Nanoscience and

technology
Contents:

FIB and
CHARACERIZATION ANALYSIS
(MICROSCOPIC ANALYSIS)
SEM,TEM, STEM, STM,AFM
(FLUROSENCE/OPTICAL)
UV/VIS,RAMAN SPECTROSCOPY
(STRUCTURAL ANALYSIS)
XRD AND EDX
CONTENTS - Surface Analysis Laboratory Techniques
1.SEM
2.TEM
3.STM
4.AFM
5.UV/VIS
SPECTROSCOPY
6.RAMAN
SPECTROSCOPY
7.XRD
8.EDX
There are two basic techniques to characterize metal nanoparticles: Spectroscopic
methods (e.g., UV–visible absorption and Raman scattering) and Microscopy (e.g.,
transmission electron and scanning atomic force). The most common tool to provide
reliable diagnostic of size distribution is electron microscopy. But this still is an
expensive, time consuming and mostly inaccessible method in terms of required
materials and infrastructure (Martínez et al., 2012). On the other hand, UV–visible
spectroscopy is an accessible and therefore widely used in the study of nanomaterials,
from a basic diagnostic of nanoparticle formation to the assessment of their complex
interactions with other elements.
 Although techniques to be used would depend upon the type of material and
information one needs to know, usually one is interested in first knowing the size,
crystalline type, composition, thermal, chemical state and properties like optical or
magnetic properties
 FIB

 FIB stand for “ Forced Ion Beam”.

 It is define as:
“ A technique used particularly in semiconductor
Industry, material science and increasingly in biological
field for site specific analysis deposition and ablation
Material”
 It resembles a scanning electronicroscope.
 SEM uses a focused beam of electron to image the sample while
FIB setup used a focused beam of ions.
 Principle of FIB

 The Gallium primary ion beam hits the sample and sputters
a small amount of material which leaves the surface as
either secondary ions (i+ or i-) ion or neutral atom(no).

 At low primary beam currents very little material sputtered.

 At higher primary currents a great de of material can be

removed by sputtering allowing precise milling of
speciman down to sub mirometer or even a nano scale.
 Working and
Diagram
• Instruments are using liquid ion source
specially Gallium ion source.
• Gallium metal is placed in contact with a
tungsten needle.
• Heated gallium wets the tungsten and flows to
the tip of needle.
• The opposing forces surface tension and
electric field form the Gallium into cusp
shaped tip called a Taylor cone.
• The huge electric field at the small tip causes
ionziation and field emission of Gallium atom.
• Accelerated to an energy of 1-50 kev.
 Applications

 Circuit modification
 Photomask repair
 Transmission electron microscope(TEM).
 Defect analysis.
 Scanning electron microscope (SEM)
Scanning electron microscope (SEM) is one of the most widely used
techniques used in characterization of nanomaterials and nanostructures.
The signals that derive from electron-sample interactions reveal information
about the sample including surface morphology (texture), chemical
composition of the sample. The morphology of the Cu nanoparticles was
carried out on JEOL, JSM- 67001 and the image is shown in figure .

 SEM is a type of electron microscope that

images the sample by scanning it with high
energy beam of electrons.
 Basically, it is an improved model of Electron
Microscope.
 The signal obtained from electron-sample
interaction reveal information about the
sample including external Morphology (shape,
size), Topography (texture or surface features of
object how it looks), Chemical Composition
and Crystalline structure (3D representation of
how atoms arranged) and orientation of
materials making up the sample
GRAPHICAL REPRESENTATION
 Electron-sample Interaction (Principle):

 During SEM inspection, a beam of electrons

is focused on a spot volume of specimen
(says sample), 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 dislodge electrons are
known as secondary electrons are attracted
and collected by a positively biased grid or
detector, and then translated into a signal.
To produce SEM image, the electron beam is
swept /scan across the area being
inspected, producing many such signals.
These signals are them amplified, analyzed
and translated into images of the
topography being inspected. Finally, the
image is shown on a CRT.
 How scientist
interpret Nanomaterials for adsorption of pollutants and
results from heavy metals:
SEM?

The SEM shown in Fig. confirms the

presence of nanopores, which makes it a
nano-sized adsorbent for the adsorption
process. Similar adsorbents with nano-
sized pores are preferred because they
will provide higher surface area for
adsorption. The availability of more
surface area will help in obtaining a
higher adsorption capacity, finally
leading to a higher pollutant removal.

