Combinatorial Microscopies: Platforms for Probing Molecular and Cellular Dynamics and Structures

在从材料科学和化学到结构和细胞生物学等领域，了解并最终控制分子如何组装成功能结构是至关重要的。确定这些复杂组装过程的结构和构象要求及其动力学，最好是通过在相关的长度尺度上、实时和名义上的真实世界条件下检查这些现象来完成。我们的研究重点是组合显微镜的设计和优化，组合显微镜是将不同的成像模式整合到一个公共框架中的工具，以便能够直接询问分子和细胞的自组装。例如，我们最新的工作使我们能够以单分子分辨率绘制位于细胞表面的膜受体的分布和结合状态，并在单个细胞的分辨率下实时跟踪发育过程。在这些创新的基础上，我们现在提议开发平台，使我们能够在整个细胞内跟踪这些现象，尤其是在细胞之间。这样的洞察力对于理解分子是如何运输到细胞表面的，它们在运输过程中的状态和结构，以及是什么相互作用驱动这些动态过程至关重要。开发工具，允许我们跟踪这些过程，然后关联细胞之间随后的细胞间相互作用，将为研究人员提供一套强大的方法，以了解细胞之间如何发生信号传递以及这些事件的功能后果。这一高度跨学科的研究项目包括光子学、生物物理学、化学、计算机科学、细胞和分子生物学以及工程学。它吸引了来自不同背景的所有级别(本科生、研究生、博士后)的HQP进入高度协作和跨学科的研究环境。由于我们强调定制设计，HQP积极参与本地和全球的合作，这为创新和创造力提供了丰富的环境。HQP直接参与独特和创新的成像平台的设计、优化、实施和修改，特别关注开放式硬件和软件设计。通过全面参与这些成像平台对复杂生物学问题和现象的应用，HQP不仅仅是在“使用”黑匣子成像策略，而是提供了一个独特而强大的机会来全面参与研究的各个方面。我们的研究工作具有高度的应用性，与最终用户和制造商密切合作，并公开分发我们实验室的协议、计划和代码职位HQP，适用于各种职业，从初创公司的科学家到学术职位和业务开发专业人员。

Understanding and ultimately controlling how molecules assemble into functional structures is critical in fields ranging from materials science and chemistry to structural and cell biology. Determining the structural and conformational requirements for these complex assembly processes, and the dynamics thereof, is best accomplished by examining these phenomena on relevant length scales, in real-time, and under nominally real-world conditions. Our research focuses on the design and optimization of combinatorial microscopies, tools that integrate different imaging modalities into a common framework, in order to enable direct interrogation of molecular and cellular self-assembly. Our most recent work has, for instance, allowed us to map the distribution and association state of membrane receptors, located on the cell surface, with single-molecule resolution, and track developmental processes in real-time at the resolution of individual cells. Building on these innovations, we now propose to develop platforms that will allow us to track these phenomena throughout the cell, and importantly between cells. Such insights are critical for understanding how molecules are transported to the cell surface, their state and structure during transport, and what interactions drive these dynamic processes. Developing tools that allow us to track these processes and then subsequent inter-cellular interactions between associating cells will provide researchers with a powerful suite of approaches for understanding how signalling occurs between cells and the functional consequences of these events. This highly interdisciplinary research program embraces photonics, biophysics, chemistry, computer science, cell and molecular biology, and engineering. It draws in HQP from all levels (undergraduate, graduate, post-doctoral) from diverse backgrounds into a high collaborative and interdisciplinary research environment. Due to our emphasis on bespoke design, HQP are heavily involved in collaborations, both locally and globally, which provides a rich environment for innovation and creativity. HQP are directly involved in the design, optimization, implementation, and modification of unique and innovative imaging platforms, with a specific focus on open hardware and software design. By being fully involved in the application of these imaging platforms to complex biological questions and phenomena, HQP are not simply "using" black box imaging strategies but are afforded a unique and powerful opportunity to be fully engaged in all aspects of the research. The highly applied nature of our research efforts and the close collaborations with end-users and manufacturers as well as the open distribution of our protocols, plans, and code positions HQP from our lab for a diverse suite of careers, ranging from scientists in start-up companies to academic positions and business development professionals.