Professor Dr HANS SCHUESSLER By Ansam Jameel Talib Molecular Physics Course History of Raman Scattering 1923 Inelastic light scattering predicted by A ID: 468320
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Slide1
Raman Spectroscopy and some experimental results
ProfessorDr. HANS SCHUESSLER
By
Ansam Jameel Talib
Molecular Physics Course
Slide2
History of Raman Scattering1923
– Inelastic light scattering predicted by A. Smekel1928 – Landsberg and Mandelstam saw unexpected frequency shifts in scattering from quartz
1928
– C.V. Raman and K.S. Krishnan saw “feeble fluorescence” from neat solvents 1930 – C.V. Raman wins Nobel Prizehttp://www.aps.org/publications/apsnews/200902/physicshistory.cfm
C. V. Raman
http://bwtek.com/raman-theory-of-raman-scattering/Slide3
Raman Spectroscopya spectroscopic technique used to observe vibrational, rotational, and other low-frequency modes in a system.
Raman spectra are similar to infrared spectra .
Useful
for functional group detection and fingerprint regions that permit the identification of specific compounds.
The advantages:
small sample requirement, minimal sensitivity toward interference by water, and high conformational and environmental sensitivity.http://www.inphotonics.com/raman.htmSlide4
Can Raman spectra be obtained from solids, liquids and gasesSolids LiquidsGels
Slurriespowders films, etc. Raman spectra can even be obtained from some metals.
It is possible to obtain Raman spectra of gases.
However, since the concentration of molecules in gases is generally very low, this typically requires special equipment, such as long
pathlength
cells.https://depts.washington.edu/ntuf/facility/docs/NTUF-Raman-Tutorial.pdfSlide5
Types of Raman Spectroscopy
1- Surface-enhanced Raman spectroscopy (SERS)2- Resonance Raman spectroscopy 3- Angle-resolved Raman spectroscopy 4- Hyper Raman5- Spontaneous Raman spectroscopy (SRS)6- Optical tweezers Raman spectroscopy (OTRS) 7- Stimulated Raman spectroscopy8- Spatially offset Raman spectroscopy (SORS)
9- Coherent anti-Stokes Raman spectroscopy (CARS)
10- Raman optical activity (ROA)
11. Transmission Raman 12. Inverse Raman spectroscopy13- Tip-enhanced Raman spectroscopy (TERS)14- Surface plasmon polariton enhanced Raman scattering (SPPERS) 15- Stand-off Remote Raman 16- Confocal Ramanhttp://en.wikipedia.org/wiki/Raman_spectroscopySlide6
Tip-enhanced Raman spectroscopy (TERS)Tip-Enhanced Raman (TERS or
nanoRaman): chemical imaging at the nanoscaleTERS (or nano-Raman) brings you the best of both worlds: the chemical specificity of Raman spectroscopy with imaging at spatial resolution typically down to 10nm. This technique can be demonstrated on various samples ranging from nanotubes to DNA. TERS has been shown to have sensitivity down to the single molecule level and holds some promise for bioanalysis applications
http
://www.intechopen.com/books/electronic-properties-of-carbon-nanotubes/detection-of-carbon-nanotubes-using-tip-enhanced-raman-spectroscopy
http://www.intechopen.com/books/electronic-properties-of-carbon-nanotubes/detection-of-carbon-nanotubes-using-tip-enhanced-raman-spectroscopy
http://www.asdn.net/asdn/nanotools/spm.shtmlSlide7
Confocal Raman
Couples a Raman spectrometer to a standard optical microscope, allowing high magnification visualization of a sample and Raman analysis with a microscopic laser spot. Raman microscopy is easy: simply place the sample under the microscope, focus, and make a measurement. Just adding a microscope to a Raman spectrometer does not give a controlled sampling volume - for this a spatial filter is required. Confocal Raman microscopy refers to the ability to spatially filter the analysis volume of the sample, in the XY (lateral) and Z (depth) axes.
http://www.horiba.com/us/en/scientific/products/raman-spectroscopy/raman-academy/raman-faqs/what-is-confocal-raman-microscopy/Slide8
Confocal Principle
Laser (
λ
nm): power (
mW
)
405 nm : 0.65
mW
532 nm : 10.5
mW
660 nm : 12.3
mW
785 nm: 35.5
mW
The
LabRAM
HR Evaluation is an integrated Raman system. The microscope is coupled to a 800 mm focal length spectrograph equipped with two switchable gratings
.
