Autoradiography methods

Abstract Quantitative receptor autoradiography methods have heen widely used over the last three decades to study the distribution and physiological role of a receptor in various tissues. This review provides an overview of in vivo and in vitro receptor autoradiography methods and their advantages as well as disadvantages in the study of receptors in the central nervous system. Comparison with immuno-histochemical and in situ hybridization methods is also highlighted in relation to the study of a given receptor in the nervous sytem. 

Kuhar MJ, Unnerstall JR. 1990. Receptor autoradiography. Methods in neurotransmitter receptor analysis. Yamamura HI, Enna SJ, Kuhar MJ, editors. New York Raven Press pp. 177-218. 

The distribution of impurities over a flat sihcon surface can be measured by autoradiography or by scanning the surface using any of the methods appropriate for trace impurity detection (see Trace and residue analysis). Depth measurements can be made by combining any of the above measurements with the repeated removal of thin layers of sihcon, either by wet etching, plasma etching, or sputtering. Care must be taken, however, to ensure that the material removal method does not contaminate the sihcon surface. 

The difference in rates of release of BCNU from wafers produced by the trituration or solution methods is also seen in vivo (11,14), as is shown in Fig. 6. Wafers of PCPP-SA 20 80 were prepared by either the solution or trituration methods, as described above, and were implanted into the brains of rabbits. The animals were sacrificed at various times after implantation and the brains were removed, fixed, and processed for quantitative autoradiography. To quantitate the percentage of the brain exposed to BCNU released from these wafers, the following calculation was performed. The percentage of the brain in which the radioactivity from the tritiated BCNU released from the wafers exceeded the background counts by at least two standard deviation units was plotted as a function of time following implantation in Fig. 6. A control set of rabbits had a solution of BCNU injected directly into the same location in the... 

Signal The end product observed when a specific sequence of DNA or RNA is detected by autoradiography or some other method. Hybridization with a complementary radioactive polynucleotide (eg, by Southern or Northern blotting) is commonly used to generate the signal. 

Compounds that are radioactive can be located on a preparative layer by contact film autoradiography, electronic autoradiography, and storage phosphor screen imaging [21-23]. These methods differ in terms of factors such as simplicity, speed, sensitivity, and resolution, and the method of choice depends on the available equipment, reagents, and instrumentation. All are nondestructive, and the detected compounds can be recovered without change for later studies. 

The second step in 2D electrophoresis is to separate proteins based on molecular weight using SDS-PAGE. Individual proteins are then visualized by Coomassie or silver staining techniques or by autoradiography. Because 2D gel electrophoresis separate proteins based on independent physical characteristics, it is a powerful means to resolve complex mixtures proteins (Fig. 2.1). Modem large-gel formats are reproducible and are the most common method for protein separation in proteomic studies. 

StumpfWE. Techniques for the autoradiography of diffusible compounds, in Methods in Cell Biology, Vol. 13, Academic Press, New York, 1976. 

McGuffee LJ, Hurwitz L, Little SA. A method of coating multiple slides for light microscopic autoradiography. J Histochem Cytochem 1977 25 1107-1108. 

Domer P. Photometric methods in quantitative autoradiography, in Microautoradiography and Electron Probe Analysis. Their Application to Plant Physiology (Luttge U, ed.), Springer Verlag, New York, 1972, pp. 7M8. 

PanayiGC, Neill WA. Scintillation autoradiography. A rapid technique.. 1 Immunol Methods 1972 2 115-117. 

Wharton JP. Receptor Autoradiography and Practice. Modern Methods in Pathology Services, Oxford University Press, New York, 1993. 

Bouteille M, Dupuy-Coin AM, Moyne G. Techniques of localization of proteins and nucleoproteins in the cell nucleus by high resolution autoradiography and cytochemistry, in Methods in Enzymology (O Malley BW, Hardman JG, eds.), Academic Press, New York, 1975, pp. 3-41. 

Autoradiography allows the localization of radionuclide within plant tissue, which is important for an understanding of the uptake and translocation mechanisms. It also represents a selection method for the efficiency... 

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