Aliyyah Munauwar

12017464

Experiment 3: Introduction to
Scanning Probe Microscopy
Part 1: Acquiring Images with the NanoSurf EasyScan 2
Sample 1: CD-ROM
Parameters
Image size: 50m
Time/line:
0.5s
Points/line: 256
Rotation:
0
Z-controller
Setpoint:
30%
P-gain:
1998
I-gain:
2199
D-gain:
0

Comment: The A clearer, better defined pattern is observed by reducing the

setpoint to 30% from 50% in the previous image, while maintaining the other
controller and parameters constant. Artifact is present on the CD-ROM sample as
identified in the circled ring.
Sample 2: PPMA
Parameters
Image size: 50m
Time/line:
0.5s
Points/line: 256
Rotation:
0
Z-controller
Setpoint:
50%
P-gain:
1998
I-gain:
2199
D-gain:
0

Comment: No artefact is identified in the image.

Sample 3: Graphene on silicone surface

Aliyyah Munauwar
12017464
Parameters
Image size: 50m
Time/line:
0.4s
Points/line: 256
Rotation:
0
Z-controller
Setpoint:
50%
P-gain:
1998
I-gain:
2199
D-gain:
0

Comment: No artefact is identified in the image.

Questions
a) Limiting factors to the value of force constant for the cantilever
Tip of the cantilever and surface of the sample are attracted to each other
by means of van der waals force of attraction. This force of attraction will
keep the tip of AFM close to the samples surface at a fixed distance but
not to the point that they are touching each other.
Potential difference also arises between tip of the cantilever and surface of
the sample. This is because tip of the cantilever is charged and hence,
inducing electrostatic force between the tip and the samples surface.
A very strong repulsive force is present between the tip and sample atoms
under a very minute tip-sample distance. This is due to the exchange
interactions arise from the overlapping of electronic orbitals at atomic
distances. The sample and tip of AFM are said to be in contact when this
repulsive force is dominant.
b) Set point is the main control of the Z-controller (NanoSurf AG, 2011). It can
be the controller of the tunnelling current, cantilever deflection and also
relative cantilever vibration amplitude depending on the mode of
operation. For the vibration amplitude control, if we set the Setpoint to be
50%, then the tip of the cantilever will be moved by the cotroller with the
same percentage closer to the sample until the amplitude of the cantilever
has decreased 50% of the vibration amplitude away from the sample.
There are three gains associated with the NanoSurf EasyScan2;
I-gain: the strength of the Z-controller. Increase in I-gain will
reduce error signal over time (NanoSurf AG, 2011).
P-gain: The strength of the Z-controller proportional to the
error signal. Error signal can be reduced by increasing P-gain
(NanoSurf AG, 2011).
D-gain: The strength of the Z-controller reaction proportional
to the derivative of the error signal. If we increase the D-gain,
2

Aliyyah Munauwar
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fast changes in the error signal will be decreases. However,
high frequency noise is also amplified (NanoSurf AG, 2011).
c) (i) Tapping mode is somewhere between the contact and the non-contact
modes of operation where the cantilever oscillates near its resonance
frequency. It is based on constant oscillation amplitude feedback
mechanism under the range of 10 to 100nm. The mode observed a
decrease in amplitude of the oscillation as the cantilever approaches the
sample due to the interaction of force. Adjusting the height of the
cantilever allows the amplitude to be maintained as the cantilever scans
the samples surface. Tip of the cantilever peridiocally strikes and
detaches itself from the surface; at the same time oscillating with
sufficient amplitude allows tapping mode to eliminate frictional forces.
This in turn prevents the tip of cantilever from being trapped by adhesive
meniscus forces from the surface.
(ii) Phase signal, which also known as phase imaging monitors the lag
phase between the
signal driving the cantilever to oscillate and the
cantilever oscillation output signal. Tapping mode applies the piezoelectric
driver to excite the cantilever into resonance oscillation. On
the other hand,
feedback signal for imaging is provided by the mean of changes in the
oscillation phase in the amplitude modulation.
(iii) Biological samples such as lipid bilayers are appropriate for tapping
mode AFM.
d) In order to reduce sources of noise in an AFM experiment, it is best to
conduct the
experiment in a closed room. As AFM tool is highly sensitive
to even the slightest vibration, it must be ensured that the experiment room
is covered with a high performance sound proof hood. Do not either purposely
or accidently in contact with the table holding AFM to prevent vibration. In
order to reduce mechanical or acoustic noise during the experiment, the
incubation ventilation should be turned off of reduced if theres happen to be
one presents in the room. Any object with fans should be removed from the
experimental table holding AFM.

