Functioning AFM and Its Application in Material Characterization

1 Introduction to Microscopes

Microscopes are instruments designed to produce magnified visual or photographic

images of small objects. The microscope must accomplish three tasks: produce a

magnified image of the specimen, separate the details in the image, and render the

details visible to the human eye or camera. This group of instruments includes not

only multiple-lens designs with objectives and condensers, but also very simple single

lens devices that are often hand-held, such as a magnifying glass.

1.1 History of the Microscope

The history of the microscope spans centuries, Roman philosophers mentioned

“burning glasses" in their writings but the first primitive microscope was not made until

the late 1300’s. Two lenses were placed at opposite ends of a tube which gave birth

to the modern microscope.

Grinding glass to use for spectacles and magnifying glasses was commonplace during

the 13th century. In the late 16th century several Dutch lens makers designed devices

that magnified objects, but in 1609 Galileo Galilei perfected the first device known as

a microscope. Dutch spectacle makers Zaccharias Janssen and Hans Lipperhey are

noted as the first men to develop the concept of the compound microscope. By placing

different types and sizes of lenses in opposite ends of tubes, they discovered that

small objects were enlarged.

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1.1.1 Lens Improvement

Later in the 16th century, Anton van Leeuwenhoek began polishing and grinding

lenses when he discovered that certain shaped lenses increased an image’s size. The

glass lenses that he created could enlarge an object many times. His lenses allowed

him to see the many microscopic animals, bacteria and intricate detail of common

objects for the first time in history. Leeuwenhoek is considered the founder of the study

of microscopy and played a vital role in the development of cell theory. The

microscope was in use for over 100 years before the next major improvement was

developed. Using early microscopes was difficult. Light refracted when passing

through the lenses and altered what the image looked like. When the achromatic lens

was developed for use in eyeglasses by Chester Moore Hall in 1729, the quality of

microscopes improved. Using these special lenses, many people would continue to

improve the visual acuity of the microscope.

1.1.2 Mechanical Improvements

During the 18th and 19th centuries, many changes occurred in both the housing

design and the quality of microscopes. Microscopes became more stable and smaller.

August Kohler is credited with inventing a way to provide uniform microscope

illumination that allowed specimens to be photographed. Ernst Leitz devised a way to

allow for different magnifications using one microscope by putting multiple lenses on

a movable turret at the end of the lens tube.

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1.1.3 Modern Technology Improving Microscopy

Charles Spencer demonstrated that light affected how images were seen. It took over

one hundred years to develop a microscope that worked without light. The first

Electron Microscope was developed in the 1930’s by Max Knoll and Ernst Ruska.

Electron microscopes can provide pictures of the smallest particles but they cannot

be used to study living things. Its magnification and resolution is unmatched by a light

microscope.

Scanning Probe Microscopy allows specimens to be viewed at the atomic level which

began first with the Scanning Tunneling Microscope invented in 1981 by Gerd Bennig

and Heinrich Rohrer. Later Bennig and his colleagues, in 1986, went on to invent

the atomic force microscope bringing about a true era of Nano research.

2 Introduction to STM (Scanning Tunneling Microscope)

The development of the family of scanning probe microscopes started with the original

invention of the STM in 1981. Gerd Binnig and Heinrich Rohrer developed the first working

STM while working at IBM Zurich Research Laboratories in Switzerland. This instrument

would later win Binnig and Rohrer the Nobel Prize in physics in 1986.

2.1 Working of STM

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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Fig 2.1.1 3D rendered Scanning Tunneling Microscope image of

atoms.

The STM is based on several principles. One is

the quantum mechanical effect of tunneling. It

is this effect that allows us to “see” the surface.

Another principle is the piezoelectric effect. It is this effect that allows us to precisely

scan the tip with angstrom-level control. Lastly, a feedback loop is required, which

monitors the tunneling current and coordinates the current and the positioning of the

tip. This is shown schematically below (in figure 2.2) where the tunneling is from tip to

surface with the tip moving in raster pattern with the help of piezoelectric in X and Y

axes and with the feedback loop maintaining a current set-point to generate a 3D

image of the electronic topography:

Fig 2.1.2

Schematic of

scanning

tunneling

microscopy

(STM).

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2.1.1`Tunneling

Tunneling is a quantum mechanical effect. A tunneling current occurs when electrons

move through a barrier that they classically shouldn’t be able to move through.

However, in the quantum mechanical world, electrons have wavelike properties.

These waves don’t end abruptly at a wall or barrier but tapers off quickly. If the barrier

is thin enough, the probability function may extend into the next region i.e. through the

barrier. Because of the small probability of an electron being on the other side of the

barrier, given enough electrons, some will indeed move through and appear on the

other side. When an electron moves through the barrier in this fashion, it is called

tunneling. Quantum mechanics tells us that

electrons have both wave and particle-like

properties. Tunneling is an effect of the

wavelike nature.

