Introduction to Raman

spectroscopy
second part
Prof. Antonio Agresti
Università degli Studi di Roma
“Tor Vergata”
 Quantum argumentation
Classic argumentation Ianti-Stokes < Istokes ?

NECESSARY QUANTUM TREATMENT

 Rotational energy transitions
The angular momentum J and the energy
of the levels rotationals are quantized
•I moment of inertia
•J=0,1,2…

ALLOWED TRANSITIONS: ΔJ=0;±2

 Vibrational energy transitions

Vibrational energy levels is quantized
•ω frequency of the
incident radiation
•ν=0,1,2,…
ALLOWED TRANSITIONS : Δν=±1
 Quantum argumentation
The allowed transitions are equally probable but ..….
they do not produce lines of equal intensity
RAMAN BAND INTENSITY:
 Degeneration: different energy levels
characterized by the same energy
 Population

where
B=ħ2/2I
e I moment
of inertia

 Temperature
Raman effect
 Raman spectroscopy advantages

Non-invasive and non-destructive technique that can be carried

out "in situ".
 Sample measurement by microscope.
Analysis of small samples thanks to the spatial resolution of the
micron order.
Organic and inorganic liquid solid sample.
 Any chemical or mechanical treatment of the sample to be
analyzed is
Water and glass do not cause interference.
 Analysis at room temperature and pressure.
 Measurements in temperature.
 Short measurement times: from 1 second to a few minutes.
 Raman spectroscopy advantages

 Highly sensitive analytical technique for identification

of different chemical compounds.
 Information on molecular and crystalline symmetry e
on the phase of the test sample (liquid, solid or gaseous).
 Limits of Raman spectroscopy

Weak efficiency of the Raman effect and therefore poor

sensitivity.

 Not all substances are active Raman

(eg metals are not).

 Some samples may undergo thermal degradation phenomena

induced by the laser.

 Fluorescence phenomena cover the weak Raman signal

making the spectra difficult to read.

 Poor spatial resolution in depth

(apparent limit).
 Main instrumental components

Excitation Managment
source system

Centering
detector
optics

Sample Collection
Spectrometer
optcis
 Laser sources

ArKr+
Kr+ Gas
Gas 647nm
325nm
nm

Solid Solid Solid He-Ne Solid

Ar+ Ar+ Ar+ Diodo NdYAG
State State State Gas State 1060nm
Gas Gas Gas 785nm
355nm 457nm 532nm 632nm 638nm (FTRaman)
244nm 488nm 514nm
 Instrumentation for Raman spectroscopy
Sources

High monochromaticity
Need to distinguish the Raman lines
High directionality
Monochromatic source
High coherency

High power
Raman intensity and excitation wavelenght λ
 Raman vs Fluorescence

Incidence of light radiation on the sample.

Loss of energy and decay in the state excited (thermalisation).
Decay at the ground state.

FLUORESCENCE!!!
200

180
Raman signal
intensity (cm-1)

160

140

120

100

80
Fluorescence
60
background
40

20
500 1000 1500 2000 2500 3000
Raman shift (cm-1)
 λ vs fluorescence

• Generally a Raman spectrum

532 nm
is independent of the excitation
source.
• In some cases interference
from a fluorescence background
signal may occur.
633 nm • The ability to easily change
the source is helpful for:
- Remove the fluorescence
background
- Increase the Raman signal
- having more sources,
especially in the analysis of
pigments, is of considerable
importance.
785 nm
Depth of penetration into λ
 Resonant Raman

OBJECTIVE: INCREASE IN SENSITIVITY

Raman band intensity:

It is possible to increase the intensity with a ν0 of the incident radiation

corresponding to an electronic transition of the investigated molecule.

 The electronic transition is allowed

and therefore more probable;
 Scattered intensity light increases
by a factor of 102-106.
 The Raman spectrum is simplified:
the vibrational modes contributing to
the more intense scattering are
reduced in number.

CHOICE OF THE MOST CONVENIENT LASER

 Sampling systems

REMOTE

MACRO

MICRO
 Micro sampling systems

bright spot diameter

excitation wavelength (l)

D = 1.22 l / NA
numerical aperture of the objective (NA)
where NA=n sinα

NA = 0.9
 Micro-Raman confocal sampling

•Axial resolution: about 1μm  The confocal aperture (pin-hole) is

•Lateral resolution: <1μm P' Confocal hole intended to block the Raman signal
from regions outside the focal plane.
P'

"OPTICAL SECTION" OF THE SAMPLE:

P' THE DEPTH FROM WHICH IS SELECTED
COLLECT THE DESIRED SIGNAL

The P‘ image of the laser MULTILAYER SAMPLES ANALYSIS

focused at point P coincides
exactly with the confocal
3D RAMAN IMAGES SAMPLE
aperture.

