Surface Innovations http://dx.doi.org/10.1680/si.13.

00043
Challenges in imaging of soft layers and
structures at solid surfaces using atomic force
microscopy
Krasowska, Niecikowska and Beattie
Themed Issue Review Paper
Received 18/12/2013 Accepted 25/02/2014
Published online 01/03/2014
Keywords: contact angle/imaging/nanostructures/soft layers/
surface imaging/tapping mode AFM

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Challenges in imaging of soft layers

and structures at solid surfaces
using atomic force microscopy
Marta Krasowska PhD* David A. Beattie PhD
Research fellow, Ian Wark Research Institute, University of South Australia, Associate professor, Ian Wark Research Institute, University of South
Adelaide, Australia Australia, Adelaide, Australia

Anna Niecikowska MSc

PhD student, Jerzy Haber Institute of Catalysis and Surface Chemistry,
Polish Academy of Sciences, Krakow, Poland

Imaging of soft, deformable fluid layers and structures, such as submicron droplets and bubbles or two-dimensional
fluid patches, at solid interfaces with an atomic force microscopy (AFM) is a very challenging task. In this article, the
authors describe the most common imaging AFM modes and advances in tapping mode AFM imaging of soft systems,
as well as discuss the most common artifacts occurring in such experiments during imaging of soft and deformable
features at solid interfaces. In general, for rigid and deformable systems, images recorded with the AFM are always a
convolution of the tip geometry and the shape of the features being imaged. With deformable samples, the size and
shape of the imaged features can additionally be affected by the imaging parameters such as set-point or gains. In
addition, AFM images can be easily misprocessed with image enhancement software, resulting in an image that does
not represent real features at the surface is created. This review will highlight the necessary procedures to ensure
successful and accurate image acquisition in soft features in AFM.

1. Atomic force microscopy
Atomic force microscopy (AFM) has been a major breakthrough
for physical, chemical, material and biosciences.1,2 The remarkable
feature of AFM is its ability to ‘see’ surface features with nanometer
or even close to atomic resolution (one needs to remember that
submicron resolution is only possible for smooth, flat and rigid
solids and involves advanced image postprocessing. Fourier
transform (FT) in some form is often used to obtain periodic pattern
giving the impression of ‘atomic resolution’ in AFM images. Such
images, for example, were obtained for graphite,3 molybdenum
disulphide and boron nitride.4 However, FT has some inherent
limitations, and these must be taken into account in order to avoid
errors. One of the main assumptions in FT is that the surface profile
is infinitely continuous and stationary (i.e. all frequency components
are present within the whole profile length). This is not fulfilled for
non-periodic surface features, for example cracks, random patterns,
droplets or particles and so on. Wavelet theory overcomes these
problems and is an effective tool for morphological analysis of the
samples with non-periodic features5–7). Unlike optical or electron
microscopes, which create a magnified two-dimensional image of
an object by focusing electromagnetic radiation (such as photons or
electrons) on its surface, AFM takes advantage of the interactions
occurring between a mechanical tip and the sample.8 It captures
the three-dimensional topography (in chosen operation modes
some physical properties can be determined, too) of a surface by
bringing the very sharp (usually 2–10 nm in diameter) tip into
close proximity with the scanned surface and then dragging the
tip across the surface, typically in a raster-like pattern.8 Depending
on the imaging mode, either the change in deflection, resonance
frequency or amplitude (and phase) is monitored by a position-
sensitive photodetector.1 The measured change is compared in a
feedback controller to some preset value (the so-called set-point).
If the measured signal is different, the feedback controller applies
a voltage to the piezo to raise or lower the sample with respect to
the cantilever, in order to restore the set-point value. The voltage
that the feedback controller applies to the piezo is proportional to
the height of surface features and is used to reconstruct the surface
topography.1
When the tip is approaching the sample surface at close separation,
surface forces start to operate. The magnitude and sign of these
forces (repulsive or attractive) vary with the tip–sample separation,
as it is schematically shown in Figure 1. From a physical point of
view, we can distinguish between the AFM modes depending on