Confirmation of graphene in Covid 19 Vaccine SEM OF Stomata in Leaves of Paphiopedilum

Transmission Electron Microscope (TEM)

TEM is the microscope that use a particle beam of

electrons to visualize specimens and generate a highly-
magnified image. TEM can magnify objects up to 2 million
times. In order to get better idea of just how small that is,
think of how small a cell is. It is no wonder TEMs have
become so valuable with the biological and medical fields

Principle:
Steam of electrons are produced by electron gun and
made to fall over the specimen using the magnetic
condensing lens. Electrons are made to pass through the
specimen and the image is formed on the fluorescent
screen, either by using the transmitted beam or by using
the diffracted beam. This high contrast image is called
bright field image.
● How does TEM work?

An electron gun at the top of a TEM emits electrons that travel through the microscope's vacuum tube. Rather
than having a glass lens focusing the light (as in the case of light microscopes), the TEM employs an
electromagnetic lens which focuses the electrons into a very fine beam.

● Is Tem destructive?
However, a major limitation with TEM is the time-consuming, destructive sample preparation necessary for
generating electron transparent specimens. Scanning electron microscopy (SEM) has the significant advantage
over TEM of being non- destructive and can rapidly image large areas.

● where it is preferred?

TEM is the preferred method to directly measure nanoparticle size, grain size, size distribution, and
morphology
How scientists interpret results from it?

For particles that are size and shape polydisperse, a size

distribution can be obtained by measuring each particle
diameter with eye and hand method. The ImageTool soft-
ware is used for obtaining of size distribution. The size
distri-
bution of the nanoparticles is a main parameter in synthesis.
By image analysis of TEM micrographs can predict the
histo-gram of size distribution. Figure 2 is the histogram of
the CuO nanoparticles synthesis.
This histogram for 50 of CuO nanoparticles is plotted. The
mean size distribution of the nanoparticles is almost
22.58 nm and most of the nanoparticles are in 22–24 ranges
in this histogram
 SEM and TEM
 Electron Microscopes are scientific instruments that use a beam of
highly energetic electrons to examine objects on a very fine scale.
 This examination can yield information about the topography,
morphology, composition and crystallographic information.

Transmission Electron Microscope (TEM) – allows one the

study of the inner structures.
Scanning Electron Microscope (SEM) – used to visualize the
surface of objects.
ADVANTAGES & 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.
 ADVANTAGES & DISADVANTAGES
OF SEM
Advantages
It gives detailed 3D and topographical imaging and the versatile information garnered
from different detectors.
This instrument works very fast.
Modern SEMs allow for the generation of data in digital Most SEM samples require
minimal preparation actions.
Disadvantages
SEMs are expensive and large.
Special training is required to operate an SEM. The preparation of samples can result in
artifacts. .
SEMS are limited to solid samples.
SEMs carry a small risk of radiation exposure associated with the electrons that scatter
from beneath the sample surface.
Scanning Tunneling Microscope (STM)

Scanning tunneling microscope (STM) is a high resolution

microscope for imaging electrically conducting surfaces down to
the atomic scale.
STM was devised based on the phenomenon of quantum
tunneling.

 Working Principle:
In STM, the sample is scanned by a very fine metallic tip. The
tip (tungsten) is mechanically connected to an xyz positioning
device realized by means of piezoelectric materials. The sample is
positively or negatively biased so that a small current known as the
“tunneling current” flows if the tip is in close proximity to, but not
actually touching the sample.
 Scanning transmission electron
microscopy(STEM)
Scanning transmission electron microscopy (STEM)
combines the principles of transmission electron microscopy
and scanning electron microscopy and can be performed on
either type of instrument. Like TEM, STEM requires very thin
samples and looks primarily at beam electrons transmitted by
the sample. One of its principal advantages over TEM is in
enabling the use of other of signals that cannot be spatially
correlated in TEM, including secondary electrons, scattered
beam electrons, characteristic X-rays, and electron energy
loss.
Like SEM, the STEM technique scans a very finely focused
beam of electrons across the sample in a raster pattern.
Interactions between the beam electrons and sample atoms
generate a serial signal stream, which is correlated with beam
position to build a virtual image in which the signal level at any
location in the sample is represented by the gray level at the
corresponding location in the image. Its primary advantage
over conventional SEM imaging is the improvement in spatial
resolution.
Definition:
Atomic force microscopy (AFM) or scanning force microscopy (SFM) is a very-high-
resolution type of scanning probe microscopy (SPM), with demonstrated resolution on
the order of fractions of a nanometer, more than 1000 times better than the optical
diffraction limit.
AFM provides a 3D profile of the surface of a nanoscale by measuring froces between a
sharp probe and the surface .
The AFM has three major abilities: force measurment , imaging and manipulation.

