Raman spectroscopy

INTRODUCTION
• Raman spectroscopy is the measurement of the
wavelength and intensity of inelastically scattered
light from molecules. The Raman scattered light
occurs at wavelengths that are shifted from the
incident light by the energies of molecular
vibrations.

• Raman spectroscopy is used to determine the

molecular motions, especially the vibrational
one.
2
Raman Spectroscopy is a non-destructive chemical analysis
technique which provides detailed information about chemical
structure, phase and polymorphy, crystallinity and molecular
interactions. It is based upon the interaction of light with the
chemical bonds within a material.
Raman is a light scattering technique, whereby a molecule scatters
incident light from a high intensity laser light source. Most of the
scattered light is at the same wavelength (or color) as the laser
source and does not provide useful information – this is called
Rayleigh Scatter. However a small amount of light (typically
0.0000001%) is scattered at different wavelengths (or colors),
which depend on the chemical structure of the analyte – this is
called Raman Scatter.
A Raman spectrum features a number of peaks, showing the
intensity and wavelength position of the Raman scattered light.
Each peak corresponds to a specific molecular bond vibration,
including individual bonds such as C-C, C=C, N-O, C-H etc., and
groups of bonds such as benzene ring breathing mode, polymer
chain vibrations, lattice modes, etc.
Raman spectroscopy probes the chemical structure of a material and
provides information about:
•Chemical structure and identity
•Phase and polymorphism
•Intrinsic stress/strain
•Contamination and impurity
Typically a Raman spectrum is a distinct chemical fingerprint for a
particular molecule or material, and can be used to very quickly
identify the material, or distinguish it from others. Raman spectral
libraries are often used for identification of a material based on its
Raman spectrum – libraries containing thousands of spectra are
rapidly searched to find a match with the spectrum of the analyte.
 Time lap
• 1923 – Inelastic light scattering is predicted by A. Smekel
• 1928 – Landsberg and Mandelstam see unexpected frequency shifts in
scattering from quartz

• 1928 – C.V. Raman and K.S. Krishnan see “feeble fluorescence” from
neat solvents
• 1930 – C.V. Raman wins Nobel Prize in Physics

• 1961 – Invention of laser makes Raman experiments reasonable

• 1977 – Surface-enhanced Raman scattering (SERS) is discovered
• 1997 – Single molecule SERS is possible

7
 OVERVIE
W
• A vibrational spectroscopy
- IR and Raman are the most common vibrational spectroscopes for
assessing molecular motion and fingerprinting species
- Based on inelastic scattering of a monochromatic excitation source
- Routine energy range: 200 - 4000 cm–1

• Complementary selection rules to IR spectroscopy

- Selection rules dictate which molecular vibrations are probed
- Some vibrational modes are both IR and Raman active

• Great for many real-world samples

- Minimal sample preparation (gas, liquid, solid)
- Compatible with wet samples and normal ambient
- Achilles Heal is sample fluorescence
8
 Raman spectrometer’s mechanism.
When a substances (in any state) is irradiated with a monochromatic light
of definite frequency(v),the light scattered at right angle to the
incident light contains lines of 1. Incident frequency and 2.Also of
lower frequency

Sometimes lines of higher frequency are also obtained that of the incident
beam will be scattered. It is called Raman scattering.

The line with lower frequency are called Stoke’s lines.

Also, the line with higher frequency are called Antistoke’s lines.

The line with the same frequency as that of the incident light is called
Rayleigh line.

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Frequency :-

This difference is called Raman frequency or Raman shift.

1
0
.
• It may be noted that raman frequencies for a
particular substances are characteristic of
that substances.

• The various observation made by raman are

called raman effect.

• Also the spectrum obtained is called

raman spectrum.

1
2
 Classical theory of
raman effect
• According to the classical theory of electromagnetic radiation, electric and magnetic
fields oscillating at a given frequency are able to give out electromagnetic radiation of
the same frequency. One could use electromagnetic radiation theory to explain light
scattering phenomena.
• For a majority of systems, only an induced electric dipole moment μ is taken into
consideration. This dipole moment which is induced by the electric field E could
be expressed by the power series
μ=μ(1)+μ(2)+μ(3)+⋯
where
μ(1)=α⋅E
μ(2)=12β⋅EE
μ(3)=16γ⋅EEE
α is termed the polarizability tensor.
It is a second-rank tensor with all the components in the unit of CV-1m2. Typically, orders of
magnitude for components in α, β, and γ are as follows,
α, 10-40 CV-1m2;
β, 10-50 CV-2m3; and
γ, 10-61 CV-3m4.
According to the values, the contributions of μ(2)and μ(3) are quite small unless electric field is
very high. Since Rayleigh and Raman scattering are observed quite readily with very much
lower electric field intensities, one may expect to explain Rayleigh and Raman scattering
in terms of μ(1) only.

