BCHEM 264

Lecture 10
Mar. 05, 2015

Radioisotope techniques
The nature of radioactivity
Atomic structure
An atom is composed of a positively charged nucleus that is surrounded by a cloud of
negatively charged electrons. The mass of an atom is concentrated in the nucleus, even
though the nucleus makes only a small fraction of the total size of the atom.

Electrons: negatively-charged with a mass of 9.10938188 × 10-31 kilograms, which is about

1/1850 times the mass of a proton.

Atomic nuclei are made up of two major particles: protons and neutrons.

Protons: are positively-charged particles with a mass of 1.67262158 × 10-27 kilograms,

approximately 1,850 times greater than that of an orbital electron. The number of protons
found in the nucleus of an atom is the atomic number (Z) which defines the type of atom.

The number of orbital electrons in an atom must be equal to the number of protons present in
the nucleus, since the atom as a whole is electrically neutral.

Neutrons are the uncharged particles in the nucleus with mass approximately equal to the
mass of a proton. Because protons are unable to bind with each other due to their mutual
electromagnetic repulsion being stronger than the attraction of the nuclear force, the presence
of a neutron is required within an atomic nucleus as they bind with protons via the nuclear
force.

The number of neutrons in the nucleus is given a notation N. The number of neutrons
determines the isotope of an element. For example, the abundant 12C isotope has 6 protons
and 6 neutrons, while the very rare radioactive 14C isotope has 6 protons and 8 neutrons.

The sum of protons and neutrons in a nucleus is the mass number (A).

Thus, A = Z + N

Since the number of neutrons in a nucleus is not related to the atomic number, it does not
affect the chemical properties of the atom. Atoms of a given element may not necessarily
contain the same number of neutrons. Atoms of a given element having same number of
protons but different mass numbers (i.e. different numbers of neutrons) are called isotopes.

Symbolically, a specific nuclear species is represented by a subscript for atomic number, and a
superscript for mass number, followed by the symbol of the element. For example, the
isotopes of carbon are represented below:
12 14 16 18
6 C 6 C 8 O 8 O
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However, in practice it is more conventional just to cite the mass number (e.g., 14 C ). The
number of isotopes of a given element varies: there are 3 isotopes of hydrogen, 1 H , 2 H , and
3
H , and 7 isotopes of carbon, 10 C to 16 C , and 20 or more isotopes of some of the elements of
high atomic number.

Atomic stability and radiation

In general, the ratio of neutrons to protons in the nucleus will determine whether an isotope of
a given element is stable enough to exist in nature. Stable isotopes for elements with low
atomic numbers tend to have an equal number of neutrons and protons, whereas stability for
elements of higher atomic numbers is associated with a neutron:proton ratio in excess of 1.
Unstable isotopes, or radioisotopes as they are more commonly known, are often produced
artificially, but many occur in nature. Radioisotopes emit particles and/or electromagnetic
radiation as a result of changes in the composition of the atomic nucleus. These processes,
which are known as radioactive decay, arise, either directly or as a result of a decay series, in
the production a stable isotope.

Types of radioactive decay

There are several types of radioactive decay; only those most relevant to biological scientists
are considered below. A summary of these decay events and their properties is given in Table
10.1.

Table 10.1. Properties of different types of radiation

Alpha Beta Gamma, X-rays
Heavy charged particle Light charged particle EM radiation
More toxic than other Toxicity same as electromagnetic Toxicity same as beta
forms of radiation radiation per unit of energy radiation per unit of energy
Not penetrating Penetration varies with source Highly penetrating

Decay by negatron emission (also called beta emission)

In this case a neutron is converted to a proton by the ejection of a negatively-charged beta (β)
particle called a negatron (β-):

Neutron → Proton + Negatron

A negatron is an electron, but the term negatron is preferred, although not always used, since
this particle originates from the nucleus. As a result of negatron emission, the nucleus loses a
neutron but gains a proton. The N/Z ratio therefore decreases while Z increases by 1 and A
remains constant. An isotope frequently used in biological work that decays by negatron
emission is 14 C .

14
6 C  147 N  β 

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Negatron emission is very important to biochemists because many of the commonly used
radionuclides decay by this mechanism. Examples are: 3 H and 14 C , which can be used to label
any organic compound; 35 S is used to label methionine, for example in the study of protein
synthesis; and 32 P , is a powerful tool in molecular biology when used as a nucleic acid label.

Decay by positron emission

Some isotopes decay by emitting positively-charged β-particles referred to as positrons (β+).

