Module 6: Spectroscopic, Diffraction and Microscopic

Techniques [5 h]

a) Fundamental concepts in spectroscopic and instrumental

techniques

b) Principle and applications of UV-Visible Spectroscopy

technique

c) Principle and applications of X-Ray Diffraction (XRD)

technique (including numerical)

d) Overview of various techniques: Scanning Electron

Microscopy (SEM)
a) Fundamental concepts in spectroscopic and instrumental techniques
Spectroscopy Basics:
• Spectroscopy is a branch of science that studies the interaction between
electromagnetic (EM) radiation and matter.
• Spectroscopy is used as a tool for studying the structures of atoms and molecules.
• The basic principle shared by all spectroscopic techniques is to shine a beam of EM
radiation onto a sample, and observe how it responds to such a stimulus. The
response is recorded as a function of radiation wavelength.
EM Radiation Basics:
• EM radiation is a form of energy that has both wave and particle properties as shown in the
classical sinusoidal wave model (Fig. shown below), and embodies parameters such as
Wavelength, λ (m), Frequency, υ (Hz), Velocity (m/s) and Amplitude (m).
• Compared to other wave phenomena such as sound, EM radiation requires no supporting
medium for its transmission, and thus passes readily in vacuum.
• EM radiation is represented as electric and magnetic fields that undergo in-phase, sinusoidal
oscillations at right angles to each other and to the direction of propagation.
Fig. Representation of a
single ray of plane-
polarized EM radiation.

Properties of EM radiation
• Wavelength (λ) is the spatial distance between two consecutive peaks (one cycle) in the
sinusoidal waveform and is measured in sub-multiples of meter, usually in nm.
• Maximum length of the vector is called Amplitude.
• Frequency (υ) -number of wavelength occur in onesecond. It therefore has the units of 1 s -1
= 1 Hz. Frequency is related to the wavelength via the speed of light (c = 2.998x10 8 m.s-1), υ =
c λ-1.
• Wavenumber (υ-1) describes the number number of wavelengths per unit distance and is
typically measured in 1 cm-1.
Types of EM radiation interaction with matter:
• If matter is exposed to EM radiation e.g. IR light (Fig. shown below), the radiation can be
either absorbed, transmitted, reflected, scattered or undergo photoluminescence.
• Photoluminescence is a term used to designate a number of effects including fluorescence,
phosphorescence and Raman scattering.
• Balance of the absorbed light gets transmitted.
• Color of an object that we see is due to the wavelengths transmitted or reflected. Other
wavelengths are absorbed. The more absorbed, the darker the color (more concentrated
solution).

Fig. Schematic representation

of interaction of radiation and
matter

• Interaction of EM radiation with matter is a quantum phenomenon and is dependent on both

the properties of radiation and appropriate structural parts of the samples involved.
• Origin of EM radiation is due to energy changes within matter itself.
• In spectrochemical methods, we measure the absorbed radiation.
Frequency range of each transition during the spectroscopic analysis
 Type of radiation Frequency range (Hz) Wavelength range Type of transition
excitement of nucleus to a
Radio waves <3x1011 >1 mm
higher spin state
molecular rotations,
Microwaves 3x1011-1013 1 mm-25 mm
electron spin flips
Infrared 1013-1014 25 mm-2.5 mm molecular vibrations
Near-infrared 1~4x1014 2.5 mm-750 nm outer e- molecular vibrations
Visible 4~7.5x1014 750 nm-400 nm outer electron
Ultraviolet 1015-1017 400 nm-1 nm outer electron
X-rays 1017-1020 1 nm-1 pm inner electron
Gamma-rays 1020-1024 <10-12 m Nuclear

Spectrometric Instruments:
• Ultraviolet-Visible (UV-Vis), IR, Raman, X-ray
Fluorescence (XRF), Energy-dispersive X-ray (EDX) and
Nuclear Magnetic Resonance (NMR) spectroscopy
techniques are mainly used for characterization of
substances.

