High resolution SECM for live sciences.

我们已经开发了一种新型的扫描离子电导显微镜（SICM），它使用精细的纳米吸管，可以对高度复杂的细胞样品进行高分辨率的地形成像，也可以使用纳米尺度的分析进行功能成像，使用相同的探针将试剂输送到细胞表面或进行电生理测量。这是活细胞成像的一个重大进步，我们的目标是通过将SICM与扫描电化学显微镜（SECM）相结合，对化学物种进行敏感的纳米级测绘，将其扩展到化学物种的成像。结合的SICM/SECM探针将被设计得足够小，以成像细胞结构，如微绒毛或精细神经元树突（50-200 nm），以便适用于高分辨率的SECM/SICM成像而不会损坏。基于我们已经进行的概念验证实验，我们建议实现复杂细胞结构和组织样本的高分辨率SECM/SICM成像，这目前是一个巨大的技术挑战。为此，我们需要首先设计和优化用于同时SICM/SECM的纳米探针，并修改我们当前的SICM设置和扫描协议。同时，SICM/SECM将用于显示记录神经介质释放、绘制呼吸活动和检测活细胞和组织中NO的可行性。这项新技术将在生物、医学和物理科学领域得到广泛应用。

We have developed a new form of scanning ion conductance microscopy (SICM) that uses a fine nanopipette that allows the high resolution imaging of the topography of highly complex cellular samples and also functional imaging using the nanoscale assays using the same probe to deliver reagents to the cell surface or to perform electrophysiological measurements. This is a major advance in live cell imaging and we aim to now extend this to image chemical species by combining SICM with scanning electrochemical microscopy (SECM) for sensitive nanoscale mapping of chemical species. The combined SICM/SECM probes will be designed to be small enough to image cellular structure such as microvilli or fine neuronal dendrites (50-200 nm) in order to be suitable for high resolution SECM/SICM imaging without damage. Building on proof of concept experiments that we have performed, we propose to achieve high resolution SECM/SICM imaging of complex cellular structures and tissue samples which currently present a great technical challenge. To do this we need to firstly design and optimize nanoprobes for simultaneous SICM/SECM and modify our current SICM set-up and our scanning protocols. Simultaneous SICM/SECM will then be used to show the feasibility to record neuromediator release, map respiratory activity and detect NO in live cells and tissue. This novel technology should have widespread application in the biological, medical and physical sciences.