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BIOPHYSICAL METHODS

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Title: BIOPHYSICAL METHODS


1
  • BIOPHYSICAL METHODS

2
  • Analysis of Bio molecules
  • UV and Visible Light
  • Spectroscopy NMR and ESR
  • Circular dichorism
  • X ray diffraction
  • Mass Spectroscopy
  • Surface plasmon Resonance

3
ELECTROMAGNETIC SPECTRUM
4
ELECTROMAGNETIC SPECTUM
5
UV SPECTRUM
6
VISIBLE SPECRUM
7
INFRARED SPECRTUM
8
Quantitative UV
  • Quantitative UV/vis is used to determine the
    concentration of an analyte usually in an aqueous
    solution.

9
  • In order to be able to do this, the analyte must
    absorb in the UV/vis region.

10
  • Beer's Law is a linear relationship between
    absorbance and concentration.
  • A a b c, where c is concentration, A is
    absorbance, b is path length (usually 1 cm) and a
    is the molar absorbtivity.
  • Beer's law is linear.

11
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12
UV Analysis - General Theory
  • UV ANALYZERS
  • The UV region consists of wavelengths from 200 to
    400 nanometers (nm).
  • The visible region extends from 400 to 800 nm,
    and the near IR (NIR) region covers 0.8 to 2.50
    micrometers (jm).

13
  • The UVVIS-NIR is a relatively small part of the
    electromagnetic radiation spectrum, and the
    shorter the wavelength the more penetrating the
    radiation.
  • The region where a compound absorbs radiation
    depends on the energy of the molecular
    transitions.

14
  • High-energy electronic transitions are observed
    in the low-wavelength UV/VIS regions.
  • Moderate-energy vibrational and rotational
    transitions are observed in the high-wavelength
    IR region.

15
  • The Main Components of UV Analyzers
  • 1. Source-provides radiation for the spectral
    region being measured
  • 2. Mono chromator-a device used to select narrow
    bands of wavelengths

16
  • 3. Sample cell-contains the sample at an
    appropriate path length
  • 4. Detector-a device which measures transmitted
    energy and converts it into electrical energy
  • 5. Readout device-provides a means of recording
    the measurement results

17
Radiation Sources
  • The function of the source is to provide
    radiation of sufficient energy to
    makemeasurements in the region of spectral
    interest.

18
  • The cadmium, mercury, and zinc vapor sources that
    are used in the UV region are emission line
    sources.
  • The output of these sources provides radiation
    as narrow discrete emission lines at a
    high-energy level

19
  • Mercury vapor lamps are often used because of
    their long service life.
  • Deuterium arc sources provide a broad band of UV
    radiation at all of the wavelengths in the UV
    region.

20
  • The energy of the deuterium source is relatively
    lower than the energy of the mercury source.

21
  • The two sources used in the visible and NIR
    regions are tungsten filaments and quartz-halide
    lamps.

22
  • Two types of UV energy sources are used
  • broad and discrete line emission sources.
  • The broad emission source provides energy in a
    broad wavelength band, and narrow-band filters
    are used to isolate the wavelengths of interest.

23
  • These sources provide all wavelengths in the
    region but usually have a low-emission, or
    low-energy level, at any given wavelength.
  • Sources of this type include hydrogen, or
    deuterium, discharge lamps tungsten lamps and
    tungsten-iodine lamps.

24
  • Discrete line sources use gas discharge lamps
    with narrow lines of emission.
  • These sources emit radiation energy at various
    discrete wavelengths at a high-energy level
  • The wavelengths that are not desired are
    filtered, leaving only the wavelength of interest

25
  • Tungsten-iodine cycle lamps can be used down to
    300 nm
  • Mercury vapor lamps are the most useful UV
    sources due to their high intensity and long life
  • Medium-pressure mercury lamps can operate down to
    300 nm
  • Zinc discharge lamps are useful due to their 214
    nm emission line

26
The Mono chromator
  • Dispersive and nondispersive mono chromator are
    used in photometric analysis

27
  • A monochromator is an optical device that
    transmits a mechanically selectable narrow band
    of wavelengths of light or other radiation chosen
    from a wider range of wavelengths available at
    the input. The name is from the Greek roots
    mono-, single, and chroma, colour, and the Latin
    suffix -ator, denoting an agent.