• Intensity of the radiation is mostly measured with a

photoelectric transducer.
(b). Principle and applications of UV-Visible Spectroscopy technique
• In UV-Vis spectroscopy, energy is absorbed by a molecule in the UV region (1
nm-400 nm) or visible region (400 nm-750 nm) resulting in electronic transition of
valence electrons.
• Different molecules absorb radiation of different wavelengths depending on their
structure. An absorption spectrum will show a number of absorption bands
corresponding to structural (functional) groups within the molecule.
• For ex. absorption by carbonyl group in acetone is of the same wavelength as the
absorption by carbonyl group in diethyl ketone.
Three types of electronic transitions involving:
(i). π, σ and n electrons; (ii). charge-transfer electrons and (iii). d and f electrons.
▪ Inorganic species show charge-transfer absorption and are called charge-transfer
complexes.
▪ For a complex to demonstrate charge-transfer behaviour, one of its components must
be able to donate electrons and other component must be able to accept electrons.
▪ Absorption of radiation then involves the transfer of an electron from the donor to an
orbital associated with the acceptor (ε will be very high > 10,000 dm3mol-1cm-1).
▪ Absorption of UV-Vis radiation in organic molecules is restricted to certain functional
groups (chromophores) that contain valence electrons of low excitation energy. The
spectrum of a molecule containing these chromophores is complex and broad.
Electronic excitations in UV- Visible spectroscopy

Empty
orbitals

σ to π* is forbidden transition

σ to σ* transitions: Electron in a bonding σ orbital is excited to the corresponding

antibonding σ* orbital. Energy required is large. For ex. methane having only C-H bonds
can undergo only σ to σ* transitions showing abs. maximum at 125 nm. Abs. maxima
due to σ to σ* transitions are not seen in typical UV-Vis. spectra (200-700 nm).
n to σ * transitions: Saturated compounds containing atoms with lone pairs (non-
bonding electrons) are capable of n to σ * transitions. These transitions usually need
lesser energy than σ to σ* transitions. They can be initiated by light whose wavelength
is in the range 150 - 250 nm. The number of organic functional groups with n to σ *
peaks in the UV region is small.
n to π* and π to π * transitions: need an unsaturated group in the molecule to provide
the π electrons. Most absorption spectroscopy of organic compounds is based on
these transitions, since their absorption peaks fall in the experimentally convenient
spectral region between 200 - 700 nm.
Chromophore: any isolated
covalently bonded group
that shows a characteristic
absorption in the UV-Vis.
region. The only molecular
moieties likely to absorb
light in the 200 to 800 nm
region are π-electron
functions and hetero atoms
having non-bonding
electron pairs.

~ λmax: 280 nm
Auxochrome: group of atoms
attached to a chromophore Auxochrome
which modifies the ability of ~ λmax: 255
that chromophore to absorb nm
light. Ex. COOH, -OH, -SO3H,
-NH2, -NH-R, -N-R2

~ λmax: 385
~ λmax: 320
Applications of UV-Vis spectrophotometer:
Qualitative analysis: Identification of chromophores by scanning the absorbance at each
wavelength.
Absorbance maximum
at this wavelength

Buta-1,3-diene
CH2=CH-CH=CH2

Quantitative analysis:
• Absorbance at a particular
λmax can be measured using
photometry mode.
• Determining the reaction rate and pKa values (dissociation
constants) of weak acids or bases.
▪ Determining the percentages of keto and enol forms.
▪ To analyze metals in waste water.
▪ Determining total serum protein, serum cholesterol, etc.
▪ Characterizing pharmaceuticals, food, paint, glass and metals.
Based on the functional group present and attached to chromophores…
Bathochromic shift: absorption maximum shifted

Absorbance
to longer wavelength (Blue to Red [Red shift]).
Hypsochromic shift: absorption maximum shifted
to shorter wavelength (Red to Blue [Blue shift]).
Hyperchromism: increase in molar absorptivity
Hypochromism: decrease in molar absorptivity
Components of a UV-Vis Spectrophotometer