*Corresponding author e-mail address: marta.krasowska@unisa.edu.au

1
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Tapping mode
Repulsive force
Contact mode
Non-contact mode
ON
Tip–sample separation: nm
Attractive force
Figure 1. Schematic representation of the interactions between the
atomic force microscopy tip and a scanned surface in air
the character of the forces involved in the interaction between the
AFM tip and sample, that is, whether these forces are attractive
(such as van der Waals in the order of 10−12 N) or repulsive (at
separation of 1 nm and below, the tip becomes repelled from the
surface due to Coulombic interactions, these forces are in the
order of 10−9 N). While there are currently numerous modes of
AFM operation, they can be separated into two major categories
– the static mode (e.g. contact mode) and dynamic mode (e.g.
non-contact or tapping modes). Each of these modes has certain
limitations. In the contact mode, the tip scans the sample in close
contact. The force on the tip is repulsive and in the range of 10−9
N. In contact mode, the change in deflection of the cantilever is
used to generate the topographic image of the surface.8 This mode
gives the highest lateral resolution; however, the drawback is the
high lateral force exerted on the sample. This may result in sample
damage (particularly for soft samples), removal of loosely adsorbed
species and potentially image distortion. Such high lateral force
may also result in tip damage, introducing artifacts to the AFM
image.1 Additional problems faced during contact mode imaging
under ambient laboratory conditions result from an adsorbate layer
(primarily water vapor), which is typically several nanometers
thick, forming at the sample surface. When the tip touches surfaces
with such liquid layers, capillary action forces a meniscus to form
and the cantilever is pulled toward the sample due to surface
tension.
In an attempt to avoid these problems in soft material imaging, a
new mode of operation was developed. In non-contact mode, the
total force acting between the tip and the sample is much weaker
than that used in contact mode, generally, in the range of 10−12 N.
The advantage of using such a weak force is that sample damage
is minimized (especially biological or soft samples) and the tip
2
Challenges in imaging of soft layers and
structures at solid surfaces using atomic
force microscopy
Krasowska, Niecikowska and Beattie
lifetime is extended (as the tip never contacts the sample during
the scanning process). However, due to the small force between the
tip and the sample, it is not possible to measure the deflection of
the cantilever directly for topography determination. This is why
one of the approaches is to oscillate the cantilever slightly above
its resonance frequency. Any change in the resonant frequency
can be detected by monitoring the amplitude of oscillation.8 In
the presence of attractive forces, the amplitude of oscillation
decreases. By contrast, when there is a repulsive force between
the tip and sample, the amplitude of oscillations is increased.
An alternative approach for cantilever position monitoring was
proposed by Albrecht et al.9 In this approach, often referred to
the true non-contact mode, the AFM cantilever is vibrated at
the resonant frequency and any change in resonant frequency is
tracked directly. When the tip is close enough to interact with the
sample, the force gradient acting on the tip may lead to a shift in
the resonance frequency of the oscillating cantilever. The feedback
keeps the frequency shift at a preset value, and a topographic image
of the sample surface can be obtained. Since the force sensitivity
is dependent on the quality factor Q (Q = mω0/b, with m being
the mass of the cantilever, ω0 its resonance frequency and b the
damping coefficient), of the oscillating cantilever, the frequency-
modulation detection non-contact mode provides the best images
under high vacuum, when the quality factor Q of the oscillating
cantilever is enhanced.10 In general, non-contact mode has been
believed to have worse lateral resolution (especially for imaging
performed in liquids since the quality factor Q of the cantilever is
much reduced due to the hydrodynamic interactions between the
cantilever and the liquid). However, recently, this perception has
been challenged by several groups successfully using non-contact
mode to image lipid bilayers,11 DNA molecules,12 nanometre-sized
polystyrene spheres,12 lipid-ion networks13 in a liquid environment.
Perhaps most impressive in terms of high lateral resolution non-
contact imaging is recent work on the distribution of water
molecules at mica–water interface.14,15 The major drawback of true
non-contact mode is very slow operation speed and the necessity
to use stiff cantilevers (in order to reduce the oscillation amplitude
without instabilities, such as ‘jump-in-contact’, and suppressing
cantilever thermal vibrations).
Tapping mode is probably the most popular imaging mode, and
it falls between the two modes described above. The cantilever is
oscillated at, or slightly below (usually about 5% below in order
