Chronic Intermittent Hypoxia and Hyperalgesic Priming - Administrative Supplement

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Our current extramural funding, through NINDS, R01 NS128574, entitled Chronic Intermittent Hypoxia and Hyperalgesic Priming, incorporates multiple proteomic experiments that depend on the visualization and quantification of protein expression patterns across multiple in vitro and in vivo tissue samples. Furthermore, Western blotting experiments are utilized throughout the grant proposal to support key experiments that track peripheral neuron and macrophage protein marker expression profiles. Unfortunately, our main piece of machinery for processing and visualizing Western blot membranes has come under a state of irreversible disrepair, and we require immediate replacement to continue experiments proposed in the application. We currently house an 18-year-old MiniMedical Services X-ray developer and processor to visualize Western blot membranes. This developer requires monthly service to clean and replace reservoirs with environmentally- dangerous developer and fixative solutions to produce films that are then digitally scanned and quantified for band densitometry by NIH Image 1.62 to determine protein expression patterns. Several months ago, 2 of the three necessary reservoir pumps failed. The cost to replace the broken pumps are estimated by service technicians to cost more than a brand-new machine, thereby forcing us to discontinue its use in our laboratory for proteomic research. We have since solicited bids to determine how we might replace the broken developer with a chemiluminescence imager. We received several quotes from multiple companies and submit the attached quote for your consideration. The ThermoFisher Life Technologies iBright CL750 Digital Imaging System is an economical replacement for the outdated MiniMed X-ray developer. This imaging system allows for imaging of chemiluminescent Western blots, protein gels, and DNA gels, and supersedes the requirement for multi- instrumental image processing (X-ray film to digital scanner) by integrating the image and sample analysis in one machine. This is expected to significantly improve experimental rigor and reproducibility by reducing sampling variables that can be inadvertently introduced during handling between multiple machines. Furthermore, this technology is much more environmentally friendly by avoiding the need for dangerous chemical disposal. We respectfully submit this request for Administrative Supplemental Funding to continue NINDS-funded research with new technology that will reduce our environmental footprint and increase rigor and reproducibility to conduct work vital to the completion of the proposed studies.