1
3
 Classical theory of
raman effect

ind = E

polarizability

10
Colthup et al., Introduction to Infrared and Raman Spectroscopy, 3rd ed., Academic Press,
. Raman Spectroscopy: Absorption, Scattering, and Fluorescence

2nd
Electronic
Excited State
25,00

Impurity
0
1st
Excitation Energy,  (cm–1)

Electronic
Excited State
Stoke Anti-Stokes
s

fluorescenc

emit

e
4,00 fluorescence Vib
0 .state Electronic
 ∆ Ground
s
0 Raman State
I Fluorescenc
R ∆=emit- e
 = Trouble 15
. Raman Spectroscopy: Classical Treatment
• Number of peaks related to degrees of freedom
DoF = 3N - 6 (bent) or 3N - 5 (linear) for
N atoms
• Energy related to harmonic oscillator
c k(m1
 or 
  m 2 ) 2
• Selection rules related to symmetry
m1m2

Rule of thumb: symmetric=Raman active,

asymmetric=IR active
CO2 H2O

Raman: 1335 cm–1

Raman + IR: 3657 cm–1
IR: 2349 cm–1
Raman + IR: 3756 cm–1

IR: 667 cm–1

Raman + IR: 1594 cm–1
16
 Theory of raman spectra
Two cases may arise depending upon whether a collision between a photon and molecules
In it’s ground state is elastic or inelastic in nature.
Case 1- if the collision is elastic – this lead to the appearance of unmodified lines (or
unmodified frequency of light) in the scattered beam and this explain rayleigh scattering.
Case 2 - if the collision is inelastic – there will be exchange or transfer of energy between
the scattering molecules and the incident photon.
The frequency of scattered light and the incident photon which is either higher or
lower than that of the incident photon is called raman frequency.
Total energy before collision =total energy aftercollision

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Presentation of Raman Spectra

ex = 1064 nm = 9399 cm-

1

Breathing mode:
9399 – 992 = 8407 cm-1

Stretching mode:
9399 – 3063 = 6336 cm-1

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Rayleigh Scattering:- Occurs when incident EM radiation induces
an
oscillating dipole in a molecules, which is re-radiated at the same frequency.

•Elastic ( does not change)

8 4 ( ')2 (1 cos2 0
•Random direction of emission ( E )
 )E
 4d 2
sc 


••Little
Littleenergy
energy loss Eugene Hecht, Optics, Addison-Wesley, Reading, MA,
15
 Raman Scattering
Occurs when monochromatic light is scattered light has been weakly modulated
by the characteristic frequencies of the molecules.
Raman spectroscopy measures the differences between the wavelengths of the
incident radiation and the scatted radiation.
Selection rule: v = ±1
Overtones: v = ±2, ±3, …
 z(t)  zzequil Emax cos 20t 

1 dzz max E max cos 2 (0 vib )t

2 r

dr
1 dzz max E max cos 2 (0  vib )t
2 r
dr
Must also have a change in polarizability

Classical Description does not suggest any

difference
between Stokes and Anti-Stokes intensities
N1 hvib
  kT

N0 e
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 The Raman
polarization
The Raman Polarization is a property of waves that can oscillate with more
than one orientation EMR or waves, such as light and gravitational wave
exhibit polarization.
Polarization state :- the shape traced out in a fixed plane by the electric
vector as such a plane wave passes over it is a description of the
polarization.
E.g. linear polarization
circular polarization
elliptical polarization
orthogonally polarization

Polarization changes are necessary to form the virtual state and hence the
Raman effect.

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 Condition for raman spectroscopy
Vibrational modes that are more polarizable are more Raman-active
Examples:
– N2 (dinitrogen) symmetric stretch
cause no change in dipole (IR-inactive)
cause a change in the polarizability of the bond – as the bond gets
longer it is more easily deformed (Raman -active)

– CO2 asymmetric stretch

cause a change in dipole (IR-active)
Polarizability change of one C=O bond lengthening is cancelled
by
the shortening of the other – no net polarizability (Raman-inactive)
Some modes may be both IR and Raman-active, others
may be one or the other!
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 Condition for raman
spectroscopy
 Raman spectra occurs as a result of oscillation of a dipole
moment, induced in a molecules by the oscillating electric field of
an incident wave.