What is AFM? ❖Atomic-force microscopy (AFM) is a

powerful technique that can image almost
any type of surface, including polymers,
ceramics, composites, glass, and biological
samples. AFM is used to measure and
localize many forces, including adhesion
strength, magnetic forces, and mechanical
properties.
  AFM microscopes operate on the
principle of surface sensing using an
extremely sharp tip on a
micromachined silicon probe. This tip
is used to image a sample by raster
scanning across the surface line by
Working Principle line, although the method varies
dramatically between distinct operating
modes. The two primary groups of
operating modes are widely defined as
contact mode and dynamic, or
tapping, mode.
 In contact mode, the tip is "dragged"
across the surface of the sample and the

Imaging contours of the surface are measured either

using the deflection of the cantilever directly
or, more commonly, using the feedback signal
Modes required to keep the cantilever at a constant
position.
 In tapping mode, the cantilever is driven to
oscillate up and down at or near its resonance
frequency.
 Typical
configuration
of an AFM.
(1): Cantilever, (2):
Support for
cantilever, (3):
Piezoelectric element
(to oscillate cantilever
at its eigen
frequency), (4): Tip
(Fixed to open end of a
cantilever, acts as the
probe), (5): Detector of
deflection and motion
of the cantilever, (6):
Sample to be measured
by AFM, (7): xyz drive,
(moves sample (6) and
stage (8) in x, y, and z
directions with respect
to a tip apex (4)),
and (8): Stage.
 Advantages and Disadvantages of AFM

• Advantages
Easy sample preparation
What are the limitations of the AFM?
• Accurate height information
AFM imaging is not ideally sharp.
• Works in vacuum, air, and liquids
• Living systems can be studied
The Future of Atomic Force Microscopy
• Disadvantages  •Atomic or angstrom resolution images
of live cell surfaces: development of
• Limited vertical range
more flexible cantilever springs and less
• Limited magnification range
damaging and nonsticky probes
• Data not independent of tip needed
• Tip or sample can be damaged
 Sharper tips by improved
microfabrication processes
 AFM vs STM
 STM is particularly  AFM is needed for
useful for probing insulating samples.
electrons at Since most polymers
surfaces, for and biomolecules
example the are insulating, the
electron waves in probe of choice for
quantum corrals or soft matter is often
the energy levels of AFM. This image
the electrons in shows DNA on mica,
dangling bonds an insulator.
and surface
molecules.
❖ XRD:

 XRD technique is an important tool for the

study of crystal structures and atomic
spacing.
 XRD is based on constructive interface of
monochromatic x ray on a crystalline
sample. it enables verification of the
crystallinity and structure of a sample but
gives no information of a chemical nature.
 Diffraction is the main technique used to
identify crystal structure
 History:

 In june 8,1912 during meeting of German physical society at university Berlin a

thirty three years old physicts Max von Laue announced his discovery of x ray
diffraction in crystals as in a three dimensional diffraction grating.

Principle:
 X-ray diffraction is based on constructive interference of monochromatic X-
rays and a crystalline sample. These X-rays are generated by a cathode ray
tube, filtered to produce monochromatic radiation, collimated to
concentrate, and directed toward the sample.

Significance:
XRD is a rapid analytical technique used for phase identification of a
crystalline material and also can provide information on unit cell dimensions.
The analyzed material is finely ground, homogenized and, the average bulk
composition is determined.
 Basic arrangement of atoms

The atoms areBasics

arrangedofincrystallography
a regular
pattern, and there is as the smallest
volume element that by repetition in three
dimensions describes the crystal. This
smallest volume element is called a unit
cell.
Crystals consist of planes of atoms that are
spaced a distance d apart but can be
resolved into many atomic planes, each
with a different d spacing.
The dimensions of the unit cell are
described by three axes: a, b, c, and the
angles between them α, β, and γ are
lattice constants that can be determined
by XRD.
The crystalline sample act as a 3D diffraction grating for x ray
wavelength similar to the spacing of planes in a crystal lattice.The
interaction of incident X rays crystal sample produce constructive
interference if conditions of Bragg's
These Diffrected X rays from various
plans are detected. Processed and
computed. All the possible
diffraction directions of the crystal
lattice can be attained by
scanning the sample through a
range of angles 2thrta.The
diffraction peaks are
the information of different
orientation of planes having
different Miller indices
 Peak position
 All the planes diffract
coherently at an angle where
Bragg's law holds good
Therefore the position of a
peak determine the inter
planar spacing for a family of
planes.
 d=inter planar spacing
 n=1 for the first maxima
 If d increase sin© should
decrease
 Bcz left term is constant
 XRD Benefits and Applications:

 Identify crystalline phases and orientation.