28
MONO CROMATOR
29
SPCTROPHOTOMETER
30
  • Spectrophotometers are dispersive instruments and
    photometers are non-dispersive instruments.
  • The function of the mono-chromator is to disperse
    light from a source and selectively pass a narrow
    spectral band to the sample and detector

31
  • Spectrophotometers are dispersive devices that
    are used to scan across a spectrum of
    wavelengths.
  • They can be used to make measurements at several
    wavelengths
  • This capability allows for the analysis of
    multiple components with a spectrophotometer.

32
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33
  • Photometers are non-dispersive devices which
    exclude a large amount of spectral radiation.
  • Photometers are used to make measurements at
    selected discrete wavelengths.

34
  • The measurement wavelength filter is selected to
    match the absorption band of the component being
    analyzed.

35
  • The ratio of the transmitted light at the
    reference and measured wavelengths is measured by
    the photometer.
  • Normally, photometers are used to measure a
    single component in a process stream.

36
The Sample Cell
  • The purpose of the sample cell is to contain a
    representative sample from the process stream.

37
  • Stainless steel is the material most commonly
    used for cell bodies
  • Other metals such as Monel, Hastelloy, and
    titanium are also used.
  • Plastic cell bodies made of Teflon or Kynar are
    used in some applications
  • Quartz, sapphire, and glass cell windows are
    used in the UV-VIS-NIR spectral regions.

38
Detectors
  • Several types of detectors are used in process UV
    analyzers, including phototubes, photomultiplier
    tubes, and photocells.

39
  • The photoelectric effect is used in the vacuum
    phototube to produce a current proportional to
    the energy striking the tube cathode
  • The photomultiplier tube offers very sensitive
    detection of UV and visible light but large
    radiation energy levels will damage the
    light-sensitive surface

40
  • The photocell (photovoltaic) is a semiconductor
    light detector of the barrier layer type
  • A current is developed proportional to the light
    intensity but, the current output is not linear
    with the energy level

41
  • Photomultiplier tubes (PMT) have traditionally
    been used in UV/VIS instruments.
  • The photoelectric effect is used in the PMT to
    produce a current proportional to the radiation
    striking the cathode of the tube.

42
PHOTOMULTIPLIER
43
  • A recent development in photometric analyzers is
    the use of photodiode arrays (PDA).
  • The PDA detectors are used throughout UV-VIS-NIR
    regions.
  • A large number of discrete detectors are located
    in a very close space in the PDA
  • This array of diode detectors allows for all of
    the wavelengths to be measured simultaneously.

44
PHOTODIODE ARRAY
45
Readouts
  • Analog meters, digital meters, strip chart
    recorders, and video display tubes (VDTs) are
    examples of readout devices used in photometers
    and spectrophotometers

46
ANALOGUE METER
47
DIGITAL METER
48
STRIP CHART RECORDER
49
VIDEO DISPLAY TUBE
50
Scanning Spectrophotometers
  • Scanning spectrophotometers are dispersive
    devices that normally utilize diffraction
    gratings to scan across a spectral region
  • Scanning devices can be used for multiple
    component applications
  • . Scanning spectrophotometers can be used in the
    UV, visible, and NIR regions

51
DIFFRACTION GRATING
52
DIFFRACTION GRATING
53
SCANNING SPECTROPHOTOMETER
54
Spectrometer
  • A spectrograph is an optical instrument used to
    measure properties of light over a specific
    portion of the electromagnetic spectrum,
    typically used in spectroscopic analysis to
    identify materials

55
SPECTROMETER
56
UV / VISIBLE SPECTROMETER
57
  • Spectrometer is a term that is applied to
    instruments that operate over a very wide range
    of wavelengths, from gamma rays and X-rays into
    the far infrared
  • If the region of interest is restricted to near
    the visible spectrum, the study is called
    spectrophotometry.

