Transcript Principles of Fluorescence Spectroscopy

XMUGXQ PFS0601
Principles of Fluorescence
Spectroscopy
Chemistry Department
XMU
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Chapter Six
Methods
of
Fluorometry
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Methods of fluorometry
6.1 Introduction
6.2 Conventional Fluorometry
6.3 Synchronous Fluorescence Scan
6.4 3D fluorescence spectroscopy
6.5 Derivative fluorescence spectroscopy
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6.1 Introduction
Light source
Models
Steady-state fluorescence measurement
I0
t
Time-domain fluorescence measurement
I0
Time resolved fluorescence
t
Frequency-domain fluorescence
measurement
I0
t
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6.3 Synchronous Fluorescence Scan SFS
Conventional Fluorometry
Excitation spectrum
Scan ex at a certain em
Emission spectra
Scan em at a certain ex
em
F
F
3
1 2 4
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

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Synchronous fluorescence scan SFS
Scan ex and em simultaneously
models
Constant-wavelength SFS
  em  ex  constant
Constant-energy SFS
v  vex  vem  constant
Variable-angle SFS
  em  ex  constant
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Constant-wavelength SFS
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Narrowing
spectral band
&
simplifying
emission spectrum
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Narrowing
spectral band
&
simplifying
emission spectrum
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Multi-component analysis
F
 
A
ex
B
ex

A
em

B
em

F
F
CFS

B
em
SFS
    
B
em
em
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B
ex
em
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Theory
I SFS  KclEx (em   )Em( em )
 KclEx (ex )Em( ex   )
Where Ex is the excitation wave function at a given
wavelength; Em is the emission wave function at a given
wavelength; l is the thickness of the sample; K is the
instrumental geometry and related parameters.
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Choosing the 
Generally, the stoke’s shift
F
 
A
ex
B
ex

A
em

B
em

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Choosing the 
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Choosing the 
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Constant-energy SFS
PAHs
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Variable-angle
SFS
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Variable-angle SFS
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Variable-angle SFS
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Variable-angle SFS
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Variable-angle SFS
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Variable-angle SFS
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Reference
Trends in analytical chemistry, 21(12): 787-798, 2002
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3 dimensional fluorescence spectroscopy
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Contour 3D spectra
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Finger printing
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Volume integral
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Derivative fluorescence
芘
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Derivative fluorescence
峰零法
峰距法
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切线法
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application
b
a
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application
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application
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application
Finger printing
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