to decrease the load force exerted on the cantilever), its resonance
frequency allowing the tip to ‘tap’ (e.g. touch for a very short period
of time) the sample in each oscillation cycle.16 This intermittent
contact lessens the damage done to both the soft sample and the tip
as well as overcomes problems associated with friction, adhesion,
electrostatic and capillary forces.1 In tapping mode, the cantilever
is oscillated with free (preset) high amplitude (usually greater than
30–100 nm) when the tip is not in contact with the sample. Such
an oscillating tip is then brought close to the sample surface, and
it begins to gently tap the surface. During scanning, the tip taps
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the surface at a frequency of 5–500 kHz (the choice of the correct
cantilever should be based on the cantilever resonant frequency
and the spring constant as well as sample type. For imaging in air,
cantilevers with higher resonance frequency and a spring constant
high enough to overcome adhesion force due to the water film
present on the sample surface should be used, while for imaging in
liquid, better performance is obtained with more flexible cantilevers
and at lower resonant frequency. When using the cantilevers
of lower spring constant, one needs to remember that larger
oscillation amplitudes are required for successful imaging. There
is a simple relationship describing energy stored in the cantilever
movement, ½kcA2 (with A being the oscillation amplitude and kc
the spring constant of the cantilever) showing that, for example, the
oscillation amplitude should be ten times greater if the cantilever
spring constant is 100 times smaller).17 On the intermittent contact
with the sample surface, the cantilever oscillation amplitude is
decreased as compared to free amplitude of oscillation due to
energy loss caused by the tip contacting the sample surface. Such
reduction in oscillation amplitude is used to obtain a topographic
image of the surface (see height image in Figure 2(a)). One of the
greatest advantages of tapping mode is the simultaneous recording
of the phase shift of the cantilever oscillation relative to driving
5·0 nm
Sample 2
Sample 1
2·0 µm Height image
0·0 nm
Oscillation
amplitude
Sample 1 Sample 2
(a)
Figure 2. Height (a) and phase (b) images of a receding thin film of
1-hexyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide on
a mica surface. Bottom part of (a): schematic representation of an
atomic force microscopy (AFM) tip operating in the tapping mode
in order to obtain topographic (height) image. Bottom part of (b):
Challenges in imaging of soft layers and
structures at solid surfaces using atomic
force microscopy
Krasowska, Niecikowska and Beattie
signal in a second acquisition channel during sample imaging. The
phase shift is directly related to the amount of energy dissipated
during the tip–sample contact. Variations in the phase of the tip
are recorded with the same resolution as the height image for the
sample area. This is how a so-called phase image is obtained (see
Figure 2(b)). Such an image provides a method of distinguishing
surface features of different moduli.17
2. Phase imaging in tapping mode AFM
and the choice of right parameters
Since a softer material leads to a larger contact on the tip–sample
encounter, and the duration of the tip–sample contact is longer on a
soft component of the sample in comparison to the harder component,
the sign of the phase shift (e.g. negative or positive) will depend on
the driving amplitude, A0, and the set-point amplitude ratio, Asp/A0
(where Asp is the set-point tapping amplitude). For example, the
phase image of the same sample recorded at large A0 and low Asp/A0
will give reversed phase contrast than the corresponding image
recorded at low A0 and high Asp/A0. Magonov et al. recorded images
of polydiethylsiloxane (PDES, low Young’s modulus) on silicon
wafer (high Young’s modulus) at constant A0 (~100 nm) and varying
Asp/A0. The images (height image on the left hand side and the phase
25˚
Sample 2
Sample 1
2·0 µm Phase image
–30˚
Phase shift
0˚ 180˚
90˚
Sample 1 Sample 2
(b)
schematic representation of an AFM tip operating in the tapping
mode in order to obtain phase image. Red solid line is the driving
force, black dashed line is the actual AFM tip response. Because two
components of the sample have different elastic characteristics, the
amount they offset the phase is different
3
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‘Light’ tapping, Asp/A00·8 − 0·9
Height Phase
(a)
‘Moderate’ tapping, Asp/A00·4 − 0·7
Height Phase
(b)
‘Hard’ tapping, Asp/A0< 0·3
Height Phase
(c)
Figure 3. Height (a) and phase (b) images of polydiethylsiloxane
patches on silicon surface in air recorded with constant A0 ~100 nm
but varying Asp/A0 ratios. In all height images, the z-scale is 400 nm.
In all phase images, the z-scale is 180°. The scan size is 30 µm × 15
µm. The images were recorded with silicone probe of kc ~40 N/m and
resonance frequency of 150–180 kHz. Data reprinted from Ref. 18 with
the permission from Elsevier, Copyright 2013
