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Electron Microscopy...
14
20
Introduction
What is Electron This booklet is written for those who
know little or nothing about electron
Microscopy? microscopy and would like to know
how an electron microscope works,
why it is used and what useful results
it can produce.
A publication of
FEI Electron Optics
FEI Electron Optics, a division of FEI
Company, is one of the world’s lead-
The Transmission ing suppliers of transmission and
scanning electron microscopes.
Electron
Our commitment to electron
Microscope microscopy dates back to the mid-
1930s, when we collaborated in EM
research programmes with universi-
ties in the UK and the Netherlands.
In 1949, the company introduced its
first EM production unit, the EM100
transmission electron microscope.
Additional
Techniques c o n t e n t s
The word is derived from the Greek mikros (small) and skopeo
(look at). Ever since the dawn of science there has been an interest
Nobody knows for certain who Why use electrons instead the objective lens in a medium with
invented the microscope. The light of light? a high refractive index (oil) gave
microscope probably developed A modern light microscope (often another small improvement but
from the Galilean telescope during abbreviated to LM) has a magnifica- these measures together only
the 17th century. One of the earliest tion of about 1000x and enables brought the resolving power
instruments for seeing very small the eye to resolve objects separated of the microscope to
objects was made by the Dutchman by 0.0002 mm (see box A). In the just under 100 nm.
Antony van Leeuwenhoek (1632- continuous struggle for better
1723) and consisted of a powerful resolution, it was found that the
convex lens and an adjustable holder resolving power of the microscope
for the object being studied (speci- was not only limited by the number
men). With this remarkably simple and quality of the lenses but also by
microscope (Fig. 1), Van Leeuwenhoek the wavelength of the light used for
may well have been able to magnify illumination. It was impossible to
objects up to 400x and with it he resolve points in the object which
discovered protozoa, spermatozoa were closer together few hundred
and bacteria and was able to classify nanometres – see box B). Using light
red blood cells by shape. with a short wavelength (blue or
ultraviolet) gave a small improve-
The limiting factor in Van ment; immersing the
Leeuwenhoek’s microscope was the specimen and
quality of the convex lens. The prob- the front of
lem can be solved by the addition of
another lens to magnify the image Resolution and
produced by the first lens. This com- Magnification (1)
pound microscope – consisting of Box A
Given sufficient light, the unaided
an objective lens and an eyepiece human eye can distinguish two points 0.2
mm apart. If the points are closer together,
together with a means of focusing,
only one point will be seen. This distance is
a mirror or a source of light and a
called the resolving power or resolution of the
specimen table for holding and eye. A lens or an assembly of lenses (a micro-
positioning the specimen – is the scope) can be used to magnify this distance and
basis of light microscopes today. enable the eye to see points even closer together
than 0.2 mm. Try looking at a newspaper
photograph or one in a magazine through a
magnifying glass for example.
4
In the 1920s it was discovered that
accelerated electrons (parts of the
atom – see box C) behave in vacuum
just like light. They travel in straight
lines and have a wavelength which is
about 100 000 times smaller than
that of light. Furthermore, it was
found that electric and magnetic
fields have the same effect on elec-
trons as glass lenses and mirrors have
on visible light. Dr Ernst Ruska at the
University of Berlin combined these
characteristics and built the first
transmission electron microscope
(often abbreviated to TEM) in 1931.
For this and subsequent work on the The Nanometre
Box D
subject, he was awarded the Nobel As distances become shorter, the
Prize for Physics in 1986. The first number of zeros after the decimal
electron microscope used two mag- point becomes larger, so microscopists
netic lenses and three years later he use the nanometre (abbreviated to nm)
added a third lens and demonstrated as a unit of length. One nanometre is a
a resolution of 100 nm (see box D), millionth of a millimetre (10 –9 metre). An
twice as good as that of the light intermediate unit is the micrometre (abbreviated
to µm) which is a thousandth of a millimetre
microscope. Today, using five
or 1000 nm.
magnetic lenses in the imaging
system, a resolving power of 0.1 nm Some literature refers to the Ångström unit (abbre-
at magnifications of over 1 million viated to Å) which is 0.1 nm and the micron for
times can be achieved. micrometre.
5
Transmission Electron Not all specimens can be made thin
Microscope (TEM) enough for the TEM. Additionally,
The transmission electron microscope there is considerable interest in
can be compared with a slide projec- observing surfaces in more detail.
tor (Fig. 2). In the latter, light from a Early attempts at producing images
light source is made into a parallel from the surface of a specimen
beam by the condenser lens; this involved mounting the specimen
passes through the slide (object) and nearly parallel to the electron beam
is then focused as an enlarged image which then strikes the surface at a
onto the screen by the objective lens. very small angle. Only a very narrow
In the electron microscope, the light region of the specimen appears in Box F
Fluorescent Screen
Specimen
Aperture (thin) Electron Beam
Electron Source
Objective Condenser
Lens Lens
Fig. 3 A modern transmission electron
microscope the Tecnai 12.
