Definition of Autoradiography

Autoradiography: A technique using X- ray film to visualize molecules or fragments of molecules that have been radioactively labeled. Autoradiography has many applications in the laboratory. Autoradiography can, for example, be used to analyze the length and number of DNA fragments after they are separated from one another by a method called gel electrophoresis.

A process by which radioactive materials, often though not exclusively incorporated into cell structures, are located by exposure to a photographic emulsion forming a pattern on the film corresponding to the location of the radioactive compounds within the cell. A technique in which radioactive molecules make their location known by exposing photographic films or emulsions.

An autoradiograph is an image on an x-ray film or nuclear emulsion produced by the pattern of decay emissions (e.g., beta particles or gamma rays) from a distribution of a radioactive substance. Alternatively, the autoradiograph can also be available as a digital image (digital autoradiography), thanks to the recent development of scintillation gas detectors[1] or rare earth phosphorimaging systems[2]. In biology, this technique may be used to determine the tissue localization of a radioactive substance, either introduced into a metabolic pathway, bound to a receptor[3][4] or enzyme, or hybridized to a nucleic acid[5]. The film or emulsion is apposed to the labeled tissue section to obtain the autoradiograph (also called an autoradiogram). The auto- prefix indicates that the radioactive substance is within the sample, as distinguished from the case of historadiography or microradiography, in which the sample is X-rayed using an external source. The use of radiolabeled ligands to determine the tissue distributions of receptors is termed either in vivo or in vitro receptor autoradiography if the ligand is administered into the circulation (with subsequent tissue removal and sectioning) or applied to the tissue sections, respectively. The ligands are generally labeled with 3H (tritium) or 125I. The distribution of RNA transcripts in tissue sections by the use of radiolabeled, complementary oligonucleotides or ribonucleic acids ("riboprobes") is called in situ hybridization histochemistry. Radioactive precursors of DNA and RNA, 3H (thymidine) and 3 (uridine) respectively, may be introduced to living cells to determine the timing of several phases of the cell cycle. RNA or DNA viral sequences can also be located in this fashion. These probes are usually labeled with 32P, 33P, or 35S.

This autoradiographic approach contrasts to techniques such as PET and SPECT where the exact 3-dimensional localization of the radiation source is provided by careful use of coincidence counting, gamma counters and other devices.