This occurs when a proton is converted to a neutron:

Proton → Neutron + Positron

Positrons are extremely unstable and have only a transient existence. Once they have
dissipated their energy they interact with electrons and are completely destroyed. The mass
and energy of the two particles are converted to two γ-rays emitted at 180o to each other. This
phenomenon is frequently described as back-to-back emission.

As a result of positron emission the nucleus loses a proton and gains a neutron, the N/Z ratio
increases, Z decreases by 1 and A remains constant. An example of an isotopes decaying by
positron emission is 22 Na :
22
11 Na  22
10 Ne  β 

Positron emitters are detected by the same instruments used to detect γ-radiation. They are
used in a clinical technique known as Positron Emission Tomography (PET scanning) to scan
the brain to identify active and inactive areas.

Decay by alpha particle emission

Isotopes of elements with high atomic numbers frequently decay by emitting alpha (α)
particles. An α-particle is a helium nucleus without electrons; it consists of two protons and two
neutrons ( 4 He2 ). Emission of α-particles results in the following:

- a considerable lightening of the nucleus,

- a decrease in atomic number of 2
- a decrease in the mass number of 4.

Isotopes that decay by α-emission are not frequently encountered in biological work. Radium-
226 ( 226 Ra ) decays by α-emission to radon-222 ( 222 Rn ), which is itself radioactive, thus,
beginning a complex decay series, which culminates in the formation of 206 Pb :

226
88 Ra  42 He2  222
86 Rn  82 Pb
206

Alpha emitters are extremely toxic if ingested, due to the large mass and the ionizing power of
the atomic particle.
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Electron capture

In this decay process, a proton captures an electron orbiting in the innermost K shell:

proton + electron → neutron + X-ray

The proton becomes a neutron and electromagnetic radiation (X-ray) is given out. Example

125
53 I  12552Te  X  ray

Decay by emission of γ-rays

In contrast to emission of α- and β-particles, γ-emission involves electromagnetic radiation

similar to, but with a shorter wavelength than X-rays. These γ-rays result from a transformation
in the nucleus of an atom (in contrast to X-rays, which are emitted as a consequence of
excitation involving the orbital electrons of an atom) and frequently accompany α- and β-
particle emission. Emission of γ-radiation only, leads to no change in atomic number or mass.

γ-radiation has low ionizing power but high penetration. For example, the γ-radiation from
60
Co will penetrate 15 cm of steel. The toxicity of γ-radiation is similar to that of X-rays.

Example:


131
53 I  131
54 Xe  β  γ

Radioactive decay energy

The usual unit used in expressing energy levels associated with radioactive decay is the
electron volt. One electron volt (eV) is the energy required by one electron in accelerating
through a potential difference of 1 V and is equivalent to 1.6 x 10 -19J. For the majority of
isotopes, the term million or mega electron volts (MeV) is more applicable. Isotopes emitting α-
particles are normally the most energetic, falling in the range of 4.0 – 8.0 MeV, whereas β- and
γ- emitters generally have decay energies of less than 3.0 MeV.

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Rate of radioactive decay

Fig. 10.1. Demonstration of exponential

nature of radioactive decay

Radioactive decay is a spontaneous process and it occurs at a definite rate characteristic of

the source. This rate always follows an exponential law. Thus the number of atoms
disintegrating at any time is proportional to the number of atoms of the isotope (N) present at
that time (t). When we express this mathematically, the exponential curve in Fig. 10.1 gives the
equation below.

dN
 N or
dt
dN
  λN...(1)
dt

where λ is the decay constant, a characteristic of a given isotope. It is defined as the fraction of
an isotope decaying in unit time (t-1). By integrating equation (1), it can be converted to a
logarithmic form:

Nt
ln   λt....(2)
N0

where Nt is the number of radioactive atoms present at time t, and N0 is the number of
radioactive atoms originally present. In practice, it is more convenient to express the decay
constant in terms of half-life (t½).

The half-life of a radioactive isotope is defined as the time taken for the activity to fall from any
value to half that value (See Fig. 10.1).

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If Nt in equation (2) is equal to half of N0, then t will equal the half-life of the isotope. Thus

ln½ = -λt½ ….(3)

or

2.303 log (½) = - λt½ …(4)

or

t½ = 0.693/ λ …(5)

The values of t½ vary widely from 3 × 10-7 s for polonium-212 ( 212 Po ) to over 1019 years for
lead-204 ( 204 Pb ). The half-lives of some isotopes frequently used in biological work are shown
in the Table 10.2.