image on the right-hand side) recorded at ‘light’ tapping mode
(Asp/A0 ~0·8–0·9) are presented in Figure 3(a). As can be seen, the
topography of the surface is not well resolved (compare circled PDES
patches in Figure 3(a) and 3(b) with the same region in Figure 3(b)).
In addition, the phase image gives no contrast. Figure 3(b) presents
the same area imaged in ‘moderate’ tapping mode (Asp/A0 ~0·4–0·7).
The topographical features of the height image are well resolved.
The phase image exhibits bright (positive phase angle shift) silicon
surface with dark (negative phase angle shift) PDES patches. Figure
3(c) shows the same area as in Figure 3(a) and 3(b) imaged in ‘hard’
tapping mode (Asp/A0 < 0·3). The height image exhibits slightly smaller
features in height image (compare squared PDES patches in Figure
3(c) with those in Figure 3(b)). The phase image presents a reversed
contrast of the phase image in Figure 3(b). Magonov et al.18 stressed
the right choice of the cantilever parameters (the resonance frequency
and the spring constant), as well as the driving amplitude and the set-
point amplitude ratio, in order to obtain reliable phase images.
3. Imaging of nanobubbles and
nanodroplets on rigid surfaces
One of the first AFM images documenting the presence of air
submicroscopic bubbles (the so-called nanobubbles) at a hydrophobic
surface was published by Ishida et al.19 and Lou et al.20 These first
4
Challenges in imaging of soft layers and
structures at solid surfaces using atomic
force microscopy
Krasowska, Niecikowska and Beattie
observations of nanobubbles nucleated at the hydrophobic surfaces
were met with skepticism. The presence or absence of nanobubbles
is still disputed in the literature for a number of reasons – the first
being the predicted lifetime, for dissolution of nanobubbles should
be in tens of milliseconds range due to extremely high pressure
inside such a nanobubble (the difference in pressure, ΔP, inside the
bubble is a ratio between the surface tension at liquid–gas interface,
γ, and the radius of such bubble, r, and is described by the Young-
Laplace equation, ΔP = 2γ/r. Since the ΔP is reversibly proportional
to the radius of the bubble, the nanobubbles will have high internal
pressures. For example, for air nanobubbles in water at T 22°C, this
extra internal pressure is 1·448 MPa for a bubble of radius 100 nm,
and 14·448 MPa for a bubble of radius 10 nm. At the same time,
the theoretically predicted bubble lifetimes for such nanobubbles
are ~1 × 10−4 and ~1 × 10−6 s, respectively.21). Another controversy
is the possibility of cavitation of these bubble formation induced
by the close proximity of the AFM tip. Although an explanation of
abnormal lifetime of the nanobubbles is still absent, nanobubbles are
a topic of considerable interest. A large part of nanobubble research
focuses on the size and shape of such features as well as on their
contact angles. Since all these parameters are easily determined
from tapping mode AFM images, great care must be taken not to
induce imaging artifacts, for example ‘hard’ tapping (smaller Asp/A0
ratio) leads to a larger contact on the tip–nanobubble contact, and
the duration of the tip–sample contact is prolonged, too. This can
result in a compression of the nanobubble, and, assuming lateral
size of such nanobubble is less affected or is significantly larger than
the nanobubble height, lead to an underestimation of the nanobubble
height and an overestimation of the contact angle. Surprisingly,
there are only a very few articles discussing the correct imaging
parameters. Zhang et al.22 carefully analyze the effect of various
Asp/A0 on height images of nanobubbles formed on the highly
ordered pyrolytic graphite (HOPG) and octadecyltrichlorosilane-
modified silicon surface under a wide range of solutes (including
monovalent and multivalent salts, cationic, anionic and non-ionic
surfactants, as well as solution pH).
Figure 4 presents height images of the same nanobubbles imaged
under different Asp/A0 ratio. In Figure 4(a)–4(g), the Asp/A0 ratio is
decreased from 0·968 (for nanobubbles presented in Figure 4(a))
to 0·774 (nanobubbles presented in Figure 4(g)) and, as a result,
the height image changes quite dramatically. It is important to
note that it is the height of the nanobubbles that is changed more
significantly (since the z-scale is the same in Figure 4(a)–4(g), the
height change manifests in less bright (thus lower) nanobubble
appearance in Figure 4(g) in comparison to Figure 4(a)) than that
of the nanobubble lateral size. This is even more evident in Figure
5(a), where the cross-sections of the nanobubble imaged in Figure
4(a)–4(g) are compared. The height of the same nanobubble
imaged using the Asp/A0 ratio 0·968 is 35 nm, and this height is then
decreased to 17 nm, that is, by the factor of two, on decreasing
the Asp/A0 ratio to 0·774. A less drastic change is detected in the
measured lateral size of the nanobubble. The measured diameter
Surface Innovations Challenges in imaging of soft layers and
structures at solid surfaces using atomic
force microscopy
Krasowska, Niecikowska and Beattie

