Autoradiography principles

Keen M, Macdermot J. 1993. Analysis of receptors by radioligand binding. Receptor autoradiography principles and practice. Wharton J, Polak JM, editors. Oxford, England Oxford University Press pp. 23-55. 

Huang, S.C., Phelps, M.E. Principles of tracer kinetic modeling in positron emission tomography and autoradiography. In Phelps. M.E.. Mazziota. J., Schelbert H. (eds) Positron Emission Tomography and Autoradiography Principles and Applications for the Brain and Heart. Raven Press, New York, 1986. 

Hoffman EJ, Phelps ME (1986). Positron emission tomography principles and quantitation. In Phelps ME, Mazziotta J, Schelbert H (eds) Positron emission tomography and autoradiography principles and applications for the brain and heart. Raven Press, New York, p 237... 

Phelps M, Mazziotta J and Schelbert H (eds) Positron Emission Tomography and Autoradiography Principles and Applications for the Brain and Heart. New York Raven Press. 

Fig. 4.12. Principle of the dideoxynucleotide chain-termination procedure. Primer ( ) is annealed to the single-stranded template at a site adjacent to the cloned sequence. Chain extension in the presence of the competing dideoxynucleotide results in the random incorporation of that nucleotide at the appropriate sites in the extended product. The mixture of labelled chain-terminated products are fractionated according to size by electrophoresis on a denaturing acrylamide gel and the ladder of products revealed by autoradiography.

Zanzonico, P. Principles of Nuclear Medicine Imaging Planar, SPECT, PET, Multi-modality, and Autoradiography Systems. Radiat. Res. 177, 349-364 (2012)... 

Autoradiography is particularly well suited for determining the distribution of radioactivity in tissue. In principle, the distribution of radioactivity in a tissue could be assessed by gas flow counting, if the tissue was dissected, its parts weighed and uniformly spread as a dry film on a degassed planchet. However, very often it is difficult to identify exactly the part of the tissue that has been dissected. Furthermore, the fluids which surround the tissue in the body may often be very highly labelled and will contaminate the dissected specimen. The use of autoradiography readily overcomes these difficulties . 

The principle in vitro types of application are for radioimmuno- (or related) assay and pre-clinical development studies prior to future in vivo use, but antibodies may also be labelled for use as probes in immunocytochemistiy. Western blots, or for immunoprecipitation. The choice of radionuclide will depend upon such factors as ease of tracer preparation, ease and efficiency of detection (counting or autoradiography), shelf-life of radiotracer, cost, and safety. A list of some of the most common radionuclides which might be employed for preparation of radiotracers with applications in vitro, together with their physical decay properties is shown in Table 1. 

Clones containing genomic DNA of a particular sequence may be identified by plaque hybridization, which is similar in principle to colony hybridization for detection of particular plasmid recombinants. A lawn of E. coli containing several thousand plaques of independent recombinants is blotted on to a nitrocellulose filter. The DNA is released from the blotted recombinant phage, denatured by alkali, and neutralized. A radioactive probe of nucleic acid containing sequences of the gene of interest is then hybridized to the DNA plaques on the filter. Positive plaques, detected by autoradiography as radioactive DNA-RNA or DNA—DNA hybrids, are then picked off and amplified at lower plaque density (approx. 100 per plate). 

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