 As the induced dipole moment is directly proportional to the

polarisability of the molecules, the molecules must possess
anisotropic polarisability which should change during molecular
rotation or vibration for vibrational or rotational-vibrational raman
spectra.

 Anisotropic polarisability depends upon the orientation of the

molecules.
 In the presence of an electric field, the electron cloud of an atom
or molecules is distorted or polarised.
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 Mutual Exclusion
Principle
For molecules with a center of symmetry, no IR active
transitions are Raman active and vice versa

Symmetric molecules
IR-active vibrations are not Raman-active.
Raman-active vibrations are not IR-
active.

O=C=O O=C=O
Raman active Raman inactive
IR inactive IR active
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 Raman
Instrumentation

25
 There are following component
involves.

1. Laser or source of light

2. Filter
3. Sample holder
4. detector

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 The block design dispersive Raman scattering
system:

Wavelengt DDeeet teccot t Recorder

Recorde
Recorde
Sample h rr
orr
selector
InInGGaaA
90· Assoorr

GGee
Radiation
sources

Block diagram

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Flow diagram dispersive Raman scattering
system: 28
Schematic diagram dispersive raman scattering
system

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 1. Laser or source of light

• Lasers are generally the only source strong enough to scatter lots of light and
lead to detectable raman scattering.

• Lasers operate using the principle of stimulated emission.

• Electronic population inversion is required to achieve gain via

stimulated
emission (before the fluorescence lifetime is reached)

• Population inversion is achieved by “pumping” using lots of photons in a

variety of laser gain media

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List of Various laser source
S.No. Laser wavelength
01 Nd:YAG 1064nm

02 He:Ne 633nm

03 Argon ion 488nm

04 GaAlAs diode 785nm

05 Co2 10600nm

06 Ti-Sapphire 800nm

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A :- He:Ne laser
• Filled with 7:1 He & Ne gas optimum output of 6328 Å

• High voltage excitation is preferred

B :- Nd:YAG System
• A typical laser system –the neodymium-doped yttrium aluminum garnet or
Nd+3

• YAG is a cubic crystalline material

• Crystal field splitting causes electronic energy level splitting

• Nd:YAG laser are optically pumped using a flash tube or laser diodes.

• These are the one of the most common type of laser.

• It emits 1064 nm wavelength

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 2.Filter
• It is therefore essential to have monochromatic radiations.

• For getting monochromatic radiations filters are used.

• They may be made of nickel oxide glass or quartz glass.

• Sometimes a suitable colored solution such as an aqueous

solution of ferricyanide or iodine in CCl2 may be used as a
monochromator.

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 3.Sample holder
• For the study of raman effect the type of sample holder to be used
depends upon the intensity of sources ,the nature and availability
of the sample.

• The study of raman spectra of gases requires samples holders

which are generally bigger in size than those for liquids.

• Solids are dissolved before subjecting to raman spectrograph.

• Any solvents which is suitable for the ultraviolet spectra can be
used for the study of raman spectra.

• Water is regarded as good solvents for the study of inorganic

compounds in raman spectroscopy. 34
 4.detector
• Researchers traditionally used single points detectors such as
photocounting, photomultiplier(PMT), not because of the weakness of a
typical raman signal, longer exposure times were often required to
obtains raman spectrum of a decent quality.

• Now days multichannel detectors like photodiode arrays(PDA),

charged couple devices(CCD)

• Sensitivity & performance of modern CCD detectors are high.

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 APPLICATION
Pharmaceuticals and Cosmetics:-
• Compound distribution in tablets
• Blend uniformity
• High throughput screening
• API concentration
• Powder content and purity
• Raw material verification
• Polymorphic forms
• Crystallinity
• Contaminant identification
• Combinatorial chemistry
• In vivo analysis and skin depth profiling
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• Geology and Mineralogy
• Raman spectra of (top to bottom) olivine, apatite, garnet and
gypsum illustrating how Raman can be used for fast mineral ID.

