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 is also available as a digital image (digital autoradiography), due to the recent development of scintillation gas detectors or rare-earth phosphorimaging systems. 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 marked using an external source. Some autoradiographs can be examined microscopically for localization of silver grains (such as on the interiors or exteriors of cells or organelles) in which the process is termed micro-autoradiography. For example, micro-autoradiography was used to examine whether atrazine was being metabolized by the hornwort plant or by epiphytic microorganisms in the biofilm layer surrounding the plant.
Applications
In biology, this technique may be used to determine the tissue (or cell) localization of a radioactive substance, either introduced into a metabolic pathway, bound to a receptor or enzyme, or hybridized to a nucleic acid. Applications for autoradiography are broad, ranging from biomedical to environmental sciences to industry.
Receptor autoradiography
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. Once the receptor density is known, in vitro autoradiography can also be used to determine the anatomical distribution and affinity of a radiolabeled drug towards the receptor. For in vitro autoradiography, radioligand was directly applying on frozen tissue sections without administration to the subject. Thus it cannot follow the distribution, metabolism and degradation situation completely in the living body. But because target in the cryosections is widely exposed and can direct contact with radioligand, in vitro autoradiography is still a quick and easy method to screen drug candidates, PET and SPECT ligands. The ligands are generally labeled with 3H (tritium), 18F (fluorine), 11C (carbon) or 125I (radioiodine). Compare to in vitro, ex vivo autoradiography were performed after administration of radioligand in the body, which can decrease the artifacts and are closer to the inner environment.
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 [3H]-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. In the realm of behavioral endocrinology, autoradiography can be used to determine hormonal uptake and indicate receptor location; an animal can be injected with a radiolabeled hormone, or the study can be conducted in vitro.
Rate of DNA replication
The rate of DNA replication in a mouse cell growing in vitro was measured by autoradiography as 33 nucleotides per second. The rate of phage T4 DNA elongation in phage-infected E. coli was also measured by autoradiography as 749 nucleotides per second during the period of exponential DNA increase at .
Detection of protein phosphorylation
Phosphorylation means the posttranslational addition of a phosphate group to specific amino acids of proteins, and such modification can lead to a drastic change in the stability or the function of a protein in the cell. Protein phosphorylation can be detected on an autoradiograph, after incubating the protein in vitro with the appropriate kinase and γ-32P-ATP. The radiolabeled phosphate of latter is incorporated into the protein which is isolated via SDS-PAGE and visualized on an autoradiograph of the gel. (See figure 3. of a recent study showing that CREB-binding protein is phosphorylated by HIPK2.)