Microscopy: Scanning

Electron Microscopy and

Transmission Electron
Microscopy

Dr. Rohan K. Kunkalekar

School of Chemical Sciences,
Goa University – Goa
Email: [email protected]
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 Scanning Electron Microscopy (SEM)
Electron microscope that produces images of a sample by scanning
the surface with a focused beam of electrons.

The electrons interact with atoms in the sample, producing various

signals that contain information about the surface topography and
composition of the sample.
The electron beam is scanned in a raster scan pattern, and the
position of the beam is combined with the intensity of the detected
signal to produce an image.

Data are collected over a selected area of the surface of the sample,
and a 2-dimensional image is generated.

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Areas ranging from ~1 cm to 5 microns in width can be imaged in a
scanning mode,
(magnification ranging from 20X to approximately 30,000X, spatial
resolution of 50 to 100 nm).
The SEM is also useful in qualitatively or semi-quantitatively
determining chemical compositions of selected point locations on
the sample.

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Principle
The SEM 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.

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Secondary electrons have very low energies on the order of 50 eV,
which limits their mean free path in solid matter.
They can only escape from the top few nanometers of the surface of
a sample.
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.
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.

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Back-scattered electrons (BSE) are beam of 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.

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.

BSE images can provide information about the distribution, but not
the identity, of different elements.

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Characteristic X-rays are emitted when the electron beam removes
an inner shell electron from the sample, causing a higher-energy
electron to fill the shell and release energy.

The energy or wavelength of these characteristic X-rays can be

measured by Energy-dispersive X-ray spectroscopy or Wavelength-
dispersive X-ray spectroscopy and used to identify and measure the
abundance of elements in the sample and map their distribution.

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SEM samples have to be small enough to fit on the specimen stage,
and may need special preparation to increase their electrical
conductivity and to stabilize them, so that they can withstand the
high vacuum conditions and the high energy beam of electrons.
Samples are generally mounted rigidly on a specimen holder or
stub using a conductive adhesive.

Nonconductive specimens collect charge when scanned by the

electron beam, and especially in secondary electron imaging mode,
this causes scanning faults and other image artifacts.

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Non-conducting materials are usually coated with an ultrathin
coating of electrically conducting material, deposited on the sample
either by low-vacuum sputter coating, electroless deposition or by
high-vacuum evaporation.
Conductive materials in current use for specimen coating include
gold, gold/palladium alloy, platinum, iridium.

Vacuums are needed to prevent electrical discharge in the gun

assembly (arcing), and to allow the electrons to travel within the
instrument unimpeded.

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Instrumentation
An electron beam emitted from an electron gun fitted with a
tungsten filament cathode.
Tungsten is normally used in thermionic electron guns
because it has the highest melting point and lowest vapor
pressure of all metals and because of its low cost.
The electron beam, with an energy ranging from 0.2 keV to 40
keV, is focused by one or two condenser lenses to a spot
about 0.4 nm to 5 nm in diameter.

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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.

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. Emission of secondary electrons,
backscattered electrons, and
characteristic X-rays from atoms
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The energy exchange between
the electron beam and the
sample results in:
Reflection of high-energy
electrons by elastic
scattering,
Emission of secondary
electrons by inelastic
scattering, and
Emission of electromagnetic
radiation, each of which can
be detected by specialized
detectors.
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Applications:
external morphology
(texture), chemical
composition, and
crystalline structure
and orientation of
materials.

Materials
Biological
Minerals
Food Industries
Forensic etc.
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 Transmission electron microscopy (TEM)
Beam of electrons is transmitted through a specimen to
form an highly-magnified image.
The specimen is most often an ultrathin section less than
100 nm thick or a suspension on a grid.

Image is formed from the interaction of the electrons with

the sample as the beam is transmitted through the specimen.

Image is then magnified and focused onto an imaging

device, such as a fluorescent screen, a layer of
photographic film.
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Instrumentation
TEM has three essential systems:
(1) An electron gun, which produces the electron beam, and the
condenser system, which focuses the beam onto the object,
Tungsten filament use as cathode.

The intensity and angular aperture of the beam are controlled by

the condenser lens system between the gun and the specimen.
A single lens may be used to converge the beam onto the object,
but, more commonly, a double condenser is employed.

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(2) Image-producing system, consisting of the objective lens,
movable specimen stage, and intermediate and projector
lenses, which focus the electrons passing through the specimen to
form a real, highly magnified image, and

(3) Image-recording system, which converts the electron image

into some form perceptible to the human eye.
It consists of a fluorescent screen for viewing and focusing the
image and a digital camera for permanent records.

In addition, a vacuum system, consisting of pumps and their

associated gauges and valves, and power supplies are required.

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The specimen holders hold a
standard size of sample grid or
self-supporting specimen.
(3.05 mm diameter, with a
thickness and mesh size ranging
from a few to 100 μm)
The sample is placed onto the
meshed area having a diameter of
approximately 2.5 mm.
Usual grid materials are copper,
molybdenum, gold or platinum.

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Transmission electron microscopes are capable of imaging at a
significantly higher resolution.
This enables the instrument to capture fine detail—even as small as a
single column of atoms, which is thousands of times smaller than a
resolvable object seen in a light microscope.
It is a major analytical method in the physical, chemical and
biological sciences.
TEMs find application in cancer research, virology, and materials
science as well as pollution, nanotechnology and semiconductor
research, but also in other fields such as paleontology and
palynology.

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