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Molecular Fluorescence Spectros

Molecular fluorescence spectroscopy involves exciting molecules to higher energy levels using electromagnetic radiation like UV light from a lamp. The excited molecules then emit fluorescence which can be analyzed using instrumentation containing an excitation source, sample cell, and fluorescence detector. Applications include quantitative analysis of molecules in solution, fluorescence detection in liquid chromatography, and studying electronic structure of atoms. Laser induced fluorescence spectroscopy uses fluorescent dyes like HpD localized in tumors for cancer imaging. Fluorescence spectroscopy can also detect oil slicks by analyzing fluorescing hydrocarbons and determine glucose levels using fluorescent Au nanoclusters.

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700 views10 pages

Molecular Fluorescence Spectros

Molecular fluorescence spectroscopy involves exciting molecules to higher energy levels using electromagnetic radiation like UV light from a lamp. The excited molecules then emit fluorescence which can be analyzed using instrumentation containing an excitation source, sample cell, and fluorescence detector. Applications include quantitative analysis of molecules in solution, fluorescence detection in liquid chromatography, and studying electronic structure of atoms. Laser induced fluorescence spectroscopy uses fluorescent dyes like HpD localized in tumors for cancer imaging. Fluorescence spectroscopy can also detect oil slicks by analyzing fluorescing hydrocarbons and determine glucose levels using fluorescent Au nanoclusters.

Uploaded by

leo mark
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© © All Rights Reserved
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MOLECULAR FLUORESCENCE

SPECTROSCOPY
1. Introduction
2. Instrumentation
3. Types
4. Some applications
 Molecular fluorescence is the optical emission from
molecules that have been excited to higher energy
levels by absorption of electromagnetic radiation.

 Analytical applications include quantitative


measurements of molecules in solution and
fluorescence detection in liquid chromatography.
 Atoms, molecules, or solids that are excited to high
energy levels can decay to lower levels by emitting
radiation (emission or luminescence).
 For atoms excited by a high-temperature energy source
this light emission is commonly called atomic or optical
emission and for atoms excited with light it is called
atomic fluorescence.
 A typical fluorimeter contains an excitation
source, sample cell, fluorescence detector.
 Molecules in solution are usually excited by UV light and
the excitation source is usually a deuterium or xenon
lamp.
 Broad-band excitation light from a lamp passes through
a monochromator, which passes only a selected
wavelength.
 The fluorescence is dispersed by another
monochromator and detected by a photomultiplier tube.
 Scanning the excitation monochromator gives the
excitation spectrum and scanning the fluorescence
monochromator gives the fluorescence spectrum.
Simple instruments sometimes use only a bandpass
filter to select the excitation wavelength.
ATOMIC-FLUORESCENCE
SPECTROSCOPY (AFS)
 Atomic fluorescence is the optical emission from gas-
phase atoms that have been excited to higher energy
levels by absorption of electromagnetic radiation.
 The main advantage of fluorescence detection compared
to absorption measurements is the greater sensitivity
achievable because the fluorescence signal has a very
low background.
 AFS is useful to study the electronic structure of atoms
and to make quantitative measurements.
 nalytical applications include flames and plasmas
diagnostics, and enhanced sensitivity in atomic analysis.
Laser induced fluorescence spectroscopy of
human tissues for cancer diagnosis
(hematoporphyrin derivative--HpD)
Laser Induced Fluorescence (LIF) has been
used for diagnosing cancer in two ways. One
approach involves systemic administration of a
drug like hematoporphyrin derivative (HpD)
which is selectively retained by the tumour.
When photo excited with light of appropriate
wavelength the drug localized in the tumour
fluoresces. This fluorescence is used for
detection and imaging of the tumour.
 Study of Marine Petroleum Pollutants
Fluorescence spectroscopy is one of the good technique to
detection of oil slicks on the water surface, determination of petroleum
contaminants in seawater and determination of particular petroleum
derivative compounds as well as identification of pollution sources.
Main components of any oil are hydrocarbons. The other components
are primarily derivatives of hydrocarbons containing single atoms of
sulfur, oxygen or nitrogen. Only a few of hydrocarbons fluoresce, while
the major of them show no ability to luminescence. The content of
compounds able to fluorescence rarely exceeds 10% of the oil mass.
At the same time the petroleum strongly absorbs radiation, especially
the ultraviolet and blue light.
 Accurate determination of glucose
Glucose is considered as a major component of animal and
plant carbohydrates in biological systems. Furthermore, blood glucose
levels are also an indicator of human health conditions: the abnormal
amount of glucose provides significant information of many diseases
such as diabetes or hypo glycemia. Fluorophotometry was used
widely owing to its operational simplicity and high sensitivity. Recently
bio-molecule-stabilized Au nanoclusters were demonstrated as a
novel fluorescence probe for sensitive and selective detection of
glucose.

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