Autoradiography in Biology and Medicine
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The radioactive part of the sample activates the silver halide crystals near by. The slide is then washed away by fixers to get insoluble Ag atom only. The autoradiogram can further be viewed and observed under the microscope. A glass slide to which the sample is fixed is dipped into the molten emulsion. A layer adheres to the glass and solidifies at room temperature. A commercially available film is cut and floated on water, flip down and is allowed to spread.
A glass slide is placed in water under the spread film and is lifted out and kept for drying. Figure 2. Source: Physical biochemistry, applications to biochemistry and molecular biology, David Freifelder, W. Freeman and Company 7. Further very low energy radiation also creates a poorer resolution image on the film. Therefore weak b-emitting isotopes 3H, 14C and 35S are most suitable because the energy of radiation is in between g and a radiations.
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Distance and Thickness of sample : If either the sample is very thick or the sample is far away from the emulsion film, resolution will be lost. Grain size and amount of silver halide crystals : The grain size should be smaller so that there is more availability of AgX crystals. Also concentration of gelatin should be less in emulsion as comapred to AgX crystals.
Thickness of emulsion: The emulsion thickness affects the efficiency of autoradiography with different emitters. For b-emitters the thickness of the emulsion should be less. Exposure time : An autoradiogram must be exposed for a sufficiently long time for proper exposure to view pattern of the track length.
Autoradiography of Fungi through a Layer of Soil and in Agar Culture
Figure 3: The figure shows a minority variant mixed sample exposed to autoradiography film for differing intervals. Lane A shows a 5 minute exposure where the mutant band is not visible. Lane B is a 10 minute exposure where the mutant band see arrow becomes visible. Lanes C and D show 15 and 20 minute exposures, respectively. In these lanes the background intensity begins to obscure the minority variant mutant band.
Source: Juliano et al. This relates the tissue activity to the track density on the plastic. This enables both the absolute abundance and the microdistribution of alpha -active nuclides present to be determined.
Autoradiography in Biology and Medicine. George A. Boyd. New : JBJS
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Diz , , Springer Protocols less. Eur J Biochem —88 Towbin H, Staehelin T, Gordin J Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
Autoradiography and Fluorography
Clin Lab. Elsevier Amsterdam. The biotechnology report. London UK. Campden Publishing Ltd, UK, pp.