Table 10.2. Half-lives of some isotopes used in biological studies

Isotope Half-life
3H 12.26 years
14C 5760 years
22Na 2.58 years
32P 14.20 days
33P 25.4 days
35S 87.20 days
42K 12.40 hours
45Ca 165 days
59Fe 45 days
125I 60 days
131I 8.05 days
135I 9.7 hours

In this table, we do not have the two important elements, oxygen and nitrogen. This is because
the half-lives of the radioactive isotopes of these elements are too short for most biological
studies. 15O has a t½ of 2.03 min, and 13N has a t½ of 10.00 min.

The advantages and disadvantages of working with isotopes of short half-lives are given in the
table below.
Advantages and disadvantages of working with isotopes of short half-lives
Advantages Disadvantages
High specific activity Experimental design must be appropriate as isotope
decays during time of experiment
Lower doses likely (e.g., in diagnostic Cost of replacement for further experiments
testing of human subjects)

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Worked example

Given ln (Nt/N0) = -λt and that the half-life of 32P is 14.2 days, how long would it take a solution
containing 42,000 d.p.m. of 32P to decay to 500 d.p.m.?

Solution
Use equation (5) to calculate the value of λ. This gives a value of 0.0488 days-1. Then use
equation (2) to calculate the time taken for the counts to decrease (both equations are
presented below0. In this equation, N0 = 42,000 and Nt = 500. This gives a value of 90.8 days.

t½ = 0.693/ λ …(5)

Nt
ln   λt....(2)
N0

Units of radioactivity

The Système International d’Unités (SI system ) uses Becquerel (Bq) as the unit of
radioactivity. Becquerel is defined as one disintegration per second (1 d.p.s.).
However, an older unit not in the SI system and still often used is the curie (Ci).

Curie is defined as the quantity of radioactive material in which the number of nuclear
disintegrations per second is the same as that in 1 g of radium, namely 3.7 x 10 10 (or 37 GBq).

For biological purposes, this unit is too large and the microcurie (μCi) and millicurie (mCi) are
used. It is important to realize that the curie refers to the number of disintegrations actually
occurring in a sample and not to the disintegrations detected by the radiation counter, which
will generally be only a fraction of the disintegrations occurring and are referred to as counts
per second (c.p.s.).

Normally, in experiments with radioisotopes, a carrier of the stable isotope of the element is
added. It therefore becomes necessary to express the amount of radioisotope present per unit
mass. This is the specific activity. Specific activity may be expressed in a number of ways
including disintegration rate (d.p.s. or d.p.m.), count rate (c.p.s. or c.p.m.) or curies (mCi or
μCi) per unit of mass of mixture (mass expressed in either moles or grams). An alternative
method of expressing specific activity, which is not frequently used is atom percentage excess.
This is defined as the number of radioactive atoms per total of 100 atoms of the compound.

Interaction of radioactivity with matter

α-particles

α-particles have quite an amount of energy (4 -8 MeV) and all the particles from a given
isotope have the same amount of energy.

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Alpha particles react with matter in two ways:

1. They may cause excitation: here, energy is transferred from the α-particles to orbital
electrons of neighboring atoms and the electrons are elevated to higher orbitals. The α-
particle continues on its path with its energy reduced by a little more than the amount
transferred to the orbital electron. The excited electron eventually falls back to its original
orbital, emitting energy as photons of light in the visible or near visible range.

2. α-particles may cause ionization of atoms in their path. When this occurs, the elevated
orbital electron is removed completely. Thus, the atom becomes ionized and forms an ion-
pair, consisting of a positively charged ion and an electron. Because of their size, their slow
movement, and double positive charge ( 4 He2 ), α-particles frequently collide with atoms in
their path. Therefore, they cause intense ionization and excitation and their energy is rapidly
dissipated. Because of this, though they have initial high energy, α-particles are not very
penetrating.

Negatrons

Negatrons are very small and rapidly moving particles, and carry a single negative charge,
compared to α-particles. They interact with matter to cause ionization and excitation in the
same way as α-particles do. However, due to their speed and size, they are less likely than α-
particles to interact with matter and therefore are less ionizing and more penetrating than α-
radiation.

Another difference between negatrons and α-particles is that, whereas for a given α-emitter all
the particles have the same energy, negatrons are emitted over a range of energy so that
negatron emitters have a characteristic energy spectrum. The maximum energy level (Emax)
varies from one isotope to the other, and ranges from 0.018 MeV for 3H to 4.81 MeV for 38Cl.
The difference in Emax affects the penetration of the radiation.