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Asp/A00·968 Asp/A00·935 Asp/A00·903 Asp/A00·871 Asp/A00·839

(a) (b) (c) (d) (e)

Asp/A00·806 Asp/A00·774 Asp/A00·839 Asp/A00·903 Asp/A00·935

(f) (g) (h) (i) (j)

Figure 4. Height images of nanobubbles formed on the
octadecyltrichlorosilane-modified silicon surface in a 0·5 CMC Tween
20 solution. The same spot of the surface was imaged under different
Asp/A0 ratio; set-point ratio was first decreased from image (a) to
image (g), and then increased from image (h) to image (j). In all height
images, the z-scale is 40 nm. The scan size for all images is 700 nm
× 700 nm and the scan rate was 3 Hz. The images were recorded
with silicon nitride probe of kc ~0·8 N/m and resonance frequency
of 6·58 kHz. Data reprinted from Ref. 22 with the permission from
American Chemical Society, Copyright 2013

Cross-section of the nanobubble Cross-section of the nanobubble

0·968
30 0·935 30 0·935
0·903
Height: nm

Height: nm

0·871
20 20 0·903
0·839
0·806
10 0·774 10 0·839

0 0
–180 –90 0 90 180 –180 –90 0 90 180
Lateral size: nm Lateral size: nm

(a) (b)

Figure 5. Cross-section of the nanobubble is shown in Figure 4; (a)
height against lateral size data obtained while decreasing the Asp/A0
ratio, which corresponds to the images in Figure 4(a)–4(g); (b) height
against lateral size data obtained while subsequently increasing the
Asp/A0 ratio, 0·935 data set corresponds to the images in Figure 4(b)
and 4(j), 0·903 data set corresponds to the images in Figure 4(c)
and 4(i) and 0·839 data set corresponds to the images in Figure 4(e)
and 4(h). Data reprinted from Ref. 22 with permission from American
Chemical Society, Copyright 2013

