An Integrated Cross-Correlative Environmental Specimen Holder Platform for Optica

描述(申请人提供)：为了全面了解细胞的结构和功能，相关的光学和电子显微镜已被广泛应用于获取整个生物体的宏观视图和单分子或大分子复合体的定位。然而，目前相关的显微解决方案需要大量的样品准备，然后才能使用EM研究生物样品，这可能会改变相关结构并产生图像伪影。因此，本项目旨在开发一种通用的连续流动环境样品夹持器系统，用于光学显微镜(LM)、具有生物功能样品基板的扫描和透射电子显微镜(SEM和TEM)以及用于相关显微镜的微米精度位置参考系统，以利用跨越一定空间尺度和频率范围的多种成像方式。这一功能强大且多功能的相关显微镜样品架平台可实现环境光学显微镜和电子显微镜、精确的温度调节(20：C至80：C)，并支持在室内进行的微流控实验，生物样品在此孵化或药物处理。它还允许跨光学显微镜、扫描电子显微镜和透射电子显微镜平台进行样品成像，而无需额外的操作。拟议的交叉相关标本夹具平台提供了将任何现有的单个显微镜改造成最完整的相关显微镜系统的能力，允许整个显微镜领域联合起来不仅发现大量见解和研究现代医学最紧迫的进展，更重要的是为整个生命和健康科学打开新的大门和机会。 公共卫生相关性：用于光学、扫描和透射电子显微镜的集成交叉相关环境标本保管器平台相关性本项目中将开发的环境交叉相关光学和电子显微镜表征技术将直接影响对生物过程的理解。它的目标是在自然液体环境中对生物结构进行相互关联的纳米级成像，以及在生物过程发生时以高分辨率观察它们的能力。这一新的相关方法与目前使用低温冷冻的相关显微技术相比是一个巨大的飞跃，将被用来全面了解细胞的结构和功能。

DESCRIPTION (provided by applicant): To gain a comprehensive understanding of cellular structures and function, correlative light and electron microscopy has been widely utilized to facilitate the acquisition of both macroscopic view of the whole organism and the localization of single molecules or macromolecular complexes. However, current correlative microscopy solutions require extensive sample preparation before biological specimens can be investigated using EM, which potentially altering the relevant structures and inducing image artifacts. Therefore, this project is aimed to develop a universal continuous-flow environmental specimen holder system for the light microscope (LM), the scanning and transmission electron microscope (SEM and TEM) with biofunctionable sample substrate and micron- precision location reference system for correlative microscopy to take advantage of multiple imaging modalities spanning a range of spatial scales and frequencies. This powerful and versatile correlative microscopy specimen holder platform enables environmental LM and EM, accurate temperature regulation (20:C to 80:C), and empowers microfluidic experiments being conducted inside the chamber, where biological samples are incubated or drug treated. It also allows sample imaging across LM, SEM and TEM platforms without additional manipulation. The proposed cross- correlated specimen holder platform offers the capability to transform any existing individual microscopes into the most complete correlative microscopy system that permits the entire microscopy field joining forces to not only uncover a great deal of insights and research the most urgent advances in modern medicine, but more importantly open new doors and opportunities to the entire life and health sciences. PUBLIC HEALTH RELEVANCE: An Integrated Cross-Correlative Environmental Specimen Holder Platform for Optical, Scanning and Transmission Electron Microscopes Relevance The environmental cross-correlative light and electron microscopy characterization techniques that will be developed in this project will have a direct impact on the understanding of biological processes. It targets the cross-correlative nanometer-scale imaging of biological structures in their native liquid environment, as well as the ability to observe biological processes with high resolution as they are occurring. This new correlative approach, which represents a giant leap forward compared to current correlative microscopy techniques using cryogenic freezing, will be used to gain a comprehensive understanding of cellular structures and function.