Tracking collective cell migration by confocal microscopy

细胞采用不同的形态来执行不同的功能。因此，了解形态动力学变化的机制对于理解包括细胞迁移、细胞分裂或细胞信号传导在内的基本细胞过程非常重要。研究形态动力学的一种有效方法是使用强大的显微镜实时监测细胞形状的变化和分子成分的分布。这可以通过使用各种细胞荧光标记来完成，从而实现对细胞成分的实时动态跟踪。在过去20年左右的时间里，这种细胞蛋白的实时可视化使我们对细胞迁移、细胞分裂和细胞信号传导机制的理解取得了巨大进展，但为了理解这些过程的工作原理，我们仍然需要学习很多东西。4位申请人的研究项目正是针对研究这些复杂过程的机械细节。为此，他们依靠强大的显微镜，使他们能够可视化某些蛋白质在体内和培养细胞内的行为，从而深入了解这些蛋白质的功能和细胞形态动力学机制。然而，他们监控这些动态事件的能力需要能够长时间地跟踪细胞迁移和细胞分裂等过程。目前，他们的设备是有限的，因为它缺乏通过DIC与荧光同时成像细胞的能力，并且无法控制焦平面的位置以进行长时间的记录。这项提案的目标是获得升级，使这种特定的实时成像技术成为可能，从而使申请人能够开展他们的研究项目，并进一步了解细胞形状是如何调节的。

Cells adopt different morphologies to perform various functions. Understanding the mechanisms involved in morphodynamical changes is thus important to understand fundamental cellular processes including cell migration, cell division or cell signalling. One potent approach to study morphodynamics is to use powerful microscopes to monitor changes in cell shape and the distribution of molecular components in real time. This can be done by using various cellular fluorescent markers, thus enabling the dynamic tracking of cellular constituents in real time. In the past 20 years or so, such visualization of cellular proteins in real time has enabled enormous progress in our understanding of the mechanisms of cell migration, cell division and cell signalling, yet there is still much we need to learn in order to comprehend how these processes works. The research programs of the 4 applicants are aimed precisely at studying the mechanistic details of these complex processes. To this end, they rely on powerful microscopes that enable them to visualize how certain proteins behave inside cells in vivo and in culture, and thus gain insight into the function of these proteins and into the mechanisms of cell morphodynamics. Yet their ability to monitor these dynamic events would require to be able to track processes such as cell migration and cell divisions for long period of time. Currently, their equipment is limiting as it lacks the ability to image cell by DIC simultaneously with fluorescence and to control the position of the focal plane for long time-lapse recordings. The goal of this proposal is to acquire upgrades that will enable such specific live imaging techniques, thus enabling the applicants to carry out their research programs and further our mechanistic understanding of how cells shape is regulated.