Source Lamp Sample Holder Photometer/Detector

Monochromator
Signal Processor and Readout
Source lamp
▪ Tungsten filament incandescent lamp used in Visible and adjacent parts of UV and
IR regions.
▪ Hydrogen or deuterium discharge lamps are used in 160~360 nm (UV region).
▪ Deuterium lamps provide maximum intensity.
▪ Source used in UV- Vis spectroscopy should meet the following criteria: (i). Beam
produced should be in the detectable and measurable range, (ii). Should serve as a
continuous source of energy and (iii). Should be stable.
Monochromator
o Filter the energy source so that a limited portion is allowed to be incident on the
sample.
o Gratings are normally used as monochromators.
o A particular wavelength can be selected using monochromator.
Sample holder
• The selection of material used for constructing the cuvette is based on the selected
range of measurement.
• Cuvette thickness depends on the absorption intensity. Cuvettes with varied shapes
are used (rectangular, cylindrical or cylindrical with flat ends).
• Cell thickness: 1, 2 and 5 cm.
• Main factor is that the cuvette window should be normal to the beam direction.
• Requirement of cuvettes in terms of its make and thickness: UV region – quartz and
Visible region – glass or quartz cells.
Photometer/Detector
Mechanism behind the photoelectric devices is the conversion of radiant energy to
electrical signal. Basically, 3 types of photometers are used: (a). Photovoltaic cells, (b).
Phototubes and (c). Photoconductive cells.
Signal processing
Electrical signal generated by the transducer is sent to the signal processor, where it is
displayed in a more convenient form for the analyst.
Wave Behaviors
(c). Principle and applications of X-Ray Diffraction (XRD) technique
• XRD is a versatile, non-destructive characterization technique widely used in materials
science and engineering for identifying unknown crystalline materials.
• XRD works by irradiating a material with incident X-rays, and then measuring the intensities
and scattering angles of the X-rays that leave the material.
• XRD is used to study the structure and function of many biological molecules, including
vitamins, drugs, proteins and nucleic acids such as DNA.
• XRD is also used to determine structural properties like lattice parameters, strain, grain size,
epitaxy, phase composition, preferred orientation, to measure thickness of thin films and
multi-layers and to determine atomic arrangement.
• XRD also yields information on how the actual structure deviates from the ideal one, owing to
internal stresses and defects.
Selected Nobel Prize Winners involving X-ray crystallography
Year Laureate(s) Prize Rationale
Discovery of diffraction of X-rays by crystals, an
1914 Max von Laue Physics important step in the development of X-ray
spectroscopy.
1915 William Henry Bragg Physics Analysis of crystal structure by means of X-rays.
Chemistr Determination of the structures of important biochemical
1964 Dorothy Hodgkin
y substances.
Ada E. Yonath, T.A.
Chemistr
2009 Steitz, R. For studying the structure and function of the ribosome.
y
Venkatraman
 Incident-beam optics • X-ray tube: source of X-rays
Receiving-side optics
• Incident-beam optics:
condition the X-ray beam before
it hits the sample.
• Goniometer: platform that
holds and moves the sample,
optics, detector, and/or tube.
• Sample holder
• Receiving-side optics: condition the X-ray
beam after it has encountered the sample.
• Detector: count the number of X-rays
scattered by the sample.
How XRD pattern produced?
• Crystalline atoms are a periodic array of
coherent scatterers and can diffract X-rays.
• The wavelength of X-rays are similar to the
distance between atoms.
• Diffraction from different planes of atoms
ce pattern)
(Interferen