58
Circular dichroism
  • Circular dichroism (CD) is the differential
    absorption of left- and right-handed circularly
    polarized light.
  • A CD Spectrometer is an instrument that records
    this phenomenon as a function of wavelength

59
CIRCULAR DICHORISM
60
CIRCULAR DICHORISM
61
  • CD can be used to help determine the structure
    of macromolecules (including the secondary
    structure of proteins and the handedness of DNA).
  • CD was discovered by the French physicist Aimé
    Cotton in 1896.

62
Interaction of circularly polarized light with
matter
  • The electric field of a light beam causes a
    linear displacement of charge when interacting
    with a molecule, whereas the magnetic field of it
    causes a circulation of charge
  • These two motions combined result in a helical
    displacement when light impinges on a molecule

63
  • The two types of circularly polarized light are
    absorbed to different extents
  • In a CD experiment, equal amounts of left and
    right circularly polarized light of a selected
    wavelength are alternately radiated into a
    (chiral) sample
  • One of the two polarizations is absorbed more
    than the other one, and this wavelength-dependent
    difference of absorption is measured, yielding
    the CD spectrum of the sample.

64
Application to biological molecules
  • In general, this phenomenon will be exhibited in
    absorption bands of any optically active
    molecule.
  • As a consequence, circular dichroism is exhibited
    by biological molecules, because of their
    dextrorotary and levorotary components

65
  • Even more important is that a secondary structure
    will also impart a distinct CD to its respective
    molecules.
  • Therefore, the alpha helix of proteins and the
    double helix of nucleic acids have CD spectral
    signatures representative of their structures
  • The far-UV (ultraviolet) CD spectrum of proteins
    can reveal important characteristics of their
    secondary structure

66
  • CD spectra can be readily used to estimate the
    fraction of a molecule that is in the alpha-helix
    conformation, the beta-sheet conformation, the
    beta-turn conformation, or some other (e.g.
    random coil) conformation

67
  • It can reveal important thermodynamic information
  • CD a valuable tool for verifying that the protein
    is in its native conformation
  • Visible CD spectroscopy is a very powerful
    technique to study metalprotein interactions

68
  • CD gives less specific structural information
    than X-ray crystallography and protein NMR
    spectroscopy
  • for example, which both give atomic resolution
    data
  • However, CD spectroscopy is a quick method that
    does not require large amounts of

69
  • CD can be used to survey a large number of
    solvent conditions, varying temperature, pH,
    salinity, and the presence of various cofactors.

70
Nuclear magnetic resonance
  • Nuclear magnetic resonance (NMR) is the name
    given to a physical resonance phenomenon
    involving the observation of specific quantum
    mechanical magnetic properties of an atomic
    nucleus in the presence of an applied, external
    magnetic field

71
NMR OVER VIEW
72
MAGNETIC RESONANCE
73
  • Many scientific techniques exploit NMR phenomena
    to study molecular physics, crystals and
    non-crystalline materials through NMR
    spectroscopy
  • NMR is also routinely used in advanced medical
    imaging techniques, such as in magnetic resonance
    imaging (MRI).

74
  • All nuclei that contain odd numbers of nucleons
    have an intrinsic magnetic moment and angular
    momentum, in other words a spin gt 0.
  • The most commonly studied nuclei are 1H
  • A key feature of NMR is that the resonance
    frequency of a particular substance is directly
    proportional to the strength of the applied
    magnetic field

75
  • If a sample is placed in a non-uniform magnetic
    field then the resonance frequencies of the
    sample's nuclei depend on where in the field they
    are located
  • The principle of NMR usually involves two
    sequential steps

76
  • The alignment (polarization) of the magnetic
    nuclear spins in an applied, constant magnetic
    field H0.
  • The perturbation of this alignment of the nuclear
    spins by employing an electro-magnetic, usually
    radio frequency (RF) pulse

77
  • The required perturbing frequency is dependent
    upon the static magnetic field (H0) and the
    nuclei of observation.

