An accessible framework to achieve multi-dimensional live-cell super-resolution high-content screening

想象一下，我们可以看到细胞内部的分子机器是如何运作的，如何形成网络，如何发展成复杂的组织！几个世纪以来，人类的梦想通过多项技术进步变得越来越接近可能。这一发展始于17世纪的光学显微镜，它使细胞和细菌得以可视化，20世纪50年代的共聚焦显微镜和电子显微镜使更小的结构得以可视化，2000年代的超分辨率显微镜使我们能够以高分辨率看到特定的蛋白质复合物，这一发展达到了顶峰。显微镜是一项突破性的技术，大多数学术实验室和制药公司都利用它来检测药物和可视化疾病模型。这种技术的一个限制是成像所需的时间，这使得获取大量图像变得不切实际-这是筛选数千种药物的先决条件。因此，高内容成像平台已经开发出来，可以在相对较短的时间内获取数千张图像。然而，由高含量筛选显微镜生成的图像通常缺乏分辨率，例如用于可视化小的亚细胞结构。在这里，在这个提案中，我们的目标是结合两种方法来产生第一个超分辨率高含量筛选平台（SR-HCS）。这个平台将真正改变细胞生物学的研究，并以前所未有的速度和分辨率实现发现。

Imagine we could see how the molecular machineries inside cells behave, form networks and develop into complex tissues! What has been a dream for humanity for centuries has become ever closer to possibility through multiple technical advances. This development started with the optical microscope in the 17th century that enabled the visualisation of cells and bacteria, confocal and electron microscopy in the 1950's to visualise smaller structures, and culminated in the advent of super-resolution microscope in the 2000's that now enable us to see specific protein complexes at high resolution. Microscopy has been a ground-breaking technology that most academic labs and pharma companies make use of to assay drugs and visualise disease models. One limitation of such techniques has been the time required for imaging, which made it impractical to acquire a large number of images - a pre-requisite for screening thousands of drugs. Therefore, high-content imaging platforms have been developed that enable the acquisition of thousands of images in a relatively short time frame. Yet, images generated by high-content screening microscopes often lack the resolution, for instance for visualising small sub-cellular structures. Here, in this proposal we aim to combine two methods in order to generate the first super-resolution high-content screening platform (SR-HCS). This platform will truly transform research in cell biology and enable discoveries at an unprecedented speed and resolution.