CHARACTERIZATION OF MATERIALS

Class Notes
 LIGHT MICROSCOPY

Reference: ZEISS microscopy, Education in Microscopy and Digital

Imaging
The human eye as a visual detector (in combination with the brain) is the
most efficient image-processing system that has ever been encountered.
Together with the muscle-adjusted lens, the curved surface of the cornea
projects an optical image onto the retina (the detector).
The level of incident brightness is controlled via the variable diameter of an
iris (much like an optical diaphragm) under the control of specific muscles.

A sharp image is produced by the flexible lens, the focal length of which is
changed by another set of muscles so that focusing is possible on any object at a
distance between approximately 20 centimeters and infinity.

The image itself is detected on the retina by approximately 130 million

photoreceptor rod cells (responsible for recognition of grey levels) and 7 million
photoreceptor cone cells (color recognition), and is then transferred to the brain
along the shortest possible path through the optic nerve.
Simple Microscope: Magnification with a Simple Lens
• Increases the visual angle on the retina
 Lens equation
A converging lens is characterized by collecting parallel light rays in a focal point F

1 1 1
 
Focal Length Object distance Image distance
 Lens Magnification
A converging lens is characterized by collecting parallel light rays in a focal point F

Image Size  Image Distance 

Magnification    
Object Size  Object Distance 
 Magnification with Multiple Lenses
Total Magnification = Objective Magnification x Eyepiece Magnification
Magnification with Multiple Lens: Compound Microscope
Total Magnification = Objective Magnification x Eyepiece Magnification

The range of useful total magnification for an objective and eyepiece

combination is defined by the numerical aperture of the system.
 Magnification with Multiple Lens

There is a minimum magnification necessary for the detail present in

an image to be resolved by the eye, and this value is typically set at
500 times the numerical aperture (500 x NA).

At the other end of the spectrum, the maximum useful

magnification of an image is usually set at 1000 times the numerical
aperture (1000 x NA)

This limit is set by the wave nature of light imposed on the objective
by diffraction.

Exceeding the limit of useful magnification causes the image to

suffer from empty magnification, where increasing magnification
through the eyepiece or intermediate tube lens only causes the
image to become more magnified with no corresponding increase
in detail resolution.
Numerical Aperture and Resolution
 Numerical Aperture (NA)
Numerical aperture describes the light-gathering ability and resolution of
an objective (resolving power)

The numerical aperture of a

microscope objective is the
measure of its ability to gather light
and to resolve fine specimen
detail while working at a fixed
object (or specimen) distance.

Numerical Aperture ( NA)  n  sin 

n is the index of refraction of the medium in which the lens is working
θ is the maximal half-angle of the cone of light that can enter or exit the lens

A lens with a larger numerical aperture will be able to visualize finer

details than a lens with a smaller numerical aperture.
 The Concept of NA for Objectives and Condensers

The numerical aperture of a microscope objective is the measure of its

ability to gather light and to resolve fine specimen detail while working
at a fixed object (or specimen) distance.
 Microscope Resolution and the Airy Disk
When light from the various points of a specimen
passes through the objective and is reconstituted as
an image, the various points of the specimen appear
in the image as small patterns (not points) known as
Airy patterns.

This phenomenon is caused by diffraction or scattering

of the light as it passes through the minute parts and
spaces in the specimen and the circular rear aperture
of the objective.
An Airy disk is the region enclosed by the first minimum of
the airy pattern and contains approximately 84 percent
of the luminous energy.
 0.61
d0 
N . A.

The limit up to which two small objects are still seen as separate
entities is used as a measure of the resolving power of a microscope.
dmin ~ 0.3m for a mid-spectrum of 0.55m
Common Optical Defects in Lens Systems
(Aberrations)
 Spherical Aberration
 Peripheral rays and axial rays have different focal points (caused by
spherical shape of the lens surfaces.
 Causes the image to appear hazy or blurred and slightly out of focus.
 Very important in terms of the resolution of the lens because it affects
the coincident imaging of points along the optical axis and degrade
the performance of the lens.
 Chromatic Aberration

 Axial - Blue light is refracted to the greatest extent followed by

green and red light, a phenomenon commonly referred to as
dispersion
 Lateral - chromatic difference of magnification: the blue image of
a detail was slightly larger than the green image or the red image
in white light, thus causing color ringing of specimen details at the
outer regions of the field of view
Aberrations Correction
 Polarization of Light

When the electric field vectors of light are restricted to a single plane by
filtration, then the the light is said to be polarized with respect to the direction
of propagation and all waves vibrate in the same plane.
 Bright field
Only direct
light falls on
the sample
surface,
where it is
absorbed or
reflected. The
quality
parameters of
the image are
brightness,
resolution,
contrast and
field depth.
 Dark field

Only refracted,
diffracted or
reflected light falls
on the sample
surface. Dark field is
suitable for all
samples with
structured surfaces
and can also be
used to visualize
structures below the
resolution limit. The
surface structures
appear bright on a
dark background.
Typical examples of
applications
 Cutting

Mounting
 Polishing

Microscopy
29
 Grain Size Examination

Objective Lens

x100
Optical Emission Spectroscopy

Reference: Oxford Instruments

 Optical Emission Spectroscopy

To determine the elemental composition of a broad range of metals

 Optical Emission Spectroscopy

OES can analyze a wide range of elements from Lithium to Uranium

in solid metal.
How does
Optical Emission Spectroscopy work?