78
  • The two fields are usually chosen to be
    perpendicular to each other as this maximises the
    NMR signal strength
  • The resulting response by the total
    magnetization (M) of the nuclear spins is the
    phenomenon that is exploited in NMR spectroscopy
    and magnetic resonance imaging

79
  • NMR phenomena are also utilized in low-field NMR,
    NMR spectroscopy and MRI in the Earth's magnetic
    field (referred to as Earth's field NMR), and in
    several types of magnetometers.

80
NMR spectroscopy
  • NMR spectroscopy is one of the principal
    techniques used to obtain physical, chemical,
    electronic and structural information about
    molecules due to either the chemical shift Zeeman
    effect, or the Knight shift effect, or a
    combination of both, on the resonant frequencies
    of the nuclei present in the sample

81
  • It is a powerful technique that can provide
    detailed information on the topology, dynamics
    and three-dimensional structure of molecules in
    solution and the solid state
  • Thus, structural and dynamic information is
    obtainable

82
NMR SPECTROPHOTOMETER
83
High magnetic field (800 MHz, 18.8 T) NMR
spectrometer being loaded with a sample.
84
  • Nuclear magnetic resonance spectroscopy, most
    commonly known as NMR spectroscopy, is the name
    given to a technique which exploits the magnetic
    properties of certain nuclei
  • Many types of information can be obtained from an
    NMR spectrum

85
  • It can, among other things, be used to study
    mixtures of analytes, to understand dynamic
    effects such as change in temperature and
    reaction mechanisms
  • It is an invaluable tool in understanding protein
    and nucleic acid structure and function. It can
    be applied to a wide variety of samples, both in
    the solution and the solid state.

86
The NMR sample is prepared in a thin-walled glass
tube - an NMR tube.
87
  • When placed in a magnetic field, NMR active
    nuclei (such as 1H or 13C) absorb at a frequency
    characteristic of the isotope.
  • The resonant frequency, energy of the absorption
    and the intensity of the signal are proportional
    to the strength of the magnetic field

88
  • For example, in a 21 tesla magnetic field,
    protons resonate at 900 MHz. It is common to
    refer to a 21 T magnet as a 900 MHz magnet,
    although different nuclei resonate at a different
    frequency at this field strength.

89
  • In the Earth's magnetic field the same nuclei
    resonate at audio frequencies. This effect is
    used in Earth's field NMR spectrometers and other
    instruments.

90
Chemical shift
  • Depending on the local chemical environment,
    different protons in a molecule resonate at
    slightly different frequencies

91
  • Since both this frequency shift and the
    fundamental resonant frequency are directly
    proportional to the strength of the magnetic
    field, the shift is converted into a
    field-independent dimensionless value known as
    the chemical shift

92
  • By understanding different chemical environments,
    the chemical shift can be used to obtain some
    structural information about the molecule in a
    sample

93
Correlation spectroscopy
  • Correlation spectroscopy is one of several types
    of two-dimensional nuclear magnetic resonance
    (NMR) spectroscopy
  • This type of NMR experiment is best known by its
    acronym, COSY

94
CORRELATION SPECTROSCOPY
95
  • Other types of two-dimensional NMR include
    J-spectroscopy, exchange spectroscopy (EXSY),
    Nuclear Overhauser effect spectroscopy (NOESY),
    total correlation spectroscopy (TOCSY) and
    heteronuclear correlation experiments, such as
    HSQC, HMQC, and HMBC

96
Solid-state nuclear magnetic resonance
  • A variety of physical circumstances does not
    allow molecules to be studied in solution, and at
    the same time not by other spectroscopic
    techniques to an atomic level

97
  • Applications in which solid-state NMR effects
    occur are often related to structure
    investigations on membrane proteins, protein
    fibrils or all kinds of polymers, and chemical
    analysis in inorganic chemistry, but also include
    "exotic" applications like the plant leaves and
    fuel cells.

