Fluorescence photobleaching/photoactivation imaging system upgrade

生物分子的荧光标记已经成为使用光学显微镜分析细胞事件的强大工具。几种使用荧光标记的定量显微镜技术已经开发出来，可以研究活细胞中的蛋白质、核酸和脂类。光漂白后荧光恢复(FRAP)和光激活(PA)是两种用于测量生物分子扩散和跟踪其运动的技术。我们的5个研究小组正在要求FRAP/PA单位升级最先进的旋转盘共聚焦激光显微镜系统。FRAP/PA装置能够准确定位和跟踪生物分子在三维(3D)中的快速运动，随着时间的推移荧光损失有限，细胞损伤减少。因此，此次升级显著提高了现有基础设施的能力。FRAP/PA单位将支持我们5个小组的研究计划，其中包括一系列基础科学研究，包括：研究细胞如何对DNA损伤做出反应；大脑中的神经细胞如何发育；脂质如何调节细胞过程；胶原等蛋白质如何形成复杂结构；以及推动平滑肌肉收缩的生物机械如何发挥作用。一旦安装完成，这一升级模块将使CFI资助的显微镜成为加拿大大西洋地区独一无二的仪器，为学生提供一个学习先进定量光学显微镜技术的独特培训机会。除本申请书中点名的研究小组外，还将向其他研究小组开放所要求的设备，从而便利达尔豪西大学许多其他调查人员的研究。

Fluorescent-labeling of biological molecules has become a powerful tool for the analysis of cellular events using the light microscope. Several quantitative microscopy techniques that use fluorescent labels have been developed that allow the study of proteins, nucleic acids and lipids within living cells. Fluorescence Recovery After Photobleaching (FRAP) and Photoactivation (PA), are two techniques used to measure the diffusion of biological molecules and to track their movement. Our 5 research groups are requesting FRAP/PA Unit to upgrade a state-of-the-art spinning-disk confocal laser microscopy system. The FRAP/PA Unit provides the ability to accurately localise and track the rapid movement of biological molecules in 3 dimensions (3D), with limited loss of fluorescence over time, and with reduced cellular damage. As such, this upgrade significantly increases the capabilities of the existing infrastructure. The FRAP/PA Unit will support the research programs of our 5 groups, which encompass a range of fundamental studies in basic science, including: the study of how cells respond to DNA damage; how nerve cells in the brain develop; how lipids regulate cellular processes; how proteins such as collagen form complex structures; and how the biological machinery that drives smooth muscle contraction functions. Once installed, this upgrade module will make the CFI funded microscope a one of kind instrument in Atlantic Canada, providing a unique training opportunity for students to learn advanced quantitative light microscopy techniques. The requested equipment will also be accessible to research groups other than those named in this application thereby facilitating the research of many other investigators at Dalhousie University.