Basic Photophysics

by Chris Lambert and Linda Jones

The Nature of Light: Light Is Energy

The phenomenon that is commonly known as light is only one small part of the electromagnetic
spectrum, which includes radiowaves, microwaves and X-rays. The part of the spectrum that we
call light is that part that our eye can detect - the wavelengths that we can see. Photophysics is
concerned with those processes, which occur when light, ultraviolet radiation or infrared
radiation interact with matter. Photophysical processes include scattering, absorption and excited
state formation.

1. How does the energy of the electromagnetic spectrum vary with wavelength?
The electromagnetic spectrum correlates the wavelength of light and its energy. Depending on
the spectral region of interest, the units used to describe the light may be frequency, energy,
wavelength or inverse wavelength. A representative table to compare different spectral regions is
given below.
Spectral Region VHF UHF microwave Infrared Visible ultraviolet X-ray -ray
Common Usage nmr epr rotational
transitions vibrational
transitions electronic transitions ionization nuclear
effects
Frequency (Hz) 5 x 108 3 x 1010 3 x 1011 3 x 1013 6 x 1014 1.2 x 1015 3.0 x 1017 1.5 x 1019
Wavelength 0.6m 1cm 1mm 10 m 500 nm 250 nm 1 nm 20 pm
Wave number (cm-1) 0.017 1.0 10.0 1000 20,000 40,000 1.0 x 107 5.0 x 108
Single photon energy (eV) 2.07 x 10-6 1.24 x 10-4 1.24 x 10-3 1.24 x 10-1 2.5 5.0 1.24 x 103 6.2
x 104
Photon energy
(kJ mol-1) 2.03 x 10-4 1.20 x 10-2 1.20 x 10-1 12.0 239 479 1.2 x 105 6 x 106
Table 1. Characteristics of the electromagnetic spectrum. Wavelength ranges of the
electromagnetic spectrum have been defined, in part, by the effects or uses of those ranges. For
example, the table shows that VHF radiation is used in nuclear magnetic resonance (nmr) while
UHF radiation is useful for electron paramagnetic resonance (epr). The other rows of the table
provide information on frequency, wavelength, wave number (reciprocal of wavelength) and
energy for a single wavelength in the middle of each spectral range. The last row gives the energy
provided by one mole of photons of the given wavelength, that is the energy per einstein.

2. Does the speed of light depend on its energy?

Documento de Estudio
VIAVIDA S.A.

While energy varies throughout the electromagnetic spectrum the speed of all types of
electromagnetic radiation is a constant. Light travels at 2.98 x 108 mos-1 in a vacuum which
implies that, in one nanosecond, it travels about 30 cm. Light travels so quickly through
materials, that most experiments do not have sufficient time resolution to record the change in
light intensity at the start or end of illumination. That is, most experiments record only the
steady-state level of light intensity. Recording on shorter time scales to capture the intensity
changes at the start or end of illumination (for example, to investigate short-lived intermediate
states) requires special equipment, such as pump-probe techniques that measure ultrafast kinetics
(lifetimes).

3. What is the quantum nature of light?

Light is absorbed and emitted in indivisible packets or "quanta," called photons.
The First Law of Photobiology states that light must be absorbed by an atom or molecule in order
to initiate a physical or chemical process. Light is absorbed when there is a transition within a
molecule (or other species) that is resonant with (or equal to) the energy of the incoming photon.
The most important region of the spectrum for photobiology extends from the infrared to the
ultraviolet. It is the most important because the light energy in this range can be absorbed by
biological molecules. The energies associated with these wavelengths are sufficient at one
extreme to set a molecule vibrating and at the other to cause photoionisation and the breaking of
covalent bonds.

Spectroscopic Methods
Studies in photochemistry and photobiology are often carried out using spectroscopic methods.
Absorption and fluorescence spectroscopy is used to identify and quantify substances. The BeerLambert law describes the absorption of light in a transparent material. This law cannot be used
for turbid (cloudy or opaque) materials where the path of the light greatly exceeds the thickness
of the container because of multiple scattering events.