produces a diffraction pattern, which contains

information about the atomic arrangement
within the crystal.
• When X-ray hits the crystal planes at specific
angles and the diffracted waves are in the
same phase, then constructive interference (a
peak) will be produced.
• Based on structure of the crystal (and crystal planes), the angles at which diffraction occurs may vary.
What is diffraction?
• Diffraction refers to different phenomena which occur when a
wave encounters an obstacle.
• In classical physics, the diffraction phenomenon is described as
the apparent bending of waves around small obstacles and the
spreading out of waves past small openings.
Interference of diffracted waves
• Interaction between diffracted waves is called interference.
• Constructive Interference: Waves are in-phase when each of their crests and troughs
occur exactly at the same time. Those type of waves stack together to produce a resultant
wave that has a higher amplitude. For constructive interference, path difference should
be multiples of n*λ.
• Destructive Interference: If the waves are out of phase by multiples of (n/2)*λ, then
destructive interference occurs and the amplitude of the resultant wave will be reduced.

path difference = multiple of path difference = multiple of

XRD patterns for 3 different forms of SiO2

▪ These three phases of SiO2 are

chemically identical.
▪ The amorphous glass does not
have long-range atomic order and
therefore produces only broad
pattern.
▪ Cristobalite form polycrystalline
structure.
▪ Quartz form single crystal structure.
Bragg model of diffraction
• Crystals are regular arrays of atoms, whilst X-rays are waves of EM radiation. Crystal atoms
scatter incident X-rays, primarily through interaction with the atom’s electrons. This
phenomenon is known as elastic scattering; the electron is known as the scatterer.
• A regular array of scatterers produces a regular array of spherical waves. In the majority of
directions, these waves cancel each other out through destructive interference, however,
they add constructively in a few specific directions, as determined by Bragg’s law:
• nλ = 2dsinθ , where “n” is an integer, and “λ” is the beam wavelength, “d” is the spacing
between diffracting planes and “θ” is the incident angle.
• X-rays scattered from adjacent planes will
combine constructively (constructive
interference) when angle θ between plane and
X-ray results in path-length difference that is
integer multiple “n” of X-ray wavelength “λ”.

Constructive interference Destructive interference

• In this model, a given reflection is associated with a set of evenly spaced sheets running
through the crystal, usually passing through the centres of the atoms of the crystal lattice.
• Orientation of a particular set of sheets is identified by its three Miller indices (h, k, l), and let
their spacing be noted by “d”.
• Specific directions appear as spots on the diffraction pattern called reflections. Consequently,
XRD patterns result from EM waves impinging on a regular array of scatterers.
• X-rays are used to produce the diffraction pattern because their wavelength, λ, is often the
same order of magnitude as the spacing “d” between the crystal planes (1-100 Ǻ).

• The path difference between Ray 1

and 2: 2a = 2d sinθ
• For constructive interference
2a = nλ = 2d sinθ
• Because crystal structures contain planes of atoms, each plane will reflect incident X-rays
differently. For ex. let two monochromatic X-ray beams (of a specific wavelength) strike a
crystal structure at an incoming angle of θ.
• Ray 1 will reflect off of the top atomic plane while Ray 2 will reflect from the second atomic
plane. However, because Ray 2 has to cross deeper into the atomic plane, it travels a
distance “2a” farther than Ray 1.
• If the distance “2a” is equal to the integral number (n*λ) of wavelength of 2 waves, then Ray
1 and 2 will be in-phase (constructively interfere) when they both exit the crystal.
• If we know wavelength λ of X-rays going into crystal and also measure angle θ of diffracted X-
rays coming out of crystal, then we can determine d-spacing between atomic planes.
• If we now reorient the crystal to a different atomic plane, we can measure d-spacing in other
planes. By doing multiple x-ray diffractions at different crystal orientations, we can
determined crystal structure and size of crystal unit cell.

