope

➢ The transmitted electrons are used to create an image of the sample

➢ Scattering occurs when electron beam interact with matter. Interaction of the

electron with the sample produces elastic (no energy change) and in elastic

scattering (energy change)

Two kind of Image collection

(a) Bright field :Transmitted beam is used for imaging

(b) Dark field : Diffracted beam is used for imaging by either using a movable

aperture or shifting the incident beam, keeping the aperture constant

Components of TEM
 Components of TEM

✓ Electron Gun- Thermionic emission Gun or Field emission Gun which produces
high energy –e beams.

✓ Vacuum system- The electron-optical column and sample chamber must

always be at a vacuum.

✓ Condenser lens- Controls the beam diameter and convergence angles of the
beam incident on a specimen.

✓ Condenser Aperture- Used to restrict the electron beams and filter out
unwanted scattered electrons before image formation.

✓ Objective lens- Focuses the transmitted electron from the sample into an
image

✓ Objective Aperture- Enhances the contrast by blocking out high-angle

diffracted electrons
 Components of TEM

✓ Intermediate lens- Used to switch the TEM between an image mode and a
diffraction mode.

✓ Projector lens- Further magnifies the image or diffraction pattern and projects
it onto the fluorescent screen

✓ Specimen holder and metal mesh disc: Supporting small foil pieces of
specimen
 Working Principle of TEM
The TEM operates on the same basic principles as the light microscope
but uses electrons instead of light to get a much better resolution.
In TEM a beam of electrons from the electron gun is focused into a
small, thin, coherent beam by the use of the condenser lens. This beam is
restricted by the condenser aperture, which excludes high angle electrons. The
beam then strikes the specimen and parts of it are transmitted depending
upon the thickness and electron transparency of the specimen. This
transmitted portion is focused by the objective lens into an image on phosphor
screen or charge coupled device (CCD) camera. Optional objective apertures
can be used to enhance the contrast by blocking out high-angle diffracted
electrons. The image then passed down the column through the intermediate
and projector lenses, is enlarged all the way.
 Working Principle of TEM

The image strikes the phosphor screen and light is generated,

allowing the user to see the image. The darker areas of the image represent
those areas of the sample that fewer electrons are transmitted through while
the lighter areas of the image represent those areas of the sample that more
electrons were transmitted through.
Chemical analysis can been done in a TEM microscope by X-rays
energy dispersive spectrometry EDS, or electron energy loss spectrometry
EELS.
 Advantages and Disadvantages of TEM
Advantages
✓TEMs offer very powerful magnification and resolution
✓TEMs have a wide-range of applications and can be utilized in a variety of
different scientific, educational and industrial fields
✓TEMs provide information on element and compound structure
✓Images are high-quality and detailed

Disadvantages
✓TEMs are large and very expensive
✓Laborious sample preparation
✓Operation and analysis requires special training
✓Samples are limited to those that are electron transparent.
✓TEMs require special housing and maintenance
✓Images are black and white
 Applications of TEM

➢ TEM is ideal for a number of different fields such as life sciences,

nanotechnology, medical, biological and material research, forensic analysis,
gemology and metallurgy as well as industry and education

➢ TEMs provide topographical, morphological, compositional and crystalline

information

➢ The images allow researchers to view samples on a molecular level, making

it possible to analyze structure and texture

➢ TEMs can be used in semiconductor analysis and production and the

manufacturing of computer and silicon chips

➢ Technology companies use TEMs to identify flaws, fractures and damages to

micro-sized objects; this data can help fix problems and/or help to make a
more durable, efficient product.
 TEM
High-Resolution Transmission Electron Microscopy (HRTEM)

An imaging mode of the transmission electron microscope (TEM) that allows for
direct imaging of the atomic structure of the sample

Environmental Transmission Electron Microscopy(ETEM)

ETEM is a unique instrument allows us to study the materials in their native

environment. In contrast to the traditional operation of transmission electron
microscopes under high vacuum, ETEM allows imaging within high-temperature
and controlled gaseous environments. It offers sample operating temperatures up
to 1,100 oC, and is equipped with a controlled gas-inlet system for introducing up
to four gases into the system. This microscope allows us to analyze atomic-level
microstructure during exposure to gases. ETEM enables to investigate atomic-
resolution imaging, spectroscopic studies of materials under dynamic operating
conditions and the fundamental atomic mechanisms of gas–solid reactions.
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