Chemistry for Everyone

Turning on the Light: Lessons from Luminescence

Patricia B. OHara*
Department of Chemistry, Amherst College, Amherst, MA 01002; *pbohara@amherst.edu
Carol Engelson
West Springfield High School, 425 Piper Rd., West Springfield, MA 01089
Wayne St. Peter
Hall High School, 975 N. Main St., West Hartford, CT 06117

How many of us have walked the shores at night and

been mystified by the eerie green glow of marine organisms
in the surf? Perhaps you paused by a meadow on a summer
evening to watch the staccato flashes of light from fireflies
seeking to attract their mates. What are the energy sources
for these mysterious lights that seem to glow without the input of electricity or heat? Are the phenomena that cause these
cold lights the same as those responsible for the light emitted from glow-in-the-dark stickers, or emergency light sticks?
Are they related to incandescent light bulbs and fluorescent
lights? The goals of this article and related laboratory exercises are to give secondary school teachers a context in which
all of these phenomena, broadly grouped under the heading
of luminescence, can be explored and explained with their
students (1). Five laboratory exercises are provided in the
Supplemental MaterialW so that students can perform their
own examinations of some of the underlying principles of
luminescence spectroscopy in a laboratory setting.
Macroscopic Energy
Fluorescent jellyfish, fireflies, stickers, light sticks, fluorescent and incandescent lights, fireworks, and even shattered
wintergreen hard candies all emit light when atoms and
molecules in an excited energy state return to a ground energy state. Most of us are familiar with the macroscopic world
in which energy is divided into kinetic and potential energy.
Kinetic energy is the energy possessed by an object because
of its motion, which is why mobiles are called kinetic sculptures. Potential energy is the energy stored in a system and
is often thought of as the potential that this system has to do
work owing to its position. Water tumbling over Niagara Falls
has potential energy owing to gravity and, when it falls, that
potential energy is converted (eventually) into electrical energy used to light our homes and cities. In that same way an
individual atom or molecule can possess potential energy that
can be converted into light. Here we describe some of the
major processes by which light is created in these systems,
and how the wonder inspired by these processes can be used
to motivate secondary school students to learn some basic
ideas of science.
Microscopic Energy
In chemical terms, energy is stored in bonds, and whenever bonds are broken or formed, there is a potential for some
energy to be released and for the system to do work. The
potential energy stored in gasoline is converted into mechanical energy every time you drive your car. In the world of large
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systems and many, many particles, energy seems to be continuous: it is theoretically possible for your car to go 20, 53.5,
or 74.12434 mph and every speed in between. In the world
of small, small systems, at the molecular level, things are a
bit different. Unlike the macroscopic world, a molecule can
only exist in certain discrete energy states, which are also referred to as quantum states. The lowest of these is called the
ground state, followed by a first excited state and so on, but
the energy is not continuous. Therefore, it is not possible for
a molecule to contain a quantity of energy that is between
the ground and the first excited state, or between the first
and second excited states. In addition, we find it useful to
subdivide the types of energy states a molecule can have. For
the purposes of this discussion, we like to think separately
about the energy of a single atom or molecule as electronic
(where the electrons that are part of the atom change without being removed from the atom), vibrational (where atoms bonded to one another move relative to one another),
and rotational (where the whole molecule tumbles in space).
The electrons in atoms and molecules can move between different energy states by either absorbing energy (moving to a
higher energy state) or emitting energy (moving to a lower
energy state). The energy difference between any two quantum states is fixed, and therefore the energy required to move
between them is also fixed. That energy is said to be quantized.
Light Energy

Incandescence and Luminescence

When an atom or molecule absorbs energy, it enters an
excited state. To return to its ground (or resting) state, it must
find a way to divest itself of the excess energy. Sometimes
the energy needed to excite or relax an atom or molecule takes
the form of electromagnetic energy, or light. When light is
emitted by an atomic or molecular system, photons (quanta
of electromagnetic radiation) carry off the extra energy of the
system. The exact nature of how the system gets excited, and
what excited energy level the system is in, will determine
whether the process of light emission is classified as incandescence or luminescence. Incandescent processes are always
caused by heating a material until it glows. Thus ordinary
light bulbs derive the energy to excite the tungsten metal from
the heat produced by electricity flowing through the tungsten filament. The remainder of this article deals with processes by which light is emitted without a simultaneous
change in temperature, and is classified as luminescence or
cold light. Luminescent processes include triboluminescence,
fluorescence (resonant emission and redshifted emission),

