BIONANOTECHNOLOGY

CH-320 (e)
Open Elective

Dr. Kalyan Sundar Ghosh

Dept. of Chemistry
 Characterization of nanomaterials
Study of the features such as size, crystalline type, composition,
chemical state, and optical, magnetic and other properties

Why characterization of nanomaterials is important?

Properties of nanomaterials vary significantly with size and shape.
Accurate measurement of size and shape is, therefore, critical for
applications

Tools used to characterize nanomaterials

Microscopic Methods
Morphology, size, structure, surface characteristics even composition
of the solid depending on the type of microscope

Scanning Electron Microscope (SEM), Transmission Electron

Microscope (TEM), Scanning Tunneling Microscope (STM), Atomic
Force Microscope (AFM), Scanning Near-field Optical Microscope
(SNOM)
Diffraction Method
Average particle size analysis as well as structural determination

X-ray diffraction (XRD), Electron Diffraction, Neutron Diffraction,

Small Angle X-ray Diffraction (SAXS), Small Angle Neutron
Scattering (SANS)

Spectroscopic Method
Chemical state analysis (bonding or charge transfer), electronic
structure (energy gaps, impurity level, band formation, transition
etc.)

UV-Vis absorption (transmission and reflection mode), FT-IR,

Atomic Absorption Spectroscopy (AAS), Raman Spectroscopy,
Luminescence Spectroscopy, Electron Spectroscopy for Chemical
Analysis (ESCA), X-ray Photoelectron Spectroscopy (XPS)

Electrical and Magnetic properties Measurement

Electron Microscopes
In optical microscopes, electromagnetic waves of appropriate
wavelength scattered from the sample are detected using a system of
focusing lenses. In Electron Microscopes, electrons are used in place
of elctromagnetic radiation and electrostatic or magnetic lenses are
used instead of glass lenses. As electrons have both particle and wave
nature, they can be used to image objects.
Advantage of using electrons is that their
wavelength can be tuned to a very small
value, just by changing their energies so
that the resolution can be enhanced. The
interactions between electrons and solid
are quite complicated due to the charge on
electrons and subsequent interaction with
electrons and ions in solid. This
interactions result into backscattering of
electrons, productions of auger electrons,
visible/UV light, X-rays etc. depending upon
the energy of electrons, type of sample and
thickness of sample. 4
 Scanning Electron Microscope
Generates image by scanning the surface of the sample in a raster
pattern, using an electron beam

Source of electron
Hot filament or

Cold cathode (cathode

emit electrons without
heating and electrons are
emitted under very high
electric field) ----- Field
Emission SEM (FE-SEM
and better image than hot
filament SEM.
Secondary electrons or backscattered electrons are detected. Due
to interaction of focused beam with solids, the backscattered
electrons are somewhat defocused resulting into lowered
resolution. To focus the electron-beam to a very small spot size,
electrostatic lens are used. Fine beam is scanned or rastered on
the sample surface by a scan generator and backscattered
electrons are collected by a detector. Signal from scan generator
along with the amplified signal from electron-collector generates
the image of sample surface. In order to avoid oxidation and
contamination of filament as well as to reduce the collisions
between air molecules and electrons, filament and sample are kept
in high vacuum chamber (~10-5 torr).

Advantages: Bulk-samples can be observed and larger sample area

can be viewed, generates photo-like images, high-resolution images
are also possible.

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Disadvantages: Samples must have
surface electrical conductivity, non-
conductive samples cannot be
analyzed directly as they get
charged due to incident electrons
and images become blurred/faulty.
So they need to be coated with a
conductive layer, like a thin metal
film of gold or platinum (<10 nm)

Sample composition determination

by Energy Dispersive Analysis of
X-rays (EXDAS)
High energy electrons striking the
sample produce characteristic X-rays of
atoms with which they interact. When
intensities of such characteristic X-rays
are compared, one can obtain the
composition analysis of the sample under
investigation.
 Transmission Electron Microscope
High resolution image is generated based on the interaction pattern
of electrons that transmit through the specimen. Very high energy
electrons (>50kV) pass through a series of electromagnetic lenses in
vacuum 10-5 torr

The various components of TEM are

electron source, condenser lens,
sample, object lens, a fluorescent
screen in the given order. The lens are
electromagnetic whose focal lengths
are varied to obtain optimized images.
Variation: Scanning Transmission Electron Microscope (STEM)
Advantages: Additional analysis techniques like X-ray spectrometry
are possible with STEM, which gives diffraction pattern and crystal
structure analysis; high-resolution,
Limitations: Needs high-vacuum chamber, sample preparation
necessary

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Sample Preparation for Electron Microscopy
For SEM, insulating samples are coated with some noble metals like
Au or Pd

Sample thickness is a limitation for TEM. Need to make thin films

deposited on solid substrate. Powder samples (particle size <300
nm) can be held on some metal (usually Cu) grid coated with a thin
(~5 nm) carbon film

Difference between SEM and TEM

SEM uses backscattered electrons from a sample for imaging
whereas TEM utilizes electrons transmitted through a sample

SEM can be used to image the surface of a thick sample but

TEM requires a thin sample (maximum thickness ~300 nm) so
that high energy electrons can transmit through it

In SEM, usually much lower electron energy is used as compared

to TEM
 Scanning Probe Microscopes
Scanning Probe Microscopy (SPM) is a common name given to a
family of microscopy in which a sharp tip of metal is scanned across
a sample surface in a raster mode to produce the image at high
resolution.