For example :
β-particles from 3H can travel only a few millimeters in air
β -particles from 32P can penetrate over 1 m of air.

W. Pauli in 1931 gave an explanation for the reason why negatrons of a given isotope are
emitted within an energy range. He postulated that each radioactive event occurs with an
energy equivalent to Emax but that the energy is shared between a negatron and a neutrino. A
neutrino is an electrically neutral particle with negligible mass. It is produced in many nuclear
reactions such as in beta decay and do not interact with matter. The proportion of the total
energy taken by the negatron and the neutrino varies for each disintegration.

γ-Rays and X-rays

For simplicity, we will refer to these rays collectively as γ-rays. These rays are electromagnetic
radiation and therefore have no charge or mass. They have the following properties:

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- They rarely collide with neighboring atoms and travel great distances before dissipating
all their energy, that is, they are highly penetrating.
- If they collide with neighboring atoms, they react with matter in many ways. The three
most important ways by which they react with matter lead to the production of
secondary electrons, which in turn cause excitation and ionization. These are

a. Photoelectric absorption: here, low energy γ-rays interact with orbital electrons,
transferring all their energy to the electron which is then ejected as a photoelectron. The
photoelectron subsequently behaves as a negatron.

b. Compton scattering: this is caused by medium energy γ-rays, and results in only part of
their energy being transferred to the target electron which is ejected. The γ-ray is
deflected and moves on with reduced energy. Again the ejected electron behaves as a
negatron.

c. Pair production: this results when very high energy γ-rays react with the nucleus of an
atom and all the energy of the γ-ray is converted to a positron and a negatron.

When high atomic number materials absorb high energy β-particles, the absorber gives out
a secondary radiation, an X-ray called Bremsstrahlung. They are X-rays produced when
energetic electrons are deflected in the field of an atomic nucleus. Thus, Bremsstrahlung
radiation is present during all beta decay processes as the emitted beta particles (both
negatrons and positrons) slow down in matter. For this reason, shields of 32P use low
atomic number materials such as Perspex.

Detection and measurement of radioactivity

There are three commonly used methods for detecting and quantifying radioactivity. The
methods are based on
1. Ionization of gases
2. Excitation of solids or solutions
3. The ability of radioactivity to expose photographic emulsion, i.e. autoradiography.

Methods based on gas ionization

A radioactive substance produces charged particle ( 42 He 2 , β  , β  ). As a charged particle

passes through gas, its electrostatic field dislodges orbital electrons from atoms of the gas
which are sufficiently close to its path and causes ionization (Fig. 10.1).

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 Fig. 10.1. Detection based on ionization.

The ability to induce ionization decreases in the order α > β > γ (10,000 : 100 : 1). Because
of this, α- and β-particles may be detected by gas ionization methods, but these methods
are poor for detecting γ –radiation. If ionization occurs between a pair of electrodes
enclosed in a suitable chamber filled with gas, ionization of the gas creates surge in the
current (called a pulse), whose magnitude depends on

1. The applied potential, that is, the voltage across the electrodes
2. The number of radiation particles entering the chamber.

Fig. 10.2 shows the effect of voltage on pulse flow.

Fig. 10.2. Effect of voltage on pulse flow.

A rise in the curve demonstrates that as voltage across the electrodes increases, the
number of ions produced from the passage of charged particles through the chamber
increases, hence increase in pulse to create five characteristic regions.

1. Ionization chamber region: The voltage here is low. Each radioactive particle produces
only one ion-pair per collision. Hence the currents are low and one needs very sensitive
measuring device to detect them. This range of voltage is little used in quantitative work,
but various types of electroscopes, which operate on this principle, are useful in
demonstrating the properties of radioactivity.
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2. Proportional counter region: At a higher voltage level than those present in simple
ionization chambers, electrons resulting from ionization move toward the anode much more
rapidly. In their movement, they cause secondary ionization of gas in the chamber,
resulting in the production of secondary ionization electrons, whose fast movement leads to
further ionization, and so on. Hence, from the original event, a torrent of electrons reaches
the anode. This is the principle of gas amplification and is known as the Townsend
Avalanche Effect, named after its discoverer. As a consequence of this gas amplification,
current flow is much greater and more pulses are created. In the proportional counter
region, we can see that the number of ion-pairs collected is directly proportional to the
applied voltage.