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phosphorescence, chemiluminescence, and bioluminescence

(2, 3). The conditions under which each of these processes
occurs are summarized in Figure 1.

emission of light
with heating?
yes

no

incandescence

luminescence
from mechanical forces?
yes

no

with excitation by light?

triboluminescence
yes

no

immediately?
yes

inanimate?
yes

no

fluorescence

no

chemiluminescence

phosphorescence

bioluminescence

Figure 1. Possible luminescent fates for an atom or molecule after

electronic excitation.

iii

S1
i

ii

ii

4
3
2
1

S1

iv

S0

S0

fluorescent emission
in atoms

fluorescent emission
in molecules

C
v

S1

T1
vi

S0
phosphorescent
emission from molecules

Figure 2. What can happen after absorption of light? (A) Up arrow (i) indicates excitation light. Down arrow represents emission
of light from atoms that occurs when an electron moves from state
S1 to S0 and is called direct or resonant fluorescence (ii). (B) Up
arrows indicate excitation with photons of increasingly short wavelengths (higher energy). Emission of light in molecules can occur
as simple resonant fluorescence (ii), or alternatively, some of the
excited energy can be lost through internal conversion (iii) between
vibronic levels 1, 2, 3, 4 in S1 and then emitted as fluorescent
light which is red-shifted (iv). (C) Up arrow indicates excitation light.
Emission in some molecules shows intersystem crossing (v) that results in phosphorescence (vi).

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Triboluminescence
You can visualize the process of triboluminescence by
breaking a wintergreen (methyl salicylate) Lifesavers candy
either with a hammer, with a pliers, or in the mouth of a
volunteer in a darkened room. While this effect can be seen
by cracking virtually any hard candy confection, the presence of the methyl salicylate in the sugar crystal makes for a
particularly bright emission. Triboluminescence occurs when
emission of light is caused by a mechanical shock or strain
in certain highly ordered crystalline systems. The effect was
first observed by Sir Francis Bacon in the early 1600s from
the abrasion of hard sugar with a knife (4). The reasons why
some systems are more prone to triboluminescence than others remain mysterious and the ultimate cause of the effect is
still debated. A recent patent application cites a process for
impregnating gift wrap with a triboluminescent material
(menthyl 9-anthracene carboxylate) so that unwrapping the
gift would be accompanied with the emission of sparks of
light (5).
Fluorescence
Fluorescent emission is the most straightforward of the
various emission processes. When atoms and some molecules
are excited to higher electronic energy states (either directly
by absorption of light or indirectly through some other process) they return to the ground state by emission of a photon of exactly the same energy as that used to excite them.
This type of fluorescence, called resonant fluorescent emission, is shown in Figure 2A.
Light in fireworks, flame tests, and stars occurs as a result of this type of resonant emission. While it is also possible to observe resonant emission in molecules, one unique
feature of molecules as shown in Figure 2B is redshifted fluorescent emission of light. This occurs when the photon that
is emitted from the system has a wavelength that is longer
than the wavelength of excitation (lower in energy). A redshift in the fluorescence, first characterized by Sir George
Stokes in 1852, occurs when the molecule that is electronically excited undergoes a two-part relaxation. In the first
phase, in a process known as internal conversion, the molecule undergoes rapid thermal relaxation (1 picosecond or
less than 1 trillionth of a second) down to the ground vibrational state of the still excited electronic state as shown in
Figure 2B. In the second phase, the molecule emits a photon within nanoseconds (one billionth of a second) of excitation to return it finally to the ground state. The photon
emitted is always of lower energy (longer wavelength or redshift) than the excitation photon because of the energy lost
in the excited state.
Detailed experimental protocols that illustrate these aspects of luminescence spectroscopy are provided in the
Supplemental Materials.W Both absorption (transmission) and
emission of light by a sample can be measured with a very
simple spectrometer that can be built using directions included in the Supplemental Material.W It is possible to interface this device with a personal computer (6). In
experiment 1, Differentiating Between Light Sources, stu-

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Chemistry for Everyone

dents are introduced to the interaction of light and matter.