• Gemstone and mineral identification

• Fluid inclusions

• Mineral and phase distribution in rock sections

• Phase transitions

• Mineral behavior under extreme conditions

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 Carbon
•Materialss
Peak fitting of the D and G bands in a DLC spectrum
• Single walled carbon nanotubes (SWCNTs)
• Purity of carbon nanotubes (CNTs)
• Electrical properties of carbon nanotubes (CNTs)
• sp2 and sp3 structure in carbon materials
• Hard disk drives
• Diamond like carbon (DLC) coating properties
• Defect/disorder analysis in carbon materials
• Diamond quality and provenance

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Semiconductors

• Photoluminescence image of a 3” MQW semiconductor

wafer, showing variation of emission peak width
• Characterisation of intrinsic stress/strain
• Purity
• Alloy composition
• Contamination identification
• Superlattice structure
• Defect analysis
• Hetero-structures
• Doping effects
• Photoluminescence micro-analysis

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Life
Sciences
• Multivariate clustering of spectra acquired from three bacterial
species, illustrating how Raman can be used to characterise and
distinguish bacteria at the single cell level.
• Bio-compatibility
• DNA/RNA analysis
• Drug/cell interactions
• Photodynamic therapy (PDT)
• Metabolic accretions
• Disease diagnosis
• Single cell analysis
• Cell sorting
• Characterisation of bio-molecules
• Bone structure
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 Differences between IR and Raman methods
S.No Raman IR

01 It is due to the scattering of light It is the result of absorption

by the vibrating molecules. of light by vibrating molecules.
02 The vibration is Raman active if it Vibration is IR active if there
causes a change in polarisability. is change in dipole moment.
03 The molecule need not possess a The vibration concerned should have a
permanent dipole moment. change in dipole moment due to that
vibration.
04 Water can be used as a solvent. Water cannot be used due to
its intense absorption of IR.
05 Sample preparation is not Sample preparation is elaborate
very Gaseous samples can rarely be
elaborate, it can be in any state. used.
06 Gives an indication of Gives an indication of
covalent character in the ionic character in the molecule.
molecule.
07 Cost of instrumentation is Comparatively inexpensive.
Advantages of Raman over IR

• Water can be used as solvent.

• Very suitable for biological samples in native state (because water
can be used as solvent).
• Although Raman spectra result from molecular vibrations at IR
• frequencies, spectrum is obtained using visible light or NIR
• radiation.
• =>Glass and quartz lenses, cells, and optical fibers can be used.
• Standard detectors can be used.
• Few intense overtones and combination bands => few
spectral overlaps.
• Totally symmetric vibrations are observable.
• Raman intensities a to concentration and laser power.
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Advantages of IR over Raman
• Simpler and cheaper instrumentation.

• Less instrument dependent than Raman spectra because IR spectra

are based on measurement of intensity ratio.

• Lower detection limit than (normal) Raman.

• Background fluorescence can overwhelm Raman.

• More suitable for vibrations of bonds with very low polarizability

(e.g. C–F).

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 Several variations of Raman
spectroscopy
1. Surface-enhanced Raman spectroscopy (SERS) – Normally
done in a silver or gold colloid or a substrate containing silver or
gold. Surface plasmons of silver and gold are excited by the laser,
resulting in an increase in the electric fields surrounding the metal.
• Given that Raman intensities are proportional to the electric field,
there is large increase in the measured signal (by up to 1011).
• This effect was originally observed by Martin Fleischmann but the
prevailing explanation was proposed by Van Duyne in 1977.
• A comprehensive theory of the effect was given by Lombardi and
Birke.

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 2. Resonance Raman spectroscopy
The excitation wavelength is matched to an
electronic transition of the molecule or crystal, so
that vibrational modes associated with the excited
electronic state are greatly enhanced. This is
useful for studying large molecules such as
polypeptides, which might show hundreds of
bands in "conventional" Raman spectra. It is also
useful for associating normal modes with their
observed frequency shifts.

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3.Surface-enhanced resonance Raman
spectroscopy (SERRS) – A combination of SERS
and resonance Raman spectroscopy that uses proximity to
a surface to increase Raman intensity, and excitation
wavelength matched to the maximum absorbance of the
molecule being analysed.

4.Coherent anti-Stokes
Raman spectroscopy
(CARS) –
Two laser beams are used to generate a
coherent anti-Stokes
frequency beam, which can be
enhanced by resonance.
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5.Ramanopticalactivity (ROA) – Measures
vibrational optical activity by means of a small difference in the
intensity of Raman scattering from chiral molecules in right-
and left-circularly polarized incident light or, equivalently, a
small circularly polarized component in the scattered light.

6. Spatially offset Raman spectroscopy (SORS)

7. Spontaneous Raman spectroscopy (SRS)

8. Optical tweezers Raman spectroscopy (OTRS)

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9. Angle-resolved Raman
spectroscopy 10. Inverse Raman
spectroscopy.
11.Tip-enhanced
Raman spectroscopy
(TERS)
12.Surface plasmon polariton enhanced
Raman scattering (SPPERS)
13. Stand-off Remote Raman –
14.Fourier-transform Rama spectroscopyapplied
to photobiological systems.
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