• Incident angle (ω) is defined between the X-ray

tube
X-ray source and sample.
Detect
• Diffracted angle (2θ) is defined between
or
the incident beam and the detector angle.
ω θ 2
• Incident angle (ω) is always ½ of the
detector angle 2θ i.e. θ. θ
• In a typical XRD instrument, the X-ray tube
is fixed, the sample rotates at θ°/min and
detector rotates at 2θ°/min.
 11
11
11
11
11
11
11
11
11
11
11
11
11
11
11
11
11
11
1
1111111 1
1111111 1
1111111 1
1111111 1
1111111 1
1111111 1
1111111 1
111111 1

1111111 1
1111111 1
Powder X-ray Diffraction

Single Crystal X-ray Diffraction

• Powder diffraction data consists of a record of photon intensity vs detector angle 2θ

• Diffraction data can be reduced to a list of peak positions and intensities.
• Diffraction patterns are best reported using dhkl and relative intensity rather than 2θ and
absolute intensity.
• Each dhkl corresponds to a family of atomic planes {hkl}
• Individual planes cannot be resolved. This is a limitation of p-XRD vs single crystal diffraction.
Calculation of Crystallite Size using p-XRD data
Crystallites smaller than ~120 nm create broadening of diffraction peaks. This peak broadening
can be used to quantify the average crystallite size of nanoparticles using the Scherrer equation -
used for nanoparticles characterization. Must know the contribution of peak width from the
instrument by using a calibration curve

Scherrer equation can be written as:

o τ is mean size of ordered (crystalline) domains, which may be smaller or equal to grain size;
o K is a dimensionless shape factor, with a value close to unity. The shape factor has a typical
value of about 0.9, but varies with the actual shape of the crystallite;
• β is the line broadening at half the maximum intensity (FWHM), after
subtracting the instrumental line broadening, given in radians
• λ is the X-ray wavelength
• θ is the Bragg angle.
• This quantity is also
denoted as Δ(2θ)
Fig. Simulated p-XRD
patterns for wurtzite CdS
spherical particles of different
sizes from 1 μm to 1 nm. The
inset shows the 1, 2, and 5
nm XRD patterns on an
expanded y-axis scale for
clarity.
 XRD Numericals
Q. 1. In a NaCl crystal, there is a family of planes 0.252 nm apart. If the first-order
maximum is observed at an incidence angle of 12.8823°, what is the wavelength of the
X-ray scattering from this crystal?
Ans.: Use the Bragg equation nλ = 2dsinθ to solve for θ.
For the first-order, n=1. Then λ = 2dsinθ/n = 2dsinθ/1
λ = 2(0.252×10−9 m)sin(12.8823°)/1
= 0.504*0.31068 = 1.57×10−10m or 0.157 nm.

Q. 2. Estimate the crystallite size of the given nanomaterial using p-XRD data:
Peak position 2θ = 21.61o, FWHM of sample = 2.51o, k = 0.9 and λ = 1.5406 Å (degree
to radian = Degree × π/180).
Ans.: 2θ = 21.61o (θ = 10.805o) and FWHM = 2.51o (0.043825 radian)
Crystalline grain size calculation by Scherrer’s equation: k*λ/β*cosθ
k = 0.9, λ = 1.5406 Å (0.15406 nm), β = FWHM in radian and 2θ = Bragg’s angle in o obtained
from p-XRD data.
Crystallite size = (0.9*0.15406)/(0.043825*0.982257) nm = 3.22 nm
Overview of Scanning Electron Microscopy (SEM)
Limitations of using Light Microscope (LM):
▪ LM has a magnification of 1000x with a resolution of 200 nm.
▪ Resolving power of LM is limited by the number and quality of the lenses but also by the
wavelength of the light used for illumination.
▪ Using light with a short wavelength (blue or ultraviolet) gave a small improvement.
▪ Immersing the specimen and the front of the objective lens in a medium with a high refractive
index (oil) gave another small improvement leading upto 100 nm.
SEM basics
o Imagine yourself alone in an unknown darkened room with only a fine beam torch. You
might start exploring the room by scanning the torch beam systematically from side to
side gradually moving down so that you could build up a picture of the objects in the
room in your memory.
o SEM uses an electron beam instead of a torch, an electron detector instead of eyes and
a fluorescent screen and camera as memory.
▪ Accelerated electrons 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, electric and magnetic fields have the same effect on electrons as glass
lenses and mirrors have on visible light.
▪ The first electron microscope used two magnetic lenses and later added a third lens to
achieve a resolution of 100 nm, twice as good as that of the light microscope.
▪ Now, the electron microscope uses five magnetic lenses in the imaging system, a
resolving power of 0.1 nm at magnifications of over 1 million times.