of the nanobubble imaged at the Asp/A0 ratio 0·968 is 315 nm; this
dimension is decreased to 306 nm, and on decreasing the Asp/A0 ratio
to 0·774, the decrease by 9 nm is not very significant (less than 3%).
Zhang et al.22 determined the contact angle for nanobubbles imaged
for different Asp/A0 ratio. Since the lateral size of the bubble is by
the order of magnitude higher than the nanobubble height, the
changes in height do not exhibit significant effect for the measured
contact angle. The contact angle of the nanobubble imaged using
the Asp/A0 ratio 0·968 equals to 151°, and a slight decrease (by 4°)
is observed on the Asp/A0 ratio decrease to 0·774. The authors also
investigated the effect of the further increase in the Asp/A0 ratio and
proved that returning to a higher values restore the height image
(compare Figure 4(h) and 4(e) for the Asp/A0 ratio 0·839, Figure 4(i)
and 4(c) for the Asp/A0 ratio 0·903 and Figure 4(j) and 4(a) for the
Asp/A0 ratio 0·935) as well as the cross-section of the nanobubble
perfectly (see Figure 6(b)).
Contact angles of the nanobubbles are of great interest as the observed
nanoscopic contact angles are significantly different (tens of degrees)

5
Surface Innovations Challenges in imaging of soft layers and
structures at solid surfaces using atomic
force microscopy
Krasowska, Niecikowska and Beattie

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50

20 100 nm 4
40
100

Height: nm
2
30

0
20
(a)
20 0 2 4 6 8
100 nm 10
100

0
0 10 20 30 40 50 60

Lateral size: nm
(b) (c)

Figure 6. (a and b) Droplets of C60 of different sizes; (c) cross- idealized (sphericap approximation) droplet profiles. Data reprinted
sections extracted from the height image for smaller (blue solid line) from Ref. 27 with the permission from American Physical Society,
and larger (red solid line) droplet. The dotted lines in (c) show the Copyright 2013

that the measured macroscopic ones. One of the reasons for such a
discrepancy may be the necessity of the modification of the Young-
Laplace equation to account for the influence of line tension.23 In
order to extract reliable line tension values, the contact angle must
be measured with a very high degree of accuracy (better than ±2°).
Although it is still arguable whether tapping mode AFM can provide
high-accuracy nanoscopic contact angle data (particularly when the
Asp/A0 ratio is randomly chosen and not kept constant as well as when
the tip convolution is not been addressed), some groups investigated
this issue. Yang et  al.24 used tapping mode AFM to determine the
line tension of the air nanobubbles at the trimethylochlorosilane-
modified silicon surfaces imaged in water. Although the authors do
not give the exact Asp/A0 ratio value, they stress the importance of
the right choice of the imaging parameters: ‘ . . . the selection of Asp
is critical to obtain a high-quality image. However, the optimal Asp
is not a constant value but varies according to the substrate. Once
the optimal Asp was selected for an individual sample, it remained
stable for several hours . . .’ Yang et al.24 scanned multiple regions on
multiple samples. The cross-sections of the nanobubbles from such
height images were extracted in order to measure nanobubble contact
angle, θ, and the radius. Then, the cos(θ) against 1/r dependency was
plotted, and the line tension was calculated from the gradient of the
regression line. Herminghaus and coworkers25,26 are the precursors of
line tension measurement using tapping mode AFM. Surprisingly,
the first experiment was carried out on small droplets (nano- and
submicron-droplets, due to lower surface tension at liquid–liquid
interface, are more difficult to image. The choice of the Asp/A0 ratio
is even more crucial than in systems containing nanobubbles, as on
‘hard’ tapping, the AFM tip may entrain some of the liquid (which
is not the case for nanobubbles and gaseous features) resulting in
serious artifacts (that will be discussed later in the manuscript)) rather
than nanobubbles. The authors successfully imaged hexaethylene
(a) (b)
Figure 7. (a and b) Height images of nanobubbles on HOPG in water;
(a) initial height image; (b) height image taken after several scans of
the area in image (a). Image size is 10 µm × 10 µm. Data reprinted
from Ref. 20 with the permission from American Vacuum Society,
Copyright 2013
glycol on a modified26 silicon wafer. However, they do not discuss
the influence of the imaging parameters so it is hard to judge what
is the error in the height and lateral size of the submicro- and micro-
droplets extracted from the AFM height images. Berg et al.27 were
the first to image the true nanodroplets (radius of the droplets ranging
between 10 and 100 nm) of fullerene (fullerene (C60) on ‘molecularly
smooth silica surface’ using tapping mode AFM (see Figure 6(a)
and 6(b)). The nanodroplets of such small spatial dimensions need
to be carefully imaged, and the authors paid particular attention
to ensure that no spurious imaging artifacts occur. They took an
alternative approach to that discussed above, and in order to avoid
irreversible deformation of the droplets, the imaging parameters were
optimized to maintain a signal phase lag of less than 1° (minimizing
the energy dissipation between the tip and the droplet and, as a result,