TEM
6
Light Source
(Reflected Light)
Light Beam
Specimen
Light Source
(Transmitted Light)
Electron Source
Specimen (thin)
Vacuum
(lens supplies for focusing and deflecting the beam and the high voltage
generator for the electron source), and software. A TEM from the Tecnai
The Transmission
Electron Microscope
The column is the crucial item. It The electron gun The beam emerging from the gun
comprises the same elements as The electron gun comprises a fila- is condensed into a nearly parallel
the light microscope as can be seen ment, a so-called Wehnelt cylinder beam at the specimen by the con-
from the ray paths of light and elec- and an anode. These three together denser lenses and, after passing
trons (Fig. 6). The light source of the form a triode gun which is a very through the specimen, projected as
light microscope is replaced by an stable source of electrons. The tung- a magnified image of the specimen
electron gun which is built into the sten filament is hairpin-shaped and onto the fluorescent screen at the
column. The glass lenses are replaced heated to about 2700 OC. By apply- bottom of the column.
by electromagnetic lenses and the ing a very high positive potential
eyepiece or ocular is replaced by a difference between the filament and If the specimen were not thin, the
fluorescent screen. The entire electron the anode, electrons are extracted electrons would simply be stopped
path from gun to screen has to be from the electron cloud round the and no image would be formed
under vacuum (otherwise the elec- filament and accelerated towards the (see box E "Penetration" on page 6).
trons would collide with air molecules anode. The anode has a hole in it so Specimens for the TEM are usually
and be absorbed) so the final image that an electron beam in which the 0.5 micrometres or less thick. The
has to be viewed through a window electrons are travelling at several higher the speed of the electrons, in
in the projection chamber. Another hundred thousand kilometres per other words, the higher the acceler-
important difference is that, unlike second (see box G) emerges at the ating voltage in the gun, the thicker
glass lenses, electromagnetic lenses other side. The Wehnelt cylinder the specimen that can be studied.
are variable: by varying the current which is at a different potential,
through the lens coil, the focal length bunches the electrons into a finely
(which determines the magnification) focused point (Fig. 7).
can be varied. (In the light micro- Other electron sources exist and
scope variation in magnification is these are discussed briefly under
obtained by changing the lens or by "Additional Techniques" on page 21.
mechanically moving the lens).
8
Electron Velocity
The higher the accelerating Box G
voltage, the faster the electrons.
80 kV electrons have a velocity of
Filament
150 000 km/second (1.5 x 10 8 m/s) source
which is half the speed of light. This
rises to 230 000 km/second for 300 kV
electrons (2.3 x 10 8 m/s – more than
Filament
three-quarters the speed of light).
Wehnelt
cylinder
Electron beam
High Voltage
Generator
Anode
Lamp Electrons
“Illumination”
Electro-
magnetic
Glass lens lens
Condenser lens
Box H
Electron Density
Specimen How many electrons impinge on
Electro-
magnetic the specimen? A typical electron
Glass lens lens beam has a current of about 1
Objective lens
picoampere (10 –12 A). One ampere is 1
coulomb/sec. The electron has a charge of
1.6 x 10 –19 coulomb. Therefore approxi-
First image
mately 6 million electrons per second
impinge on the specimen.
Electro-
magnetic
Glass lens lens
Projector lens
Final image
Ocular Fluorescent
screen
9
What happens in e. The impinging electrons can The electromagnetic lenses
the specimen during the cause the specimen itself to Figure 10 shows a cross-section of
electron bombardment? emit electrons (these are called an electromagnetic lens. When an
Contrary to what might be expected, secondary electrons). electrical current is passed through
most specimens are not affected by f. The impinging electrons cause the coils (C), an electromagnetic field
the electron bombardment as long the specimen to emit X-rays is created between the pole pieces
as it is kept under control. When whose energy and wavelength (P) which form a gap in the magnet-
electrons impinge on the specimen, are related to the specimen’s ic circuit. By varying the current
a number of things can happen: elemental composition. through the coils, the magnification
a. Some of the electrons are absorbed g. The impinging electrons cause of the lens can be varied. This is the
as a function of the thickness and the specimen to emit photons essential difference between the
composition of the specimen; these (or light); this is called magnetic lens and the glass lens.
cause what is called amplitude cathodoluminescence. Otherwise they behave in the same
contrast in the image. h. Finally, electrons which have lost way and have the same types of
b. Other electrons are scattered over an amount of energy because of aberration: spherical aberration (the
small angles, depending on the interaction with the specimen magnification in the centre of the
composition of the specimen; can be detected by an Energy lens differs from that at the edges),
these cause what is called phase Loss Spectrometer which is the chromatic aberration (the magnifica-
contrast in the image. equivalent of a prism in light optics. tion of the lens varies with the
c. In crystalline specimens, the elec- wavelength of the electrons in the
trons are scattered in very distinct In a standard TEM the first two beam) and astigmatism (a circle in
directions which are a function of phenomena contribute to the the specimen becomes an ellipse
the crystal structure; these cause formation of the normal TEM image in the image).
what is called diffraction contrast for non-crystalline (biological)
in the image. specimens, while for crystalline Spherical aberration is a very
d. Some of the impinging electrons specimens (most non-biological important characteristic which is
are reflected (these are called materials), phase contrast and largely determined by the lens
backscattered electrons). diffraction contrast are the most design and manufacture. Chromatic
important factors in image formation. aberration is reduced by keeping
It is necessary to add accessories or the accelerating voltage as stable as
peripheral equipment to the basic possible and using very thin speci-
microscope in order to exploit the mens. Astigmatism can be corrected
additional information which can be by using variable electromagnetic
obtained by studying the last five compensation coils.
interactions listed above.
