Characterization Techniques
for Nanoparticles
Chapter TWO
Characterization Techniques
Two types of nanomaterial characterization:
• Spectroscopic methods
• i.e. UV-VIS, DLS
• Imaging methods
• i.e. TEM, SEM, AFM
UV-VIS spectrum SEM image
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Characterization Techniques
• Spectroscopic methods: is the general field that measures and interprets
the electromagnetic spectra that result from the interaction between
electromagnetic radiation and matter as a function of the wavelength or
frequency of the radiation.
Characterization Techniques
• Imaging methods: Microscopy techniques can provide the necessary
information by acquiring an image of the material and the morphology of
the sample is analyzed with the help of this image at micro or nanoscale.
• Different microscopic techniques have been developed over the years
ranging from optical microscopes to electron microscopes and scanning
probe microscopes for the analysis of material morphology on varying
length scales.
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Nanoparticles:SEM
Color of a nanoparticle solution is dependent on
nanoparticle size.
UV-Vis Absorption
Gives quantitative measure of color.
What wavelengths are absorbed?
What wavelengths are transmitted?
Gold Nanoparticles (left) A = sphereical, B = urchin shaped
UV-Vis spectrum (right) red solution absorbs green light and allows red light to be transmitted to our eyes while blue
solution absorbs red light and allows the blue end of the spectrum to be transmitted to our eyes.
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UV-Vis Spectrometer
UV-Vis Spectrometer
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.
When matter absorbs ultraviolet radiation, the electrons present in it undergo excitation. This causes them to
jump from a ground state to an excited state.
It is important to note that the difference in the energies of the ground state and the excited state of the
electron is always equal to the amount of ultraviolet radiation or visible radiation absorbed by it.
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Dynamic Light Scattering (DLS)
DLS measures the Brownian motion of the nanoparticles
and correlates this to particle size
Dynamic Light Scattering (DLS)
Dynamic light scattering (DLS), which is also referred to as photon
correlation spectroscopy (PCS) is used to determine the nanoparticle
size in the colloidal solution.
DLS measures the hydrodynamic size of particles, by the mechanism
of light scattering from a laser that passes through colloidal solution
and analyzes modulation of the intensity of scattered light as a
function of time.
Brownian motion of particles correlates with their hydrodynamic
diameter.
The smaller the particle, the faster it will diffuse than a larger one
and the DLS instrument will generate a correlation function that is
mathematically linked with particle size and its time-dependent light
scattering capacity.
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Dynamic Light Scattering (DLS)
DLS instrument
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Imaging Methods
SEM micrograph
Light (Optical) Microscopy
Electron Microscopy
• TEM
• SEM
Scanning Probe Microscopy
• STM
• AFM
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Resolution Limit
Light microscopes
• 500 X to 1500 X magnification
• Resolution of ~0.2 µm
• Limits reached by early 1930’s
Resolution dependent on:
• wavelength of illumination () 0.612
d
• Numerical Aperture (NA) of lens system NA
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Electron Microscopes
Wavelength of the electron dependent on:
• Electron mass (m)
• Electron charge (q)
• Potential difference to accelerate electrons (V)
h
2mqV
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Transmission Electron Microscope
Transmission Electron Microscope
(TEM)
1. e-beam strikes sample
and is transmitted through
the sample
2. Scattering occurs
3. Un-scattered electrons
pass through sample and
are detected
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Transmission Electron Microscope
Transmission electron microscopy
(TEM) is the best technique to
determine the morphology of NPs.
It is a microscopy technique in which a
beam of energetic electrons is
transmitted through a sample and the
interaction of electrons with the sample
forms an image.
The image is then magnified and
focused onto an imaging device, such
as a fluorescent screen, a layer of
photographic film, or a sensor like a
charge-coupled device.
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Transmission Electron Microscope
TEM
Copper Grid Sample Holder
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TEM Images
Carbon Naotubes Core-Shell
Polymer
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Scanning Electron Microscope
Scanning Electron Microscope (SEM)
1. e- beam strikes sample and electron
penetrate surface
2. Interactions occur between electrons and
sample
3. Electrons and photons emitted from
sample
4. Emitted e- or photons detected
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Scanning Electron Microscope (SEM)
SEM Blood cells
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Scanning Electron Microscope (SEM)
SiO2 spheres CNT
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Scanning Electron Microscope (SEM)
A hair “split end” A “bug” eye
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Scanning Probe Microscopy
Measure feedback from atomically defined tip
Many types of feedback (dependent on tip)
AFM – Forces between sample and tip
STM – Tunneling current between sample and tip
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Scanning Tunneling Microscope
The scanning tunneling microscope (STM)
works by scanning a very sharp metal wire
tip over a surface.
By bringing the tip very close to the
surface, and by applying an electrical
voltage to the tip or sample, we can image
the surface at an extremely small scale –
down to resolving individual atoms.
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Scanning Tunneling Microscope
• Tip scans just above
surface of stage
• Electrons have a small
probability of escaping
material to tip creating
tunneling current
• Tunneling current is
depends on distance
between tip and
sample
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STM Images
• “See” individual atoms
• Must have high vacuum , low temp (4 K)
Pt and Ni atoms on an alloy surface STM at University of Wisconsin
STM is actually measuring electrons. Pt which has more electrons than Ni appears
darker in the STM image 26
Atomic Force Microscope (AFM)
Atomic force microscopy (AFM) is
a kind of scanning probe
microscopy, where a probe or tip
is used to map the contours of the
sample.
During operational mode, the tip
connected to a cantilever is
scanned over the surface of the
sample, with a small repulsive
force present between the sample
and the tip.
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Atomic Force Microscope (AFM)
1. Tip scans across surface
2. Laser reflects off of
cantilever to a
photodetector
3. Feedback loop changes
tip to sample distance
4. Height changes recorded
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Atomic Force Microscopy (AFM)
AFM Cantilevers and tips
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Atomic Force Microscopy (AFM)
2D and 3D atomic force microscopy and scanning electron microscopy
A) Protein-free chitosan nanoparticles,
B) Protein-loaded chitosan
nanoparticles at a concentration of
10% and
C) Protein-loaded chitosan
nanoparticles at a concentration of
20%.
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Atomic Force Microscopy (AFM)
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