Radioactivity is generally used in life sciences for highly sensitive and direct measurements of biological phenomena, and for visualizing the location of biomolecules radiolabelled with a radioisotope.

All atoms exist as stable or unstable isotopes and the latter decay at a given half-life ranging from attoseconds to billions of years; radioisotopes useful to biological and experimental systems have half-lives ranging from minutes to months. In the case of the hydrogen isotope tritium (half-life = 12.3 years) and carbon-14 (half-life = 5,730 years), these isotopes derive their importance from all organic life containing hydrogen and carbon and therefore can be used to study countless living processes, reactions, and phenomena. Most short lived isotopes are produced in cyclotrons, linear particle accelerators, or nuclear reactors and their relatively short half-lives give them high maximum theoretical specific activities which is useful for detection in biological systems.

Radiolabeling is a technique used to track the passage of a molecule that incorporates a radioisotope through a reaction, metabolic pathway, cell, tissue, organism, or biological system. The reactant is 'labeled' by replacing specific atoms by their isotope. Replacing an atom with its own radioisotope is an intrinsic label that does not alter the structure of the molecule. Alternatively, molecules can be radiolabeled by chemical reactions that introduce an atom, moiety, or functional group that contains a radionuclide. For example, radio-iodination of peptides and proteins with biologically useful iodine isotopes is easily done by an oxidation reaction that replaces the hydroxyl group with iodine on tyrosine and histadine residues. Another example is to use chelators such DOTA that can be chemically coupled to a protein; the chelator in turn traps radiometals thus radiolabeling the protein. This has been used for introducing Yttrium-90 onto a monoclonal antibody for therapeutic purposes and for introducing Gallium-68 onto the peptide Octreotide for diagnostic imaging by PET imaging. (See DOTA uses.)

Radiolabeling is not necessary for some applications. For some purposes, soluble ionic salts can be used directly without further modification (e.g., gallium-67, gallium-68, and radioiodine isotopes). These uses rely on the chemical and biological properties of the radioisotope itself, to localize it within the organism or biological system.

Molecular imaging is the biomedical field that employs radiotracers to visualize and quantify biological processes using positron emission tomography (PET) and single-photon emission computed tomography (SPECT) imaging. Again, a key feature of using radioactivity in life science applications is that it is a quantitative technique, so PET/SPECT not only reveals where a radiolabelled molecule is but how much is there.

Radiobiology (also known as radiation biology) is a field of clinical and basic medical sciences that involves the study of the action of radioactivity on biological systems. The controlled action of deleterious radioactivity on living systems is the basis of radiation therapy.

Tritium (hydrogen-3) is a very low beta energy emitter that can be used to label proteins, nucleic acids, drugs and almost any organic biomolecule. The maximum theoretical specific activity of tritium is 28.8 kCi/mol (1,070 TBq/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.

For tritium detection, liquid scintillation counters have been classically employed, in which the energy of a tritium decay is transferred to a scintillant molecule in solution which in turn gives off photons whose intensity and spectrum can be measured by a photomultiplier array. The efficiency of this process is 4–50%, depending on the scintillation cocktail used. The measurements are typically expressed in counts per minute (CPM) or disintegrations per minute (DPM). Alternatively, a solid-state, tritium-specific phosphor screen can be used together with a phosphorimager to measure and simultaneously image the radiotracer. Measurements/images are digital in nature and can be expressed in intensity or densitometry units within a region of interest (ROI).