6
Surface Innovations Challenges in imaging of soft layers and
structures at solid surfaces using atomic
force microscopy
Krasowska, Niecikowska and Beattie

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deformation of the droplet interface). Since, in some cases, the droplet
radius is equal/close to the tip diameter, the tip convolution effects
were tested as well. Images taken with two tip sizes: 2 and 10 nm were
compared. The authors observed that there was a slight decrease in a
contact angle obtained from the cross-sections on droplets (especially
significant for droplets of small radius) for the 10 nm tip.
Figure 6(c) presents the cross-sections of smaller (blue solid
line) and larger (red solid line) C60 droplets. These profiles
were fitted with a sphericap (dotted lines in Figure 6(c)), and
a small discrepancy between the real (solid lines) and the
idealized (dotted lines) profiles can be noted close to the three
phase contact line (see insert in Figure 6(c)). This difference was
attributed to a tip size artifacts; however, one needs to remember
that in order to get negligible phase shift a ‘light’ tapping must
be used. As discussed above, ‘light’ tapping may suffer from
insufficient topography resolution (due to poor tracking of the
sample surface by the AFM tip).
4. Further artifacts due to
high loading force
As already discussed above, the Asp/A0 ratio being set to too low value
results in ‘hard’ tapping (e.g. high loading force), similar effects may
come from using the cantilever of too high spring constant. In some
of the cases, it leads to the distortion (clearly seen in the profile
shapes recorded at the Asp/A0 ratio 0·968 and 0·774 in Figure 5(a)) in
the height image. However, there are some more extreme situations

3·0 nm

200 nm 400 nm 2·0 µm

0·0 nm
(a) (b) (c)

Figure 8. Example of typical artifacts due to strong tip–liquid coupling
in the AFM height images of 1-butyl-3-methyl-imidazolium bis-
(trifluoromethylsulfonyl)imide precursor film on mica surface; (a) 1 µm
× 1 µm height image obtained with low Asp/A0 ratio; (b) 2 µm × 2 µm
height image, the white square shows the area previously imaged and
displayed in image (a); (c) 10 µm × 10 µm height image, the white
square shows the area previously imaged and displayed in image
(b). Date reprinted from Ref. 28 with the permission from American
Chemical Society, Copyright 2013

Height image

Cross-section across the white line

160

110
Height: nm

60

10

Height ∼235 nm
–40
Length ∼4 µm

0 5
Lateral size: nm
(a) (b)
Figure 9. (a) Height image of gas layers and nanobubbles on the from Ref. 29 with the permission from Royal Society of Chemistry,
hydrophobic stripes 40 min after water injection; (b) cross-section Copyright 2013
of the gaseous features across the white line in (a). Data reprinted