The condenser lens system focuses
the electron beam onto the speci-
men under investigation as much
as necessary to suit the purpose.
The objective lens produces an
image of the specimen which is
then magnified by the remaining
imaging lenses and projected onto
the fluorescent screen.
p p
p p
C C
Difraction
Box I
When a wave passes through a
periodic structure whose periodicity is of
the same order of magnitude as the wavelength,
the wave emerging is subject to interference which
produces a pattern beyond the object. The phenomenon
can be observed when ocean waves pass through a regular
line of posts or when a street lamp is seen through an
umbrella. The street lamp appears as a rectangular pattern of
spots of light, bright in the centre and then getting fainter. This is Fig. 12 A special electron
caused by diffraction of light by the weave of the umbrella fabric and diffraction technique (HAADF)
the size and form of the pattern provide information about the struc- produced this spectacular image.
ture (closeness of weave and orientation). In exactly the same way
electrons are diffracted by a crystal and the pattern of spots on the
screen of the microscope gives information about the crystal lattice
(shape, orientation and spacing of the lattice planes) – see Fig. 11.
11
Vacuum
Normal atmospheric air pressure is around 760 mm of
mercury. This means that the pressure of the atmos-
phere is sufficient to support a column of mercury 760 The electronics monitor and record the operating
mm high. Modern physicists use the Pascal (Pa). To obtain the very high conditions of the microscope. This
Normal air pressure = 100 000 Pa; residual pressure in resolution of which modern results in a dramatic reduction in
the microscope = 2.5 x 10 –5 Pa. At this pressure, the TEMs are capable, the the number of control knobs
number of gas molecules per litre is about 7x10 12 and accelerating voltage and the compared with earlier models and
the chance of an electron striking a gas molecule
current through the lenses a microscope which is very easy to
while traversing the column is almost zero.
must be extremely stable. The use. Furthermore it allows special
power supply cabinet contains a techniques to be embedded in the
number of power supplies whose instrument so that the operator can
output voltage or current does not carry them out using the same con-
deviate by more than one millionth trols. The PC can be attached to a
of the value selected for a particular network to allow automatic back-ups
Box J purpose. Such stabilities require very to be made and results to be down-
sophisticated electronic circuits. loaded to other workstations. The
microscope can always be brought
Improved electron optical design up to date by installing new software
has made possible a number of or by replacing the computer with
Vacuum increasingly more complicated the latest model.
Electrons behave like light only when electron-optical techniques. This in
they are manipulated in vacuum. turn has created the need for more
As has already been mentioned, the ease of operation. Digital electronic
whole column from gun to fluores- techniques in general and micro- Fig. 16
cent screen and including the camera processor-based techniques in Chromosome Fig. 17
is evacuated. Various levels of vacuum particular play an important role from a human Woven fabric
are necessary: the highest vacuum is in this respect. Modern electron cancer cell. (SEM image).
around the specimen and in the gun; microscopes employs a fast, powerful
a lower vacuum is found in the PC (personal computer) to control,
projection chamber and camera
chamber. Different vacuum pumps
are used to obtain and maintain
these levels. The highest vacuum Fig. 15 Arrangement of
attained is of the order of a ten atoms in ceramic.
millionth of a millimetre of mercury
(see box J).
12
Specimen orientation and manip- an air lock. It is the specimen holder 0.1 micrometre) to permit electrons
ulation rod which provides the extra tilt axis to pass through. In a semiconductor
With most specimens it is not or the rotation or heating, cooling or it is sometimes desired to cut out a
sufficient to move them only in the straining, a special holder being need- section of material perpendicular to
horizontal plane. Although the ed for each purpose. the surface in order to investigate
specimen is thin, there is nevertheless a defect. This is done by ion-beam
information in the image coming Specimen preparation etching. We shall return to this in
from various depths within the speci- A TEM can be used in any branch a later section.
men. This can be seen by tilting the of science and technology where
specimen and taking stereo pho- it is desired to study the internal A technique which is gaining impor-
tographs. In order to define the axis structure of specimens down to the tance in the field of structural biology
of tilt, it is necessary to be able to atomic level. It must be possible to is that of freezing the specimen and
rotate the specimen. Crystalline make the specimen stable and small observing it in the frozen state.
specimens need to have a second tilt enough (some 3 millimetres in
axis perpendicular to the first tilt axis diameter) to permit its introduction It is beyond the scope of this booklet
in order to be able to orient a part of into the evacuated microscope to describe the many specimen
the specimen so as to obtain the column and thin enough (less than preparation techniques that are used
required diffraction pattern. These about 0.5 micrometres) to permit nowadays. What can be said is that
requirements can be fulfilled in a the passage of electrons. the preparation of a tiny specimen for
device called a goniometer. TEM is often relatively complicated
Every branch of research has its own and certainly more complicated than
The goniometer is a specimen stage specific methods of preparing the the operation of present-day TEMs.
