Raman

Spectroscopy
 

Submitted by:
Ms.Bushra Qamar
Ms63-10-815
• Raman spectroscopy provides information
about molecular vibrations that can be used
for sample identification and quantitation.
• Raman spectroscopy is a spectroscopic
technique based on inelastic scattering of
monochromatic light, usually from a laser
source.
• Inelastic scattering means that the frequency
of photons in monochromatic light changes
upon interaction with a sample.
Raman effect
The concept of origin
 Raman effect
• Photons of the laser light are absorbed by
the sample and then reemitted.
• Frequency of the reemitted photons is
shifted up or down in comparison with
original monochromatic frequency, which
is called the Raman effect.
• This shift provides information about
vibrational, rotational and other low
frequency transitions in molecules.
• The Raman effect is based on molecular
deformations in electric field E determined by
molecular polarizability α.
• The laser beam can be considered as an
oscillating electromagnetic wave with electrical
vector E.
• Upon interaction with the sample it induces
electric dipole moment P = αE which deforms
molecules.
• Because of periodical deformation, molecules
start vibrating with characteristic frequency υm
• Amplitude of vibration
is called a nuclear
displacement.
• monochromatic laser
light with frequency υ0
excites molecules and
transforms them into
oscillating dipoles.
• Such oscillating
dipoles emit light of
three different
frequencies
1. Elastic Rayleigh scattering.: where a molecule with no
Raman-active modes absorbs a photon with the frequency
υ0 and resturns back to the same basic vibrational state
,emits light with the same frequency υ0 as an excitation
source.
2. “Stokes”. Where a photon with frequency υ0 is absorbed by
Raman-active molecule which at the time of interaction is in
the basic vibrational state. Part of the photon’s energy is
transferred to the Raman-active mode with frequency υm
and the resulting frequency of scattered light is reduced to
υ0 - υm.
3. AntiStokes frequency .A photon with frequency υ0 is
absorbed by a Raman-active molecule, which, at the time of
interaction, is already in the excited vibrational state.
Excessive energy of excited Ramanactive mode is released,
molecule returns to the basic vibrational state and the
resulting frequency of scattered light goes up to υ0 + υm.
• About 99.999% of all incident photons in
spontaneous Raman undergo elastic
Rayleigh scattering.
• This type of signal is useless for practical
purposes of molecular characterization.
• Only about 0.001% of the incident light
produces inelastic Raman signal with
frequencies υ0 ± υm.
 Raman Sample
• It can be obtained from most molecular Samples
– solids, liquids, mixtures, aqueous solutions, gels,
slurries, powders, films, etc. and even with some
metals.
• Through many containers such as glass bottles,
Pyrex®, reaction vessels, plastic containers,
blister packs, bags
• Raman is usually not destructive,
• Temperatures up to 500 ˚C and pressures of up
to 3000 PSI can be easily accommodated.
 Raman Spectra

• Plotting the intensity of "shifted" light

versus frequency results in a Raman
spectrum of the sample.
• Generally, Raman spectra are plotted with
respect to the laser frequency such that
the Rayleigh band lies at 0 cm-1.
• On this scale, the band positions will lie at
frequencies that correspond to the energy
levels of different functional group
vibrations
Example of RAMAN Spectra
Example of RAMAN Spectra
 Instrumentation

• A Raman system typically consists of four

major components:
• 1. Excitation source (Laser).
• 2. Sample illumination system and light
collection optics.
• 3. Wavelength selector (Filter or
Spectrophotometer).
• 4. Detector (Photodiode array, CCD or
PMT).
Typical Instrumentation
Layout
 Excitation sources