35
 Sample
Electrode

The difference in electrical potential between the sample and

electrode produces an electrical discharge, this discharge
passes through the sample, heating and vaporizing the material
at the surface and exciting the atoms of the material, which
then emits the element-characteristic emission lines.
Optical Emission Spectroscopy
Iron emits just over 8000 different wavelengths so choosing the
optimum emission line for a given element in a sample is important.
The characteristic light emitted by the atoms in the sample is
transferred to the optical system where it is split into its spectral
wavelengths by the high-tech grading, the grading contains up to
3600 grooves per millimeter.
Next the individual spectral line peak signals are collected by
detectors and processed to generate a spectrum showing the light
intensity peaks versus their wavelengths.

The peak wavelength identifies the element, and its peak area or
intensity gives an indication of its quantity in the sample.

The analyzer then uses this information to calculate the sample’s

elemental composition based on a calibration with certified
reference material.

Advantages
•It’s fast and relatively easy to use.
•It measures a wide range of elements and concentrations in many
different types of materials, including important elements such as
carbon, sulfur, phosphorous, boron and nitrogen.
•It’s extremely accurate when measuring low levels of trace and
tramp elements.
•It’s fairly low-cost compared to other techniques.
X-Ray Fluorescence (XRF)

40
 X-ray fluorescence (XRF)

X-rays are on the high energy side of ultraviolet, and are expressed in terms of
their energy in kilo electron volts (keV), or wavelength in nanometers (nm).
 X-ray fluorescence (XRF) can typically analyse elements from sodium
to uranium, in concentrations ranging from parts per million to high
percents, in solids, liquids, and powders.
How does
X-ray fluorescence work?

43
Primary x-rays are generated by the source and directed at the
sample’s surface, sometimes passing through a filter to modify the x-
ray beam.
When the beam hits the atoms in the sample, they react by
generating secondary x-rays that are collected and processed by a
detector.
The x-rays emitted by the atoms in the sample are collected by a
detector, and processed in the analyser to generate a spectrum
showing the x-rays intensity peaks versus their energy.
The peak energy identifies the element and its peak area (or
intensity) gives an indication of its amount in the sample.
What happens to the atoms in the
sample during the analysis?

46
 A stable atom is made
of a nucleus and
electrons orbiting it. The
electrons are arranged
in energy levels or shells
(K, L, M, N) and different
energy levels can hold
different numbers of
electrons.
 When a high energy
primary X-ray collides
with an atom, it disturbs
its equilibrium.
An electron is ejected from
a low energy level, and a
vacancy is created,
making the atom unstable.
To restore stability, an electron
from a higher energy level falls
into this vacancy and the
excess energy released as the
electron moves between the
two levels is emitted in the
form of a secondary X-ray.
Atomic Force Microscopy (AFM)

52
The atomic force
microscope (AFM)
is one kind of
scanning probe
microscopes (SPM).
SPMs are designed
to measure local
properties, such as
height, friction,
magnetism, with a
probe. To acquire
an image, the SPM
raster-scans the
probe over a small
area of the
sample, measuring
the local property
simultaneously.
Imaging Mode in AFM
 Imaging Modes in AFM
Contact Mode:
Feedback: Lever deflection
The feedback system adjusts the height of the
cantilever base to keep this deflection constant as
the tip moves over the surface.
(Friction force microscopy, conductive probe AFM)

Non Contact Mode:

Feedback: Oscillation Amplitude
The cantilever oscillates close to the
sample surface without making contact
with the surface.
(Electrostatic/ magnetic AFM)

Intermittent Contact Mode:

Feedback: Oscillation Amplitude
The cantilever oscillates and the tip makes repulsive
contact with the surface of the sample at the lowest
point of oscillation.
(Tapping Mode AFM)
Raman Spectroscopy

58
Raman Scattering
Radiation scattered by molecules
contains photons with the same
frequency as the incident radiation, but
may also contain photons with changed
or shifted frequency. This effect is very
weak—approximately one photon out of
a million (0.0001%) will scatter from that
sample at a wavelength slightly shifted
from the original wavelengths.