In experiment 2, Relative Quantum Yield, common dyes are
used to introduce students to quantitative transmission and
fluorescence measurement in the computation of a quantum
yield using this instrument.
Many applications of fluorescence exist in the real world.
Vanishingly small quantities of protein or nucleic acid can be
detected in medical diagnostic by complexation with
fluorescent dye molecules or naturally fluorescent proteins.
Some detergents that claim to be whiter than white not only
reflect incident light but also cause white light to be emitted
and therefore appear whiter than the original garment. The
detergent contains colorless dye molecules that absorb invisible
ultraviolet light and then fluoresce white light when they
absorb sunlight. Chlorophyll, one of the principal biological
molecules responsible for capturing the energy of sunlight by
green plants, is a fluorescent molecule. It is easily extracted
from plants, and its bright red fluorescence can be observed
in a darkened room by shining an ultraviolet lamp on a
solution of chlorophyll and water. Instructions for this simple
experiment are given in the experiment 3, Chlorophyll
Fluorescence.

Phosphorescence
Many students will be familiar with glow-in-the-dark
stickers or stars, in which an initial charging of the system
with a light bulb can cause material to glow for hours. What
you are observing is phosphorescence, a process very similar
to fluorescence but long lived (7). When light shines on a
phosphorescent material, there is a slight delay in the emission of light from the sample. This delay in which the system loses energy and returns to the ground state might be as
long as minutes to hours. Rocks containing the element phosphorus are naturally phosphorescent, hence the name of this
process. At the molecular level, what has happened is the same
kind of excited electronic state that was generated in fluorescence, however in phosphorescence, the species undergoes a
further transformation before it returns to the ground state.
A ground state for an atom or molecule can have either one
or two electrons in each level, but if two electrons are in the
same energy level, they must have opposite magnetic spins;
we say that their spins are paired. In phosphorescence, as
shown in Figure 2C, the excited electron changes its magnetic spin by a process known as intersystem crossing. The
new state for the system in which the remaining ground-state
electron and excited-state electron now have the same magnetic spin is called a triplet state. Once this happens, the
molecule is still excited, and it is very difficult to get back
down to the ground state because to do so would require an
additional spin flip and a change in electronic energy level.
Very difficult to a molecule means it might take the molecule as much as a millisecond (one thousandth of a second)
to do this. Experiment 4, Understanding The Differences
Between Fluorescence And Phosphorescence, enables students
to get hands-on experience with these two related processes.
Chemiluminescence
Often light is emitted from a system without the input
of electrical, mechanical, or electromagnetic energy. Here, the
energy to excite the system is derived from a chemical reaction. Glow sticks, bracelets, and necklaces exhibit chemiluwww.JCE.DivCHED.org

minescence, a process in which an inanimate system is excited by the energy of a chemical reaction (8). In these devices, two reactants are separated by a membrane. The
reaction is initiated by breaking the membrane between the
chambers, thereby mixing the reactants. The reaction
progresses with the spontaneous emission of red, blue, violet, or green light and over time when the reaction is complete, the mixture no longer emits light. Chemiluminescence
derived from the oxidation of luminol has been a staple of
chemical demonstrations for introductory students in chemistry (9, 10). Luminol has gained attention from the print
media and television, where it is now in widespread use at
many crime scene investigations. Our procedure to use
luminol to detect the presence of simulated blood (a dilute
solution of bleach) via chemiluminescence is described in
experiment 5, Use Of Luminol In Forensic Analysis.