Fig 2.1.1.1 Classical Mechanics and Quantum Tunneling

Fig 2.1.1.2 Schematic of electron wave function.

The top image shows us that when an electron (the

wave) hits a barrier, the wave doesn’t abruptly end, but

tapers off very quickly – exponentially. For a thick

barrier, the wave doesn’t get past.

The bottom image shows the scenario if the barrier is quite thin (about a nanometer).

Part of the wave does get through and therefore some electrons may appear on the

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other side of the barrier. Because of the sharp decay of the probability function through

the barrier, the number of electrons that will actually tunnel is very dependent upon

the thickness of the barrier. The current through the barrier drops off exponentially

with the barrier thickness.

To extend this description to the STM: The starting point of the electron is either the

tip or sample, depending on the setup of the instrument. The barrier is the gap (air,

vacuum, liquid), and the second region is the other side, i.e. tip or sample, depending

on the experimental setup. By monitoring the current through the gap, we have very

good control of the tip-sample distance.

Fig 2.1.1.3 Feedback loop for scanning tunneling microscopy (STM)

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2.1.2 Piezoelectric Effect

The piezoelectric effect was discovered by Pierre Curie in 1880. The effect is created

by squeezing the sides of certain crystals such as Quartz or Barium Titanate. The

result creates opposite charges on both the sides. The effect can be reversed as well

by applying a voltage across a piezoelectric crystal as a result it will elongate or

compress. These materials are used to scan the tip in a scanning tunneling

microscopy (STM) and most other scanning probe techniques. A typical piezoelectric

material used in scanning probe microscopy is PZT (Lead Zirconium Titanate).

2.1.3 Feedback Loop

Electronics are needed to measure the current, scan the tip, and translate this

information into a form that we can use for STM imaging. A feedback loop constantly

monitors the tunneling current and makes adjustments to the tip to maintain a constant

tunneling current. These adjustments are recorded by the computer and presented as

an image in the STM software this type of setup is called a constant current image. In

addition, for very flat surfaces, the feedback loop can be turned off and only the current

is displayed this is called constant height image.

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2.2Advantages of STM:

 It is capable of capturing much more detail than other microscopes. This helps

researchers better understand the subject of their research on a molecular

level.

 STMs are also versatile. They can be used in ultra-high vacuum, air, water and

other liquids and gasses.

 They will operate in temperatures as low as zero Kelvin up to a few hundred

degrees Celsius.

2.3 Disadvantages of STM

 It has limitation to use conductive and semi conductive samples.

 STMs can be difficult to use effectively. There is a very specific technique that

requires a lot of skill and precision. STMs require very stable and clean

surfaces, excellent vibration control and sharp tips.

 STMs use highly specialized equipment that is fragile and expensive.

The limitation to conducting sample led the inventors to immediately think about a new

instrument that would be able to image insulating samples. In 1986 Binning, Quate

and Gerber published a paper entitled ‘Atomic Force Microscope’.

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3 Atomic Force Microscope

Analogous to how a Scanning Tunneling Microscope works, a sharp tip is raster-

scanned over a surface using a feedback loop to adjust parameters needed to image

a surface. Instead of using the quantum mechanical effect of tunneling, atomic forces

are used to map the tip-sample interaction. Often referred to as scanning probe

microscopy (SPM), there are Atomic Force Microscopy techniques for almost any

measurable force interaction – van der Waals, electrical, magnetic, and thermal. For

some of the more specialized techniques, modified tips and software adjustments are

needed. In addition to Angstrom-level positioning and feedback loop control, there are

2 components typically included in Atomic Force Microscopy: Deflection and Force

Measurement.

3.1 AFM Probe Deflection

Traditionally, most Atomic Force Microscopes use a laser beam deflection system

where a laser is reflected from the back of the reflective AFM lever and onto a position-

sensitive detector.

Fig 3.1.1 Laser beam deflection for

atomic force microscopes

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3.3 Force Measurement

Because the 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.

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.

3.4 Feedback loop

Atomic Force Microscopy has a feedback loop using the laser deflection to control the

force and tip position. As shown, a laser is reflected from the back of a cantilever that

includes the AFM tip. As the tip interacts with the surface, the laser position on the photo

detector is used in the feedback loop to track the surface for imaging and measuring.

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Fig 3.4.1 Schematic for contact mode Atomic

Force Microscopy

4 Contact Modes

Atomic force microscopy offers a multitude of different measurement modes that

enables researchers to characterize the properties of any given samples, ranging from

high resolution surface topography to electrical or magnetic properties.

