Luminescence

Prepared and Presented by :

Amany Mohamed Elshamy
Department of Medical Laboratories
Badr Academy
 Outlines

Introduction Definition Principle Components

Application Summary Question

2
 • Luminescence is the emission of visible or
near-visible (λ = 300–800 nm) radiation
which is generated when an electron
transitions from an excited state to ground
state.
Luminescence
• Instruments or measuring this type of light
emission are known generically as
luminometers.
Excitation event
 Luminescence
• Chemiluminescence,
bioluminescence, and
electrochemiluminescence
are types of luminescence in
which the excitation event is
caused by a chemical,
biochemical, or
electrochemical reaction,
and by photo illumination.
Chemiluminescence
Basic concept of
Chemiluminescence
• The physical event of light
emission in chemiluminescence
occurs from an excited singlet
state, and the light is emitted
when the electron returns to the
ground state.
• Chemiluminescence is the
emission of light when an
electron returns from an excited
or higher energy level to a lower
energy level.
Basic concept of
Chemiluminescence
• The excitation event is caused by a
chemical reaction with compound such
as
(1) Luminol
(2) Isoluminol
(3) acridinium esters
(4) Luciferin
(5) Hydrogen peroxide
(6) Hypochlorite
(7) Qxygen
Luminol
• Luminol is an organic
compound
which, when
oxidized, emits
light — a
phenomenon known
as
chemiluminescence
 Light is emitted from the excited product
formed in the oxidation reaction.
These reactions occur in the presence of
catalysts such as
Basic concept of
• (1) Enzymes (e.g., alkaline phosphatase,
Chemiluminescence
horseradish peroxidase, microperoxidase).
• (2) Metal ions or metal complexes (e.g.,
Cu2+ and Fe3+ phthalocyanine complex).
• (3) Hemin.
Principle of Chemiluminescence
Chemiluminescence
Chemiluminescence

• Chemical Rx that
produces light.
• Usually involves
oxidation of luminol,
acridinium esters,…..etc
Components
 • Reagent probes
• Sample
Components • Reagent cuvette
parts • Photomultiplier tube
• Readout device.
Chemiluminescence • Chemiluminescence • Light will be
immuno-assays is the production of emitted when the
based on
light following a/an enzyme-labeled
• Immune Rx (Ag-Ab molecule reacts
Rx). with the
substrate.
• Chemical Rx
(Luminol). • Doesn’t require
monochromators
like fluorometry.
Method
Applications
• Chemiluminescence assays are
ultrasensitive and have wide
dynamic ranges.
• They are now widely used in
automated immunoassay and DNA
probe assay systems with no
radiation.
• Use in environmental monitoring,
clinical diagnosis, food safety and
pharmaceutical analysis.
Clinical laboratory
• Hormones and vitamins (ex; thyroid
profile test, B-HCG, PRL, Vit B12, Vit
D….)
• Viruses' antibodies test.
• Tumor markers (ex; CA15.3,CEA,
PSA…).
• Ferritin.
• Toxoplasma, Rubella.
• Cardiac assays (Troponin, CK…)
 • High cost.
• Limited Ag detection.
Limitations • Limited tests panel
• Closed analytical systems.
Question
• Which of the following components is mostly not needed in a
chemiluminescent immunoassay analyzer?
• A. Source lamp
• B. Monochromator
• C. Photodetector
• D. Wash station
• Oxidation of an organic compound with resultant light emission is known as:

• a. nephelometry.

• b. turbidimetry.

• c. chemiluminescence.

• d. fluorescence
• What is Chemiluminescence?
• A.Light emitted by the sun
• B.Light given off by mammals
• C.Photons released at the initial reactant energy level
• D.Production and emission of light
• Which substance is used to generate the light signal in
electrochemiluminescence?
• A. Acridinium
• B. Luminol
• C. Dioxetane phosphate
• D. Ruthenium
• The light formed from a chemical reaction is called
• Luminescence
• Bioluminescence
• Chemiluminescence
• All the above
Luciferin are an example of
• Bioluminescence
• Photolysis
• Photosynthesis
• Photoisomerisation
All bioluminescent chemical reactions include which chemical?
1.Luciferin
2.Luciferase
3.Photoproteins
4.Calcium
In which environment do most bioluminescent organisms live?
1.Ocean
2.Freshwater rivers
3.Freshwater lakes
4.Forests
• Chemiluminescence:
a. Has excellent sensitivity and dynamic range
b. Does not require sample radiation
c. Uses unstable chemiluminescent reagents and conjugates.
d. Both a and b
• Which of the following best characterizes chemiluminescent assays?

a. Only the antigen can be labeled.

b. Tests can be read manually.
c. These are only homogeneous assays.
d. A chemical is oxidized to produce light.