High resolution microscopy techniques

Electron microscopy uses a beam of accelerated electrons as a source of illumination to create an

image. The wave-particle duality of electrons is exploited.

Types of electron microscopy

 Transmission Electron Microscopy (TEM)
Parallel electron beams pass through the whole sample and the transmitted electrons produce the
magnified image. TEM provides information on the inner structure.
 Scanning transmission electron microscope (STEM)
The electron beam is focused on a specific point of the sample and the intensity of the transmitted beam is
detected as a function of the beam location. In general, if you need to look at a relatively large area and
only need surface details, SEM is ideal. If you need internal details of small samples at near-atomic
resolution, TEM will be necessary.
 Scanning electron microscope (SEM)
The electron beam is focused on a specific point of the sample and the emitted secondary electrons are
detected in the most common SEM mode. The number of secondary electrons depends on the sample
surface. The number of secondary electrons is determined at various points to create the image of the
sample surface. Thick samples can be measured. TEM has much higher resolution than SEM. SEM
allows for large amount of sample to be analysed at a time whereas with TEM only small amount of
sample can be analysed at a time. SEM also provides a 3-dimensional image while TEM provides a 2-
dimensional picture. SEM images a larger area of the sample than TEM.

Scanning Probe Microscopy (SPM) is a branch of microscopy that forms images of

surfaces using a small tip (the probe) that scans across the sample surface. This type of methods can be
used in air and liquid and provides 3-dimensional image.
 Atomic Force Microscopy (AFM) detects forces present between the surface and the tip
 Scanning Tunneling Microscope, (STM) detects the amount of electron tunneling between probe
and surface
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Transmission electron microscopy (TEM) is a high-resolution imaging technique in which
a beam of electrons passes through a thin sample to produce an image. The electron beam is impacted by
the thickness, density, composition and in some cases, crystallinity of the sample. A TEM specimen must
be thin enough to transmit sufficient electrons to form an image with minimum energy loss.
The TEM operates on the same basic principles as the light microscope or slight projector but uses
electrons instead of light.

The electrons have wave-particle duality. According to the de Broglie relationship the wavelength (λ)
of the electron is given by
ℎ
𝜆 = 𝑚𝑣
where h is the Planck constant, m denotes the electron mass and v means the velocity of the electron.
Faster moving electron has shorter wavelength. The maximal resolution of microscopy is similar to
the wavelength of light or electron. Therefore, accelerated electrons must be employed in electron
microscopy to gain an excellent resolution. For example, the electron accelerated by 1 kV has a
wavelength of ~0.039 nm. Hence, the wavelength of electrons is much smaller than that of visible
light. The optimal resolution attainable for TEM images is many orders of magnitude better than that
obtained by a light microscope.

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Atomic Force Microscope (AFM) traces the topography of samples with extremely high (up
to atomic) resolution by recording the interaction forces between the surface and a sharp tip mounted on a
tiny probe called a cantilever. AFM provides spatial information parallel and perpendicular to the surface.
In addition, local material properties such as adhesion and stiffness can be investigated by analysing
tipsample interaction forces. The forces can be classified into attractive and repulsive forces. For
attractive forces, van der Waals interaction, electrostatic force and chemical force are included. The
repulsive forces (for example electron–electron Coulomb interaction) have very short range.

The sample is mounted on a piezoelectric scanner, which ensures three-dimensional positioning with high
resolution. The force is monitored by attaching the probe to a pliable cantilever, which acts as a spring.
The forces that are exerted between the tip and the sample are measured by the amount of bending (or
deflection) of the cantilever. The amount of deflection can be correlated with a height. A laser beam is
focused on the free end of the cantilever, and the position of the reflected beam is detected by a position-
sensitive detector. AFM can operate in gas and fluid environments. It can measure physical properties
including elasticity, adhesion, hardness, friction.

AFM cantilevers and tips typically made of silicon or silicon nitride by micro fabrication techniques.

Advantages of AFM over scanning electron microscope (SEM):

 Unlike the SEM which provides a two-dimensional image of a sample, the AFM provides a three-
dimensional surface profile.
 Samples viewed by AFM do not require any special treatments that would irreversibly change or
damage the sample.
 While SEM needs an expensive vacuum environment, AFM can work well in ambient air or
liquid environment.
 AFM can provide higher resolution than SEM.

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Disadvantages of AFM over scanning electron microscope (SEM):
 SEM can image an area on the order of millimetres by millimetres with a depth on the order of
millimetres. AFM can only image a maximum height on the order of micrometres and a maximum
scanning area of around 150 by 150 micrometres.
 At high resolution, the quality of an AFM image is limited by the radius of curvature of the probe
tip.
 An incorrect choice of tip for the required resolution can lead to image artifacts.
 AFM could not scan images as fast as an SEM

Constant-force mode is generally preferred for most applications.