designed to provide, in addition to X specimen for electron microscopy.
and Y translation of the specimen, tilt In biology for example, tissues are Scanning Transmission Electron
about one or two axes and rotation sometimes treated as follows: first Microscopy (STEM)
as well as Z movement (specimen there is a chemical treatment to The technique can be applied in a
height) parallel to the beam axis. remove water and preserve the tissue SEM but there is much more interest
It is usual also to provide for as much as possible in its original in applying it in TEM because of the
heating, cooling and straining state; it is then embedded in a lenses beneath the specimen which
of the specimen for spe- hardening resin; after the resin has greatly expand the number of possi-
cialised experiments in hardened, slices (sections) with an bilities for gathering information. This
the microscope. The average thickness of 0.5 micrometres technique was first demonstrated by
goniometer is mounted are cut with an instrument called an FEI in 1969. Today most TEMs can be
very close to the objec- ultramicrotome equipped with a glass equipped with this facility and what
tive lens; the specimen is or diamond knife. The tiny sections was once an attachment is now an
actually located in the thus obtained are placed on a speci- integral part of the instrument.
objective lens field between men carrier – usually a 3 mm diame-
the pole pieces because it is ter copper specimen grid which has The latest TEM from FEI offers a
there that the lens aberrations been coated with a structureless resolution of 1 nm at a magnification
are smallest and the resolution carbon film 0.1 micrometre thick. of more than 1 million times.
is highest.
In metallurgy the following method Once a scanning facility has been
The goniometer itself provides of preparation is sometimes applied: introduced into the TEM column, it
motorised X, Y and Z move- a 3 mm diameter disc of material is possible to take advantage of the
ment and tilt about one (thickness say 0.3 mm) is chemically backscattered electrons as well as of
axis. The specimen is treated in such a way that in the the secondary electrons which are
mounted near the tip of a centre of the disc the material is emitted during specimen bombardment.
rod-shaped holder which fully etched away. Around this hole
in turn is introduced into there will usually be areas that are
the goniometer through sufficiently thin (approximately
Fig. 13 A site-specific specimen from a 200 mm semiconductor wafer is thinned to about 300 nm.
Cuts are made to partially detach the section. The wafer is tilted to 60° and FIB cuts made at the
base and sides. From here, milling to final thickness will be done at low beam currents and final
"release" cuts will be made at 0° tilt.
13
A scanning electron microscope, like the TEM, consists of an electron
erably shorter because there are only three lenses to focus the electrons
into a fine spot onto the specimen; in addition there are no lenses
larger because the SEM technique does not impose any restriction on
specimen size other than that set by the size of the specimen chamber.
All the components of a SEM are in sympathy. Both the beam in the The most important differences
usually housed in one unit (Fig. 5 microscope and the one in the CRT between TEM and SEM are:
page 7). On the right is the electron- are scanned at the same rate and a. the beam is not static as in the
optical column mounted on top of there is a one to one relationship TEM: with the aid of an electro-
the specimen chamber. In the cabinet between each point on the CRT magnetic field, produced by the
below this is the vacuum system. screen and a corresponding point on scanning coils, the beam is
On the left is the display monitor, the specimen. Thus a picture is built scanned line by line over an
the keyboard and a "mouse" for up (see Fig. 18). The ratio of the size extremely small area of the spec
controlling the microscope and the of the screen of the viewing monitor imen’s surface (see box K);
camera. All the rest is below the desk (CRT) to the size of the area scanned b. the accelerating voltages are
top which gives the whole instrument on the specimen is the magnification much lower than in TEM because
its clean appearance. (see box K). Increasing the magnifi- it is no longer necessary to
cation is achieved by reducing the penetrate the specimen; in a
The electron gun at the top of the size of the area scanned on the SEM they range from 200 to
column produces and electron beam specimen. Recording is done by 30 000 volts.
which is focused into a fine spot less photographing the monitor screen c. specimens need no complex
than 4 nm in diameter on the speci- (or, more usually, a separate high preparations.
men. This beam is scanned in a resolution screen), making videoprints Box L
rectangular raster over the specimen. or storing a digital image.
Apart from other interactions at the
specimen, secondary electrons are The electron gun
produced and these are detected by The electron gun consists of a fila-
a suitable detector. The amplitude of ment and Wehnelt cylinder and is
the secondary electron signal varies the same as that in a TEM. Also the
with time according to the topogra- illumination system, consisting of
phy of the specimen surface. The sig- electron gun, anode and condenser
nal is amplified and used to cause lenses is not much different. The
the brightness of the electron beam final lens focuses the beam onto the
in a cathode ray tube (CRT) to vary surface of the specimen to be studied.
