Chronic Intermittent Hypoxia and Hyperalgesic Priming - Administrative Supplement

我们目前的外部资金，通过NINDS，R01 NS128574，题为慢性间歇性低氧和 过敏性痛觉启动，结合了依赖于可视化的多个蛋白质组实验 多个体外和体内组织样本中蛋白质表达模式的量化。此外， 在整个拨款提案中都使用了Western blotting实验来支持跟踪 外周神经元和巨噬细胞蛋白标记物的表达谱。不幸的是，我们的主要部分 处理和可视化蛋白质印迹膜的机器已经处于不可逆转的状态 年久失修，我们需要立即更换以继续申请中提出的实验。 我们目前拥有一名18岁的微型医疗服务公司的X光显像师和处理器，用于可视化WESTERN 印迹膜。该开发商要求每月提供服务，以清洁和更换水库与环保- 危险的显影剂和定影剂溶液，用于生产胶片，然后进行数字扫描和量化 用NIH图像1.62进行条带密度测定以确定蛋白质的表达模式。几个月前，其中两个 三台必要的储油泵都失灵了。更换损坏的泵的费用是按服务估算的 技术人员的花费超过了一台全新的机器，从而迫使我们停止在实验室使用它 用于蛋白质组学研究。 自那以后，我们已经进行了招标，以确定如何用化学发光来取代损坏的显影剂 成像仪。我们收到了来自多家公司的多个报价，并将随附的报价提交给您 对价。ThermoFisher Life Technologies iBright CL750数字成像系统经济实惠 取代过时的MiniMed X光显影剂。该成像系统允许对 化学发光蛋白质印迹、蛋白质凝胶和DNA凝胶，取代了对多个 仪器图像处理(从X射线胶片到数字扫描仪)，将图像和样品分析集成在 一台机器。预计这将显著提高实验的严密性和重复性，因为它减少了 在多台机器之间进行处理时可能会无意中引入的采样变量。 此外，这项技术通过避免使用危险化学品，对环境更加友好。 处理完了。 我们谨此提交行政补充资金申请，以继续NINDS资助的研究 新技术将减少我们的环境足迹，并提高进行的严密性和可重复性 对完成拟议研究至关重要的工作。

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.