Modes of operation:
 Contact mode (also called static mode): The cantilever tip is dragged over the substrate
maintaining the deflection. The force (F) can be estimated according to Hooke’s law (F = k s),
from the detected variations of the cantilever bending. The displacement being the only
controllable parameter. Tip can damage soft samples and distort image data.
 Tapping mode is a type of an amplitude modulation technique, in which large-amplitude
vibrations of the AFM probe are applied for imaging. It allows for measurements to be made at
ambient conditions. When the oscillating AFM probe hits the sample, its short-time interactions
with minimal shear are less destructive than the tip-sample forces in contact mode.
 Non-contact mode The tip of the cantilever does not hit the sample surface because its oscillation
has smaller amplitude than in the tapping mode. It provides low resolution and contaminant (e.g.,
water) on the surface can interfere.

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Force modulation spectroscopy records how the force magnitude changes as a function of tip-sample
distance. In the static mode the force spectroscopic data are acquired by moving the tip along z –
direction (approach and retract), while the cantilever deflection is observed. Force spectroscopic results in
amplitude modulation mode are acquired by recording the evolution of the cantilever amplitude with the
variation of the tip-sample distance.
It gives information on the local variations of the elastic properties on the sample surface.
Lateral Force Microscopy (LFM) measures lateral deflections (twisting) of the cantilever that arise
from forces on the cantilever parallel to the plane of the sample surface.

Variations in surface friction arising from inhomogeneity in surface material.

Slope variations of surface feature. Two sharp up and down changes at both sides.

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Scanning Tunneling Microscope (STM): Tunneling current between sample and tip is
measured
A conductive very sharp tip with applied voltage is brought near the surface of a conductive sample, and
an image can be obtained based on the tunneling probability of an electron from the tip to the sample
since it is a function of distance. The current intensity vs. tip location gives the topology of the surface.
Real space imaging with atomic resolution, high vacuum and low temperature (4 K) is needed. STM is
applicable only to conducting samples. The resolution is better for STM than for AFM.

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 is the creation of opposite charges
on the sides. The effect can be reversed as well; by applying a voltage across a piezoelectric crystal, it
will elongate or compress.
These materials are used to scan the tip in an scanning tunneling microscopy (STM) and most other
scanning probe techniques. A typical piezoelectric material used in scanning probe microscopy is lead
zirconium titanate.
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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. In classical terms, if you don’t have
enough energy to move “over” a barrier, you won’t. However, in the quantum mechanical world,
electrons have wavelike properties. These waves don’t end abruptly at a wall or barrier, but taper off
quickly. If the barrier is thin enough, the probability function may extend into the next region, 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.

Scheme of electron wavefunction

The top image shows 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 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 scanning tunneling microscope: 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 can obtain the tip-sample distance.

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Scanning electron microscopy
It produces images of a sample by scanning the surface with a focused beam of electrons. The signals
used by an SEM to produce an image result from interactions of the electron beam with atoms at various
depths within the sample. Various types of signals are produced including secondary electrons (SE),
reflected or back-scattered electrons (BSE), characteristic X-rays and light (cathodoluminescence) (CL),
absorbed current (specimen current) and transmitted electrons. Secondary electron detectors are standard
equipment in all SEMs, but it is rare for a single machine to have detectors for all other possible signals.
The signal from secondary electrons tends to be highly localized at the point of impact of the primary
electron beam, making it possible to collect images of the sample surface with a resolution of below 1
nm. Back-scattered electrons (BSE) are beam electrons that are reflected from the sample by elastic
scattering. Since they have much higher energy than SEs, they emerge from deeper locations within the
specimen and, consequently, the resolution of BSE images is less than SE images. However, BSE are
often used in analytical SEM, along with the spectra made from the characteristic X-rays, because the
intensity of the BSE signal is strongly related to the atomic number (Z) of the specimen.
When the primary electron beam interacts with the sample, the electrons lose energy by repeated random
scattering and absorption within a teardrop-shaped volume of the specimen known as the interaction
volume, which extends from less than 100 nm to approximately 5 µm into the surface. The size of the
interaction volume depends on the electron's landing energy, the atomic number of the specimen and the
specimen's density. The energy exchange between the electron beam and the sample results in the
reflection of high-energy electrons by elastic scattering, emission of secondary electrons by inelastic
scattering and the emission of electromagnetic radiation, each of which can be detected by specialized
detectors. The beam current absorbed by the specimen can also be detected and used to create images of
the distribution of specimen current.

The most common imaging mode collects low-energy (<50 eV) secondary electrons that are ejected from
conduction or valence bands of the specimen atoms by inelastic scattering interactions with beam
electrons. Due to their low energy, these electrons originate from within a few nanometers below the
sample surface. The spatial resolution of the SEM depends on the size of the electron spot, which in turn
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depends on both the wavelength of the electrons and the electron-optical system that produces the
scanning beam. The resolution is also limited by the size of the interaction volume, the volume of
specimen material that interacts with the electron beam. The spot size and the interaction volume are both
large compared to the distances between atoms, so the resolution of the SEM is not high enough to image
individual atoms, as is possible with a transmission electron microscope (TEM). The SEM has
compensating advantages, though, including the ability to image a comparatively large area of the
specimen.