Development and application of automated multiwell plate fluorescence microscopy

该项目涉及基于MicroManager的模块化开源自动荧光显微镜平台的开发，并集成了光学自动聚焦模块。这将使自动化荧光寿命成像(FLIM)能够分析蛋白质相互作用和细胞代谢过程的变化。将探索一种基于自发荧光的多光束多光子激发方案，用于光学切片和无标记对比度。使用单分子定位显微镜技术的自动超分辨率成像也将被实施。该平台将在适当的情况下使用开放源码软件和低成本组件开发，以扩大对这些先进显微镜技术的获取。将探索这些成像方式在医学研究中的应用，包括COPD巨噬细胞宿主病原体的异质性，与胚胎干细胞分化和癌症相关的细胞代谢变化的研究，以及细胞核内激活诱导的脱氧胞苷脱氨酶(AID)的时间和空间调节。

This project involves the development of a modular open source automated fluorescence microscope platform based on Micromanager and incorporating an optical autofocus module. This will enable automated fluorescence lifetime imaging (FLIM) for assays of protein interactions and changes in cell metabolic processes. A multibeam multiphoton excitation scheme will be explored for optical sectioning and label-free contrast based on autofluorescence. Automated super-resolved imaging with single molecule localisation microscopy techniques will also be implemented. This platform will be developed with open source software and low-cost components where appropriate to widen access to these advanced microscopy techniques. Applications of these imaging modalities to medical research will be explored including to heterogeneity in the host-pathogen of macrophages for COPD, to studies of cellular metabolic changes associated with the differentiation of embryonic stem cells and with cancer and to the temporal and spatial regulation of activation-induced deoxycytidine deaminase (AID) in the cell nucleus.