Dynamic Bioimaging Facility for Cells and Cell Surfaces

这项研究提案将创造一种新的仪器，它将结合物理学和生物学，并使其能够同时记录来自生物细胞的几种类型的信息。原子力显微镜是一种纳米分辨率的显微镜，它在物理上“接触”细胞的表面。它对细胞外层的形状和结构很敏感，并产生细胞表面的三维地图。另一方面，共焦显微镜是一种强大的光学显微镜技术，其中激光的焦点可以在细胞的三维体内可控地移动，因此我们可以通过在细胞内移动这个焦点来建立细胞内部的图像。如果我们能够同时记录这两个显微镜的信息，我们就可以直接将细胞表面的特征与细胞内部的特性联系起来。这方面的一个例子是在获得表面图的同时记录来自膜的荧光。另一个应用将是有目的地用AFM探针的尖端缩进细胞，并记录细胞的反应，因为它的内部骨骼适应外表面的刺激。我们还将把这两个显微镜与一种名为电生理学的技术结合起来，这种技术将提供穿过细胞表面的电信号的信息。最终，我们获得的信息将使我们更深入地了解细胞过程和特殊功能，细胞之间如何相互沟通，以及它们如何相互作用和适应环境。

This research proposal would create a novel instrument which would combine Physics and Biology, and which would make it possible to simultaneously record several types of information from biological cells. The Atomic Force Microscope is a nanometre-resolution microscope, which physically 'touches' the surface of the cell. It is sensitive to the shape and structure of the outer layer of the cell, and produces a 3-dimensional map of the cell surface. Confocal microscopy on the other hand is a powerful light microscope technique, in which the focal point of a laser beam can be controllably moved throughout the 3-dimensional body of the cell, and therefore we can build up a picture of the inside of the cell by moving this focal point around inside the cell. If we are able to record information from these 2 microscopes at the same time, we can correlate directly features on the cell surface with internal cellular properties. One example of this would be to record fluorescence from membranes whilst obtaining a surface map. Another application would be to purposefully indent the cell with the tip of the AFM probe, and record the response of the cell, as its internal skeleton adapts to the stimulus on the outside surface. We will also combine these two microscopes with a technique called electrophysiology which will give information on the electrical signals crossing the surface of the cell. Ultimately the information we obtain will give us more insights into cell processes and specialised functions, how cells communicate with one another, and also how they interact and adapt to their environment.