MadSci Network: Molecular Biology |
Basically, phosphoimaging is autoradiography but instead of using X-ray film and developing an image photographically, the radioactive pattern is transfered to a phosphoimaging plate which is read by scanning with a laser. When the radioactive membrane is exposed to the phosphoimaging plate, (which contains fine crystals of BaFBr:Eu(2+)), the ionising radiation excites an electron of Eu(2+) into the conduction band, which results in the oxidation of Eu(2+) to Eu(3+) and the reduction of BaFBr to BaFBr- BaFBr:Eu(2+) --> BaFBr-:Eu(3+) The BaFBr- complex has an absorbtion around 600 nm. Exposing the excited complex to a helium-neon laser, red light (633 nm), results in the release of an electron, reducing Eu(3+) to Eu(2+*). The Eu(2+*) (excited state) then releases a photon at 390 nm (blue light) as it returns to its ground state. Phosphoimaging plates can be reused after a 10-minute exposure to visible light i.e. the stored radioactive image is scrubbed by exposure to visible light. http://www5.amershambiosciences.com/aptrix/upp01077.nsf/Content/gel_blot_s torm_storage_phosphor http://analytical.chem.wisc.edu/524class/Folders/Bailey/524topic.html
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