Abstract
The detection of free radicals in vivo is very important for the study of many physiologic and pathologic conditions. Free radicals have been implicated in a number of diseases such as ischemia, inflammation, kidney damage, and cancer. Proton-electron double-resonance imaging (PEDRI) allows the indirect detection of free radicals via the Overhauser effect. Nitroxide free radicals used for in vivo PEDRI studies present spectra with two or three lines, but most PEDRI experiments performed to date have used only single-line electron paramagnetic resonance (EPR) irradiation. There is theoretical evidence that simultaneous irradiation of multiple EPR transitions could increase the maximum achievable PEDRI enhancement. From the experimental point of view, this requires the combined use of a suitable multiple-frequency EPR source and a multiple-tuned EPR resonator. A novel radiofrequency (RF) triple-tuned loop-gap resonator for use in PEDRI has recently been developed, and dynamic nuclear polarization (DNP) data were reported. In the present study we describe a new PEDRI apparatus, equipped with a triple-tuned resonator, that is suitable for simultaneous double- or triple-EPR irradiation of nitroxide free radicals. In particular, the details of the EPR hardware used to generate the two or three EPR frequencies are given, and PEDRI images obtained with simultaneous multiple EPR irradiation are shown. Moreover, DNP experimental results showing the increase of the enhancement as a function of the EPR power for single and simultaneous double EPR irradiation are presented. The main goal of this apparatus is to improve the sensitivity and/or to reduce EPR irradiation power in a PEDRI experiment. This is likely to be particularly important in future biologic applications of PEDRI where the applied power must be optimized to reduce sample heating.
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