4.1 Static Mode

Static mode, or contact mode, is the original and simplest mode to operate an AFM.

In this mode, the probe is in continuous contact with the sample while the probe raster

scans the surface. In other words, the probe "drags" across the sample. The most

common configuration of static mode is to operate it in constant force or deflection

feedback mode. In this mode, the cantilever deflection is the feedback parameter. The

cantilever deflection is set by the user and is related to how hard the tip pushes against

the surface so that the user controls how gentle or aggressive the interaction between

the probe and the sample is. Static mode can also be operated in constant height

mode where the probe maintains a fixed height above the sample. There is no force

feedback in this mode.

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4.2 Lateral force mode

Lateral force mode or frictional force mode is a form of static mode (contact mode). In

lateral force mode, the imaging is exactly as it is in static mode except the cantilever

scanning motion is generally performed perpendicular to the axis of the cantilever, as

opposed to a freedom of scan rotation for conventional static mode. A schematic of

the lateral force mode scan configuration is shown below.

Fig 4.2.1

Lateral Force

Mode

This mode is particularly effective for measuring the friction of a surface as the side to

side twisting of the cantilever by torque, measured as the probe raster scans along

the surface. Lateral force measurements can be converted to frictional force through

calibration of the torsional spring constant of the cantilever.

4.3 Dynamic Mode

It is also known as tapping mode, intermittent-contact mode or acoustic AC mode or

oscillating mode.

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A stiff cantilever is oscillated closer to the sample than in noncontact mode. The tip

intermittently touches or taps the surface. Very stiff cantilevers are typically used, as

tips can get stuck in the water contamination layer. Lateral forces such as drag,

common in contact mode, are virtually eliminated. For poorly adsorbed specimens on

a substrate surface, the advantage is clearly seen.

Fig 4.3.1 Schematic of dynamic AFM mode (DFM)

operation

The user should tweak 3 parameters during operation of dynamic mode:

 Cantilever spring constant:

The stiffness of the lever must be appropriately suited to image the material.

Often some empirical trial and error is required to find a suitable cantilever. If

the cantilever is too stiff, the result may be destructive to the sample or cause

tip wear. If the cantilever is too soft, it may not be able to interact with the

sample to generate any contrast or it stays in contact with the surface.

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 Free vibration amplitude:

This is the amplitude of the oscillation by the cantilever when the cantilever is

vibrating in free space away from the sample. This parameter is set in units of

voltage. Rougher samples need a larger free vibration amplitude.

 Setpoint:

This is the reduced target amplitude, which results after the tip is in intermittent

contact with the sample. The setpoint is expressed as a percentage of the free

vibration amplitude. Lower amplitude setpoints will favor a more aggressive tip-

sample interaction or a more repulsive tip-sample interaction.

4.1 Phase Contrast

In Phase Contrast mode, the phase shift of the oscillating cantilever relative to the

driving signal is measured. This phase shift can be correlated with specific material

properties that effect the tip/sample interaction. The phase shift can be used to

differentiate areas on a sample with such differing properties as friction, adhesion and

viscoelasticity. The technique is used simultaneously with Dynamic Force Mode

(DFM) mode so topography can be measured as well.

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4.2 Force Modulation

Force modulation refers to a method used to probe properties of materials through

sample/tip interactions. The tip (or sample) is oscillated at a high frequency and

pushed into the repulsive regime. The slope of the force-distance curve is measured

which is correlated to the sample’s elasticity. The data can be acquired along with

topography which allows comparison of height and material properties.

Fig 4.2.1 Force

Modulation Mode

applies a dynamic

force while in contact

mode

5 Electrical Modes for AFM

5.1 Electric Force Microscopy

EFM oscillates a conducting AFM tip to sense electric force gradients. Usually, this is

done in 2 passes – one to measure the topography in a standard dynamic mode and the

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second to “lift” a set amount above the recorded

profile to measure electric field strength. Using the

phase signal, a map of the gradient of the electric

field is created.

Fig 5.1.1Schematic of EFM operation in lift mode with a second pass

at a defined height above the surface to sense electric field strength

5.2 Kelvin Probe Force Microscopy

Like EFM, KFM also is an oscillating technique that requires a conducting

probe. Kelvin Probe Force Microscopy (KFM) mode measures the work function of the

surface. It is also known as surface potential microscopy. This mode can be done as

a lift technique (dual pass) or as a single-pass method. The single-pass method,

referred to as High Definition KFM (HD KFM), offers higher sensitivity for measuring

the surface potential plus higher spatial resolution. HD KFM requires a second lock-in

amplifier as shown below. Because the single pass does not require tip to “lift” above

the surface, resolution of the electrical signal is much stronger and provides the added

resolution.