98
Electron paramagnetic resonance
  • Electron paramagnetic resonance (EPR) or electron
    spin resonance (ESR) spectroscopy is a technique
    for studying chemical species that have one or
    more unpaired electrons, such as organic and
    inorganic free radicals or inorganic complexes
    possessing a transition metal ion

99
  • The basic physical concepts of EPR are analogous
    to those of nuclear magnetic resonance (NMR), but
    it is electron spins that are excited instead of
    spins of atomic

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  • EPR was first observed in Kazan State University
    by a Soviet physicist Yevgeny Zavoisky in 1944,
    It was developed independently at the same time
    by Brebis Bleaney at Oxford University.

102
EPR spectrometer
103
  • In principle, EPR spectra can be generated by
    either varying the photon frequency incident on a
    sample while holding the magnetic field constant,
    or doing the reverse

104
EPR applications
  • EPR spectroscopy is used in various branches of
    science, such as chemistry and physics, for the
    detection and identification of free radicals and
    paramagnetic centers

105
  • EPR is a sensitive, specific method for studying
    both radicals formed in chemical reactions and
    the reactions themselves
  • For example, when frozen water (solid H2O) is
    decomposed by exposure to high-energy radiation,
    radicals such as H, OH, and HO2 are produced.
    Such radicals can be identified and studied by
    EPR

106
  • Organic and inorganic radicals can be detected
    in electrochemical systems and in materials
    exposed to UV light
  • Medical and biological applications of EPR also
    exist
  • Specially-designed nonreactive radical molecules
    can attach to specific sites in a biological
    cell, and EPR spectra can then give information
    on the environment of these so-called spin-label
    or spin-probes.

107
  • EPR also has been used by archaeologists for the
    dating of teeth.
  • Radiation damage over long periods of time
    creates free radicals in tooth enamel, which can
    then be examined by EPR and, after proper
    calibration, dated

108
  • Radiation-sterilized foods have been examined
    with EPR spectroscopy, the aim being to develop
    methods to determine if a particular food sample
    has been irradiated and to what dose.

109
X-ray scattering techniques
  • This is an X-ray diffraction pattern formed when
    X-rays are focused on a crystalline material, in
    this case a protein
  • Each dot, called a reflection, forms from the
    coherent interference of scattered X-rays passing
    through the crystal.

110
X RAY SCATTERING
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  • X-ray scattering techniques are a family of
    non-destructive analytical techniques which
    reveal information about the crystallographic
    structure, chemical composition, and physical
    properties of materials and thin films

113
  • These techniques are based on observing the
    scattered intensity of an X-ray beam hitting a
    sample as a function of incident and scattered
    angle, polarization, and wavelength or energy.

114
X-ray diffraction techniques
  • X-ray diffraction finds the geometry or shape of
    a molecule using X-rays.
  • X-ray diffraction techniques are based on the
    elastic scattering of X-rays from structures that
    have long range order

115
X RAY DIFFRACTION
116
  • Single-crystal X-ray diffraction is a technique
    used to solve the complete structure of
    crystalline materials, ranging from simple
    inorganic solids to complex macromolecules, such
    as proteins.