 A proper compromise between

"depth of fire" and Raman signal
P strength must be reached.
P
P
 Dispersive systems

They allow to separate the different spectral

components through the use of diffraction gratings
Dispersive system: monochromator

 Entrance slit
 Collimator element (lens
or mirror)
 Prism or stepped grating
(as a dispersing element)
 Focusing element (lens or
mirror)
 Exit split
 Diffraction Gratings
In optics, a diffraction grating is an optical component with a
periodic structure that splits and diffracts light into several
beams travelling in different directions. The directions of these
beams depend on the spacing of the grating and the
wavelength of the light so that the grating acts as
the dispersive element. Because of this, gratings are commonly
used in monochromators and spectrometers.
For practical applications, gratings generally have ridges
or rulings on their surface rather than dark lines. Such gratings
can be either transmissive or reflective.

The relationship between the grating spacing and the angles of

the incident and diffracted beams of light is known as the
grating equation. According to the Huygens–Fresnel principle,
each point on the wavefront of a propagating wave can be
considered to act as a point source, and the wavefront at any
subsequent point can be found by adding together the
contributions from each of these individual point sources.
 Diffraction Gratings
An idealised grating is made up of a set of slits of spacing d (from here the importance of
number of line/mm), that must be wider than the wavelength of interest to cause
diffraction. Assuming a plane wave of monochromatic light of
wavelength λ with normal incidence (perpendicular to the grating), each slit in the grating
acts as a quasi point-source from which light propagates in all directions (although this is
typically limited to a hemisphere). After light interacts with the grating, the diffracted light is
composed of the sum of interfering wave components emanating from each slit in the
grating. At any given point in space through which diffracted light may pass, the path length
to each slit in the grating varies. Since path length varies, generally, so do the phases of the
waves at that point from each of the slits. Thus, they add or subtract from each other to
create peaks and valleys through additive and destructive interference.
When the path difference between the light from adjacent slits is
equal to half the wavelength, λ/2, the waves are out of phase, and
thus cancel each other to create points of minimum intensity.
Similarly, when the path difference is λ, the phases add together
and maxima occur. The maxima occur at angles θm, which satisfy the
relationship d sinθm/λ = | m |, where θm is the angle between the
diffracted ray and the grating's normal vector, and d is the distance
from the center of one slit to the center of the adjacent slit, and m is
an integer representing the propagation-mode of interest.
 Dispersive systems: spectral resolution
Low spectral resolution

CCD
size

To increase the spectral resolution, the

lines of the diffraction grating must be
increased.

CCD
size
Recording of the entire
spectrum (4000 cm-1) requires
rotation of the diffraction
grating
Average spectral
resolution
 Notch filter

The notch filter has the task of separating the Rayleigh emission
from that diffused for Raman effect, because otherwise the Rayleigh
spread, 106 to 1012 times more intense than Raman, would totally
cover the latter. The filter blocks the light with the same frequency
as used laser, while it allows the passage of other frequencies. It
worth to note that light with a shorter and larger wave number shift
of about 100 cm-1 with respect to that of the laser is equally
reflected, not allowing the analysis of the Raman light of such
frequencies.
Consequently:
Every laser needs its own notch
 Notch Filter
Transmission curve for a notch filter, an Holographic Super-Notch-Plus Filter
produced by Kaiser Optical System Inc. and centered at 632.8 nm.

The notch actually has a narrow filtering band that prevents light
transmission with a frequency close to that of the incident light and
instead guarantees an excellent transmission (~ 80%) of the other
frequencies.
 Multi-channel detectors

Pixels Features:
•High sensitivity
•High acquisition speed
CCD adapter
(charge-coupled device)
•Reduced background noise
(temperature function)
Cooling down •Possibility of two-dimensional
system
acquisition
•Limited linearity of response
Sensor •Resolution limited by the size of the
To the Acquisition System pixels
 Raman Imaging
• The laser is focused on the sample (spot size ~ 1 μm)
• The sample moves in X and in Y thanks to a motorized stage
• For each new position of the stage a spectrum is acquired
• The software reconstructs the multispectral image.

Aspirin 48.36%

Acetaminophen 35.07%

Cellulose 3.59%

Caffeine 12.35%

The stage moves in X and in Y

 Raman applications

Nanotechnology Biosciences and biotechnologie

Geoscience and gemology

Art and cultural heritag

Pharmaceutical industry Environmental Sciences

ScieMaterials science and semiconductors Forensic analysis