Updated Infrastructure for the Carleton University Biomolecular Radiation Facility

这项提案寻求资金，以更换损坏和过时的设备，并扩大卡尔顿大学生物分子辐射设施的研究能力。这是校园里唯一有放射性同位素工作许可证的潮湿实验室空间。因此，生物分子辐射设备服务于全校研究人员的不同需求，包括生物系(Bruin、Bigga、Willmore、McKay、MacMillan)、神经科学系(Chee、Abizaid、McQuaid)和健康科学系(Holcik、Cassol)。这一设施迫切需要更新和扩展，以满足科学系研究人员日益增长的需求。对于职业生涯早期的调查人员詹妮弗·布鲁因和凯尔·比格来说，情况尤其如此。我们迫切需要两个关键设备：一个自动伽马计数器和一个生物分子成像仪。布鲁因实验室研究环境因素，如污染物，如何影响胰腺内分泌细胞的功能和生存。为了彻底评估内分泌细胞的激素分泌，Bruin实验室进行灌流实验，每次实验产生多达600个流出样本，这一点用ELISA法测量是不现实的。Bruin实验室迫切需要一个高容量的自动伽马计数器，以可持续的方式通过放射免疫分析来测量动态激素分泌。比格实验室研究赖氨酸甲基转移酶(KMT)酶如何识别甲基化的特定蛋白质底物。这些研究将重组KMT应用于定制的多肽阵列，其反应条件允许体外甲基化介导从~3H-S-腺苷-蛋氨酸(~H-SAM)甲基供体转移标记的~3H。目前基于胶片的荧光照相方法通常需要长达两周的曝光时间，而且往往仍不够灵敏，无法检测到极低水平的~3H-β衰变。生物分子成像仪为放射性标记的多肽提供了更灵敏的检测，并将实验时间从几周缩短到几个小时。因此，对于紧急项目，比格实验室目前正在将标记的阵列运送给以色列的合作者进行磷成像，但这种方法不是长期可持续的。生物分子辐射设备将被校园内的研究人员大量使用，其中包括每年至少79名实习生。HQP将接受卡尔顿大学辐射安全官员的培训，并接受生物系一名全职实验室技术人员的特定仪器培训。这一新的基础设施将使我们的设施现代化，并极大地扩大我们总部可用技术的数量，并使他们的技能多样化。这一基础设施对于布鲁因和比格实验室的HQP完成他们的论文并继续在高质量的期刊上发表他们的研究来说尤其关键。

This proposal seeks funds to replace broken and outdated equipment and to expand the research capacity of the Carleton University Biomolecular Radiation Facility. This is the only wet lab space on campus with a maintained license for radioisotope work. Therefore, the Biomolecular Radiation Facility serves the diverse needs of researchers across the university, including within the Departments of Biology (Bruin, Biggar, Willmore, McKay, MacMillan), Neuroscience (Chee, Abizaid, McQuaid), and Health Sciences (Holcik, Cassol). This facility desperately needs to be updated and expanded to meet the growing needs of researchers in the Faculty of Science. This is particularly true for early career investigators, Drs Jennifer Bruin and Kyle Biggar. We urgently require two key pieces of equipment: an automated Gamma Counter and a Biomolecular Imager. The Bruin Lab studies how environmental factors, such pollutants, impact pancreatic endocrine cell function and survival. To thoroughly assess hormone secretion by endocrine cells, the Bruin Lab performs perifusion experiments that generate up to 600 effluent samples per experiment, which is not practical to measure by ELISA. The Bruin Lab urgently requires a high capacity, automated Gamma Counter to measure dynamic hormone secretion in a sustainable manner by radioimmunoassay. The Biggar Lab investigates how lysine methyltransferase (KMT) enzymes identify specific protein substrates for methylation. These studies apply recombinant KMTs to custom peptide arrays in reaction conditions that permit in vitro methylation-mediated transfer of radiolabeled 3H from 3H-S-adenosyl-methionine (3H-SAM) methyl-donor. Current film-based fluorography methods regularly require exposure times of up to 2 weeks and are often still not sensitive enough to detect the extremely low levels of 3H beta-decay. A Biomolecular Imager offers far more sensitive detection of radiolabeled peptides and dramatically reduces the experimental timeframe from weeks to hours. Thus, for urgent projects the Biggar lab is currently shipping labeled arrays to a collaborator in Israel for phosphorimaging, but this approach is not sustainable long-term. The Biomolecular Radiation Facility will be heavily used by researchers across campus, including at least 79 trainees per year. HQP will receive training from the Carleton University Radiation Safety Officer and instrument-specific training by a full-time lab technician with the Department of Biology. This new infrastructure will modernize our facility and greatly expand the number of techniques available to our HQP and diversify their skill set. This infrastructure is particularly critical for HQP in the Bruin and Biggar labs to complete their theses and continue to publish their research in high quality journals.