Bioluminescence
A related phenomenon is the emission of light from biological organisms such as fireflies and jellyfish, a process called
bioluminescence. We know that the function of this light in
fireflies, derived from the oxidation of luciferase, is for the
female to attract a mate. In the Pacific jellyfish, Aequoria
victoria, blue light is first emitted via bioluminescence
through the oxidation of the protein, aequorin, and then these
photons are absorbed by green fluorescent protein (GFP),
which subsequently fluoresces green. It is unclear in the jellyfish whether the emission of light is intended to attract a
mate or food. Molecular geneticists have devised a clever
means to use the GFP fluorescence to tell whether or not
they have been successful in inserting a new gene into a host
organism and inducing that organism to express (make) the
protein coded for by the gene under investigation. They do
this by attaching the gene for GFP onto the end of the gene
under investigation. If, after inserting the new gene into the
host cell and inducing protein expression, the host cells glow
green, then you can be sure that the experiment has succeeded
(11) and you have made a transgenic organism. For teachers, two excellent resources that can be used to introduce students of biology or biochemistry to these processes are
provided through the Educational Division of Bio-Rad. One
educational resource kit, pGLO Bacterial Transformation Kit,
provides necessary chemicals and equipment for a class of 24
students to transform harmless bacteria with the gene for GFP.
With the GFP-producing colonies in hand, it is then possible, with the Green Fluorescent Protein Chromatography
Kit, for a similarly sized class to perform the isolation and
purification of the GFP with almost all of the supplies and
chemicals provided in the kit. These kits have a high success
rate for our students and complementary curricular support
materials are provided from Bio-Rad.
Conclusion
Light emission, in all of its many forms and manifestations, has the capacity to excite the imagination of students.
Some of the processes discussed here are part of the technological revolution that is taking place all around us. DNA
microarrays have the capacity to use fluorescence to display
thousands to tens of thousands of genes, proteins, or other
markers on a tiny microchip. The presence or absence of a

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particular gene, piece of DNA or RNA, or protein is most

often detected by the fluorescence of a complementary molecule that binds to the molecules of interest. Prolume Ltd., a
company whose primary focus is cancer research, is exploring the use of the harmless GFP to create glow-in-the-dark
drinks and food, including icing for birthday cakes. With a
future that has the potential to include glowing birthday
cakes, sparking gift wrap, personalized fluorescent DNA identification cards, and transgenic green animals and plants, we
would be wise to educate both ourselves and our students in
the background processes that make this all possible.
W

Supplemental Material

Instructions for the students and notes for the instructor for the five experiments are available in this issue of JCE
Online.
Literature Cited
1. Harvey, E. N. A History of Luminescence; American Philosophical Society Press: Philadelphia, PA, 1957.

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Journal of Chemical Education

2. Principles of Fluorescence Spectroscopy, 2nd ed.; Lakowitz, J. R.,

Ed.; Plenum Publishing Corp.: New York, 1999.
3. Weidemann, E. Ann. Phys. Chem. 1888, 34, 446.
4. Bacon, F. Advancement and Proficience of Learning; Oxford
University Press: Oxford, United Kingdom, 1605.
5. Geddes, N. J.; Sage, I. C.; Rozelaar, C. F.; Mason, I. R.; Grant,
H. Manufacture of Triboluminescent Materials in Paper Products for Wrapping or Gift Papers (Qinetiq Limited, United
Kingdom) PCT Int. Appl., 2002, WO 2002062914.
6. Riley, S. A.; Nishimura, A. M. J. Chem. Educ. 1997, 74, 1243.
7. Lisensky, G. C.; Patel, M. N.; Reich, M. L. J. Chem. Educ.
1996, 73, 1048.
8. Carlson, R.; Lewis, S. W.; Lim, K. F. Aust. J. Chem. Ed. 2000,
14, 51.
9. Schreiner, R.; Testen, M. E.; Shakhashiri, B. Z.; Dirreen, G.
E.; Williams, L. G. Chemical Demonstrations: A Handbook for
Teachers of Chemistry; The University of Wisconsin Press:
Madison, WI, 1983; Vol. 1.
10. Lister, T. Classic Chemical Demonstrations; Royal Society of
Chemistry: London, 1996.
11. Chan, A. W. S.; Chong, K. Y.; Martinovich, C.; Simerly, C.;
Schatten, G. Science 2001, 291, 309.

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