4. What is the Beer-Lambert Law?

Lambert's law states that the proportion of light absorbed by a material is independent of the
intensity of the incident radiation. Beer's law states that the absorption is proportional to the
concentration of the sample. The combination of these two laws gives the expression:
where A is the absorbance, is the molar absorption coefficient, C is the concentration and is the
pathlength. The absorbance is measured experimentally and is given by:

Documento de Estudio
VIAVIDA S.A.

where Io is the intensity of the incident light and It is the intensity of the light transmitted through
the sample. It is sometimes more convenient to express the molar absorption coefficient as an
absorption cross section :

Where N is the number of absorbing species per unit volume.

There is a common confusion with absorbance that arises from terminology. It must be made
clear that absorbance is the log of the ratio of incident and transmitted light. The absorbed light is
the number of photons absorbed. The number of photons absorbed can be calculated by
rearranging the Beer-Lambert equation given above:
5. How is light absorption determined in turbid materials?
Turbid materials contain particles that scatter light. For example, milk is turbid because it
contains fat particles. Skim milk is not as cloudy as whole milk. Light flow through turbid
materials is described by the equation of radiative transfer:
The left hand side of the equation represents light that travels in direction . The light flux in
direction is decreased through absorption and scattering, represented by the first term on the
right hand side. The light flux in direction is increased by light that is scattered into direction
from all other directions, represented by the second term of the right hand side. The diffusion
approximation of the radiative transfer equation is commonly used to describe light flux for
turbid media in which scattering dominates over absorption. Light propagation will be described
in detail in an Advanced Photophysics Light Scattering module.
Energy is conserved.
When light energy is absorbed, it can be converted to thermal energy, kinetic energy or chemical
energy. [No matter what conversion process takes place, the initial absorption always creates an
excited state of the absorbing molecule.]
6. Can we predict the outcome of energy conversion?
The fate of the excited state is determined by a number of factors, which control whether the
atom or molecule loses the energy by emission of a photon (fluorescence or phosphorescence),
heat, or some photochemical process. (Intermediate states can also occur if the atom or molecule
undergoes intersystem crossing or internal conversion.)
The Jablonski diagram
The photophysical processes that occur following absorption of a photon can be shown
graphically in a Jablonski diagram. This is perhaps the most important diagram in photobiology
for the understanding of photoexcited states. It gives a one dimensional picture of the relative
energy levels of the excited states of a molecule and describes the fate of the absorbed energy.
Documento de Estudio
VIAVIDA S.A.

