VIETNAM NATIONAL UNIVERSITY HO CHI MINH CITY
HO CHI MINH CITY OF TECHNOLOGY
CHAPTER 4
RAMAN SPECTROSCOPY
ENGINEERING OPTICS AND APPLICATION
1. Raman scattering
Dr. Pham Thi Hai Mien
2. Raman spectrophotometer
Faculty of Applied Science
3. Application of Raman spectroscopy
1. RAMAN SCATTERING
• Raman spectroscopy is based on the Raman effect, according to which when the
laser light interacts with molecular vibrations in the analyte, the energy of the
laser photons is shifted down (Stokes shift) or up (anti-Stokes shift). The shift
in photon energy gives information about the chemical bonds in the analyte.
• Most of the scattered light is at the same frequency ( or wavelength) as the laser
source and does not provide any useful information. This is called elastic or
Rayleigh scattering and is the dominant process.
• A small amount of the incident light (typically 1 in 10 million photons) is
scattered at different frequencies (or wavelengths), which depend on the
chemical structure of the analyte. This is called inelastic or Raman scattering.
• If the molecule gains energy from the photon during the scattering (excited to a
higher vibrational level) then the scattered photon loses energy and its
wavelength increases, which is called Stokes Raman scattering.
• Inversely, if the molecule loses energy by relaxing to a lower vibrational level,
the scattered photon gains the corresponding energy and its wavelength
decreases; which is called anti-Stokes Raman scattering.
• A Raman spectrum is a plot of the intensity of Raman scattered radiation as a function of
Stokes Raman scattering is always more intense and probable than the anti-Stokes its frequency difference from the incident radiation (usually in units of wavenumbers, cm-
1). This difference is called the Raman shift.
and for this reason, it is nearly always the Stokes Raman scatter that is measured in
Raman spectroscopy. • The center of the spectrum is the Rayleigh scatter peak at the laser wavelength. This peak
is millions of times more intense than the Raman scatter peaks. Symmetrically placed on
either side of the Rayleigh peak are the three Stokes and three Anti-Stokes peaks
corresponding to the three most intense Raman active vibrations. The Anti-Stokes lines are
much weaker than the Stokes due to the larger population of molecules in the ground
vibrational level of each mode.
Raman Spectrum of CCl4 measured using a 532 nm laser
CRITERIA OF A COMPOUND TO BE RAMAN ACTIVE A COMPARISON OF RAMAN AND IR SPECTROCOPY
• In order for a vibrational transition to be Raman active, the molecule must
Raman IR
undergo a change in polarizability during the vibration.
Vibrational spectroscopy
• In contrast, for a molecule to be infrared active, the vibration must cause a change
in the dipole moment due to electric field of the radiation. Based on inelastic scattering process Based on absorption process
• For example: Detects vibrations involving a change Detects vibrations involving a change
- The symmetric stretch of CO2 is Raman active but invisible in infrared in polarizability Symmetric in dipole moment Materials with
spectra since the molecule has no permanent dipole. The asymmetric vibrations are observable low polarizability are observable
stretches of CO2, which induce a dipole, are infrared active but Raman Fluorescence can contaminate Raman Background fluorescence can
inactive. spectra overwhelm Raman
- Diatomic molecules are observed in the Raman spectra but not in the IR More expensive Simpler and cheaper instrumentation
spectra. Water can be used as solvent suited Strong infrared absorption of water in
• As a result of this fundamental difference, the IR and Raman spectra provide for in vivo tissue analysis tissues
complementary information about many of the vibrations of molecular species. Measured in UV, Vis and IR regions Measured in IR region
Raman spectra are usually less complex than IR spectra.
Monochromatic radiation sources Polychromatic radiation sources
(lasers)
2. RAMAN SPECTROPHOTOMETER
Main parts of an Raman spectrophotometer:
• A radiation source
• A sample cell
• Optical filters
• A spectrometer
Typically, a sample is illuminated with a laser beam. Elastic scattered radiation at RADIATION SOURCES
the wavelength corresponding to the laser line (Rayleigh scattering) is filtered out
by either a notch filter, edge pass filter, or a band pass filter, while the rest of the • Lasers are used as photon sources due to their highly monochromatic nature and
collected light is dispersed onto spectrometer. high intensity.
• Laser wavelengths ranging from UV through Vis to NIR can be used for Raman
spectroscopy.
• UV: 244 nm, 257 nm, 325 nm, 364 nm.
• Vis: 457 nm, 473 nm, 488 nm, 514 nm, 532 nm, 633 nm, 660 nm.
• NIR: 785 nm, 830 nm, 980 nm, 1064 nm.
SAMPLE CELLS
• Sample cells must be transparent to either UV or Vis or IR radiation, depending on the
radiation sources.
OPTICAL FILTERS SPECTROMETER
• A notch filter is used to selectively block the laser line (Rayleigh scattering) while allow • A Raman spectrometer typically uses a holographic grating and a CCD in order to detect
the Raman scattered wavelength through to the spectrometer, since a notch filter has a Raman scattered wavelengths.
sharp and discrete absorption. • Typical gratings vary from 300 gr/mm (low resolution) to 1800 gr/mm (high resolution).
• Each laser wavelength requires an individual filter. • A CCD (Charge coupled device) is a silicon based multichannel array detector of UV, Vis
or NIR light.
• CCD are used because they are extremely sensitive to light and thus suitable for analysis
of the weak Raman signals, and allow multichannel operation (which means that the entire
Raman spectrum can be detected in a single acquisition).
Transmission spectrum of notch filter
3. APPLICATION OF RAMAN SPECTROSCOPY
• IR spectroscopy is used in both qualitative and quantitative analyses.
• Qualitative analysis: The general spectrum profile (peak position and relative • Normally blood glucose levels are measured invasively
peak intensity) provides a unique chemical fingerprint which can be used to by a finger prick; a droplet of blood is applied to a
identify a material, and distinguish it from others. Often the actual spectrum is glucometer that uses a disposable test strip to measure
quite complex, so comprehensive Raman spectral libraries can be searched to the glucose concentration in the blood. There is no
find a match, and thus provide a chemical identification. doubt that electrochemical glucose sensor technology
• Quantitative analysis: The intensity of a spectrum is directly proportional to will continue to be most accurate, affordable and
concentration. Typically, a calibration procedure will be used to determine the suitable for in-home use by patient for self-monitoring
relationship between peak intensity and concentration, and then routine and management of diabetes. However, patients
measurements can be made to analyze for concentration. suffering from so-called brittle fingers may have
• Typical examples of where Raman is used today: serious problems measuring blood glucose as
- Art: characterization of pigments, ceramics and gemstones. frequently as needed.
- Carbon materials: structure and purity of nano-tubes, defect characterization. • Noninvasive glucose monitors are not yet available,
- Chemistry: structure, purity, and reaction monitoring. but patients would benefit highly from such a device.
- Geology: mineral identification and distribution, fluid inclusions. Raman spectroscopy has been investigated previously
- Life sciences: single cells and tissue, drug interactions, disease diagnosis. for its applicability on noninvasive glucose measuring.
- Pharmaceutics: content uniformity and component distribution.
Typical Raman spectrum of Glucose The schematic of non-invasive Raman spectroscopy for
continuous and pain-free glucose monitoring