59
 Principle and Applications
When light meets particles that are
smaller than the light's wavelength,
the light spreads in different directions.
This occurs, for example, when light
packets - photons - encounter
molecules in a gas. In 1928 Venkata
Raman discovered that a small portion
of the scattered light acquires other
wavelengths than that of the original
light. This is because some of the
incoming photons' energy can be
transferred to a molecule, giving it a
higher level of energy. Among other
things, the phenomenon is used to
analyze different types of material.

Source:
https://www.nobelprize.org/nobel_prizes/physics/laureates/1930/raman-
facts.html
60
 Raman Energy Levels

The different possibilities of light scattering: a) Rayleigh scattering (no exchange

of energy: incident and scattered photons have the same energy), b) Stokes
Raman scattering (atom or molecule absorbs energy: scattered photon has less
energy than the incident photon) and c) anti-Stokes Raman scattering (atom or
molecule loses energy: scattered photon has more energy than the incident
photon)
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62
Infrared (IR) Spectroscopy

63
 Functional groups present in the molecules.

Wagging

Stretching
vibration

Formaldehyde
Covalent bonds behave like a spring

A functional group will absorb light if the frequency of the light matches the frequency of
stretching, bending or wagging.
64
1-Propanol
 Principle and Applications
Infrared Spectroscopy is the analysis of infrared light interacting
with a molecule.
This can be analyzed in three ways by measuring absorption,
emission and reflection.

IR Spectroscopy measures the vibrations of atoms, and based

on this it is possible to determine the functional groups.
Generally, stronger bonds and light atoms will vibrate at a high
stretching frequency (wavenumber).

The main use of this technique is in organic and inorganic

chemistry. It is used by chemists to determine functional groups
in molecules.

It has great significance to scientific researchers in many fields

such as protein characterization, nanoscale semiconductor
analysis and space exploration.
 Principle and Applications
Stronger the bond, the higher the wave number/frequency.
The lighter the atom, the higher the wave number/frequency.
The broadening of an OH absorption band is due to hydrogen
bonding.
Electron delocalization (resonance) can affect the wave number of
a functional group.

IR Spectroscopy measures the vibrations of atoms, and based on this

it is possible to determine the functional groups. Generally, stronger
bonds and light atoms will vibrate at a high stretching frequency
(wavenumber).

The main use of this technique is in organic and inorganic chemistry.

It is used by chemists to determine functional groups in molecules.

It has great significance to scientific researchers in many fields such

as protein characterization, nanoscale semiconductor analysis and
space exploration.
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73
Important IR Stretching Frequencies

74
75
Mass Spectrometry
 CH3CH2CH3
What is this substance/molecule?

What is the Molecular Weight?

 CH3CH2CH3

Mass Spectrometer
CH3CH2CH3

Electron Beam
Core Electrons
Bonding Electron
Bonding Electron
Bonding Electron
Bonding Electron
Thermal Analysis

Reference: METTLER TOLEDO

Differential scanning calorimetry (DSC)
Heat flow changes v/s temperature or time

Differential thermal analysis (DTA)

Temperature difference v/s temperature or
time

Thermo gravimetric analysis (TGA)

Mass change versus temperature or time
Differential scanning calorimetry (DSC)
DSC measures enthalpy changes in samples due to changes in their
physical and chemical properties as a function of temperature or
time.
Typical Information That Can Be Derived from DSC Measurements:

Characteristic temperatures (melting, crystallization, polymorphous

transitions, reactions, glass transition)

Melting, crystallization, transformation and reaction heats

(enthalpies)

Crystallinity of semi-crystalline substances

Decomposition, thermal stability

Measurement principle

DSC measures the difference between the heat flows from the
sample and reference sides of a sensor as a function of temperature
or time.
Differential scanning calorimetry (DSC)
 Steel Material — Phase Transitions

Change in crystal structure

Magnetic Transition Melting Point

Aluminium Alloy — Melting & Solidification
Polymers — Polyethylene terephthalate (PET)

Melting

Amorphousness

Crystallanity
 Differential thermal analysis (DTA)

In this technique, it is the heat flow to the sample and reference that
remains the same rather than the temperature. When the sample and
reference are heated identically, phase changes and other thermal
processes cause a difference in temperature between the sample
and reference.
 DSC v/s DTA

Both DSC and DTA provide similar information.

DSC measures the energy required to keep both the reference and
the sample at the same temperature whereas

DTA measures the difference in temperature between the sample

and the reference when the same amount of energy has been
introduced into both.
Thermo gravimetric analysis (TGA)
TG is a technique that measures mass
change in a sample, and it is used to detect
evaporation, decomposition, oxidation and
other effects of temperature change that
cause mass changes.
Porcelain Raw Material — Mass-Loss Steps

Evaporation of humidity
burn-up of organic content

dehydration of kaolin

solid-solid transition
 Dolomite — Thermal Decomposition

The mass loss steps during the thermal decomposition of

dolomite [CaMg(CO3)2
Rubber Compound for Tires — Decomposition