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Medical Laboratory Instrumentation

Lecture 5

By FAWAD MAHMOOD
• M.Phil. Medical Laboratory Sciences
 Bioluminescence
Bioluminescence is the production and emission of light by a living
organism

• Way fireflies glow

Luciferase
Luciferin + O2 …………… ……………Oxyluciferin + Light
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Electro-chemiluminescence (ECL)
• “ Electro” refers to electrical stimulation. “Chem” indicates a
chemical reaction. “Luminescence”means “produces light.”
=Electrochemiluminescence (ECL)

• The chemiluminescent reactions that lead to the emission of

light from the complex are initiated electrically rather than
chemically.
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Light
• A light wave is an electromagnetic wave

Light waves are produced by vibrating electric charges

Electromagnetic wave is a transverse wave that has both an

electric and a magnetic component.
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History

Fluorescence polarization was first observed 1926 by Perrin

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Light
• A light wave that is vibrating in more than one plane is referred
to as unpolarized light

• Polarized light waves are light waves in which the vibrations

occur in a single plane

• The process of transforming unpolarized light into polarized

light is known as polarization.
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Fluorescence polarization immunoassay

(FPIA)

Fluorescence polarization immunoassay (FPIA) is a class of in

vitro biochemical test used for rapid detection
of antibody or antigen in sample

The technique of fluorescence polarization (FP) is based on the

observation that when a fluorescently labeled molecule is excited
by polarized light
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Fluorescence polarization immunoassay

Applications
• FPIA has emerged as a viable technique for quantification of

small molecules in mixtures, including: Pesticides, Mycotoxins

in food,

• Pharmaceutical compounds in wastewater,

• Metabolites in urine,

• The analysis of hormone-receptor interactions etc.