7
Surface Innovations
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in which, for example, the features can be completely removed from
the surface. Figure 7(a) and 7(b) present the images of the same area
of HOPG. The height image in Figure 7(a) was recorded initially; the
same spot has been imaged several times prior recording the height
image in Figure 7(b). It is clearly seen that the nanobubbles present
in the central part of the height image in Figure 7(a) are removed by
the tip during the second scan, and they do not appear in Figure 7(b).
Moreover, some of the nanobubbles appear larger.
The effect of too high loading force is even more evident in case
of the liquid nanodomains. Recently, Beattie et al.28 studied the
so-called precursor films formed by imidazolium-based ionic liquids
on molecularly smooth mica surface. The authors paid particular
attention, and thorough cross-checks were performed, to ensure that
no spurious imaging artifacts occured. The images were recorded
with two different tips, the Asp/A0 ratio was controlled and the images
of the precursor films were taken for small area (300 nm × 300 nm;
500 nm × 500 nm), followed by enlarging the scan area (keeping the
center of the height image fixed) to 1 µm × 1 µm, followed by 2 µm
× 2 µm. The 1 µm × 1 µm scans were then accepted as artifacts free,
only if (a) the next scan in the enlarging sequence was not showing
any visible differences from the preceding one and (b) none of the
scans showed visible signs of liquid features merging. The authors
investigated the effect of low Asp/A0 ratio (‘hard’ tapping), and the
example of typical artifacts caused by wrongly chosen set-point ratio
in the AFM topography images of 1-butyl-3-methyl-imidazolium
bis-(trifluoromethylsulfonyl)imide, precursor film on mica surface
is presented in Figure 8(a)–8(c).
The 1 µm × 1 µm scan shown in Figure 8(a) was taken first. This
was followed by collecting images over larger areas: 2 µm × 2
µm (shown in Figure 8(b)) and 10 µm × 10 µm (shown in Figure
8(c)). The white squares marked in Figure 8(b) and 8(c) indicate
the previously scanned areas shown in Figure 8(a) and 8(b),
respectively. It can be clearly seen that the imaged ionic liquid
features inside the squares, that is, inside the areas previously
imaged, look significantly different from those observed right
outside the squares (i.e. in the regions that are scanned for the first
time). The authors attribute this effect to ‘a strong coupling between
the AFM tip and the liquid leading to an ‘Ostwald-ripening’-like
effect in which the smaller patches of liquid are entrained by the
AFM tip and forcefully coalesced with larger ones.’
5. Artifacts associated with
scanners imperfections
The final group of artifacts we would like to mention is the so-called
drift in AFM height and phase images. Such artifact is caused
by ‘creep’ in the piezoelectric scanner when the scan position is
changed (i.e. when applying a set-point voltage to the piezoelectric
scanner and then trying to maintain it, in order to move to a certain
location, the piezoelectric scanner tends to continue to move in
the same direction for a certain period of time). Essentially, this
8
Challenges in imaging of soft layers and
structures at solid surfaces using atomic
force microscopy
Krasowska, Niecikowska and Beattie
artifact is easy to avoid, usually it is enough to wait until the image
starts to appear more normal over time. An example of such an
image distortion is presented in Figure 9(a). Belova et  al.29 were
investigating a time effect on the evolution of thin gaseous films on
patterned silicon surfaces. The authors were aware of the distorted
top part of the height image due to scanner creep. However, since
they needed high time resolution, they decided to proceed with the
imaging process and use undistorted parts of image to obtain height
and lateral size of thin films and gaseous features (see Figure 9(b),
where the cross-section of the undistorted gas feature is presented).
6. Conclusion
AFM is unparalleled in its ability to image and visualize soft
deformable material, whether it be droplets and bubbles, ultrathin
layers of fluid or molecular layers/structures of surfactants,
polymers and biomolecules. The successful image acquisition of
these systems requires a careful attention to surface forces and the
nature of tip–surface interactions in modern instruments. Without
correct parameter setting for tip deflection and tip oscillation
frequency feedback, image artifacts can dominate the acquired
data. This review has highlighted the most critical aspects of
parameter setting for contact and non-contact imaging, in addition
to tapping mode (including phase imaging). Finally, the importance
of incorrect loading force on the observation of image artifacts has
been illustrated with examples of tip–sample interactions altering
the appearance of liquid droplets and air bubbles on solid surfaces.
Acknowledgements
The authors would like to thank Prof. Kazimierz Malysa for his
inspiring discussions. Marta Krasowska acknowledges the financial
support from the Swiss National Science Foundation Sinergia scheme,
grant ‘Designing interactions across interfaces in ionic liquids’,
Anna Niecikowska acknowledges Polish Ministry of Sciences and
Higher Education grant 1794/B/H03/2010/39 and David A. Beattie
acknowledges the financial support from the Australian Research
Council (ARC) Future Fellowship scheme, grant FT 100100393.
This study was performed in part at the South Australian node of the
Australian National Fabrication Facility, a company established under
the National Collaborative Research Infrastructure Strategy to provide
nano- and micro-fabrication facilities for Australia’s researchers.
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