Radioactivity in the life sciences

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

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 specific activities. This lowers the detection time compared to radionuclei with longer half-lives, such as carbon-14. In some applications they have been substituted by fluorescent dyes.

Examples of radionuclei

A good example of the difference in energy of the various radionuclei is the detection window ranges used to detect them, which are generally proportional to the energy of the emission, but vary from machine to machine: in a Perkin elmer TriLux Beta scintillation counter , the H-3 energy range window is between channel 5–360; C-14, S-35 and P-33 are in the window of 361–660; and P-32 is in the window of 661–1024.

Detection

Quantitative

Qualitative

Microscopy

Scientific methods

Main article: Protein methods
Main article: Nucleic acid methods

Radioactivity concentration

A vial of radiolabel has a "total activity". Taking as an example γ32P ATP, from the catalogues of the two major suppliers, Perkin Elmer NEG502H500UC or GE AA0068-500UCI , in this case, the total activity is 500 μCi (other typical numbers are 250 μCi 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 μL.

Not all molecules in the solution have a P-32 on the last (i.e., gamma) phosphate: the "specific activity" gives the radioactivity concentration and depends on the radionuclei's half-life. If every molecule were labelled, the maximum theoretical specific activity is obtained that for P-32 is 9131 Ci/mmol. Due to pre-calibration and efficiency 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 vial’s activity is the same as on the label. "Pre-calibration" is when the activity is calibrated in a future date to compensate for the decay occurred during shipping.

Comparison with fluorescence

Prior to the widespread use of fluorescence in the past three decades radioactivity was the most common label.

Advantages are:

Note: a channel is similar to "colour" but distinct, it is the pair of excitation and emission filters specific for a dye, e.g. agilent microarrays are dual channel, working on cy3 and cy5, these are colloquially referred to as green and red.

Disadvantages are:

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 significance. Nevertheless the effects of low doses are mostly unknown so many regulations exist to avoid unnecessary risks, such as skin or internal exposure. Due to the low penetration power and many variables involved it is hard to convert a radioactive concentration to a dose. 1 μCi of P-32 on a square centimetre of skin (through a dead layer of a thickness of 70 μm) 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

References

  1. Biochemic methods. Sample for medicine Students. 2nd ed 2008, by Birgitte Lüttge. Aarhus University.
  2. NCRP. 160. Missing or empty |title= (help)
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