117
  • Powder diffraction (XRD) is a technique used to
    characterize the crystallographic structure,
    crystallite size (grain size)
  • Powder diffraction is commonly used to identify
    unknown substances, by comparing diffraction data
    against a database maintained by the
    International Centre for Diffraction Data

118
  • Thin film diffraction and grazing incidence X-ray
    diffraction may be used to characterize the
    crystallographic structure and preferred
    orientation of substrate-anchored thin films

119
  • High-resolution X-ray diffraction is used to
    characterize thickness, crystallographic
    structure, and strain in thin epitaxial films. It
    employs parallel-beam optics
  • X-ray pole figure analysis enables one to analyze
    and determine the distribution of crystalline
    orientations within a crystalline thin-film
    sample

120
Compton scattering
  • Compton scattering or the Compton effect is the
    decrease in energy (increase in wavelength) of an
    X-ray or gamma ray photon, when it interacts with
    matter
  • Compton scattering usually refers to the
    interaction involving only the electrons of an
    atom

121
COMPTON SCATTERING
122
  • The Compton effect was observed by Arthur Holly
    Compton in 1923
  • Arthur Compton earned the 1927 Nobel Prize in
    Physics for the discovery.

123
X-ray Raman scattering
  • X-ray Raman scattering (XRS) is non-resonant
    inelastic scattering of x-rays from core
    electrons
  • .
  • It is analogous to Raman scattering, which is a
    largely-used tool in optical spectroscopy, with
    the difference being that the wavelengths of the
    exciting photons fall in the x-ray regime and the
    corresponding excitations are from deep core
    electrons

124
Mass spectrometry (MS)
  • Mass spectrometry (MS) is an analytical technique
    for the determination of the elemental
    composition of a sample or molecule
  • It is also used for elucidating the chemical
    structures of molecules, such as peptides and
    other chemical compounds

125
MASS SPECTROMETRY
126
MASS SPECTROMETER
127
  • The MS principle consists of ionizing chemical
    compounds to generate charged molecules or
    molecule fragments and measurement of their
    mass-to-charge ratios

128
Typical MS procedure
  • 1) a sample is loaded onto the MS instrument, and
  • 2) the components of the sample ionized by one of
    a variety of methods (e.g., by impacting them
    with an electron beam), which results in the
    formation of charged particles (ions),

129
  • 3) directing the ions into a electric and/or
    magnetic fields
  • 4) computation of the mass-to-charge ratio of the
    particles based on the details of their motion of
    the ions as they transit through electromagnetic
    fields

130
  • 5) detection of the ions, which in step 4) were
    sorted according to m/z.

131
  • MS instruments consist of three modules 1.An ion
    source, which can convert gas phase sample
    molecules into ions
  • A mass analyzer, which sorts the ions by their
    masses by applying electromagnetic fields

132
  • A detector, which measures the value of an
    indicator quantity and thus provides data for
    calculating the abundances of each ion present

133
  • The technique has both qualitative and
    quantitative uses
  • These include identifying unknown compounds,
    determining the isotopic composition of elements
    in a molecule, and determining the structure of a
    compound by observing its fragmentation

134
Main steps of measuring with a mass spectrometer
135
  • Other uses include quantifying the amount of a
    compound in a sample or studying the fundamentals
    of gas phase ion chemistry
  • MS is now in very common use in analytical
    laboratories that study physical, chemical, or
    biological properties of a great variety of
    compounds.

136
Tandem mass spectrometry
  • A tandem mass spectrometer is one capable of
    multiple rounds of mass spectrometry, usually
    separated by some form of molecule fragmentation

137
TANDEM MASS SPECTROMETER
138
  • For example, one mass analyzer can isolate one
    peptide from many entering a mass spectrometer.
  • A second mass analyzer then stabilizes the
    peptide ions while they collide with a gas,
    causing them to fragment by collision-induced
    dissociation (CID).
  • A third mass analyzer then sorts the fragments
    produced from the peptides

139
  • There are various methods for fragmenting
    molecules for tandem MS, including
    collision-induced dissociation (CID), electron
    capture dissociation (ECD), electron transfer
    dissociation (ETD), infrared multiphoton
    dissociation (IRMPD) and blackbody infrared
    radiative dissociation (BIRD).

