ATOMIC FORCE MICROSCOPY

Under Guidance of

Dr. Vandana Soni

By

Surbhi Dubey

CONTENTS

SCANNING PROBE MICROSCOPY (SPM) ATOMIC FORCE MICROSCOPE COMPONENTS OF THE MICROSCOPE Scanner Probe

Optical lever
Feedback TYPES OF FORCES TIP SURFACE INTERACTION

FORCE VS. DISTANCE CURVE

MODES OF OPERATION Contact Mode Non contact Mode Tapping mode APPLICATIONS

Scanning Probe Microscopy (SPM)

One of the powerful modern research techniques that allow

to investigate the morphology and the local properties of the solid body surface with high spatial resolution at the atomic and molecular level. In all SPM techniques a tip interacts with the sample surface through a physical phenomenon. Measuring a local physical quantity related with the interaction, allows constructing an image

of the studied surface.

Two types of SPM :

1.Scanning Tunneling Microscope (STM) first in the probe microscopes family invented in 1981 by the Swiss scientists Gerd Binnig and Heinrich Rohrer measures tunnelling currents between a probing tip and sample. limitation : requirement of sample conductivity.

2. Atomic force microscope (AFM) invented by Binning and Quate in 1986. Overcome STM limitation : can be used for conductive as well as to insulating

materials
measure forces or interactions between a sharp probing tip and sample surface .

Atomic Force Microscope

Principle :
AFM measures the forces acting between a fine tip and

a sample using special probes made by an elastic cantilever with a sharp tip on the end. Attractive or repulsive forces resulting from interactions between the tip and the surface will cause a positive or negative bending of the cantilever. The bending is detected by means of a laser beam, which

is reflected from the back side of the cantilever.

Components of the microscope

1. Scanner :
The AFM scanners are tubular piezoelement made from piezoelectric material, which expands/contracts proportional with an applied voltage and

conversely, they develop an electrical potential in response to mechanical

pressure. In this way, movements in x, y and z direction are possible. . The scanner moves the probe over the sample (or the sample under the probe) and must be able to control the position extremely accurately.

They allow obtaining large enough movements with rather small control
voltages. Tubular piezoelements are hollow thin-walled cylinders with electrodes (thin metal layers), plated on the external and internal surfaces, and the end tube faces remain uncovered tube

 The scanner itself is constructed by stacking three independent piezoelectric crystals, each of them being responsible for movements on one axis x, y or z.
There are two types of scanner configurations: 1. Scanned Tip AFM : where the piezoelectric scanner is rigidly attached to the probe and is moved over the sample surface which stands still. 2. Scanned Sample AFM : where the scanner is attached to the sample and it is moved under the tip. The diagram shows a typical scanner arrangement, with a hollow tube of piezoelectric material and the controlling electrodes attached to the surface.

2. Probe :
The probe represents a micromachined cantilever with a sharp tip at one end, which is

brought into interaction with the sample surface.

The tip is generally pyramidal or tetrahedral in shape, and usually made from silicon or

silicon nitride. In most AFMs the motion of the tip is detected by reflecting a laser off the back surface

of the cantilever. For contact mode AFM imaging, it is necessary to have a cantilever which is soft
Therefore it should have a low spring constant, this is achieved by making it thin (0.32

enough to be deflected by very small forces without damaging the sample surface or tip. m). It also needs a high resonant frequency to avoid vibrational instability, so is typically

short (100200 m). Silicon Nitride tips are used for contact mode.

Basically, two types of probes are used in AFM cantilever shaped as a beam of rectangular section (A) and triangular cantilever formed by two beams (B),.

(A)

(B)

Probes with triangular cantilever have higher stiffness and, hence, higher resonant frequencies. They are usually used in oscillating AFM techniques. V-shaped cantilevers are often used for contact mode as these can provide low resistance to vertical deflection, whilst resisting lateral torsion.

For tapping mode AFM a high spring constant is required to reduce noise and instabilities.
Rectangular cantilevers are often used for tapping mode.

 One end of the cantilever is firmly fixed on the silicon base - the holder, and the tip is located on the free cantilever end.

Cantilevers typically range from 100 to 200 m in length (l), 10 to 40 m in

width (w), and 0.3 to 2m in thickness (t), produced by photolithography and etching of silicon, SiO2 or SiN4 layers onto a silicon wafer.

The probe is placed on the end of a cantilever (which one can think of as a spring). The amount of force between the probe and sample is dependant on the spring

constant (stiffness )of the cantilever and the distance between the probe and the
sample surface. This force can be described using Hookes Law: F = k. Z F = Force k = spring constant (cantilever elastic constant) Z = the tip displacement corresponding to the bending If the spring constant (typically 0.1-1Nm) is less than the surface, cantilever bends and the deflection is monitored.

The cantilever resonant frequency is important

during AFM operation in

oscillating modes. Self frequencies of cantilever oscillations are determined by

the following formula :

l E J

is the cantilever length; the Youngs modulus; the inertia moment of the cantilever cross-section;

the material density; S the cross section;

a numerical coefficient (in the range 1100), depending on the oscillations mode.