3. Limited proportional region: as voltage is increased and more ion-pairs are produced, a
point is reached where increase in voltage does not result in further ionization - a plateau
occurs. Before the plateau is reached, there is a region known as the limited proportion
region, which is not often used for detection and quantification of radioactivity.

The main drawback of counters that are manufactured to operate in the proportional region is
that they require very stable voltage supply because small fluctuations in voltage result in
significant changes in amplification. Proportional counters are useful for detecting and
quantifying α-emitting isotopes, but relatively few such isotopes are used in biological work.

4. In the Geiger-Müller region: at this voltage level, all radiation particles, including weak β-
particles, induce complete ionization of the gas in the chamber. Thus, the size of the
current is no longer dependent on the primary ions produced. Since maximal gas
amplification is produced in this region, the size of the output pulse from the detector will
remain the same over a considerable voltage range (the Geiger- Müller plateau).
Therefore, it is not possible to distinguish between different isotopes using this type of
counter.

Since it takes a finite time for the ion-pair to travel to the electrodes, other ionizing particles
entering the tube during this time fail to produce ionization and hence are not detected, thereby
reducing the counting efficiency. This is referred to as the dead time of the tube and is
normally 100 to 200 μs.

5. Continuous discharge region: when the ions reach the electrode, they are neutralized.
Inevitably, some escape and produce their own ionization avalanche. Because of this, if
unchecked, a Geiger-Müller tube would tend to give continuous discharge. To overcome
this, the tube is quenched by the addition of a suitable gas, which reduces the energy of
the ions. Common quenching agents are ethanol, ethyl formate and the halogens.

Instrumentation

Counters based on gas ionization were previously the main methods employed in
quantification of radioisotopes in biological samples. Currently, scintillation counting is
commonly used. However, all labs use small hand-held radioactivity monitors based on gas
ionization, the end-window design being the most popular.
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 Fig. 10.3. (a) The Geiger-Müller (G-M) tube. (b) Effect of applied voltage on count rate.

The Geiger counter

A Geiger counter is the most common radiation detector based on gas ionization. This device
uses a Geiger-Müller tube as the sensor. This tube is filled with an inert gas, usually argon,
helium or neon with halogens added, and voltage in the chamber is over 100 V. When a
radioactive particle passes the tube, the gas becomes conductive and produces pulse which
can be amplified by a cascade effect and outputs a magnified current or pulse, which is
displayed visibly as glows of light or as audible clicks.

Fig. 10.4. A hand-held Geiger counter.

A large amount of radiation passing through the tube produces a higher reading and more
clicks because of the greater amount of electric current generated inside the tube.

Geiger counters usually contain a sealed, gas-filled cylinder and a fine axial wire. The
cylinder is operated at ground potential and the wire at a positive potential that is higher
than that in the proportional counter. In this region, the discharge due to ionization spreads
along the whole length of the wire. Total number of electrons produced does not depend on
the amount of primary ionization, and all of the discharge pulses become equal in
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 amplitude. In other words, each particle crossing the cylinder produces a gas ionization that
is roughly independent of the particle's nature and energy.
The number of pulses (clicks) over a certain time (count rate) is a quantitative measure of
radiation. The count rate is referred to as counts per minute (c.p.m.)

Geiger counters are useful for detecting the ionizing radiations: alpha, beta and gamma
rays. With gamma radiation, sensitivity can be lower. Most hand-held Geiger counters are
at their best with alpha and beta rays. The density of the gas within the tube is usually
sufficient for these two rays but not for high-energy gamma rays.

These counters have a thin end-window made of aluminum and can detect β-radiation
from high energy (32P) and weak emitters (14C) but are incapable of detecting weaker 3H
because the radiation cannot penetrate the end-window. For the same reason, they are not
very efficient detectors of α-radiation.
End-window ionization counters are used for routine monitoring of the radioactive
laboratory to check for contamination. They are also useful in experimental situations
where the presence or absence of radioactivity needs to be known rather than the absolute
quantity, for example, quick screening of radioactive gels prior to autoradiography, or
checking of chromatographic fractions for labeled components.

The inability of end-window counters to detect β-emitters presents a problem in biosciences

because 3H is a commonly used radioisotope. This problem can be overcome by using a
windowless counter where a gas flow is used. These instruments are cumbersome and
need to be conveyed by trolleys. They are useful for mass screening of premises for 3H
contamination but are rarely use routinely. Most labs monitor for 3H by doing a wipe test
regularly. This test uses wet paper towels or cotton wool to take swabs for scintillation
counting.

Methods based on excitation

Radioactive isotopes interact with matter in two ways- they cause (1) ionization, which forms
the basis of Geiger-Müller counting, and (2) excitation.