http://
www.horiba.comSlide9
Ansam J. Talib1
, Sandra C. Bustamante1 , Zachary N. Liege 1,2
, Sarah Ritter
1
, Alexander Sinyukov1
, Dmitri V. Voronine1,2, Alexei V. Sokolov1,2,
Kenith Meissner
1
and
Marlan
O. Scully
1,2,3
1
Texas A&M University,
2
Baylor University,
3
Princeton University
Raman
Spectroscopy
and Imaging of Red Blood CellsSlide10
MotivationsToday, more than 340 million people
suffer from diabetes and this number doubles every 15 years.*
In the USA, 30 million people have diabetes and 87 million more have pre-diabetes.
Common diabetic monitoring procedures include
checking blood sugar levels multiple times a day by finger pricks using
a glucose meter or an implantable glucose biosensor.**
*Danani
G, Finucane
MM, et al. National,
regional,and
global trends in fasting plasma
gluocose
and diabetes prevalence since 1980: systematic analysis of health examination surveys and epidemiological studies with 370 country- years and 2.7 million participants. The Lancet 2011; 378:31-40.
**
Vashist
SK. Non-invasive glucose monitoring
technologyin
diabetes management: A review. Anal
Chim
Acta
2012;750:16-27.
http://
www.medicalmalpracticeinquirer.com/assets_c/2011/01/iStock_000004641088Large-thumb-300x200-6230.jpgSlide11
Goals of the projectOur long-term goal is to develop red blood cells (erythrocytes) loaded with a fluorescent dye (
erythrosensors), which can be detected with a light source through the skin and can be used as biocompatible glucose bio-sensors (because such cells will stay in circulation instead of triggering an immune response, and getting out of circulation
).
The specific aim is to measure
the difference between FITC-ghost glygly and normal red blood cells.Slide12
Red blood cells (RBCs)RBCs (also called erythrocytes) are the most common type of blood cells
and are the vertebrate organism's principal means of delivering oxygen to body tissues.RBCs are the most abundant cells in blood, with a shape of a biconcave disk with a flattened center (look like donuts).
RBCs
contain a special protein called
hemoglobin, which helps carrying oxygen.http://bio662.dyndns.info/s3b/b3n/b3n02BloodCirculation/b3n02eBldC112BloodCells.htmSlide13
Cell MembranesRBC membrane plays many roles that aid in regulating their surface deformability, flexibility, adhesion to other cells and immune
recognition.They can be squeezed to 3 microns.
Membranes of RBC
100x Slide14
Fluorescein isothiocyanate (FITC)
Fluorescein (mistakenly abbreviated by its commonly-used reactive isothiocyanate form, FITC) is a small organic molecule, and is typically conjugated to proteins via primary amines. Usually, between 3 and 6 FITC molecules are conjugated to each antibody
In cellular biology, FITC is often used to label and track cells
Excitation: max = 495 nm
Emission: max = 525
nm http://www.sigmaaldrich.com/content/dam/sigmaaldrich/docs/Sigma/Product_Information_Sheet/f7250pis.pdfSlide15
RBC
Membranes
of RBC
FITC ghost Slide16
Raman Spectrum of Hemoglobin
Laser 532 nm
Slit 200
Grating 600 gr/mm
Objective 50xSlide17
Optical image and Raman spectrum of RBC
Laser 532 nm
Slit 200
Grating 600 gr/mm
Objective 50
xObjective 50xSlide18
Raman Images of RBC
Bimolecular class: Unknown
Trp
: tryptophan
C-N stretch
Cytochrome- like moiety
(resonance enhanced)
Protein
Slide19
Raman Spectrum of FITC Ghost Cells
Laser 532 nm
Slit 200
Grating 600 gr/mm
Objective 50xSlide20
Optical and Raman Images of FITC Ghost Cells
Laser 532 nm
Slit 200
Grating 600 gr/mm
Objective 50xSlide21
RBC Slide22
ConclusionsConfocal microscopy will allow us to investigate the vesicle size and shape of RBCs and FITC ghost.We can get a clear understanding for the distribution of the hemoglobin inside the RBCs and the FITC-ghost.
TERS of RBCsSlide23
Thanks For Your ATTENTION !!!!