140
  • An important application using tandem mass
    spectrometry is in protein identification

141
  • An important type of Tandem mass spectrometry is
    Accelerator Mass Spectrometry (AMS), which uses
    very high voltages, usually in the mega-volt
    range, to accelerate negative ions into a type of
    tandem mass spectrometer.
  • One of the most important applications of this
    technique is radiocarbon dating.

142
Mass spectrum analysis
  • Since the precise structure or peptide sequence
    of a molecule is deciphered through the set of
    fragment masses, the interpretation of mass
    spectra requires combined use of various
    techniques

143
  • Usually the first strategy for identifying an
    unknown compound is to compare its experimental
    mass spectrum against a library of mass spectra

144
  • Computer simulation of ionization and
    fragmentation processes occurring in mass
    spectrometer is the primary tool for assigning
    structure or peptide sequence to a molecule

145
  • Another way of interpreting mass spectra involves
    spectra with accurate mass
  • A computer algorithm called formula generator
    calculates all molecular formulas that
    theoretically fit a given mass with specified
    tolerance.

146
Applications
  • Isotope dating and tracking
  • Mass spectrometer to determine the 16O/18O and
    12C/13C isotope ratio on biogenous carbonate

147
  • Pharmacokinetics
  • Pharmacokinetics is often studied using mass
    spectrometry because of the complex nature of the
    matrix (often blood or urine) and the need for
    high sensitivity to observe low dose and long
    time point data

148
  • Protein characterization
  • Mass spectrometry is an important emerging method
    for the characterization of proteins. The two
    primary methods for ionization of whole proteins
    are electrospray ionization (ESI) and
    matrix-assisted laser desorption/ionization
    (MALDI).

149
  • Space exploration
  • As a standard method for analysis, mass
    spectrometers have reached other planets and
    moons. Two were taken to Mars by the Viking
    program

150
CHANDRAYAN
151
High Resolution Mass Spectrometer
152
  • Respired gas monitor
  • Mass spectrometers were used in hospitals for
    respiratory gas analysis beginning around 1975
    through the end of the century

153
Surface plasmon resonance
  • The excitation of surface plasmons by light is
    denoted as a surface plasmon resonance (SPR) for
    planar surfaces or localized surface plasmon
    resonance (LSPR) for nanometer-sized metallic
    structures.

154
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  • This phenomenon is the basis of many standard
    tools for measuring adsorption of material onto
    planar metal (typically gold and silver) surfaces
    or onto the surface of metal nanoparticles.
  • It is behind many color based biosensor
    applications and different lab-on-a-chip sensors.

158
  • Surface plasmons, also known as surface plasmon
    polaritons, are surface electromagnetic waves
    that propagate in a direction parallel to the
    metal/dielectric (or metal/vacuum) interface
  • Since the wave is on the boundary of the metal
    and the external medium , these oscillations are
    very sensitive to any change of this boundary,
    such as the adsorption of molecules to the metal
    surface.

159
  • In order to excite surface plasmons in a resonant
    manner, one can use an electron or light beam
    (visible and infrared are typical
  • The incoming beam has to match its impulse to
    that of the plasmon

160
  • In the case of p-polarized light (polarization
    occurs parallel to the plane of incidence), this
    is possible by passing the light through a block
    of glass to increase the wavenumber and achieve
    the resonance at a given wavelength and angle

161
SPR Emission
  • When the surface plasmon wave hits a local
    particle or irregularity -like on a rough
    surface-, part of the energy can be reemitted as
    light
  • This emitted light can be detected behind the
    metal film in various directions

162
Applications
  • Surface plasmons have been used to enhance the
    surface sensitivity of several spectroscopic
    measurements including fluorescence, Raman
    scattering, and second harmonic generation

163
  • in their simplest form, SPR reflectivity
    measurements can be used to detect molecular
    adsorption, such as polymers, DNA or proteins,
    etc

164
Magnetic Plasmon Resonance
  • Recently, there has been an interest in magnetic
    surface plasmons
  • These require materials with large negative
    magnetic permeability, a property that has only
    recently been made available with the
    construction of metamaterials.
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