Essential components of SEM

1. Electron gun as Source

2. Electron Lenses
3. Sample Stage
4. Detectors for all signals of interest
5. Display/Data output devices
Vacuum
1. Electron gun as Source:
Electron gun consists of 3 important
components:
(i). Filament made from hair-pin shaped
Tungsten (or Lanthanum Hexaboride)
functioning as Cathode. Voltage applied
to the filament heats it up to 2700 oC;
(ii). Wehnelt cylinder (also known as
grid cap or cylinder) made from
platinum or tantalum foil. It is used for
focusing and controlling the electron
beam and has the shape of a hollow
cylinder. The bottom side of the cylinder
has an opening at its centre with 200 to
1200 μm diameter; and
(iii). Anode, which is positive with
respect to the filament, forms powerful
attractive forces for electrons. This
causes electrons to accelerate (several
hundred thousand km/sec) toward the
anode. Some accelerate right by the
anode and down by the column onto These 3 components together form a triode electron
the sample. gun which is a very stable source of electrons.
2. Electron lenses - Magnification and resolution
o Magnification is entirely determined by electronic circuitry that scans the beam over the
specimen (and simultaneously over fluorescent screen of monitor where the image appears).
o Magnification can be as high as 3,00,000x. Increasing the magnification is achieved by
reducing the size of the area scanned on the specimen.
o Resolution of 1 nm can be attained depending on:
a) Diameter of electron beam on the specimen surface
b) Specimen properties
c) Specimen preparation technique
d) Instrumental parameters such as
i. beam intensity
ii. accelerating voltage
iii. scanning speed
iv. distance from the last lens to specimen (referred to as the working distance) and
v. angle
3. Sample of What
stage: the specimen
happens surface
to thewith respectduring
specimen to the detector.
electron bombardment?
Electron beam scan the specimen in a rectangular spot less than 4 nm in diameter.
a) Specimen itself emits secondary electrons.
b) Some of the primary electrons are reflected (back-scattered electrons).
c) Electrons are absorbed by the specimen.
d) Specimen emits X-rays, and sometimes emits photons (light).
All these phenomena are interrelated and depends on the specimen topography, its chemical
state and atomic number. The number of backscattered electrons, secondary electrons and
absorbed electrons at each point of the specimen depends on the specimen’s topography to a
much greater extent than the other properties mentioned above.
Specimen orientation and manipulation
o SEM image quality depends on specimen orientation and distance from detectors and lens.
o Specimen stage allows the specimen to be moved in a horizontal plane (X and Y direction)
and up and down (Z direction) and rotated and tilted as required. These movements are
often motorized and controlled by the PC.
o SEM models differ in size of their specimen chambers allowing various sizes of specimens to
be introduced and manipulated. Maximum specimen size also determines the price because
larger the specimen chamber, larger the goniometer movement needed and larger the
pumping system needed to maintain a good vacuum.
o The simplest model accepts specimens of a few cm in diameter and can move them 50 mm
in the X and Y directions. Largest chamber accepts samples up to 200 mm in diameter and
can move them 150 mm in each direction.
o All models allow samples to be tilted up to high angles and rotated through 360 o.
o There are special stages for heating, cooling and straining specimens.
4. Detectors for all signals of interest
▪ Detectors for backscattered electrons and secondary electrons are either a
scintillation detector or a solid state detector.
▪ In the case of scintillation detector, electrons strike a fluorescent screen, which then
emits light that is amplified and converted into an electrical signal by
photomultiplier tube.
▪ In the case of solid state detector, it works by amplifying the minute signal produced
by the incoming electrons in a semiconductor device.
▪ Amplified signal of the electron beam is also impacted on a cathode ray tube. Both
the beam in the microscope and the one in the CRT are scanned at the same rate
and the 1-to-1 relationship between each point on the CRT screen and the
corresponding point on the specimen is used to build the SEM image.
5. Display/Data output devices
o SEM is usually equipped with two image monitors, one for observation by operator and
other a high resolution monitor, equipped with ordinary photo camera (or Polaroid)
o To facilitate the observation and correct choice of the parameters mentioned above, SEM
have an image store in which the image is built up scan by scan and displayed at TV speed so
that there is a steady, flicker-free image on the viewing monitor.
o Digital images are stored electronically for subsequent enhancement and analysis.
o Since the SEM image is electronically produced, it can be subjected to contrast
enhancement, inversion (black becomes white), mixing of images from various detectors,
subtraction of the image from one detector from that produced by a different detector,
colour coding and image analysis.
o All these techniques may be applied if it suits the primary aim of extracting the best possible
information from the specimen.
6. Importance of Vacuum in SEM column:
• In SEM, the column through which electron beam passes through must always be at
vacuum.
• If the sample is in a gas filled environment, the electron beam cannot be generated or
maintained because of a high instability in the beam. Also, gases could react with the
electron source, causing it to burn out, or cause electrons in the beam to ionize, which
produces random discharges and leads to instability in the beam.
• Transmission of beam through electron optic column would also be hindered by the
presence of other molecules from the sample or the microscope itself, and could form
compounds and condense on the sample. This would lower the contrast and obscure
detail in the image.
• Sufficiently low vacuum in SEM column is produced by either an oil diffusion pump or a
turbo-molecular pump backed by a rotary pre-vacuum pump.
• These combinations provide reasonable exchange times for specimen, filament and
aperture (less than 2 minutes) without the need to use vacuum airlocks.
• SEM vacuum system is fully automatically controlled and protected against operating
failures.
Strengths of SEM