• Before the development of laser, mercury

lamps were user to tranmist a single or
limited wavelength range of light.
• Now Laser beam is used because of
following properties:
– Monochromatic
– small diameters
– high focusing radiant flux
– linearly polarized
Sample illumination
• Focusing nature of laser
illuminate the sample
• Excitation and the
collection from the sample
is obtained by using many
optical settings such as 90
and 180 scattering
• It consists of an achromatic
lense system with a
collecting lense and a
fosuing lense
• The monochromator must disperse the scattered
light across a slit for sequential presentation to
the detector or across a solid-state diode array.
• The standard dispersing device in modern
Raman spectrometers is the double
monochromator or the dual grating system.
• The disadvantage of this system is the
transmission loss and, as a consequence, the
Diode Array system is now the more popular
 Detection
•Raman spectrum can be photographed with an ordinary
spectrograph.
•Basically there are two different ways to detect and record
Raman lines.
•The easiest way is to gather the scattered light emerging
through a glass window at the end of the Raman sample
tube.
•It is passed through a .prism or grating and then focused on
a photographic plate.
• The plate is then developed and both the line frequencies
and intensities can be measured using external equipments
• Modern spectrometers have photo
multiplier tubes are direct measurements
and automatic scanning of a spectrum.
• The spectrum produced by the
monochromator is passed through a slit
which allows a narrow wavelength region
to pass through which is focused on to a
photo multiplier type detector.
 Direct current amplification
• The electron pulsed from the pm tube are
averaged over time and the resulting dc
current is amplified directly and measured
by picometer.
 Photon counting
• The electron pulses caused by individual
photons reaching the photocathode are
measured
• A substantial portion of dark signal is
electronically discriminated from photon
pulses making detector more sensitive.
• but the maximum signal is limited to a
photon count rate at which phonon event
do not overlap
 CCD detectors
• The CCD (charge coupled device )are
extensively now used in RS
• CCD is a silicon bases array of
photosensitive elements , each one of
which generates photoelectrons and
stores them as a small charge
• Major advantage of CCD is low readout
noise, high quantum efficieny and
sensitivity in wide range
 Wave length selector
• They can be classified into several
categories.
• Interference filter
– Simple
– Variable
– wedge shaped
– Acoustic
– liquid crystal tunable filters
• Prism and grating monochromators and
spectrographs
 Ways to improve
Raman signal intensity
 • Very strong laser pulse with electric
field strength > 109 V·cm-1
Stimulated transforms up to 50% of all laser
Raman
pulse energy into coherent beam at
Stokes frequency υ0 - υm .
• The Stokes beam is unidirectional
with the incident laser beam.
Strongest mode υm in the regular
Raman spectrum is greatly
amplified.
• Weaker Raman active modes are
eliminated
• High frequency acts a secondary
excitation source generating the
second Stokes line with frequency
υ0 - 2υm.
• The second Stokes line generates
the third one with the frequency υ0
- 3υm etc.
• 4-5 orders of magnitude
enhancement
 Coherent
Anti-Stokes Raman, CARS
• It is non-linear” Raman spectroscopy
• Instead of the traditional one laser, two
very strong collinear lasers irradiate a
sample.
• Frequency of the first laser is constant,
while the frequency of the second one can
be tuned in a way that the frequency
difference between the two lasers equals
exactly the frequency of some Raman-
active mode of interest
 CARS(continued)
• If the frequency difference between
two lasers υ1 - υ2 is equal to the
frequency um of a Raman-active
rotational, vibrational or any other
mode then a strong light of frequency
υ1 + υm is emitted.
• To obtain strong Raman signal the
second laser frequency should be
tuned in a way that υ2 = υ1 – υ
• Then the frequency of strong scattered
light will be 2υ1 - υ2 = 2υ1- (υ1 - υm) =
υ1 + υm, which is higher then the
excitation frequency u1 and therefore
considered to be Anti-Stokes
frequency.
 Resonance Raman (RR)
• Some colored substance may absorb laser
beam energy and generate strong fluorescence
which contaminates Raman spectrum
• But that under certain conditions some types of
colored molecules can produce strong Raman
scattering instead of fluorescence.
• The Resonance Raman effect takes place when
the excitation laser frequency is chosen in a way
that it crosses frequencies of electronic excited
states and resonates with them
• Intensity of Raman enhanced 3-5 orders
• Chromophoric group(having the has the
highest level of light absorption), responsible
for the molecule’s coloration, experiences
the highest level of enhancement.
• The highest intensity of Resonance Raman
signal is obtained when laser frequency
equals to the first or the second electronic
excited state
 Surface-Enhanced Raman
Spectroscopy
• Raman signal from molecules adsorbed
on certain metal surfaces can be 5-6
orders of magnitude stronger then the
Raman signal from the same molecules in
bulk volume.
• As intensity of Raman signal isproportional to the square
of electric dipole moment P = αE, there are two possible
reasons
– - the enhancement of polarizability α
– the enhancement of electrical field E.
• One disadvantage of SERS is the difficulty of spectra
interpretation.
• The signal enhancement is so dramatic that Raman
bands that are very weak and unnoticeable in
spontaneous Raman spectra can appear in SERS
• Some trace contaminants can also contribute additional
peaks
 Variations
in Raman spectroscopy
• Angle Resolved Raman Spectroscopy - Not
only are standard Raman results recorded but
also the angle with respect to the incident laser.
If the orientation of the sample is known then
detailed information about the phonon
dispersion relation can also be gleamed from a
single test
• Transmission Raman - Allows probing of a
significant bulk of a turbid material, such as
powders, capsules, living tissue, etc
• There are also medical diagnostic applications
• Optical Tweezers Raman Spectroscopy
(OTRS) - Used to study individual particles, and
even biochemical processes in single cells
trapped by optical tweezers.