3. Optical lever (beam deflection technique) for Detection of Cantilever Deflection

There are a number of ways to detect

the deflection of the cantilever in an AFM. The most common method is using a laser beam. A laser beam coming from a laser diode

is reflected off the back of the cantilever onto a photodetector. The sensitivity of such a system is 0.1 nm.

The photodetector consists of a split photodiode that senses the changes in

light intensity from the reflected beam due to the deflection of the cantilever following tip-sample interaction.

This system amplifies a very small cantilever deflection, such that the longer

the cantilever-detector beam path, the more amplification is provided at the

detector level.

 The diode is divided into four parts (Fig)

When the laser is displaced vertically along the positions top (B-A) and bottom (D-C), there exists a bending due to topography, while

if this movement is horizontal left (B-D)

and right (A-C), it produces a torsion due to friction (lateral force).

4. Feedback
When the tip contacts the surface directly the tip and/or surface may be

damaged. If the tip is blunted or damaged, then the imaging capability of the AFM is reduced. Soft surfaces (e.g. on biological samples) can also be easily damaged. In almost all operating modes, a feedback circuit is connected to the deflection sensor and attempts to keep the tipsample interaction constant by controlling the tipsample distance. This protects both the tip and the sample. Either the cantilever deflection (in static mode) or oscillation amplitude (in

dynamic mode) is monitored by the feedback circuit, which attempts to keep this at a setpoint value by adjusting the z height of the probe. The height of the probe is what is recorded to produce a topographic image.

The feedback system is affected by three main parameters:

Setpoint this is the value of the deflection or amplitude that the feedback circuit attempts to maintain. This is usually set such that the force on the cantilever is small, but the probe remains engaged with the surface. Feedback gains the higher these are set, the faster the feedback system will react. However if the gains are too high then the feedback circuit can become unstable and oscillate, causing high frequency noise in the image. Scan rate scanning the probe over the surface more slowly gives the feedback

circuit more time to react and results in better tracking, but this increases the time
needed to acquire an image.

What types of forces are measured?

When the tip is brought close to the sample, a number of

The combination of these interactions results in a force-distance curve similar to that below

forces may operate.

Typically the forces

contributing most to the movement of an AFM cantilever

are the coulombic and van der

Waals interactions. Coulombic Interaction: This strong, short range repulsive force arises from electrostatic repulsion by the electron clouds of the tip and sample. This repulsion increases as the separation decreases. Van der Waals interactions: These are longer range attractive forces, which may be felt at separations of up to 10 nm or more. They arise due to temporary fluctuating dipoles.

Tip Surface Interaction

In the contact regime, the cantilever is

held less than a few angstroms from the sample surface, and the interatomic force between the cantilever and the sample is repulsive.

Attractive forces near the surface are by a nanoscopic layer of

caused

contamination that is present on all

surfaces in ambient air. The amount of

contamination depends on the

environment in which the microscope is being operated.

Tip Surface Interaction

Repulsive forces increase as the probe begins to contact the surface. The repulsive forces in the AFM tend to cause the cantilever to bend up. In the non-contact regime, the

cantilever is held on the order of tens to

hundreds of angstroms from the sample surface, and the interatomic force

between the cantilever and sample is attractive (largely a result of the longrange Van der Waals interactions). In figure an experimental force vs.

distance curve is shown. It corresponds

to one cycle of the tip approaching to, getting into contact and separating from the sample.

Experimental force vs. distance curve It corresponds to one cycle of the tip approaching to, getting into contact and separating from the sample.
At the right side of the curve, the

scanner is fully retracted and the

cantilever is undeflected since the tip is not touching the sample (region I). As the scanner extends, the

cantilever remains undeflected until it comes close enough to the sample surface for the tip to experience the

attractive van der Waals force.

 In the point II, the cantilever suddenly bends slightly towards the surface. As the scanner continues to extend, the cantilever deflects away from the surface, approximately linearly (region III, red color). After full extension, at the extreme left of the plot (region III, black color), the scanner begins to retract. The

cantilever deflection retraces the same curve. In the point (IV), the scanner

retracts enough that the tip springs

free.

MODES OF OPERATION :
Contact Mode :
The tip apex is in direct contact with the surface, and the force (attractive or
repulsive) acting between the atoms of tip and sample is counterbalanced by the elastic force produced by the deflected cantilever.

The contact mode may be carried out either at constant force or at constant
average distance.

The detector monitors the changing cantilever deflection and the force is
calculated using Hookes law: F=kx (F = force, k = spring constant, x = cantilever deflection)

The feedback circuit adjusts the probe height to try and maintain a constant
force and deflection on the cantilever. This is known as the deflection set

point.

In the constant height mode the height of the tip is fixed, whereas in the constant-force mode the deflection of the cantilever is fixed and

the motion of the scanner in z-direction is recorded. By using contactmode AFM, even atomic resolution images are obtained.

