Updated Infrastructure for the Carleton University Biomolecular Radiation Facility

该提案寻求资金来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。这是校园里唯一一个有放射性同位素工作许可证的湿实验室。因此，生物分子辐射设施服务于整个大学的研究人员的不同需求，包括生物系（Bruin, Biggar, Willmore, McKay, MacMillan），神经科学（Chee, Abizaid, McQuaid）和健康科学（Holcik, Cassol）。这个设施迫切需要更新和扩展，以满足理学院研究人员日益增长的需求。对于早期职业研究者，詹妮弗·布鲁恩博士和凯尔·比格博士来说尤其如此。我们急需两件关键设备：一台自动伽马计数器和一台生物分子成像仪。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实验室进行了灌注实验，每次实验产生多达600个流出物样本，这是不可用ELISA测量的。布鲁因实验室迫切需要一个高容量，自动化伽马计数器，以可持续的方式通过放射免疫测定测量动态激素分泌。Biggar实验室研究了赖氨酸甲基转移酶（KMT）酶如何识别甲基化的特定蛋白质底物。这些研究将重组KMTs应用于定制肽阵列的反应条件下，允许体外甲基化介导从3H- s -腺苷-蛋氨酸（3H- sam）甲基供体转移放射性标记的3H。目前基于胶片的荧光照相方法通常需要长达2周的曝光时间，并且通常仍然不够敏感，无法检测到极低水平的3H β衰变。生物分子成像仪提供了对放射性标记肽的更灵敏的检测，并显着将实验时间从数周缩短到数小时。因此，对于紧急项目，比格实验室目前正在向以色列的合作者运送标记阵列，用于磷成像，但这种方法无法长期持续下去。生物分子辐射设施将被整个校园的研究人员大量使用，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，并由生物系的全职实验室技术人员进行仪器特定培训。这个新的基础设施将使我们的设施现代化，并大大增加我们HQP可用的技术数量，使他们的技能多样化。这种基础设施对于Bruin和Biggar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尤为重要。

This proposal seeks funds to replace broken and outdated equipment and to expand the research capacity of the Carleton University Biomolecular Radiation Facility.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.