 Determine structural properties: - Lattice parameters. - Strain. - Grain size. -
Epitaxy. - Phase composition. - Preferred orientation.
 Measure thickness of thin films and multi-layers.
 Determine atomic arrangement.
 Advantages of XRD

XRD is the most convenient, the most widely used method to determine
the crystal structure.
XRD gives also information about the structure of solids, and
arrangements of the atoms that compose solids.

Disadvantages of XRD

XRD has size limitations. It is much more accurate for measuring large
crystalline structures rather than small ones because small structures are
trace in amount and are undetected by XRD readings.
It is relatively low in sensitivity.
 Uses of XRD
It is a laboratory-based technique used for the
identification of crystalline materials.
It is also used for the analysis of unit cell dimensions.
It is also used to determine the structure of proteins.
It is also used to distinguish between different crystal
structures with identical compositions.
It is used to study rapid biological and chemical
processes.
It is also used for crystallographic applications.
Definition:
Energy dispersive X-ray spectroscopy (EDX) is an analytical method for
analytical or chemical characterization of materials.
EDX systems are generally attached to an electron microscopy instrument such
as transmission electron microscopy (TEM) or scanning electron microscopy
(SEM).
EDX is based on the emission of a specimen characteristic X-rays.
A beam of high energy charged particles (electrons or protons) are focused
into the investigated sample.

EDX → Energy-dispersive X-ray

spectroscopy (EDS, EDX, EDXS or XED
S), sometimes called energy dispersive X-
ray analysis (EDXA or EDAX) or energy
It is also referred as: dispersive X-ray
EDS: Energy Dispersive Spectroscopy microanalysis (EDXMA),
EDXS: Energy Dispersive X-ray Spectroscopy
X-EDS: X-ray Energy Dispersive Spectroscopy
EDX: Energy Dispersive X-ray analysis
 PRINCIPLE
Energy-dispersive X-ray spectroscopy (EDX) is a surface analytical technique where an
electron beam hits the sample, exciting an electron in an inner shell, causing its ejection
and the formation of an electron hole in the electronic structure of the element. When the
electron is displaced, it attracts another electron from an outer shell to fill the vacancy

A schematic describing the EDX spectroscopy method

is shown in Fig. 5.
❖ EDX can be used to confirm the composition and distribution of the nanoparticles through spectrum and
elemental mapping. This protocol outlines the procedures for compositional identification of
nanoparticles using an EDX spectrometer incorporated into a scanning electron microscopy (SEM) system.

EDX analysis gives a spectrum that displays the peaks correlated to the elemental composition of the
investigated sample.

In addition, the elemental mapping of a sample can be created with this characterization method.

The peak intensities are proportional to the elemental concentration and specimen thickness as shown in
the graph
 Applications of EDX

 Because of its many advantages, EDX analysis has become common practice across
industries ranging from manufacturing or research to energy and resource management
to consumer-packaged goods.
 Product deformulation and competitor analysis
 Adhesion, bonding, delamination investigations
 Optical appearance, haze and colour problems
 Disputed claim investigations and expert witness
 Failure investigations, identification of cause
 Catalyst quality, poisoning and elemental distribution
 Product imperfections and defect analysis
 Contamination detection, isolations and identification
 Quality control, raw material and end product
 Filler, pigment, fibre, additive distribution, orientation
 Assessment of plant particulate emissions
 Construction and maintenance monitoring
❖ Uv/vis spectroscopy

Ultravoilet and visible (UV-VIS)

absorption spectroscopy is the
measurment of the attenuation
of a beam of light after it passes
through a sample or reflection
from a sample
surface.absorption measurments
can be at a single wavelength or
over an extended spectral lines.
❖UV-Vis Spectroscopy
Ultraviolet (UV)-visible spectroscopy is a type of absorption
spectroscopy in which UV-visible light is absorbed by the
molecule. Absorption of the UV-visible radiations results in the
excitation of the electrons from lower to higher energy levels.
It is useful in the identification of pure drug compounds. Many
molecules contain chromophores which will absorb specific
wavelengths of UV or visible light.
Using the Beer Lambert law, the absorption of spectra generated
from these samples at given wavelengths can be related directly
to the concentration of the sample. Normally UV and UV-VIS
spectra are recorded at high and low pH and the results of both
for the sample under question compared with known standards.

