Atomic force microscopy (AFM)

AFM is a technique to measure surface topography on a scale from

angstroms to 100 microns. Binnig, Quate and Gerber invented the first AFM in 1986 and

Gerd Binnig with Heinrich Rohrer developed the precursor to the atomic force microscopy

(AFM), called, the scanning tunneling microscope (STM) that earned them the Nobel Prize

for Physics in 1986.

AFM is operated by measuring attractive or repulsive forces between a tip and the

sample. Depending on the situation, forces that are measured in AFM include mechanical

contact force, Van der Waals forces, capillary forces, chemical bonding, electrostatic

forces, magnetic forces [in Magnetic force microscope (MFM)], etc. A schematic view of

an AFM system is shown in Fig. 1. The technique involves imaging a sample using a

probe, or tip. The tip is located at the free end of a cantilever that is 100 or 200 μm long.

The tip is typically made of silicon or silicon nitride with a tip radius of curvature on the

order of nanometers (~ 10 – 30 nm). The tip (or the sample) is scanned using piezoelectric

tubes. Piezoelectric scanners are designed to be moved precisely in any of the three

perpendicular axes (x,y,z). When the tip is brought into proximity of a sample surface,

forces between the tip and the sample lead to a deflection of the cantilever according to

Hooke's law. This deflection is measured using the position of a laser light reflected from

the cantilever head as the angular deflection of the cantilever causes a twofold larger

angular deflection of the reflected light. A two or a four-segment position sensitive photo-

detector detects the laser spot. By measuring the difference signal of the photo-detector

segments, changes in the bending of the cantilever can be measured. The measurements of

cantilever deflections allow a computer to generate a map of surface topography. A

feedback signal of the laser spot position at the photo-detector is sent to the piezoelectric

stage, which controls the movement of the sample under the tip.

Photodetector

B
Mirror D
A
Laser C

Control Unit

Cantilever
with tip

Scanner
tube

Fig. 1: Working principle of atomic force microscopy (AFM)

The AFM can be operated in two basic modes (a) with feedback control and (b)

without feedback control. If the electronic feedback is switched on, then the positioning

piezo, which is moving the sample (or tip) up and down can respond to any change in

force, which is detected, and alter the tip-sample separation to restore the force to a pre-

determined value. This mode of operation is known as constant force or height mode. If the

feedback electronics are switched off, then the microscope is said to be operating in

constant height or deflection mode. This is particularly useful for imaging very flat samples
at high resolution. The way in which image contrast is obtained can be achieved in many

ways.

The three main classes of interaction of tip to sample surface are contact mode,

tapping mode and non-contact mode. Contact mode is the most common method of

operation of the AFM. As the name suggests, the tip and sample remain in close contact as

the scanning proceeds. By "contact” it means that the tip sample interaction is in the

repulsive regime of the inter-molecular force curve (see Fig. 2). One of the drawbacks of

remaining in contact with the sample is that a large lateral force exists on the sample and

the surface of the specimen may get damaged.

Force Intermittent
Contact

Contact

Non contact

Fig. 2: Variation of interatomic forces with tip-sample interaction

Tapping mode is the next most common mode used in AFM. When operated in

air or other gases, the cantilever is oscillated at its resonant frequency (often hundreds of

kilohertz) and positioned above the surface so that it only taps the surface for a very small

fraction of its oscillation period. This is still in contact with the sample in the sense defined

earlier, but the very short time over which this contact occurs means that lateral forces are
dramatically reduced as the tip scans over the surface. When imaging poorly immobilized

or soft samples, tapping mode may be a far better choice than contact mode for imaging.

Non-contact operation is another mode, which may be employed when imaging

by AFM. The cantilever must be oscillated above the surface of the sample at such a

distance that we are no longer in the repulsive regime of the inter-molecular force curve.

This is a very difficult mode to operate in ambient conditions with the AFM. The thin layer

of water contamination that exists on the surface on the sample will invariably form a small

capillary bridge between the tip and the sample and cause the tip to "jump-to-contact".

Even under liquids and in vacuum, jump-to-contact is extremely likely.

In addition to basic AFM, the instrument is capable of producing images in a

number of other modes, including magnetic force, electrical force and force-distance mode

etc. Magnetic force mode imaging utilizes a magnetic tip to enable the visualization of

magnetic domains on the sample. In electrical force mode imaging a charged tip is used to

locate and record variations in surface charge. In force-distance mode, the sample is

oscillated beneath the tip, and a series of force-distance curves are generated. This permits

the separation of sample topography, stiffness and adhesion values producing three

independent images or three individual sets of data simultaneously.

Scanning electron microscopy (SEM)

The scanning electron microscope (SEM) uses electrons rather than light to form an

image. There are many advantages of using the SEM instead of a light microscope. The

SEM has a large depth of field, which allows a large area of the sample to be in focus at

one time. The working principle of SEM is shown in the Fig. 3. When the

Primary Electron Beam

Secondary Electrons (SE)

Carrying Inner
Information of Specimen 一
一
一

Backscattered 一
Electrons (BSE)
一 SE
一 一 一
一
一 一
Excitation volume
for SE emission
Z
一 一

一
一 一
一 一
SE
Specimen

Fig. 3: Principle of scanning electron microscopy (SEM)

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 few nm to around few 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

secondary electrons, backscattered electrons, auger electrons, characteristic X-rays and

several other radiations are released from the specimen.

A beam of electrons is generated in the electron gun, located at the top of the

column. This beam is attracted through the anode, condensed by a condenser lens, and

focused as a very fine point on the sample by the objective lens. The electron beam, which

typically has an energy ranging from a few hundred eV to 40 keV, is focused by one or two

condenser lenses to a spot about 0.4 nm to 5 nm in diameter. The beam passes through

pairs of scanning coils or pairs of deflector plates in the electron column, typically in the

final lens, which deflect the beam in the x and y axes so that it scans in a raster fashion over

a rectangular area of the sample surface.

Generally, the secondary electrons are collected to form the image in the SEM

mode. The beam current absorbed by the specimen can also be detected and used to create

images of the distribution of specimen current. Electronic amplifiers of various types are

used to amplify the signals which are displayed as variations in brightness on a cathode ray

tube (CRT).