In the diagram as drawn, the molecule may be in a singlet or a triplet state. This is the multiplicity
of a species and describes how that species interacts with a magnetic field. Transitions between
states are allowed if they are of the same multiplicity (i.e., from one singlet state to another or
from one triplet state to another) and formally forbidden if a change of multiplicity occurs (i.e.,
from singlet to triplet or vice versa). Formally forbidden transitions are sometimes called
relatively forbidden because they can occur, but are often less frequent (take a longer time) than
transitions that are not forbidden.
The ground state is designated So which is the lowest energy singlet state. The first excited
singlet state is designated S1. Virtually all organic and biological molecules have a singlet ground
state. An important exception to this is oxygen, which has a triplet ground state. The thickly
drawn lines in this diagram represent the lowest vibrational level of each state. The thinner lies
are higher vibrational levels of that state. Absorption of a photon occurs within 10-15 second.
The energy of the photon must match the energy of the transition. If there is not a transition of the
same energy in a molecule then absorption cannot occur. Jagged lines in the diagram represent
radiationless transitions; that is, photons are neither absorbed nor emitted. The first excited
singlet state typically has a lifetime of nanoseconds. Should intersystem crossing occur to the
triplet state, which can happen with high efficiency under certain circumstances, then the triplet
state may live for milliseconds or even longer.
Figure 1. The Jablonski Diagram. This figure depicts the total electronic energy of a molecule
that is capable of absorbing energy in the visible or ultraviolet portion of the electromagnetic
spectrum. The absorbed energy adds electronic energy to the molecule. There is an implied
vertical energy scale in this diagram such that higher electronic energy levels are above lower
energy levels. By pointing at the words just below the diagram, you can learn more about what
happens to the absorbed energy. Alternately, you can point at arrows in the diagrm to learn what
process they represent. When you point at words or arrows, you will see a "Play" and "Stop"
button. The Play buttons run simple animations describing some of the more complex process
from start to finish. (Try the Play button for Fluorescence.)
Fluorescence
The emission of light is a random spontaneous process. The emission lifetime is a measure of the
probability of a certain percentage of the excited states decaying within that time. It is a
reasonable approximation that electronic transitions originate from the lowest vibrational level of
a given state. (What happens to the rest of the absorbed light energy?) Frequently the highest
energy fluorescence band is of lower energy than the lowest energy absorption band. The
difference in wavelength observed between these two bands is called the Stokes shift. A Stokes
shift is most easily observed as the difference between the wavelength of maximum absorption
and the wavelength of maximum fluorescence.
Internal Conversion
The radiationless transitions described above that don't involve a change of multiplicity are
referred to as internal conversions. Energy is released in the form of heat when internal
conversion occurs. Internal conversion can be a very rapid process and is responsible for the
Documento de Estudio
VIAVIDA S.A.

prompt loss of vibrational energy to the lowest vibrational level (the thick lines in the Jablonski
diagram) for each electronic state. This is the reason that fluorescence is assumed to occur from
the lowest vibrational level of any state (above). No matter what level is initially populated by
light absorption, internal conversion will rapidly release vibrational energy until the lowest
vibrational level is reach - long before fluorescence occurs.
Intersystem Crossing
Radiationless transitions between two states of different multiplicity are termed intersystem
crossings. Like internal conversion, these transitions result in a loss of energy as heat, but
because they are formally forbidden, they require a longer time to occur. The electronic
configuration of some molecules favors intersystem crossing from singlet states produced by
light absorption to a triplet state of similar energy. These transitions are very important in
photobiology because the excited triplet state can have a relatively long lifetime allowing ample
time for the excited state molecule to use the absorbed energy in a reaction with another molecule
nearby. Intersystem crossing is also the typical precursor event to phosphorescence.

Phosphorescence
Phosphorescence is the emission of a photon involving a change of multiplicity, while
fluorescence does not involve a change. Because the change in multiplicity is formally forbidden
(i.e. unlikely), phosphorescence lifetimes are significantly longer than fluorescence lifetimes.
Phosphorescence is commonly observed from T1 to So. Because the lifetime of phosphorescence
is so long, molecules in T1 may lose their energy by other transitions that are more efficient or by
chemical reaction. Common methods of deactivation include solvent interactions and energy
transfer to oxygen.

Quantum yield
We can predict how much of the absorbed light energy will follow each of the possible
deactivation pathways or photophysical processes if we know the rates of the various transitions.
The efficiency of a photophysical process is normally expressed in terms of a quantum yield. The
quantum yield is given the symbol and is defined as the number of products formed per photon
absorbed. For example the quantum yield for photoionization, I can be expressed as

The quantum yield of fluorescence F may be written:

The quantum yield may also be written as a fraction of species that take one route compared to
species that take all routes, i.e.:
where kr is the radiative rate constant, knr is the non radiative rate constant, kisc is the rate
constant for intersystem crossing, and kc is the rate constant for chemical reaction. [A radiative
process involves the emission of a photon. A non-radiative process does not involve the emission
Documento de Estudio
VIAVIDA S.A.

of a photon.] The quantum yield for generation of S1 is unity since for every photon absorbed the
molecule will be promoted to S1.
Light Sources
One of the early lessons of childhood is that brightly glowing objects are hot. The temperature of
a glowing object is related to the color of light that it emits.

