Acquisition of an imaging platform for proteins and nucleic acids

收购蛋白质和核酸成像平台

• 批准号: RTI-2021-00641
• 负责人: Pearson, Angela
• 金额: $ 10.13万
• 依托单位: Institut national de la recherche scientifique
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=718202
• 关键词: Acquisition; imaging; platform; proteins; nucleic

To understand the mechanisms underlying fundamental biological processes such as microbial replication and cell differentiation, it is essential to be able to efficiently and reliably analyze samples of proteins and nucleic acids, both qualitatively and quantitatively. For example, to follow cell signaling events it is often necessary to quantify posttranslational modifications such as phosphorylation and ubiquitination. Immunoblotting is a standard approach for studying such modifications; however, to obtain high quality data, gel documentation systems are required for efficient acquisition of quantitative data that can be subjected to statistical analyses. Moreover, it is often necessary to assess both the global levels of a particular protein and levels of a specific modified form, as well as levels of loading controls. In addition to protein detection, the imaging and analysis of nucleic acids that are resolved using agarose or acrylamide gel electrophoresis form the backbone of molecular biology. This proposal is to acquire a bioimaging system for proteins and nucleic acids to support fundamental research programs in the biological sciences in a pavilion for 12 laboratories in microbiology and immunology research. The requested imaging module for proteins is a state-of-the art laser scanner module with the capacity for multiplex fluorescence imaging and chemiluminescence imaging. It will be complemented with a work-horse module to support routine nucleic acid imaging and data analysis using standard fluorescent stains. The system will be operated from a computer with proprietary software for image acquisition and analysis. Multiple software licenses will be included to ensure trainees can analyze data at their respective workstation, and thus use of the imaging system will be reserved for data acquisition. There is an urgent need for this system as the nucleic acid imager in the pavilion is past its useful life, due to outdated software and computer operating systems that cannot be updated. Moreover, for immunoblot experiments researchers in the pavilion are presently relying on a decades old film developer for which it has been impossible to order replacement parts for several years. The acquisition of a biomolecular scanner that allows researchers to conduct multiplex immunoblotting will provide higher quality data than is possible with the present infrastructure in the pavilion. It will also ensure that trainees acquire research experience using state-of-the-art approaches to quantitatively analyze specific proteins and nucleic acids in the context of their experiments, while incorporating best practices for the inclusive presentation of data in visual format, so that as many individuals as possible with vision differences can access the data.

为了了解微生物复制和细胞分化等基本生物过程的机制，必须能够有效可靠地定性和定量分析蛋白质和核酸样品。例如，为了跟踪细胞信号事件，通常需要定量翻译后修饰，如磷酸化和泛素化。免疫印迹是研究这种修饰的标准方法;然而，为了获得高质量的数据，需要凝胶记录系统来有效地获取可以进行统计分析的定量数据。此外，通常需要评估特定蛋白质的总体水平和特定修饰形式的水平，以及上样对照的水平。除了蛋白质检测，使用琼脂糖或丙烯酰胺凝胶电泳解析的核酸的成像和分析形成分子生物学的支柱。该提案旨在获得蛋白质和核酸的生物成像系统，以支持微生物学和免疫学研究的12个实验室的展馆中的生物科学基础研究计划。所要求的蛋白质成像模块是最先进的激光扫描仪模块，具有多重荧光成像和化学发光成像的能力。它将补充一个工作马模块，以支持常规核酸成像和数据分析使用标准荧光染色。该系统将通过一台配有专有软件的计算机进行操作，用于图像采集和分析。 将包括多个软件许可证，以确保受训人员可以在各自的工作站分析数据，因此将保留成像系统用于数据采集。由于过时的软件和计算机操作系统无法更新，展馆中的核酸成像仪已过使用寿命，因此迫切需要该系统。此外，对于免疫印迹实验，展馆中的研究人员目前依赖于几十年前的胶片显影剂，几年来一直无法订购更换部件。获得一个生物分子扫描仪，使研究人员能够进行多重免疫印迹将提供更高质量的数据比可能与目前的基础设施在展馆。它还将确保学员获得研究经验，使用最先进的方法在实验中定量分析特定的蛋白质和核酸，同时采用最佳实践，以视觉格式全面展示数据，以便尽可能多的人有视力差异可以访问数据。