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

观察细胞对环境的反应、分裂和死亡，给我们提供了无法用其他方式测量的重要信息。我们需要一台能够长时间拍摄详细照片同时保持细胞存活的显微镜;我们目前的设施无法支持这项工作，因为它们已经被预订了，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领域的基本分子途径和细胞结构。首先，萨拉·萨巴蒂诺斯询问DNA是如何被环境压力和药物治疗保护或突变的。第二，Roberto Botelho和Gagan Gupta观察细胞器的形成，成熟和在细胞健康中的作用。第三，Costin Antonescu测试了细胞如何感知和响应饥饿或营养，以及这在细胞生长和分裂中的作用。我们将一起使用各种蛋白质，这些蛋白质与发光的荧光蛋白质融合。使用活细胞显微镜监测这些荧光融合，以观察随时间推移的模式。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，细胞的进食机器，可以观察到它们的形成，融合和变化。细胞的纤毛是毛发状的结构，我们可以看到它的形成和生长。在这些检测中，我们将使用荧光标记来筛选未知群体，并发现新的基因相互作用和蛋白质功能，这些功能可以在以后的几年中进行测试。我们要求的显微镜是针对这些实验进行优化的;它可以拍摄高分辨率图像，并具有低亮度和环境调节室，有助于在电影制作过程中保持细胞存活。此外，它还能够长时间拍摄和跟踪多种不同的条件，这是一个“高内容”的过程。我们的实验室，我们的部门和我们的研究生课程最近的增长增加了本科生，研究生和博士后学生谁会使用这种显微镜的数量。所要求的设备将在我们描述的工作中牢固且持续地使用。我们在进行这些尖端研究的能力方面受到严重限制，因为我们缺乏所需的设备。我们目前的2台显微镜已被预订一空，无法用于长时间的电影拍摄。它们也不适合这些实验，也不能获得高含量的数据。我们的学生将从这个设备中受益匪浅-它将用于完成他们的论文，写论文，我们的实验室和国际合作。我们的学生在这项工作中获得了宝贵的显微镜技能，并帮助我们创造新的方法来分析数据。这种培训给我们的HQP一个优势，将转化为就业机会，和新的研究方向。

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.