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Radioactivity in the life sciences

This article is about radioactivity as a tool in life science.


For the eect of radiation on living organisms, see
Radiation poisoning. For organisms which harness radiation, see Radiotrophic fungus. For the bacterium highly
resistant to radiation, see Deinococcus radiodurans.

sulfur. This thiophosphate acts the same as a normal phosphate group, although there is a slight bias
against it by most polymerases. The maximum theoretical specic activity is 1,494 Ci/mmol (55.28
PBq/mol).

Radioactivity can be used in life sciences as a radiolabel


to visualise components or target molecules in a biological system. Some radionuclei are synthesised in particle
accelerators and have short half-lives, giving them high
maximum theoretical specic activities. This lowers the
detection time compared to radionuclei with longer halflives, such as carbon-14. In some applications they have
been substituted by uorescent dyes.

Phosphorus-33 is used to label nucleotides. It is less


energetic than P-32 and does not require protection
with plexi glass. A disadvantage is its higher cost
compared to P-32, as most of the bombarded P31 will have acquired only one neutron, while only
some will have acquired two or more. Its maximum
specic activity is 5,118 Ci/mmol (189.4 PBq/mol).

Phosphorus-32 is widely used for labeling nucleic


acids and phosphoproteins. It has the highest emission energy (1.7 MeV) of all common research radioisotopes. This is a major advantage in experiments for which sensitivity is a primary consideration, such as titrations of very strong interactions
(i.e., very low dissociation constant), footprinting
experiments, and detection of low-abundance phosphorylated species. 32P is also relatively inexpensive. Because of its high energy, however, a number of safety and administrative controls are required (e.g., acrylic glass). The half-life of 32P is
14.2 days, and its maximum specic activity is 9131
Ci/mmol.

Examples of radionuclei
Tritium (hydrogen-3) is a very low energy emitter
that can be used to label proteins, nucleic acids,
drugs and toxins, but requires a tritium-specic lm
or a tritium-specic phosphor screen. In a liquid
scintillation assay (LSA), the eciency is 2050%,
depending on the scintillation cocktail used . The
maximum theoretical specic activity of tritium is
28.8 Ci/mmol (1.066 PBq/mol). However, there
is often more than one tritium atom per molecule:
for example, tritiated UTP is sold by most suppliers with carbons 5 and 6 each bonded to a tritium
atom. C-14, S-35 and P-33 have similar emission
energies. P-32 and I-125 are higher energy emitters
-> inaccurate, see beta vs gamma radiation.

Iodine-125 is commonly used for labeling proteins,


usually at tyrosine residues. Unbound iodine is
volatile and must be handled in a fume hood. Its
maximum specic activity is 2,176 Ci/mmol (80.51
PBq/mol).

Carbon-14 has a long half-life of 5,73040 years.


Its maximum specic activity is 0.0624 Ci/mmol
(2.31 TBq/mol). It is used in applications such as
A good example of the dierence in energy of the various
radiometric dating or drug tests.
radionuclei is the detection window ranges used to detect
them, which are generally proportional to the energy of
Sodium-22 and chlorine-36 are commonly used to
the emission, but vary from machine to machine: in a
study ion transporters. However, sodium-22 is hard
Perkin elmer TriLux Beta scintillation counter , the H-3
to screen o and chlorine-36, with a half-life of
energy range window is between channel 5360; C-14,
300,000 years, has low activity.[1]
S-35 and P-33 are in the window of 361660; and P-32
is in the window of 6611024.
Sulfur-35 is used to label proteins and nucleic acids.
Cysteine is an amino acid containing a thiol group
which can be labeled by S-35. For nucleotides that
do not contain a sulfur group, the oxygen on one 2 Detection
of the phosphate groups can be substituted with a
1

6 SAFETY

2.1

Quantitative

In a liquid scintillation assay (LSA), or liquid scintillation counting (LSC), a small aliquot, lter or swab
is added to scintillation uid and the plate or vial
counter in a scintillation counter.
A Geiger counter is a quick and rough approximation of activity. Lower energy emitters such as tritium can not be detected.