A chromophore is the part of a molecule responsible for its color

 ★ What is the principle of UV-Vis Spectroscopy?
The Principle of UV-Visible Spectroscopy is based on the
absorption of ultraviolet light or visible light by chemical
compounds, which results in the production of distinct spectra.
Spectroscopy is based on the interaction between light and matter.
When the matter absorbs the light, it undergoes excitation and de-
excitation, resulting in the production of a spectrum.
● For what kind of materials it is used?
● What can be done?
● -Identify molecules in a solid or liquid sample
● – Determine the concentration of a particular molecule in
solution
● – Characterize the absorbance or transmittance through a
liquid or solid
● — over a range of wavelengths
● – Characterize the reflectance properties of a surface or
measuring the color of a material
● – Study chemical reactions or biological processes.
● What is the working mechanism of it?

Strengths Weakness
Non Stray light
Light
Destructive scattering
Quick Geometrical-
Easy to use Considerations
Minimal
processing
Inexpensive
The relationship between the energy difference and wavelength is described by the
Planck equation.

E=hν=hc/λ

where E is the energy required to promote an electron from the ground to excited state, h is
Planck’s constant, ν is the wavenumber, c is the speed of light, and λ is the wavelength.

Planck’s equation demonstrates that the less energy needed to excite the electrons, the longer
the wavelength of the absorption band. The absorption bands are indicative of the molecular
structure of the sample and will shift in wavelength and intensity depending on the molecular
interaction and environmental conditions. These bands are typically broad and featureless due to
the numerous molecular vibrational levels associated with the electronic energy levels.

UV-Visible/NIR spectroscopy can be divided into ultraviolet, visible, and near-infrared regions of
the spectrum. The ultraviolet region is defined as 180 to 400 nm, visible between 400 and 800
nm, and the near-infrared is from 800 to 3200 nm. Near-infrared light is generally poorly absorbed
because its photon energy is insufficient to induce electronic transitions and its frequency is
greater than the natural vibration frequency of most chemical bonds. However, since the
frequency in the NIR is close to the overtone frequency of many natural vibrations, weak
substance-specific absorption bands can be detected.
Interpreting UV/Vis Spectroscopy Data

TRANSMITTANCE AND ABSORBANCE

➢ In a typical UV/vis spectroscopy measurement, we
are measuring those photons that are not absorbed or
scattered by the sample.
➢Figure 1 illustrates the relationship between
transmittance and absorbance; the upper plot is the
absorbance spectrum of 50 nm gold nanoparticles,
and the lower plot shows the calculated transmittance.
➢In the near IR, where the sample does not absorb
strongly, the transmittance is close to 100%. In the
UV portion of the spectrum, where the sample
absorbs strongly, the transmittance drops to around
10% or less.
Example absorption spectrum. The peak around 280 nm
requires less energy to promote electrons into the excited
state than the peak around 215 nm.
 UV-Vis Spectroscopy Applications

For Analysis of the For Water Analysis using a UV- For Evaluation of UPF for
Melting Temperature and Visible Spectrophotometer Sun Protection Fabrics
Thermodynamic with a 30 cm Cell
Parameters of a Nucleic
Acid using a UV-Visible
Spectrophotometer
Beer-Lambert law

UV spectroscopy obeys the Beer-Lambert law, which states

that when a beam of monochromatic light is passed through a
solution of an absorbing substance, the rate of decrease of intensity
of radiation with thickness of the absorbing solution is
proportional to the incident radiation as well as the concentration
of the solution.
 A molecule absorb ultraviolet radiation of frequency
, the electron in that molecule undergo transition
from lower to higher energy level.
 The energy can be calculated by the equation,
 E=hv
 The expression of Beer-Lambert law is-
A = log (I0/I) = Ecl
Where, A = absorbance
I0 = intensity of light incident upon sample cell
I = intensity of light leaving sample cell
C = molar concentration of solute
L = length of sample cell (cm.)
E = molar absorptivity

From the Beer-Lambert law it is clear that greater the number

of molecules capable of absorbing light of a given wavelength,
the greater the extent of light absorption. This is the basic
principle of UV/Vis spectroscopy.
Lambert's Law
Beer's Law
 Graph
The different sizes explain the fact that these nanoparticles have different
light absorption properties. In the present case, silver nanoprisms were grown
by adding different volumes of the silver nanoseeds solution.