14
SEM Magnification
length of one line "written"
by the electron beam on
the monitor
SEM magnification =
length of one "track" of the
electron beam on the
specimen
Box K
Filament source
Filament
Wehnelt aperture Filament
Anode Vacuum
Wehnelt aperture
Electron Beam
Electromagnetic lens
Fig. 18
Schematic Vacuum
diagram of a
Scan
SEM showing Generator
the column and Deflection coil
how the image
is formed on
Electromagnetic lens
the monitor
Deflection
coil
Vacuum
Signal
Specimen Amplifier
A Image on
fluorescent screen
of TV monitor
B
Scanning C
Electron Microscope
In the introduction, the functioning SEM
of a SEM was compared, with some
restrictions, with a reflected light micro-
scope (Fig. 3). Another valid comparison is
illustrated in Fig. 18. The electron beam trajec-
tories in a SEM are very similar to those in a
TV monitor. Both devices have an electron gun, both are
evacuated, both have deflection coils.
The electrons produced when the primary beam strikes the specimen in
a SEM are detected and turned into an electrical signal; in the monitor,
the primary electrons are turned into light to produce an image on the
fluorescent screen.
15
What happens in the specimen the study of (dynamic) electrical
during electron bombardment? phenomena in electronic devices
When discussing the TEM, it was such as integrated circuits.
seen that when electrons strike
the specimen several phenomena Magnification and resolution
Electron Interactions occur. In general, five of these In the SEM, the magnification is
with Matter Box M phenomena are used in an ordinary entirely determined by the electronic
In the modern view of matter, SEM (see box M). circuitry that scans the beam over
an atom consists of a heavy
the specimen (and simultaneously
charged nucleus surrounded by a
number of orbiting electrons (see box a. The specimen itself emits over the fluorescent screen of the
C, page 5). The number of electrons is secondary electrons. monitor where the image appears).
equal to the number of protons in the b. Some of the primary electrons
nucleus, this is known as the atomic are reflected (backscattered Magnification can be as high as
number of the atom. The incoming electrons). 300 000x which is usually more than
electron can interact with the nucleus c. Electrons are absorbed by the sufficient. In principle the resolution
and be backscattered with virtually specimen. of a SEM is determined by the beam
undiminished energy (just as a space probe d. The specimen emits X-rays. diameter on the surface of the speci-
is deviated by the gravity of a planet during
e. The specimen sometimes emits men. The practical resolution how-
fly-past). Or it can interact with the orbit-
ing electrons: an electron may be ejected photons (= light). ever depends on the properties of
from the atom, (this is a secondary elec- the specimen and the specimen pre-
tron), and the atom with one electron All these phenomena are interrelated paration technique and on many
short restores the status quo by emitting and all of them depend to some instrumental parameters such as
its excess energy in the form of an extent on the topography, the atomic beam intensity, accelerating voltage,
X-ray quantum or a light photon. number (see box M) and the chemical scanning speed, distance from the
(See box P, page 20) state of the specimen. The number last lens to the specimen (usually
of backscattered electrons, secondary referred to as the working distance)
electrons and absorbed electrons at and the angle of the specimen
each point of the specimen depends surface with respect to the detector.
on the specimen’s topography to a Under optimum conditions a resolu-
much greater extent than the other tion of 1 nm can be attained
properties mentioned. It is for this (see box N).
reason that these three phenomena
are exploited primarily to image the Observation and recording of
specimen’s surface. the image
A SEM is usually equipped with two
Electron detection image monitors, one for observation
Detectors for backscattered elec- by the operator and the other, a
trons and secondary electrons are high resolution monitor, is equipped
usually either a scintillation detector with an ordinary photo camera
or a solid state detector. In the (which can be 35 mm or Polaroid).
former case, electrons strike a
fluorescent screen which thereupon To facilitate the observation and
emits light which is amplified and correct choice of the parameters
converted into an electrical signal by mentioned above, FEI SEMs have
a photomultiplier tube. The latter an image store in which the image is
detector works by amplifying the built up scan by scan and displayed
minute signal produced by the at TV speed so that there is a steady,
incoming electrons in a semi- flicker-free image on the viewing
conductor device. When the monitor. Images are digital and can
specimen is not directly connected be stored electronically for subse-
to earth but via a resistor, the quent enhancement and analysis.
electrons that are not reflected
generate a potential difference Image treatment
across the resistor. This changing Because the image in a SEM is com-
potential difference can be amplified pletely electronically produced, it can
and the resulting signal used to be subject to all kinds of treatment
produce a third sort of image on using modern electronics. This
the monitor. This facility also permits includes contrast enhancement,
16
Fig.19 Forensic investigation of a tungsten car-lamp filament
following an accident. Globules of melted glass on the wire
show that at the time of impact, the lamp was alight.
Photo courtesy of Dr Rabbit, Military School, Brussels
Vacuum
In general a sufficiently low vacuum
for a SEM is produced by either an
oil diffusion pump or a turbo-
molecular pump in each case backed
by a rotary pre-vacuum pump.
These combinations also provide
reasonable exchange times for
specimen, filament and aperture
(less than 2 minutes) without the
need to use vacuum airlocks. The SEM
vacuum system is fully automatically
controlled and protected against
operating failures.
17
Specimen orientation and ESEM
manipulation As mentioned above, samples for
As has been mentioned, the quality conventional SEM generally have to
of the image in a SEM depends on be clean, dry, vacuum compatible
the orientation and the distance of and electrically conductive. In recent
the specimen from the detectors and years the Environmental Scanning
the final lens. The specimen stage Electron Microscope (ESEM) has
allows the specimen to be moved in been developed to provide a unique
a horizontal plane (X and Y direction) solution for problematic samples.
and up and down (Z direction) and Examples of specimens which pose
rotated and tilted as required. These problems are wool or cotton tissue,
movements are often motorised and cosmetics, fats and emulsions
controlled by the PC. (e.g. margarine).
