This document summarizes ultraviolet fluorescence, including its discovery, principles, instrumentation, applications, limitations, and safety considerations. It was discovered in 1845 by Fredrick Herschel who excited tonic water with UV radiation. UV fluorescence occurs when materials absorb ultraviolet light and emit visible light of longer wavelengths. The instrumentation to produce and detect fluorescence involves a light source, monochromator, sample, detector, and recorder. Applications include imaging of amino acids, DNA quantitation, and material detections for industrial purposes. Limitations are potential misinterpretation of results and sample damage from UV light. Safety precautions like protective eyewear and clothing are needed due to risks of vision problems and skin cancer from exposure.
2. Table of Contents
UV fluorescence
Limitations Safety
1 2 3
7 8 9
4 5 6
Instrumentation
Applications
Difference in Fluorescence
and Spectroscopy
Principle
Discovery
Observation
3. Ultra Violet
Radiations
01
Short wave
electromagnetic
Waves
02 Frequency
-7.5x10^14 - -
3x10^16hz
03
Wavelength
180-400nm
04
They have shorter
wavelengths than
visible radiations
06
05 Undetectable
by human eye
Produces Fluorescence
after falling on certain
materials
5. UV Fluorescence
when UV light is absorbed by certain materials, it is
reflected back towards the eye as longer wavelength
visible radiation, or visible light. This phenomenon is
referred to as UV-induced visible fluorescence.
6. UV Fluorescence
(Discovery)
Discovered By Fredrick W. Herschel
in 1845
Excited tonic water with UV
radiation
Later on, Sir George G.Stokes
studied the fluorophores of
fluorescent materials and their
actions as well
7. 1. Light Source
2. Monochromator
3. Sample ( Material to be
fluoresced)
4. Detector ( Fluorometer)
5. Recorder
Instrumentation
8. Principle
During fluorescence imaging,
the item being inspected
absorbs ultraviolet (UV)
light) that excites a specific
fluorophore, causing it to
release photons that
fluoresce and emit light at a
longer wavelength.
9. Obervations
The colors of the observed
fluorescence will depend on
the material and the
wavelength of the UV light
that is being used.
Objects UV Lamp Results
Cellulose Long Wave-
UV
Fluoresces
bright white
Linseed Oil Short Wave Fluoresces
deep yellow
Long Wave Fluoresces
orange
Crystal/lead
glass
Short-wave UV Fluoresces
dramatic icy
blue
10. Applications
One important application is the
imaging of aromatic amino acids
including tryptophan, tyrosine, and
phenylalanine.
Another important application is DNA
quantitation. Purines and pyrimidines –
bases for nucleic acids like DNA and
RNA
Material Detections
Used for Industrial purposes (a very
selective detection method useful for
measuring many types of petroleum
contaminants.
11. Limitations
It may be easily
misinterpreted, or
produce misleading
results
UV light can
cause damage to
the material or
the sample
Further testing
may be required
to get a proper
result
12. Safety
• Long term exposure to UV radiation can
lead to serious and irreversible vision
problems
• UV radiation may also increase the risk of
skin cancer
• According to WHO in 2016 , Exposure of
unprotected skin to UV light should be
minimized
MUST HAVE PPE WHEN CONDUCTING UV
LIGHT EXPERIMENTS
• UV filtering safety glasses
• Long sleeves (e.g. lab coat)
• Nitrile gloves