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Fig 5.2.1 Single Pass Kelvin Probe Microscopy

5.3 Conductive AFM

Conducting AFM, or Scanning Spreading Resistance (SSR), is used to image a

material in contact mode with a conductive AFM tip. By biasing the tip (or sample) the

current is measured passing from one to the other and a map of the conductivity (or

inversely, resistivity) can be obtained.

Fig 5.3.1 Conducting mode AFM measuring

current between tip and sample

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5.4 Piezoresponse Force Microscopy

Piezoresponse Force Microscopy (PFM) mode is a Contact mode technique that maps

piezoelectric domains. PFM uses the inverse piezoelectric effect by applying a voltage

to use the resulting deformation in the material to map the piezoelectrical properties

of a surface. This method requires a conductive probe and an applied AC signal. The

amplitude and phase signals are used to measure piezoelectric orientation.

Fig 5.4.1Piezoresponse force microscopy

schematic

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6 Force Spectroscopy

AFM Force spectroscopy refers to single point measurements in which the

cantilever approaches and “pokes into” the sample, and then withdraws. During this

measurement the cantilever deflection vs. piezo movement is measured, and this can

ultimately be converted to a force vs. tip sample separation measurement that

provides mechanical information about the sample. This conversion requires the

calibration of the spring constant and deflection sensitivity. A model force curve of

cantilever deflection vs. z piezo looks as follows:

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The force curve can be divided in different segments where A-C (black line) refers to

motion where the tip is approaching the surface and D-F (gray line) is when the tip is

retracting from the surface.

 Cantilever is approaching the surface. If there are attractive or repulsive forces

between the tip and the sample, those could be measured in this segment by

the resp. down or up bending of the cantilever.

 “Snap-in”: the cantilever snaps into contact with the sample. This snap-in is due

to tip-surface interactions, like capillary forces.

 Repulsive portion: the tip and sample are in contact and bends up upon further

movement of the z-piezo. This section is referred to as the net-repulsive

portion.

 Repulsive portion on withdrawal: the tip is now unbending while being

withdrawn from the surface.

 Pull-out: the tip gets “stuck” in an adhesive dip before it is able to emerge from

the adhesion at the interface.

 The cantilever has returned to its unperturbed state while the z-piezo further

increases the tip sample distance.

6.1 Adhesion Measurements

Measuring the stickiness or adhesion of a sample with an atomic force microscope

can be done by measuring the adhesive dip in a force curve collected through force

spectroscopy. The adhesion was measured on the blend of polybutadiene and

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polystyrene revealing quantitatively different adhesive dips on the 2 materials, as

shown in the force curves below on the two materials.

Fig 6.1.1 Adhesion Measurements for polybutadiene and polystyrene

7. Paper Review

Nano-structured antimicrobial surfaces: From nature to synthetic analogues

Aaron Elbourne, Russell J. Crawford, Elena P. Ivanova

The scientific and industrial interest in antimicrobial surfaces has significantly

increased in recent times. This interest is largely in response to the persistent

microbial contamination of industrial and, importantly, Medical implant surfaces.

Bacterial contamination of implant surfaces often leads to infection at the implant

tissue interface, the treatment of these infections is becoming far more challenging.

This paper reviews the recent advances in understanding the basis of these

mechanical antimicrobial mechanisms, and discusses the progress being made

towards the fabrication of optimized, biocompatible, synthetic analogues.

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The types of antimicrobial surfaces covered are:

 Bactericidal or antifouling surface?

 Commonly used medical substrates.

 Surface coatings.

 Antibiotic coatings

 Polymer coatings

 Antimicrobial Nano-materials

Naturally occurring surfaces such as insect wings, which possess the ability to kill cells

once they have attached to the surface, represent an excellent prospect and template

for the development of synthetic antibacterial surfaces. Such surfaces contain high

aspect-ratio Nano-pillars that act mechanically to rupture any pathogenic cells

attaching to the surface, killing the microbe. The precise geometry of the surface

topographies varies between substrates, and hence the nature of these Nano-

architectures ultimately determines the extent of antimicrobial efficacy, adhesion

behavior, and biocidal selectivity towards a given microbial species. Unlike chemical-

based methods for preventing bacterial attachment, microbes cannot readily develop

resistance to this approach due to the mechanical basis of the biocidal mechanism.

This is particularly relevant for the development of biomedical implant materials, where

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pathogenic biofilms can form on substrate surfaces prior to, or during, surgical

procedures.

This also covers the manufacturing process to generate the surfaces to prevent the

bacteria or other contaminants to infect the surface.

References

 The Microscopy by Mortimer Abramowitz and Michael W. Davidson

 Atomic Force Microscopy by Peter Eaton and Paul West

 www.nanoscience.com

 www.nanosurf.com

 www.researchgate.net

 https://cosmosmagazine.com

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