Advantages:
Could be performed in “Spectromicroscopy” ----spectroscopy of
same sample area of which microscopy is performing. One will be
able to get not only the details of morphology and structure of the
material but also can know the chemical nature or electronic
structure of the material and study mechanical, thermal, optical,
magnetic and many other physical properties too

No sample preparation like SEM and TEM

Vacuum is not necessary

Thus it is possible to investigate not only insulating but live

biological samples also.
Applications
Scanning Tunneling Microscope (STM), Atomic Force Microscope
(AFM), Scanning Near-field Optical Microscope (SNOM)
In all these microscopy scanning probe (fine tip of ~10 nm diameter)
and raster scanning is done.

SPM scans are made over few nm to 100 μm in

the horizontal plane (x-y) and about few nm
to 10 μm in vertical plane (z). Starting at one
point the scans are made by moving the tip on
a line and then moving it to the next line.
Distance between two lines determines the
step size. In order to achieve high resolution,
it is essential that the microscopes be
shielded from mechanical vibrations, external
magnetic field/electrical noises.

Limitations: Probe tip radius, image processing speed, small image

size
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 Scanning Tunneling Microscope

Based on Tunneling Principle:

When two metals say M1 and M2 are at small distances (but larger
than 10 nm) , even though their Fermi levels do not coincide,
transfer of electrons from one metal to the other is not possible.
To transfer electrons from one metal to the other, it is necessary
to overcome the potential barrier known as work function of the
material.

Work functions of metals are generally 2-5 eV and transfer of

electrons at room temperature is forbidden. However, the metals
brought in extremely close distance (only few nm, less than 10 nm)
behave differently.

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Then electrons can be transferred
from one metal to the other to
establish a common Fermi level
without going over the potential
barrier, set by work function [Fig.
(b)]

Hence electrons can tunnel from one

metal to the other to occupy state
of lower energy. This causes Fermi
levels to coincide with a small
“Contact Potential”.

Once the Fermi levels coincide, the

electrons cannot flow from one
metal to other, but however by
raising the Fermi level of one metal
w.r.t the other, electrons can tunnel
from one to other [Fig. (c)].
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Scanning Tunneling Microscope
Operation: Based on tunneling current

Modes of operation:
i. Constant-current
ii. Constant-height

Advantages: Very high image

resolution (capable of seeing and
manipulating atoms), wide temperature
range and capable of operating in
ultra-high vacuum and other gas
environments

Limitations: Radius of curvature of

tip, extremely sensitive to ambient
vibrations
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 Atomic Force Microscopy (AFM)
Limitation of Scanning Tunneling Microscopy is that the same has
to be conductor or at least a semiconductor. This limitation can be
overcome by Atomic Force Microscopy (AFM).

When two atoms are close to each other, there are attractive and
repulsive forces, which depend upon the distances of their
separation as per the following equation-

Where F is resultant force between two atoms, A and B are the

constants and R is the distance between two atoms. First term is
repulsive and second term is attractive force. Repulsive force is
more effective at very short distance and changes rapidly with
distance.
 Scanning Near-Field Optical Microscopy (SNOM)
In conventional optical microscopy, the object is first illuminated
and then the scattered light by the object is collected so as to
form the magnified image. In an optical microscope the detector
is kept at very large distance of at least few mm to cm from the
sample, which is very large compared to the wavelength of the
visible light (~ 500 nm) normally used.

The reflected light or the light leaving from a luminous object has
two components: evanescent beam and propagating beam
Evanescent bam is related to what is known as near field and the
propagating beam is related to far filed . Near field extends to
very small distance and if the near field zone is disturbed, it
affects the propagating waves. This is used for a high resolution
image by SNOM.

In STM, electrons tunnel between tip and the sample, whereas in

SNOM, photon tunnels if the distance between the probe and the
sample is very small (< 1 to 2 nm).

A special probe with very small aperture of few nm diameter is

brought very close to the sample surface. The diameter of the
aperture as well distance between aperture and sample has to be
smaller than the wavelength of light. Under such condition, light
leaves the sample before diffracting.
Resolution in SNOM will depend upon the size of the aperture and
the distance at which the probe can be placed. Using intense source
like laser, fine movement of the probe using piezoelectric drive,
high resolution is obtained in SNOM.

Probe: Optical fibers with metal coating, optical fiber is tapered at

one end forming an aperture of few nm in size.
Reflected or transmitted light carries the signature of the
near-field changes, which would be measured in far-field
mode. Observation in both reflection and transmission
geometries are possible

CURRENT RESEARCH

Challenges: Accurate characterization of nanoparticles is very

critical to study their properties. Image analysis is very tedious
and current methods are not robust, which reduces their
compatibility. Faster, robust, and more accurate methods need
to be developed

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