 A larger amount of
Ag-nanoseeds was
used as a precursor
for making the Ag-
nanoprisms. The
shift in the peak
maximum (position
and intensity) in the
UV–vis spectra
demonstrate the
effect of size of the
Ag-NPs.
 Applications of UV-Vis spectroscopy

 Detection of functional groups

 Identification of an unknown compound
 Determination of configurations of
geometrical isomers
 Determination of the purity of a
substance
Raman spectroscopy

What is spectroscopy?
The study of molecular structure and dynamics through
the absorption emission and scattering of light .
Ramna spectroscopy
Raman spectroscopy was discovered by C.V. Raman in
1928.
•It is a spectroscopic technique used to observe vibration,
rotational and other low frequency modes in a system
•Raman spectroscopy is commonly used in chemistry to
provide a fingerprint by which molecules can be
identified.
Principle :

 When light interacts with molecules in a gas, liquid,

or solid, the vast majority of the photons are
dispersed or scattered at the same energy as the
incident photons. This is described as elastic
scattering, or Rayleigh scattering.
 A small fraction of light scattering of optical
frequencies different than the frequency of
incident photons . This described as inelastic or
raman effect .
 Emitted photon is of lower frequency than incident
photon is know as Stoke raman scattering.
 Emitted photon is of higher
frequency than incident
photons known as anti stokes
raman scattering.

Data analysis of raman

spectroscopy of cholesterol
Raman spectroscopy of
cholesterol

 The Raman spectrum of cholesterol, a representative

biological molecule.
 The spectrum is a plot of the scattered light intensity
versus its change in frequency, relative to that of the
incident light.
 The raman peak at 1440 cm−1 is Characteristics of he
CH2 and CH3 deformation vibrations, and the peak at
1670 cm−1 is due to C=C stretching vibrations.
 A sample of biological tissue contains cholesterol, these
peaks will be present in its Raman spectrum.
 The molecular structure and composition of a material
under study is encoded as a set of frequency shifts in
the Raman scattered light
Application of raman
spectroscopy

