Scribd
Upload
40 views19 pages

Spectroscopy and Its Application: Chemical and Biological Detection Professor: Nam Sun Wang Haimo Liu

This document discusses spectroscopy and its applications. It describes different types of spectroscopy including continuous spectroscopy, absorption spectroscopy, and emission spectroscopy. It explains emission and absorption processes. Applications of absorption and emission spectroscopy are discussed including deducing elemental composition of stars and building spectra of planets by comparing absorption and emission lines. Time-resolved fluorescence spectroscopy and its advantages are also summarized. Mathematical methods for spectroscopy analysis including convolution are presented. Finally, applications to living cell spectral imaging are briefly mentioned.

Uploaded by

DEEPA.V UG PHYSICS SF
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
40 views19 pages

Spectroscopy and Its Application: Chemical and Biological Detection Professor: Nam Sun Wang Haimo Liu

This document discusses spectroscopy and its applications. It describes different types of spectroscopy including continuous spectroscopy, absorption spectroscopy, and emission spectroscopy. It explains emission and absorption processes. Applications of absorption and emission spectroscopy are discussed including deducing elemental composition of stars and building spectra of planets by comparing absorption and emission lines. Time-resolved fluorescence spectroscopy and its advantages are also summarized. Mathematical methods for spectroscopy analysis including convolution are presented. Finally, applications to living cell spectral imaging are briefly mentioned.

Uploaded by

DEEPA.V UG PHYSICS SF
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
You are on page 1/ 19

Spectroscopy and its

Application
Chemical and Biological detection
Professor: Nam Sun Wang
Haimo Liu
12/04/2007
Spectrum and Spectroscopy
 Spectrum:
 (a). Different colors observed when the
white light was dispersed through the
prism
 (b). The changing of light intensity as a
function of frequency

 Spectroscopy: Study of spectrum, to


identify substances
Spectroscopy
 Types of spectroscopy:
 (a) Continuous spectroscopy
 (b) Absorption spectroscopy
 (c) Emission spectroscopy
Emission Process
Electrons ground level
Energy emission

Absorb energy

High energy level


Equipment
 Left: Equipment diagram

 Right: Schematic diagram


Applications
 Absorption spectrum: used in deducing
the presence of elements in stars and
other gaseous objects which cannot be
measured directly.

 Emission spectrum: provide a definition


of the spectrum of each atom, used to
be compared with absorption spectrum
Spectrum of planets
 Compare the
absorption spectrum
with the element’s
emission spectrum,
people can build the
spectrum of planets.
Fluorescence Spectroscopy
 Light source, self-emission
which means the electrons
transferred to the lowest
level spontaneously

 Different fluorescence:
 (a) different meta-stable
states
 (b) different various
vibrational states of the
ground state
Time-resolved fluorescence
spectroscopy
 It provides fluorescence intensity decay
in terms of lifetimes
 Advantages:
 enhance the discrimination among
fluorophores (overlapping emission
spectra )
 sensitive to various parameters of the
biological microenvironment
Time-resolved fluorescence
spectroscopy
 Time-resolved laser-induced fluorescence
spectroscopy (tr-LIFS)
Mathematical method
y[n]  T [ x[n]]
T [ x[n]  y[n]]  T [ x[n]]  T [ y[n]]
T [ x[n]]  T [ x[n]]

x[n]   x[k ] [n  k ]
k

T [ x[n]]  T [ x[k ] [n  k ]
k
Mathematical method
T [ x[n]]   x[k ]T [ [n  k ]]
k

y[n]   x[k ]T [ [n  k ]]
k

If  [ x] is an impulse, then the impulse


response will be h[ x ]  T [ [ x]]

h[n  k ]  T [ [n  k ]]
y[n]   x[k ]h[n  k ]
k
Mathematical method
 Based on the definition of convolution:

( f  g )(m)   f (n) g (m  n)
n

y[n]  ( x  h)[n]  x[n]  h[n]


Mathematical method
 For the tr-LIFS system, the impulse response
function IRF (t ) is what would be recorded as
the observed fluorescence decay F (t )

F (t )  I (t )  IRF (t )
Estimation of the intrinsic
fluorescence decay was carried out via
deconvolution of the observed
fluorescence
Spectral Imaging system
 Imaging provides
intensity at every pixel
of the image I (x, y)

 spectrometer provides
the intensity of a
single spectrum, I(λ)

 spectral image provides


a spectrum at each
pixel, I (x, y, λ)
Observation of multiple
activities
 Trying to use 5 different kinds of fluorescent
molecules to label each of the 24
chromosomes in human body
 2 to 5 minus 1=31
Observation of multiple colors
Living cell spectral imaging
 Compromise: only two kinds of cellular organ
were labeled
¿Questions?

You might also like