Not all molecules in the solution have a P-32 on the last


(i.e., gamma) phosphate: the specic activity gives the
radioactivity concentration and depends on the radionucleis half-life. If every molecule were labelled, the maximum theoretical specic activity is obtained that for P-32
is 9131 Ci/mmol. Due to pre-calibration and eciency
issues this number is never seen on a label; the values often found are 800, 3000 and 6000 Ci/mmol. With this
number it is possible to calculate the total chemical concentration and the hot-to-cold ratio.

Calibration date is the date in which the vials activity


is the same as on the label. Pre-calibration is when the
2.2 Qualitative
activity is calibrated in a future date to compensate for
Autoradiography: A membrane such as a Northern the decay occurred during shipping.
blot or a hybridised slot blot is put against a lm that
is then developed.

5 Comparison with uorescence

Phosphor storage screen: The membrane is placed


against a phosphor storage screen which is then
See also: Fluorescence in the life sciences
scanned in a phosphorimager. This is ten times
faster and more precise than lm and the result is
Prior to the widespread use of uorescence in the past
already in digital form.
three decades radioactivity was the most common label.

2.3

Microscopy

Electron microscopy: The sample is not exposed to a


beam of electrons but detectors picks up the expelled
electrons from the radionuclei.
Micro-autoradiography imager: A slide is put
against scintillation paper and in a PMT. When two
dierent radiolabels are used, a computer can be
used to discriminate the two.

Scientic methods

Main article: Protein methods


Main article: Nucleic acid methods

Advantages are:
uorescence is much safer and more convenient to
use
Several uorescent molecules can be used simultaneously (given that they do not overlap, cf. FRET),
whereas with radioactivity two isotopes can be used
(tritium and a low energy isotope, e.g. 33 P due to
dierent intensities) but require special machinery
(a tritium screen and a regular phosphor-imaging
screen or a specic dual channel detector, e.g. ).
Several properties are extremely useful (cf. next section)
Note: a channel is similar to colour but distinct, it is
the pair of excitation and emission lters specic for a
dye, e.g. agilent microarrays are dual channel, working
on cy3 and cy5, these are colloquially referred to as green
and red.

Schild regression is a radioligand binding assay. It


is used for DNA labelling (5' and 3'), leaving the
Disadvantages are:
nucleic acids intact.

Radioactivity concentration

A vial of radiolabel has a total activity. Taking as an


example 32P ATP, from the catalogues of the two major suppliers, Perkin Elmer NEG502H500UC or GE
AA0068-500UCI , in this case, the total activity is 500
Ci (other typical numbers are 250 Ci or 1 mCi). This
is contained in a certain volume, depending on the radioactive concentration, such as 5 to 10 mCi/mL (185 to
370 TBq/m3 ); typical volumes include 50 or 25 L.

the dye may be a hindrance or toxic

6 Safety
If good health physics controls are maintained in a laboratory where radionuclides are used, it is unlikely that
the overall radiation dose received by workers will be of
much signicance. Nevertheless the eects of low doses
are mostly unknown so many regulations exist to avoid
unnecessary risks, such as skin or internal exposure. Due

3
to the low penetration power and many variables involved
it is hard to convert a radioactive concentration to a dose.
1 Ci of P-32 on a square centimetre of skin (through
a dead layer of a thickness of 70 m) gives 7961 rads
(79.61 grays) per hour . Similarly a mammogram gives
an exposure of 300 mrem (3 mSv) on a larger volume
(in the US, the average annual dose is 620 mrem or 6.2
mSv[2] ).

See also
Radiation poisoning
Background radiation
Uranium Medical Research Centre (UMRC)

References

[1] Biochemic methods. Sample for medicine Students. 2nd ed


2008, by Birgitte Lttge. Aarhus University.
[2] NCRP 160.

Bruce Alberts, Alexander Johnson, Julian Lewis,


Martin Ra, Keith Roberts, Peter Walter (2007).
Molecular Biology of the Cell, Fifth Edition, Garland Science, 1268 pages. ISBN 0-8153-4105-9.

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