Nano-Biotechnology

Nano-materials- Characterization

2nd Feb., 2023

Disclaimer:- Study material including images is collected from various papers, book chapters and webpages. Dr. Kumar acknowledges the authors or IP rights holders.
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Material Characterization

• Composition
• Size
• Shape
• Surface morphology
• Surface charge/dispersion
• Surface modification
• Elemental analysis

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Size, shape and Surface morphology- Atomic Force
Microscopy (SPM-AFM)

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• STM (Scanning Tunneling Microscopy) is first type of SPM (Scanning
probe Microscopy)

• 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 a nanometer.

• Traditionally, most Atomic Force Microscopes use a LASER (Light

Amplification by Stimulated Emission of Radiation.) beam deflection system
where a LASER is reflected from the AFM lever and detected onto a
position-sensitive detector.

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• The AFM consists of a
cantilever with a sharp
tip (probe) at its end that
is used to scan the
specimen surface.

• The cantilever is typically

silicon or silicon nitride
(Si3N4) with a tip radius
of curvature on the order
of nanometers.

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• Atomic Force Microscope relies on the forces between the tip and
sample, these? forces impact AFM imaging.

• The force is not measured directly, but calculated by measuring the

deflection of the lever, knowing the stiffness of the cantilever.

• When the tip is brought into proximity of a sample surface, forces

between the tip and the sample lead to a deflection of the cantilever
according to Hooke's law.

• Hooke’s law gives:

F = kz

• Where F is the force, k is the stiffness of the lever, and z is the distance
the lever is bent.

• Softer cantilever– Lower the spring constant

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• Depending on the situation, forces that are measured in AFM include
mechanical contact force, van der Waals forces, capillary forces, chemical
bonding, electrostatic forces, magnetic forces (magnetic force
microscope, MFM), Casimir forces (arises from the quantum fluctuations
of the field- uncertainty principle), etc.

• The AFM can be operated in a number of modes, depending on the

application.

• In general, possible imaging modes are divided into-

o Static (also called contact) mode

o Dynamic modes
1. Non-contact
2. Tapping

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Static (also called contact) mode

Dynamic modes

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Contact mode

• The tip is "dragged" across the surface of the sample and the contours of
the surface are measured either using the deflection of the cantilever
directly or using the feedback signal (keeping the cantilever at a constant
position).

Non-contact mode

• In this mode AFM does not suffer from tip or sample degradation that
are sometimes observed after taking numerous scans with contact AFM.

• This makes non-contact AFM preferable to contact AFM for measuring soft
samples, e.g. biological samples and organic thin film.

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• An AFM operating in contact mode tip may penetrate the liquid layer to
image the underlying surface, whereas in non-contact mode tip will stay
above the adsorbed fluid layer to image both the liquid and surface.

Tapping mode

• In tapping mode, the cantilever is driven to oscillate up and down

• This oscillation is commonly achieved with a small piezo element in the

cantilever holder

• The amplitude of this oscillation usually varies from several nm to 200 nm

• The interaction of forces acting on the cantilever when the tip comes close
to the surface, forces cause the change in the amplitude of the
cantilever's oscillation as the tip gets closer to the sample.

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Biological applications

• Force spectroscopy is used in biophysics to measure the mechanical

properties of living material (such as tissue or cells).

• AFM is a powerful tool to study the cell surface – topography, elasticity,

viscosity, interactions and also the viscoelastic behavior of the cell.

• AFM is used for mechanically unfolding proteins. Mean unfolding forces

with the appropriate model leads to the obtainment of the information about
the unfolding rate and free energy profile parameters of the protein.

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