7. How do hot objects emit light?

Boltzmann distribution
The Boltzmann distribution helps us to understand how temperature relates to light emission. It is
derived from statistical thermodynamics. The Boltzmann distribution is given by:

where N1 is the number of species in the upper excited state, No the number in the lower excited
state, E is the energy difference in the two states, k is the Boltzmann constant and T is the
absolute temperature. Application of this equation allows the determination of the relative
populations of states of different energies.

Figure 2. The Boltzmann Distribution. This diagram shows how energy and temperature can alter
the population of energy levels. Up to 10 levels will be displayed depending on the energy gap
between levels and the temperature that you specify. Once you have entered your values, click on
the "Submit" button to see the results. Try an energy gap between levels of 20 kcal/mol and a
temperature of 4000 K to start and then experiment with other values.

The heated molecules will emit photons from the higher energy levels, producing light that
depends on the temperature of the material. This is called "blackbody radiation". The color of a
hot object gives information about its temperature. Red stars are cooler than blue stars.
(Remember from section 1 that red light has a lower energy than blue light.) The human body, at
a temperature of about 37C emits photons in the invisible infrared region. A standard light bulb
filament has a temperature of about 3000K (3373C) and emits photons in the visible and near
infrared region of the spectrum.
The Boltzmann equation gives an equilibrium distribution. Chemical reactions create nonequilibrium populations in which the excited vibrational states rapidly decay to the distribution
given by the Boltzmann relation for the existing temperature.

8. Are there any kinds of lamps that do not operate through heat?

Documento de Estudio
VIAVIDA S.A.

Incandescent lamps emit light from a heated filament. The output depends on the temperature of
the filament. Arc lamps and fluorescent lamps utilize a non-thermal mechanism. Arc lamps
contain gas that becomes excited and emits light at characteristic emission wavelengths. Some
heating is involved in the excitation of the gas, but the output spectrum of an arc lamp depends on
the type of gas it contains. Fluorescent lamps are low pressure mercury arc lamps. The inside of
the glass envelope is coated with a phosphorescent material. The phosphor absorbs the light
emitted by the mercury vapor and emits light at a longer visible wavelength.
9. What is the difference between light emitted by a hot object and light emitted by a laser?
The word "laser" is an acronym for light amplification by stimulated emission of radiation. Laser
action arises from the nonrandom decay of an excited state. All laser action arises from
fluorescence, that is the emission of a photon from an upper excited state to a lower energy state
of the same multiplicity. Fluorescence is a random process in which the time at which a given
photon decays cannot be predicted. However, laser action arises from stimulated emission from
the excited state. The stimulus is the arrival of a photon of the same energy, which causes the
excited state to relax to the ground state, emitting a photon. In a laser cavity, a large fraction of
the species in the laser medium are promoted to an excited state. This excited state is then
simultaneously deactivated by incoming photons, which promote stimulated emission. The net
result is the amplification of the incoming pulse.
Suggested Readings
Some of the following books are sadly no longer in print. However, they are still listed here
because they are so valuable.
Banwell, C.N., Fundamentals of Molecular Spectroscopy, McGraw Hill Book Co Ltd, May 1983.
Birks, J.B., Photophysics of Aromatic Molecules, Wiley-Interscience.
Calvert, J.G. and J.N. Pitts, Photochemistry
Horspool, W. M. and Pill-Soon Song, eds., Handbook of Organic Photochemistry and
Photobiology, CRC Press, June 29, 1995.
Jaff, H.H. and M. Orchin, Theory and Applications of Ultraviolet Spectroscopy, John Wiley &
Sons, June 1962.
Murrell, J.N., The Theory of the Electronic Spectra of Organic Molecules
Parker, C.A., Photoluminescence of solutions
Smith, K.C., The Science of Photobiology, Plenum Pub Corp, December 1977.
Turro,N.J., Modern Molecular Photochemistry, University Science Books, April 1991.
Documento de Estudio
VIAVIDA S.A.