Super-resolution fluorescence atomic force (SURFACE) microscopy

细菌是对人类极其重要的微生物。考虑到它们的微小尺寸，我们对它们如何生长和分裂的理解大部分是通过各种显微镜方法确定的。自从最早的显微镜问世以来，已经有了许多重要的发现，而且不断的技术发展也带来了令人兴奋的新发现。细菌细胞生物学和确定成分亚细胞定位的能力在很大程度上是由荧光显微镜驱动的，其中单个成分用荧光标记进行标记。然而，检测的限制，再加上细菌的小尺寸限制了这种方法的真正潜力。最近的新技术使荧光显微镜能够跨越这一屏障，达到接近单个分子的分辨率水平。然而，由于技术上的困难，缺乏商业可用性和一些发展的费用，它们尚未产生很大的影响。我们一直在使用一种叫做原子力显微镜的补充方法，它可以提供非常高的表面分辨率。我们已将此应用于细菌细胞壁（重要抗生素如青霉素的作用部位）的研究。这揭示了许多重要的新见解。本项目旨在建立一种耦合超分辨荧光和原子力的新型显微镜。鉴于研究团队的跨学科专业知识，我们将使用现有的显微镜作为支架，快速有效地开发新机器。新机器将在我们目前的细菌细胞壁结构和动力学研究领域进行测试和进一步开发。这将为开发一种新型显微镜提供一个理想的框架，然后可以更广泛地利用和复制，从而使一种新颖而令人兴奋的方法得以迅速采用。

Bacteria are microscopic organisms of immense importance to humans. Given their diminutive size much of our understanding of how they grow and divide has been determined by a variety of microscopy approaches. Since the earliest microscopes many important discoveries have been made and continued technological developments have permitted new and exciting revelations. Bacterial cell biology and the ability to determine the subcellular localization of components have been largely driven by fluorescence microscopy, whereby individual components are labelled with fluorescent markers. However the limit of detection, coupled with the small dimensions of bacteria has limited the true potential of this approach. Very recently new techniques have allowed fluorescence microscopy to traverse this barrier to attain a level of resolution approaching single molecules. However due to the technical difficulties, lack of commercial availability and expense of some of these developments they have not as yet made a great impact. We have been using a complementary approach called atomic force microscopy, which gives very high resolution of surfaces. We have applied this to the study of bacterial cell walls (the site of action of important antibiotics such as penicillin). This has revealed many important new insights. The aim of the project is to build a new type of microscope coupling super resolution fluorescence and atomic force. We will use our existing microscopy set up as a scaffold to rapidly and efficiently develop the new machine, given the interdisciplinary expertise of the investigator team. The new machine will be tested and further developed within our current bacterial cell wall architecture and dynamics research area. This will provide an ideal framework to develop a new microscope, which can then be utilized and duplicated more widely to allow the rapid uptake of a novel and exciting approach.