On-Chip Expansion Microscopy

摘要 通过产生精确模拟体内细胞的三维（3D）微组织或类器官， 功能，器官芯片技术在开发生物学方面发挥着越来越重要的作用 和筛选药物化合物。他们的成功效用伴随着对上的高需求- 微组织的形态、功能和分子特征的现场成像，使我们能够了解 他们的行为和反应。显然，为了对3D微组织成像，成像技术应该理想地 提供体积信息以及用于细胞甚至亚细胞分析的显微镜分辨率。 然而，微流体装置内的完整3D微组织的芯片上成像从根本上受到以下因素的阻碍： 组织内光散射因此，本提案的总体目标是创新一种方法， 将开发精心设计的微流体生物反应器平台，以实现同时扩增和 直接在芯片上清除完整的微组织，用于原位、超分辨率和体积表征， 突破了传统荧光显微镜在成像深度和光学衍射方面的限制。

Abstract By generating three-dimensional (3D) microtissues or organoids that accurately mimic the in vivo cellular functions, the organ-on-a-chip technologies have been playing an increasingly important role in exploiting biology and screening pharmaceutical compounds. Their successful utility is accompanied by the high demand for on- site imaging of the microtissues’ morphology, function, and molecular signatures that allow us to understand their behaviors and responses. Clearly, to image the 3D microtissues, the imaging technique should ideally provide volumetric information, together with microscopic resolutions for cellular or even subcellular analyses. On-chip imaging of intact 3D microtissues within microfluidic devices is however, fundamentally hindered by intratissue optical scattering. The overall goal of this proposal thus seeks to innovate a methodology, where a meticulously designed microfluidic bioreactor platform will be developed to enable simultaneous expansion and clearing of intact microtissues directly on-chip for in situ, super-resolution, and volumetric characterizations, breaking limits in both imaging depth and optical diffraction with conventional fluorescence microscopy.