Aliyyah Munauwar
12017464

Part 2: Finding Force Constants and Adhesion Forces Using

AFM
Exercise 1:

Frequency Sweep
0.35
0.3
0.25
0.2
Gaussian function
Amplitude (V) 0.15

Fitted Gaussian Function

0.1
0.05
0
162000

162500

163000

163500

164000

Frequency (Hz)

Figure 3: Frequency sweep for cantilever of the AFM

Plotting the frequency sweep as well as the Gaussian function of the frequency
on the same graph enabled near-accurate approximation of constants A, b and c
to be determined. Table 1 below summarises the values of the constants.
Table 1: Summary of the values of constant for Gaussian function of f(x)
Constants
Amplitude of the peak, A
Resonant frequency, b
Standard deviation of the
peak, c

Value from fitted Gaussian

function, f(x)
0.29
162960
205

Exercise 2:
Constant b and c are needed to calculate quality factor of the frequency sweep.
The quality factors together with all the constants are necessary in order to find
the spring constant of the cantilever, k by using Sader method. In addition to
that, the imaginary part of the hydrodynamic function

fi

is also required to

find k and this complicates the calculation to find k. To simplify the calculation,
an online calculator was used to calculate the spring constant. Table 2 shows the
summary of all the required parameters in finding k.
Table 2: Summary of the parameters to calculate k and k value
Parameter

Value
4

Aliyyah Munauwar
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Density of the fluid, (kg/m3)
Width of cantilever,b (m)
Length of cantilever, L (m)
Quality factor, Qf
Resonant frequency, f (Hz)
Spring constant, k (N/m)

1.18
50
450
447.4022
162960
58.8

The spring constant value obtained is considerably acceptable. The experimental

value of the spring constant and the resonant frequency however, could not be
compared to the manufacturers stated value as this value could not be obtained
from literature.
Exercise 3:

QPD Voltage vs. Distance Curve

12
10
8
QPD Voltage (V)

f(x) = 0.01x - 7.48

R = 1

4
2
0
5.00E+02
-2

1.00E+03

1.50E+03

Distance (nm)

Figure 4: QPD voltage against distance curve

AFM measures the deflection of the laser beam by means of the voltage
difference across the QPD. The voltage difference is related to the force on the
cantilever by a calibration constant which can be found from the slope of the
graph of QPD voltage against distance of the deflection of the cantilever. The
slope of the curve and the calibration constant value are summarised in table 3
below.
Table 3: Summary from the QPD voltage against distance curve
Spring constant, k (N/m)
Slope of the curve (V/m)
Calibration constant, C
(N/V)

58.8
12500000
4.7 x 10-6

Aliyyah Munauwar
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Exercise 4:

Force vs. Distance Curve

50000
40000
30000
Force (nN)

f(x) = 58.9x - 35165.33

R = 1

20000
10000
0
5.00E+02
-10000

1.00E+03

1.50E+03

Distance (nm)

Figure 5: Force against distance curve of the cantilever

By using the calibration curve found in exercise 3, force of the cantilever can be
calculated and hence enable a force-distance curve to be plotted as seen in
Figure 5. QPD voltage is directly proportional to force, linked by the calibration
constant, C. For a soft gel system, a well defined approach and retract curve can
be observed in the force-distance curve. As the result, the magnitude of
adhesion force as well as cohesion force could be determined from the plot.
However, the curve in Figure 5 only displays a single line. This is because the gel
samples had hardened over time, causing the approach and retract force to be
relatively the same which explains the single line observed in the force-distance
curve.

References
NanoSurf AG. (2011, July ). Nanosurf easyscan 2 AFM Operating Instructions for
SPM Control Software Version 3.0. Switzeland.