Luminescence

• Luminescence – or the emission of light from a substance that has not been
physically heated – is commonly used in science to provide visualization of
entities that are not normally visible to the human eye.
Chemical reactions can be used to create luminescence without the use of external
light sources in techniques such as bioluminescence and chemiluminescence.
Measurement of light from a chemical reaction is highly useful because the
concentration of an unknown is directly related to the amount of light emitted
and is a relative indicator of the amount of luminescent material present in the
sample of interest
• Other forms of luminescence, however, require external light to trigger a
glow within a substance.
These latter forms of luminescence are referred to as Photoluminescence and
include
• Fluorescence and
• Phosphorescence.
A brief overview of these techniques and how they are applied in life sciences
research
BioLuminescence

• The light emitted by a bioluminescent organism is produced by energy

released from chemical reactions occurring inside (or ejected by) the
organism.
Bioluminescence refers to the powerful ability to see what is going on inside of
organisms through the emission of visible light
The technique enables researchers to study biological processes in real-
time, in vivo
• It has been developed over the past few decades and used primarily in small
animals for the purposes of molecular imaging.
• The technique can be used to visualize activity within internal organs. 
 • Bioluminescence works by detecting visible light that is produced when an
enzyme oxidizes a molecular substrate.
• Most often, the enzyme, which is referred to as a bioluminescent reporter, is
firefly luciferase. 
• Luciferases from other species have also been deemed useful.
These luciferases come from organisms including jellyfish, coral, beetle, and
bacteria.
• There are a wide range of research applications for bioluminescence,
including
• Monitoring basic activity like protein interactions and intracellular motility,
as well as
• Monitoring more clinically-relevant processes like infection progression,
• Tumor growth and metastasis,
• Transgene expression, and The impact of gene therapy.
Chemiluminescence
• Chemiluminescence is the light emitted by a chemical reaction.
• Chemiluminescent reactions do not usually release much heat, because
energy is released as light instead.
• Luminol produces a light when it reacts with an oxidising agent.
Chemiluminescence, light is detected due to a reaction of a substrate with a
reactive oxygen species.
• Chemiluminescence a well-established technique with many applications.
• Advantages of chemiluminescence can be realized using both in vitro and in
vivo applications.
• For instance, the technique can help in the detection of several analytes,
including proteins, enzymes, nucleic acid and even cells.
• Chemiluminescence immunoassay (CLIA) is an assay that combine
chemiluminescence technique with immunochemical reactions, utilize
chemical probes which could generate light emission through chemical
reaction to label the antibody.
 PHOTOLUMINESCENCE
• In photoluminescence, a substance’s glow is triggered by light, in
contrast to chemiluminescence, where the glow is caused by a chemical
reaction.
• Photoluminescence requires that a molecule absorb light radiation and
for its electrons to become excited.
• When this occurs, molecules transform from the ground state to an
excited state.
Because the excited state is unstable, the molecule inevitably reverts back
to its ground state, dissipating energy.
Fluorescence and phosphorescence
• Fluorescence and phosphorescence are two forms of photoluminescence.
• Both fluorescence and phosphorescence are based on the ability of a
substance to absorb light and emit light of a longer wavelength and therefore
lower energy.
• The main difference is the time in which it takes to do so.
• In fluorescence, the emission is basically immediate and therefore
generally only visible, if the light source is continuously on (such as UV
lights); while
• Phosphorescent material can store the absorbed light energy for some
time and release light later, resulting in an afterglow that persists after
the light has been switched off.
• Depending on the material, this afterglow can last anywhere from a few
seconds to hours.