Updated Infrastructure for the Carleton University Biomolecular Radiation Facility

卡尔顿大学生物分子辐射设施的基础设施更新

• 批准号: RTI-2022-00577
• 负责人: Bruin, Jennifer
• 金额: $ 0.8万
• 依托单位: Carleton University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=745630
• 关键词: Updated; Infrastructure; Carleton; University; Biomolecular

This proposal seeks funds to replace broken and outdated equipment and to expand the research capacity of the Carleton University Biomolecular Radiation Facility.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

该提案寻求资金，以取代破碎和过时的设备，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拥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，包括生物学系（Bruin，比格尔，Willmore，McKay，MacMillan），神经科学（Chee，Abizaid，McQuaid）和健康科学（Holcik，Cassol）。该设施迫切需要更新和扩大，以满足科学学院研究人员日益增长的需求。对于早期的职业调查人员詹妮弗·布鲁因博士和凯尔比格尔博士来说，情况尤其如此。我们急需两个关键设备：自动伽马计数器和生物分子成像仪。 Bruin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存活。为了彻底评估内分泌细胞的激素分泌，Bruin Lab进行了灌注实验，每次实验产生多达600个流出物样品，这对于ELISA测量是不实际的。Bruin实验室迫切需要一台高容量的自动化伽马计数器，通过放射免疫分析法以可持续的方式测量动态激素分泌。比格尔实验室研究赖氨酸甲基转移酶（KMT）如何识别甲基化的特定蛋白质底物。这些研究在允许体外甲基化介导的放射性标记的3 H从3 H-S-腺苷甲硫氨酸（3 H-SAM）甲基供体转移的反应条件下将重组KMT应用于定制肽阵列。目前基于胶片的荧光摄影方法通常需要长达2周的曝光时间，并且通常仍然不够灵敏，无法检测极低水平的3 H β衰变。生物分子成像仪提供了更灵敏的放射性标记肽的检测，并大大减少了实验时间从几周到几小时。因此，对于紧急项目，比格尔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光成像，但这种方法不是长期可持续的。 生物分子辐射设施将被整个校园的研究人员大量使用，每年至少有79名受训人员。HQP将接受卡尔顿大学辐射安全官员的培训，并接受生物系全职实验室技术人员的仪器特定培训。这一新的基础设施将使我们的设施现代化，大大增加我们的HQP可用的技术数量，并使他们的技能多样化。这种基础设施对于Bruin和比格尔实验室的HQP完成论文并继续在高质量期刊上发表研究成果尤为重要。