Excitation of a fluorescent compound (fluor or phosphor) leads to the emission of photons of

light in the visible region of the electromagnetic spectrum which can be detected and counted.
This process is known as scintillation counting.

Scintillators are materials which fluoresce (or phosphoresce) in the presence of ionizing
radiation. Scintillation counters combine this fluorescence with another well-known
phenomenon — the photoelectric effect — to convert incident radiation from a radioisotope into
an electrical signal and quantify it.

TYPES OF SCINTILLATION COUNTING

There are two types of scintillation counting illustrated in Fig. 10.5.

1. Solid scintillation counting
2. Liquid scintillation counting
13
 Fig. 10.5.Diagrammatic illustration of A) Solid
scintillation counter. B) Liquid scintillation
counter

When a radioactive particle hits the scintillator, its energy is absorbed and immediately
reemitted at a different wavelength. The emitted photons are gathered and focused toward the
photocathode, where they are absorbed and their energy is used to emit electrons via the
photoelectric effect. The photoelectrons are in turn focused toward a series of metal electrodes
known as dynodes (A dynode is an electrode in an electron tube that produces electrons
through secondary emission).

When a photoelectron hits the first dynode, its kinetic energy is sufficient to knock several
more electrons off the dynode surface via the Auger effect. These Auger electrons proceed
through the rest of the dynode series, producing more and more electrons with each impact.
Finally, the amplified electron signal is absorbed at the anode, measured, and quantified.

The electrical pulse that results from the conversion of light energy to electrical energy in the
photomultiplier is directly proportional to the energy of the original radioactive event. The
advantage of this method is that two or more isotopes can be separately detected and
measured in the same sample, provided they have sufficiently different emission energy
spectra. The mode of action of a photomultiplier is shown in Fig. 10.6.

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Fig. 10.6. (a) The mode of action of a photomultiplier and (b) the energy spectrum of a typical β-emitter.

Scintillation counting provides two kinds of information:

1. Quantitative: the number of scintillations is proportional to the rate of decay of the
sample, i.e. the amount of radioactivity
2. Qualitative: the intensity of light given out and therefore signal from the photomultiplier
is proportional to the energy of radiation

Solid scintillation counting

In solid scintillation counting, the radioactive sample is placed adjacent to a crystal of

fluorescent material called a scintillation phosphor which is coupled to a suitable light- sensitive
photomultiplier tube which in turn is connected to a high voltage supply and a scaler (Fig.
12.6). As radiation interacts in the scintillation crystal, energy is transferred to bound
electrons of the crystal’s atoms. The following crystals are used:

For γ–emitters - sodium iodide

For α-emitters – zinc sulphide
For β-emitters – organic scintillators such as anthracene

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Solid scintillation counting is particularly useful for γ-emitters because they produce
electromagnetic radiation and collide only rarely with neighboring atoms to cause ionization or
excitation. In a crystal, the atoms are densely packed, making collisions more likely.
Conversely, solid scintillation counting is generally unsuitable for weak β-emitters such as 3H
and 14C, because even the highest energy negatrons emitted by these isotopes would have
hardly sufficient energy to penetrate the walls of the counting vials in which the samples are
placed for counting.

Because many of the isotopes used in radioimmunoassay are γ-emitters, solid scintillation
counting is frequently used in biological work.

Liquid scintillation counting

Liquid scintillation counting (Fig. 12.5) detects radioactivity via the same type of light emission
events which are used in solid scintillation. The key difference is that in liquid scintillation
counting the scintillation takes place in a solution of scintillator, rather than in a solid crystal.
This allows close contact between the isotope atoms and the scintillator, which is not possible
with solid scintillation. With liquid scintillation counting, the short path length of soft β-
emissions is not an obstacle to detection.

In this method, the sample is mixed with a scintillation cocktail containing a solvent and one or
more fluors. Liquid scintillation cocktails absorb the energy emitted by radioisotopes and re-
emit it as flashes of light. To accomplish these two actions of absorption and re-emission,
cocktails contain two basic components, the solvent and the fluor(s). The solvent carries out
the bulk of the energy absorption. The fluor, which is dissolved in the solvent, converts the
absorbed energy into light. Many cocktails contain additional materials to extend their range of
use to other sample compositions but the solvent and the fluor provide the scintillation of the
mixture. Liquid scintillation is particularly useful in quantifying weak β-emitters such as 3H, 14C
and 35S, which are frequently used in biological work.

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