 Raman spectrum can provide a ‘fingerprint’ of a

substance from which the molecular composition can
be determined.
Biological application of raman
spectroscopy
 Raman spectroscopy has been Widely for the study of
biological system .
 Small sample requirements.
 The minimal sensitivity towards interference by water .
 The spectral detail.
 The conformational and environmental sensitivity .
 Raman spectroscopy(named after Indian physicist C. V. Raman)
★Variants of RS
At least 25 variations of Raman spectroscopy have been
developed.[8] The usual purpose is to enhance the
sensitivity (e.g., surface-enhanced Raman), to improve the
spatial resolution (Raman microscopy), or to acquire very
specific information (resonance Raman).
● Spontaneous (or far-field) RS
● Enhanced (or near-field) RS
● Non-linear RS
● Morphologically-Directed RS
Raman spectroscopy is a technique that provides
highly chemical-specific information about
samples based on the fundamental vibrational
modes of the molecules
Raman spectroscopy is a valuable tool for qualitative and
quantitative polymeric membrane characterization. The
interactions among the functional groups, chain orientation,
structural changes upon treatment/modification, and
interfacial properties of the polymeric composite are some
of the interesting information that can be revealed from the
Raman spectra.
Raman spectroscopy is suitable for the microscopic
examination of minerals, materials such as polymers and
ceramics, cells, proteins and forensic trace evidence.This
technique is being used for the characterization of large-
scale devices, mapping of different compounds and
dynamics study. It has already been used for the
characterization of graphene layers ,carbon nanotubes and
multiple other 2D materials such as MoS2 and WSe2
❏ PRINCIPLE
Raman is based on A Raman microscope begins with a standard optical microscope, and adds an
scattering.The sample is excitation laser, a monochromator or polychromator, and a sensitive detector
irradiated with a coherent (such as a charge-coupled device (CCD), or photomultiplier tube (PMT)
source, typically a laser. Most
of the radiation is easily ★ Working mechanism of RS:
scattered(Rayleigh). Small Raman looks at changes in a molecular bonds polarizability. Interaction of light
portion is inelastically with a molecule can induce a deformation of its electron cloud. This
scattered (Raman scatter deformation is known as a change in polarizability. Molecular bonds have
composed of Stokes and specific energy transitions in which a change of polarizability occurs, giving
A.stokes line) rise to Raman active modes.
How to interpret results from it?
The defects and subsurface damages induced by crystal growth have a significant impact on
the functional performance of machined products. These defects and damages have a
significant impact on the mechanical, optical, or electronic performance and sustainability of
the machined workpiece Therefore, they should be reduced and eliminated as much as
possible. In fact, if these defects and damages can be accurately characterized by an efficient
and powerful method, it will be helpful to understand and deal with them.
Stacking Faults Characterization
Figure shows the Raman spectra of 3C-SiC layer, which
was grown on Si (001) and contained stacking faults,
dislocations, and antiphase boundaries at around the
interface of SiC and Si. Using the (001) face, the TO mode
is forbidden and LO mode is active Raman, while the TO
and LO bands are Raman active and forbidden using the
(110) surface. Figure 5a shows that the intensity of the TO
band was very weak at forbidden geometry. As shown in
Figure 5b, the shape of the forbidden band at the high
frequency side was almost the same as that of the allowed
band, which indicated that the asymmetric TO band
resulted from the breakdown of the wave vector and
polarization selection rules induced by the stacking faults.
Besides, broadening of the forbidden TO band at the high
frequency side was found, which revealed that the defects
and stacking faults shorten the phonon lifetime.
Fluorescence method:
Fluorescence spectroscopy is a spectroscopy method used to analyze the fluorescence properties of a sample by determining the concentration
of an analyte in a sample. This technique is widely used for measuring compounds in a solution, and it is a relatively easy method to perform
Principle:
Fluorescence describes a phenomenon where light is emitted by an atom or molecule that has absorbed light or electromagnetic radiation from
another source. In absorption, high energy light excites the system, promoting electrons within the molecule to transition from the ground state,
to an excited state
Fluorescence is used mainly for measuring compounds in solution.
In fluorescence spectroscopy, a beam with a wavelength varying between 180 and ∼800 nm passes through a solution in a cuvette. We then
measure – from an angle - the light that is emitted by the sample. Example of fluorescence occurs when the absorbed radiation is in the
ultraviolet region of the spectrum, and thus invisible to the human eye, while the emitted light is in the visible region, which gives
the fluorescent substance a distinct color that can be seen only when exposed to UV light.
What does fluoresces mean?
Fluorescence is the glow you sometimes see when an object emits visible light. Some diamonds fluoresce when they are exposed to ultraviolet
(UV) rays from sources like the sun and fluorescent lamps. This can cause them to emit a bluish light or more rarely, a yellow or orang light
Why does fluoresces happen?
Fluorescence occurs when electrons go back from a singlet excited state to the ground state. But in some molecules the spins of the excited
electrons can be switched to a triplet state in a process called inter system crossing. These electrons lose energy until they are in the triplet
ground state

What cause fluorescence in Raman spectroscopy?

The fluorescence interference in Raman spectroscopy may result from the compound analysed or from fluorescent impurities in the sample. It
is an absorption process that causes molecules to be excited to a higher electronic state, which requires high-energy photons.
Why is fluorescence rather than absorption used for high-sensitivity detection? Fluorescence is more sensitive because of the different ways of
measuring absorbance and fluorescence. Light absorbance is measured as the difference in intensity between light passing through the reference
and the sample.
 REFERENCES
1. https://www.sciencedirect.com/topics/materials-science/scan
2. https://www.researchgate.net/publication/330168803_Scanning_Electron_
Microscopy_SEM_A_Review?enrichId=rgreq-
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k2OTU4NzJAMTU0NjY5NzYzMDA3MA%3D%3D&el=1_x_2&_esc=publicationC
overPdf
3. Scanning Electron Microscopy (SEM) (carleton.edu)
4. https://www.researchgate.net/publication/267104201_Transmission_Electron_Microscopy_as_Best_Techniqu
e_for_Characterization_in_Nanotechnology

1. The Basics of UV-Vis Spectroscopy (agilent.com)

2. https://www.technologynetworks.com/analysis/articles/uv-vis-
spectroscopy-principle-strengths-and-limitations-and-applications-349865