▪ Gold as conductive coating: A heavy element like gold is preferred for use as conductive

coating as it gives a good yield of secondary electrons and thereby a good quality image. In

addition, it gives a fine grain coating and is easily applied in a sputter coater. The layer

required to ensure a conducting layer is quite thin (about 10 nm).

▪ Operational features: Most SEM models are easier to operate, with user-friendly interfaces,

minimal sample preparation and rapid data acquisition (less than 5 min/image).

▪ SEM generate data in digital formats, which are highly portable.

Limitations of using SEM:
o Sample condition: Must be solid and stable in a vacuum to order of 10-5~10-6 torr.
Samples likely to outgas at low pressures (rock sample saturated with hydrocarbons, wet
samples such as coal, organic materials or swelling clays) are unsuitable for examination
in conventional SEM. However, they can be measured using "low vacuum" SEM.
o SEM cannot measure samples that fail to withstand the electron bombardment.
o If the specimen contains any volatile components such as water, it need to be removed
using a drying process (or can be frozen solid).
o Non-conducting specimens will charge up under electron bombardment, and hence an
electrically conductive coating must be applied to electrically insulating samples for study
in conventional SEM, unless the instrument is capable of operation in a low-vacuum
mode.
o Detector type: Solid state x-ray detector (EDS) cannot detect elements with A.N. less than
11 (Na). While EDS is very fast and easy to utilize, they have relatively poor energy
resolution and sensitivity to elements present in low abundances when compared to
wavelength dispersive x-ray detectors (WDS) on most electron probe micro-analyzers.
o Power supply: Voltages and currents required for the electron gun and condenser lenses
should be sufficiently stable to attain the best resolution. Similarly, stability of electronic
circuitry associated with the detectors should be extremely well-controlled upto 1 ppm.