Dynamic Imaging: high resolution, long-term approaches to molecular mechanisms

动态成像：高分辨率、长期的分子机制方法

• 批准号: RTI-2019-00723
• 负责人: Sabatinos, Sarah
• 金额: $ 10.93万
• 依托单位: Ryerson University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=663997
• 关键词: Dynamic; Imaging; resolution; long; term

Watching cells as they react to conditions, divide, and die gives us important information that can't be measured in other ways. We request a microscope that takes detailed pictures over long periods of time while keeping cells alive; our current facilities cannot support this work because they are booked solidly and are not suitable. This equipment is important because tracking individual cells during environmental stress- such as starvation, temperature change, or drug treatments- gives information that influences Canadian agriculture, livestock and the environment.******Our labs study the basic molecular pathways and cellular structures that influence three important areas. First, Sarah Sabatinos asks how DNA is protected or mutated by environmental stress and drug treatments. Second, Roberto Botelho and Gagan Gupta observe organelle formation, maturation and roles in cell health. Third, Costin Antonescu tests how cells sense and respond to starvation or nutrition, and the role that this plays in cell growth and division. Together, we will use a variety of proteins that are fused to glowing fluorescent proteins. These fluorescent fusions are monitored using the live cell microscope to watch patterns over time. For example, tagged chromosomes can detect if DNA is altered by temperature and drug treatments. Lysosomes, the cell's eating machines, can be watched as they form, fuse and change. A cell's cilia are hair-like structures, that we watch form and grow. In these assays, we will use fluorescent tags to screen unknown populations and discover new gene interactions and protein functions that can be tested in later years.******The microscope that we request is optimized for these experiments; it takes high-resolution images, and has low-light levels and an environmental conditioning chamber which helps to keep cells alive during the movie-making process. Additionally, it is able to photograph and track multiple different conditions over long periods of time, a “high-content” process.******The recent growth of both our labs, our Department and our graduate program has increased the number of undergraduate, graduate and postdoc students who would use this microscope. This requested equipment will be used solidly and constantly with the work we describe. We are severely limited in our ability to perform these kinds of cutting edge research because we lack this requested equipment. Our 2 current microscopes are fully booked and cannot be used for long movies. They are also not appropriate for these experiments, nor can they acquire high-content data.******Our students will benefit tremendously from this equipment- it will be used to complete their theses, write papers, collaborate between our labs and internationally. Our students gain valuable skills in microscopy in this work, and help us to create new ways to analyze data. This training gives our HQP an advantage that will translate into jobs, and new research directions.

观察细胞对条件的反应、分裂和死亡为我们提供了无法用其他方式测量的重要信息。我们需要一台显微镜，可以在长时间内拍摄详细的照片，同时保持细胞的存活；我们现有的设施无法支持这项工作，因为它们被预订得很满，不适合。这种设备很重要，因为在环境压力(如饥饿、温度变化或药物治疗)期间跟踪单个细胞可以提供影响加拿大农业、畜牧业和环境的信息。*我们的实验室研究影响三个重要区域的基本分子途径和细胞结构。首先，萨拉·萨巴蒂诺斯问道，环境压力和药物治疗是如何保护或突变DNA的。其次，Roberto Botelho和Gagan Gupta观察了细胞器的形成、成熟和在细胞健康中的作用。第三，科斯廷·安东内斯库测试了细胞对饥饿或营养的感知和反应，以及这在细胞生长和分裂中所起的作用。在一起，我们将使用各种蛋白质，这些蛋白质融合成发光的荧光蛋白质。这些荧光融合是用活细胞显微镜监测的，以观察一段时间的模式。例如，标记的染色体可以检测DNA是否因温度和药物治疗而改变。溶酶体是细胞的进食机器，人们可以观察它们的形成、融合和变化。细胞的纤毛是毛发状的结构，我们观察形成并生长。在这些分析中，我们将使用荧光标签来筛选未知群体，并发现新的基因相互作用和蛋白质功能，这些功能可以在未来几年进行测试。*我们要求的显微镜是为这些实验而优化的；它可以拍摄高分辨率的图像，具有低光照水平，以及一个环境调节室，有助于在电影制作过程中保持细胞的生命。此外，它能够在很长一段时间内拍摄和跟踪多种不同的情况，这是一个“高含量”的过程。*我们的实验室、系和研究生项目最近的增长增加了使用这种显微镜的本科生、研究生和博士后的数量。我们所要求的设备将与我们所描述的工作一起坚固和持续地使用。我们进行这类尖端研究的能力受到严重限制，因为我们缺乏所需的设备。我们目前的两台显微镜都订满了，不能用于长时间的电影。它们也不适合这些实验，也不能获得高含量的数据。*我们的学生将从这种设备中获得巨大的好处--它将被用来完成他们的论文，写论文，我们的实验室和国际之间的合作。我们的学生在这项工作中获得了宝贵的显微镜技能，并帮助我们创造了分析数据的新方法。这种培训给我们的HQP带来了优势，将转化为就业机会和新的研究方向。