• Hyper Raman - A non-linear effect in which the

vibrational modes interact with the second harmonic of
the excitation beam. This requires very high power, but
allows the observation of vibrational modes that are
normally "silent". It frequently relies on SERS-type
enhancement to boost the sensitivity
• Tip-Enhanced Raman Spectroscopy (TERS) - Uses a
metallic (usually silver-/gold-coated AFM or STM) tip to
enhance the Raman signals of molecules situated in its
vicinity. With resolution apex (20-30 nm). TERS has
been shown to have sensitivity down to the single
molecule.
• Spontaneous Raman Spectroscopy (SRS) - Used to
study the temperature dependence of the Raman
spectra of molecules.
• Spatially-offset Raman spectroscopy (SORS), which
is less sensitive to surface layers than conventional
Raman, can be used to discover counterfeit drugs
without opening their packaging, and for non-invasive
monitoring of biological tissue
 Applications
• Characterize materials, measure temperature, and find
the crystallographic orientation of a sample
• Gives information on the crystal orientation
• To detect explosives for airport security
• Used in medicine for real-time monitoring of anaesthetic
and respiratory gas mixtures during surgery
Advantages
 Raman offers several advantages over mid-IR

:
and near-IR spectroscopy, including

• Little or no sample preparation is required

• Water is a weak scatterer - no special
accessories are needed for measuring
aqueous solutions
• Inexpensive glass sample holders are
ideal in most cases
• The standard spectral range reaches well
below 400 cm-1, making the technique
ideal for both organic and inorganic
species
 Raman spectra are "cleaner than mid-IR spectra

• - Raman bands are narrower, and overtone and

combination bands are generally weak
• Fiber optics (up to 100's of meters in
length) can be used for remote analyses
• Since fundamental modes are measured,
Raman bands can be easily related to chemical
structure
• Raman spectroscopy can be used to measure
bands of symmetric linkages which are weak in
an infrared spectrum (e.g. -S-S-, -C-S-, -C=C-)
 Commercially available
Raman instruments
• Princeton Instruments
• PerkinElmer instruments
• HORIBA
• bayspec
Future trends
 Raman optical activity
• Laurence D. Barron  discovered during hisdoctoral work in Oxford
with Peter Atkins of a new optical process involving interference
between light waves scattered via the polarizability and optical
activity tensors of a chiral molecule.
• This leads to a dependence of the scattered intensity on the degree
of circular polarization of the incident beam and to a small circularly
polarized component in the scattered beam.
• He subsequently developed the definitive theory of ROA with David
Buckingham in Cambridge in 1971, and made the first observations
(in the form of a small difference in the intensity of Raman scattering
in right- and left-circularly polarized incident light) with David
Buckingham and Martin Bogaard 1973.
Raman optical activity
 Raman optical activity
• 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
 References

i. Introductory Raman spectroscopy, By John R. Ferraro, Kazuo Nakamoto,

Chris W.
ii. Analytical applications of Raman spectroscopy, By Michael J.
PelletierBrownhttp://www.analyticalspectroscopy.net/spectropaedia.htm#Diode
iii. Molecular spectroscopy, By Chemical Society (Great Britain)
iv. Characterization in Compound Semiconductor Processing, By:
Gary McGuire; Yale E. Strasser
v. http://chemwiki.ucdavis.edu
vi. http://www.analyticalspectroscopy.net
vii. http://www.bayspec.com
viii. http://www.inphotonics.com
ix. www.princetoninstruments.com
x. www.horiba.com