18
Signal Amplification in the ESEM
Gaseous Cascade
environment Amplification
X-ray Analysis
To return for a moment to the analogy of a person in an
unknown darkened room with a fine beam torch (box F). While
scanning the room to determine its topography, the viewer would also be
aware of the colour of objects. Likewise by looking at the wavelengths of X-rays
Box P
emitted (their "colour") we can determine which chemical elements are present at
each spot and the intensity of the X-rays is a measure of the element concentration.
The impinging electron in the primary beam may cause an electron in an atom of the
specimen to be removed from its orbit (if the atom is near the surface, it can become a
secondary electron). The atom is left in an excited state (it has too much energy) and it
returns to its stable state by transferring an electron from an outer orbit to replace the one
removed (see box C). Finally the outermost missing electron is replaced by one from the
free electrons always present in a material. Each of these transfers results in the release of
surplus energy as an X-ray quantum of characteristic wavelength (in the case of outer to
inner orbital transitions) or a light quantum (in the case of free electron to outer orbit
transfers). We can see that different materials produce different colours by examining a
colour television screen with a magnifying glass. Different phosphors produce different
colours of light under the impact of the scanning electron beam.
20
Fig. 25 The quality of a soldered joint depends on
getting the right structure and mixture on cooling.
The coloured X-ray maps show the locations of
lead (red) and tin (green) corresponding to
the secondary electron image.
21
Ion Beam technology material. Examples of applications circuit. Used in defect review, a FIB
So far this booklet has been about are the production of thin-film heads unit can improve manufacturing
electron microscopy and what useful for reading computer storage media yields by analysing surface and sub-
information can be obtained using a and the removal of material round a surface defects on wafers from the
primary electron beam. However, defect in a semiconductor device to manufacturing (fab) line. It can also
electrons are not the only charged allow the defect to be studied in a cross-section critical structures to
particles that can be accelerated and transmission electron microscope provide three-dimensional monitoring
focused using electrical and magnetic (see Fig. 13, page 13). of process performance. As a failure
fields. It was explained earlier (see analysis tool, it speeds the diagnosis
box C) that the atom consists of a How is such a fine beam controlled of IC malfunction and the location
positively charged nucleus surrounded and how does the operator know of subsurface defects. As micro-
by electrons in orbits. Normally the when the micro-machining is finished? electronic structures get smaller,
atom is neutral because there are Quite simply by having a detector to contamination control becomes
equal numbers of protons and elec- observe the secondary electrons more important. Some of the new
trons. An atom that has lost one produced by the impact of the ions. FIB models feature robotic wafer
or more of its outermost electrons By having a scanning electron micro- handling and automatic wafer align-
has a positive charge and can be scope as an integral part of the same ment to comply with the cleanroom
accelerated, deflected and focussed instrument, the point of impact of requirements of the fab line.
in a similar way to its negatively the ion beam can be observed
charged cousin the electron. non-destructively and the operator FIBs in Materials Science
can control the ion beam process For materials science applications,
The difference lies in the mass precisely. These instruments are a FIB unit can cross-section complex
(analogous to weight – see box Q). called dual-beam instruments. alloys and layered materials and pre-
The electron is almost 2000x lighter pare extremely thin TEM specimens.
than the lightest ion and ions can FIBs in IC Manufacturing An ion beam’s submicron milling has
be up to 250x heavier still. The rela- By precisely focusing a high current uses in the manufacture and test of
tively low-mass subatomic electrons density ion beam, a FIB workstation microelectromechanical systems,
interact with a sample and release can not only remove material and such as acceleration detectors.
secondary electrons which, when expose defects, but also deposit new Other hard surface applications
collected, provide high quality images conducting paths or insulating layers, include micromachining fresnel
at near atomic-level resolution. analyse the chemical composition of lenses and sharpening scanning
The greater-mass ions dislodge a sample and view the area being tunnelling microscope tips.
surface particles, also resulting in modified, all within submicron toler-
displacement of secondary ions and ances. IC manufacturers find it a FIBs in Biological Science
electrons. The ion beam directly cost-effective tool with a versatile Used in the biological sciences, an
modifies or mills the surface with range of applications that can reduce ion beam’s precise cut can remove
submicron precision. Secondary elec- time-to-market and improve yields. the surface of a cell or prepare a
trons and ions are used for imaging During IC design and testing, these cross-section of a virus to allow for
and compositional analysis. products speed the fabrication and detailed examination under an
By carefully controlling the energy debugging of prototypes. Focused electron microscope. It is opening
and intensity of the ion beam, it is ion beam technology gives rapid up new possibilities in the field of
possible to carry out very precise access to design problems, then structural biology.