Non contact Mode :

In non-contact mode the cantilever oscillates near the surface of the sample, but does not contact it. The oscillation is at slightly above the resonant frequency where the amplitude

of oscillation is typically a few nanometers (<10 nm).

Van der Waals and other long-range forces decrease the resonant frequency

just above the surface This decrease in resonant frequency causes the amplitude of oscillation to decrease. Combined with the feedback loop system maintains a constant oscillation

amplitude or frequency by adjusting the average tip-to-sample distance to construct a topographic image of the sample surface. In ambient conditions the adsorbed fluid layer is often significantly thicker than

the region where van der Waals forces are significant. So the probe is either out
of range of the van der Waals forces it attempts to measure, or becomes trapped in the fluid layer. Therefore non-contact mode AFM works best under ultra-high vacuum conditions.

Tapping mode:
Oscillates the cantilever at its resonance frequency and lightly taps the tip on the surface during scanning. For intermittent contact (tapping) mode AFM, a stiff cantilever (k

typically 1050 N/m) with a resonance frequency of 100400 kHz is

chosen. The cantilever, is excited to vibrate by an integrated piezoactuator.

Instead of deection (contact force), the amplitude of the forced

oscillating lever is detected, analyzed, and utilized in the feedback loop The electrostatic forces increase when tip gets close to the sample

surface, therefore the amplitude of the oscillation decreases. The amplitude of cantilever oscillation is sensed and feedback loop

maintains a constant oscillation amplitude by moving the scanner vertically at every x,y data point.

Advantage
High scan speeds Rough samples with extreme
changes in vertical topography Contact Mode can sometimes be scanned more easily

Disadvantage
Lateral (shear) forces may distort features in the image In ambient conditions may get strong capillary forces
due to adsorbed fluid layer

Combination of lateral and strong normal forces

reduce resolution and mean that the tip may damage the sample, or vice versa

Lateral

forces

almost

Slower scan speed than in contact mode

Tapping Mode

eliminated
Higher lateral resolution on most samples Lower forces so less damage to soft samples or tips

Lateral
Non-contact Mode eliminated

forces

almost

In ambient conditions the adsorbed fluid layer may be

too thick for effective measurements

Higher lateral resolution on

most samples

Slower scan speed than tapping and contact modes to

avoid contacting the adsorbed fluid layer

Lower forces so less damage

to soft samples or tips

APPLICATIONS

AFM allows the visualization of conducting as well as non conducting samples. High lateral and vertical resolution can be achieved in vacuum, air and liquid covered surfaces. The AFM is capable of measuring nanometer scale images of insulating surfaces with little or

no sample preparation as well as measuring three dimensional images of surfaces and studying the topography. Some possible applications of AFM are:

Substrate roughness analysis. Step formation in thin film epitaxial deposition. Pin-holes formation or other defects in oxides growth. Grain size analysis. Phase mode is very sensitive to variations in material properties, including surface stiffness, Comparing the tip-samples forces curves for materials to study the ratio of Youngs Modulus

elasticity and adhesion.

(graphite as a reference for measure of the indentation). Obtaining information of what is happening under indentation at very small loads. By In situ AFM analysis with changes in temperature we can study changes in the structure.

Pharmaceutical Applications :
It allows investigation of surface properties of tablets, podwers, capsules etc., like

frictional forces,
softness, viscoelasticity , moleular forces between single ligand/receptor pairs and the charge density on a nanometer scale
Biological processes can be studied in their native form s such as aqueous enviorment in nature. Foe example : monitoring dynamics and enzyme degradation of DNA Measurement of the adhesion forces between receptor and ligand. Chromosomes, membranes, proteins and nucleic acids can also be imaged by AFM. Anlysis of surface properties of drug delivery system which can then be modified to get desired surface properties for controlled drug delivery. Example : polymeric nanoparticles and liposomal drug formulation have been studied extensively by using AFM where their size and morphology where revealed .

 Oscillating cantilever in tapping mode has been used to reveal polymeric forms
from single crystal measurements [2] to confirm phase separation of two copolymers for drug delivery, to establish the stability of the formulation on different environments and to identify formation of amorphous domains during milling of crystalline salbutamol . The ability of AFM to operate on different conditions of temperature humidity and

in liquid allows studying the kinetics of range of phenomena. Example :

the crystal growth rate of Nifedipine, dispersed in PEG polymer matrix, was
quantified .

Also, the dissolution process of aspirin on different facets of the crystal was studied
by

showing their differences in dissolution rate . Force measurements have been used to identify the encapsulation the efficiency of
liposomes as the rigidity of encapsulating liposomes is higher.

Availability in India
Indian Institute of Sciences, Banglore IIT Delhi IIT Madras IIT Mumbai IIT Kharagpur NIPER, Mohali PSG Institute of Advanced Studies , Coimbatore

Photographs of the essential components of a sample scanning AFM: (a) scanner base, (b) cantilever holder, (c) optical head, and (d) scanner of a typical scanned sample AFM