micro-machining to produce minute gas injection techniques allow fast
components or to remove unwanted cut-and-splice modifications to the
Box Q
22
g l o s s a r y
Aberration Chromatic aberration Excited atom Nanometre Scintillation detector
The deviation from perfect imaging in an See aberration. The magnification of the lens An atom which has lost one of its inner elec- Unit of length (distance). One nanometre Electron detector used in SEM or STEM in
optical system which is caused by imperfec- varies with the wavelength of the electrons trons (see also ion) has a higher energy (the (abbreviated to nm) is a millionth of a millime- which electrons to be detected are accelerated
tions in the lens or by non-uniformity of the in the beam. extra energy is that needed to remove the tre (10-9 metre) (see box D, page 5). towards a phosphor which fluoresces to pro-
electron beam. electron). It seeks to return to its ground state duce light which in turn is amplified by means
Cleanroom by rearranging its electrons and emitting the Objective lens of a photomultiplier (q.v.) to produce an
Accelerating voltage Room containing extremely low numbers of excess energy as an X-ray quantum. In a TEM, this is the first lens after the specimen electrical signal.
The potential difference in an electron gun dust particles used for the manufacture of whose quality determines the performance of
between cathode and anode over which elec- semiconductors. Fab line the microscope. In a SEM it is the last lens Secondary electrons
trons are accelerated. The higher the voltage, The semiconductor production line which is before the specimen and produces the Electrons originating in the specimen which
the faster the electrons and the more pene- Column located in a clean room (q.v.). extremely fine electron spot with which the are emitted under influence of the primary
trating power they have. Voltages usually The evacuated electron beam path, the elec- specimen is scanned. beam.
range from a few thousand volts up to several tromagnetic lenses and the specimen and FEG
hundred thousand. aperture mechanisms are accommodated in Field Emission Gun is defined on page 21. Oil diffusion pump SEM
a vertical column (see page 8). Vacuum pump where the pumping action is Scanning Electron Microscopy is defined on
Airlock FIB produced by the dragging action of a stream page 7.
A chamber within the electron microscope Condenser lens Field ion beam instrument. A beam of ions of oil vapour though an orifice.
that can be isolated from the rest to allow the Part of the illumination system between the replaces the beam of electrons. Dual beam Semiconductor detector
specimen to be inserted. The airlock is then gun and the specimen designed to concen- instruments also have an electron column. Phase Electron detector used in SEM or STEM in
pumped out and the specimen moved into trate the electron beam into a parallel or into Relative position in a cyclical or wave motion. which the electron to be detected causes a
the column vacuum. This reduces the amount a finely focussed spot on the specimen (or, in Filament It is expressed as an angle, one cycle or one small change in the semiconductor material
o
of air and other contaminants brought into the case of the scanning electron microscope, Metal wire, usually in the form of a hairpin, wavelength (q.v.) corresponding to 180 . which is amplified by making the semi-
the column. Airlocks also facilitate exchange into the objective lens). which, when heated in vacuum, releases conductor part of an electrical circuit.
of photographic material and gun emitter. free electrons and so provides the source Phase contrast
CRT of electrons in the electron microscope. Image contrast caused by the conversion of Spectrometer
Amplitude Cathode ray tube. The screen of a conventional phase differences in light leaving the object Instrument for obtaining a spectrum (q.v.).
The maximum value which a periodically TV set or a PC monitor is the front end of a Fluorescent screen into amplitude differences in the image.
changing physical magnitude can reach. cathode ray tube. Behind is an electron gun Large plate coated with a material (phosphor) Spectrum
For electromagnetic waves the intensity of producing an electron beam, lenses to focus it which gives off light (fluoresces) when bom- Phase diagram A display produced by the separation of a
the disturbance is proportional to the square and a scanning system to make the beam scan a barded by electrons). In the TEM, the image is Graph of temperature and pressure showing complex radiation into its component
of their amplitude. raster (q.v.) on the screen to produce an image. formed on the fluorescent screen (see page 11). the range of each under which agiven material wavelengths or energies.
can exist in the solid, liquid or vapour phase.
Amplitude contrast Crystal Focal length of a lens Spherical aberration
Image contrast caused by the removal of elec- A material in which the atoms are ordered into The distance (measured from the centre of Photomultiplier See aberration. The magnification in the centre
trons (or light) from the beam by absorption rows and columns (a lattice) and because of the lens) at which a parallel incident beam is Electronic tube in which light is amplified of the lens differs from that at the edges.
in the specimen. this periodicity, electrons, whose wavelength brought to a focus. without interference (noise) to produce an
(q.v.) is about the same size as the spacing electrical signal. Sputter coater
Ångström between atoms, undergoes diffraction (q.v.) Focusing Instrument for coating a non-conducting
Unit of length used in the early days of The act of making the image as sharp as possi- Photons specimen with a very thin uniform layer of a
microscopy. 1 Å = 0.1 nm (see box D, page 5). Detector ble by adjusting the objective lens (q.v.). Discrete packets of electromagnetic radiation. A conducting element such as gold to make it
A device for detecting particular electrons or light beam is made up of a stream of photons. suitable for examination in a scanning electron
Anode photons in the electron microscope. Goniometer microscope without danger of it charging up.
In an electron gun (see page 8), the negative- Specimen stage allowing linear movement of Primary electrons
ly charged electrons are accelerated towards Diffraction the specimen in two or more directions and Electrons in the beam as it leaves the gun and STEM
the anode which has a positive charge relative Deviation of the direction of light or other rotation of the specimen in its own plane and impinges on the specimen. Scanning Transmission Electron Microscopy is
to the filament (cathode) from which they wave motion when the wavefront passes the tilting about one or more axes (see page 13). defined on page 7.
emerge. In practice (for ease of construction), edge of an obstacle. Quantum
the filament has a high negative charge and Ground state A discrete packet of X-radiation. TEM
the anode is at earth (ground) potential. Diffraction contrast The lowest energy state of an atom. All elec- Transmission Electron Microscopy is defined
Image contrast caused by the removal of elec- trons in their proper places and no charge. Raster on page 6.
Aperture trons (or light) from the beam by scattering The track of the beam in a SEM or STEM. It is
A disc of metal with a small hole designed to by a periodic (e.g. crystalline) structure in the Ion analogous to eye movements when reading a Turbomolecular pump
stop those electrons which are not required specimen. An atom which has lost one or more of its book: left to right word by word and down Vacuum pump in which the centrifugal force
for image formation (e.g. scattered electrons). outer electrons. It has a positive charge. the page line by line. of fast rotating discs forces molecules from the
EDX axis to the periphery.
Astigmatism Energy Dispersive X-ray Analysis or Ion getter pump Refraction
A lens aberration (q.v.) A circle in the speci- Spectrometry. An EDX spectrometer makes a Vacuum pump in which the movement of ions Changes in speed (and sometimes direction) Vacuum
men becomes an ellipse in the image. spectrum (q.v.) of X-rays emitted by the spec- in a strong magnetic field drag molecules of gas when light or electrons pass through a Space from which (most) gases and vapours
imen on the basis of their energy. An alterna- and embed them in the cathode of the pump. specimen. have been removed (see box J, page 12).
Atom tive method of making the spectrum,
There are many ways of looking at the atom. Wavelength Dispersive Spectrometry (WDS), Lattice Refractive index Wavelength
The most useful one for electron microscopists exists but is not referred to in this book. Regular array of atoms in a crystal (q.v.). The ratio of the speed of light in a vacuum The distance on a periodic wave between
is to think of it as consisting of a positively to that in a given medium such as glass, water two successive points at which the phase is
charged nucleus (containing positively EELS Lens or oil. the same.
charged protons and uncharged neutrons) Electron Energy Loss Spectroscopy In a light microscope, a piece of transparent
surrounded by negatively charged electrons (or Spectrometry) is defined on page 20. material with one or more curved surfaces, Resolving power Wehnelt cylinder
in discrete orbits. which is used to alter systematically the direc- The ability to make points or lines which are An electrode between the cathode (filament)
Electron tion of rays of light. In an electron microscope, closely adjacent in an object distinguishable in and the anode (earth) in an electron gun
Atomic number Fundamental atomic particle rotating in an a similar effect is achieved on a beam of elec- an image. (see box B, page 5) which focuses and controls the electron
The number of protons in the atomic nucleus. orbit around the nucleus of the atom (see trons by using a magnetic (or electrostatic) current in the beam and keeps it constant
This number determines the chemical nature box C). Free electrons can easily flow in a field (see pages 5 & 10). Resolution (see Fig. 7, page 9).
of the atom. An atom of iron for example has conductor and can be extracted into a The act or result of displaying fine detail in an
29 protons, an atom of oxygen 8 and so on. vacuum by heat and or an electrical field. Micrometre image. (see box A, page 4) Working distance
Unit of length (distance). One micrometre The physical distance between the external
Backscattered electrons Electron microscope (abbreviated to µm) is a thousandth of a mil- Rotary pump metal parts of the objective lens and the speci-
Primary electrons which have been deflected A microscope (q.v.) in which a beam of limetre or 1000 nm (see box D, page 5). Vacuum pump in which the pumping action is men surface. This is the space available for
by the specimen through an angle generally electrons is used to form a magnified image produced by moving packets of air from one placing certain electron, X-ray and cathodolu-
o
greater than 180 with little or no loss of energy. of the specimen. Microscope side of a rotating cylinder to another by minescence detectors. For highest resolution,
An instrument designed to extend man's means of an eccentric drum. the working distance has to be made as small
Binocular viewer ESEM visual capability, i.e. to make visible minute as possible which leads to compromises.
A light microscope built into a TEM for viewing Environmental Scanning Electron Microscopy detail that is not seen with the naked eye. Scanning
a fine-grain fluorescent screen for critical is defined on page 18. Process of investigating a specimen by moving X-rays
focussing and astigmatism correction. Microtome a finely focussed electron beam systematically Extremely short wavelength radiation. In the
Excitation Instrument for cutting extremely thin sections in a raster over the surface. The same tech- electron microscope, an X-ray photon (q.v.) is
Cathodoluminescence The input of energy to matter leading to the from a specimen prior to examination in the nique in the display monitor leads to an image emitted when an excited atom releases its
The emission of light photons by a material emission of radiation. microscope. In electron microscopy this is in which every point corresponds to a point excess energy and returns to its ground state.
under electron bombardment (see page 11). usually referred to as an ultramicrotome. on the specimen.
23
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