An Integrated Cross-Correlative Environmental Specimen Holder Platform for Optica

描述（由申请人提供）：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被广泛应用于获得整个生物体的宏观视图和单分子或大分子复合物的定位。然而，目前的相关显微镜解决方案需要大量的样品制备，然后才能使用EM研究生物标本，这可能会改变相关结构并引起图像伪影。因此，该二期项目旨在为光学显微镜（LM）和扫描和透射电子显微镜（SEM和TEM）开发并推向市场的通用连续流环境标本支架系统，该系统具有生物功能的样品基板和用于相关显微镜的微米精度定位参考系统，以利用跨越一系列空间尺度和频率的多种成像模式。这种功能强大且用途广泛的相关显微镜标本支架平台将实现环境LM和EM，允许精确的温度调节（从低于0℃到100℃），并授权在室内进行微流体实验，其中生物样品孵育或药物处理。它还允许在LM， SEM和TEM平台上进行样品成像，而无需额外的操作。在第一阶段，我们已经成功地创建了系统的工作原型。提出的交叉相关标本支架平台提供了将任何现有的单个显微镜转换为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，不仅可以发现许多见解和研究现代医学中最紧迫的进展，更重要的是为整个生命和健康科学领域打开新的大门并提供机会。

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 alters the relevant structures and induces image artifacts. Therefore, this Phase II project is aimed to develop and bring to market a universal continuous-flow environmental specimen holder system for the light microscope (LM) and the scanning and transmission electron microscope (SEM and TEM) with a biofunctionable sample substrate and a 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 will enable environmental LM and EM, allow accurate temperature regulation (from below 0 C to 100 C), and empower microfluidic experiments conducted inside the chamber, where biological samples are incubated or drug treated. It will also allow sample imaging across LM, SEM, and TEM platforms without additional manipulation. In Phase I we have successfully created a working prototype of the system. The proposed cross-correlated specimen holder platform offers the capability to transform any existing individual microscopes into a complete correlative microscopy system that permits the entire microscopy field to join forces to not only uncover many insights and research the most urgent advances in modern medicine, but more importantly to open new doors and offer opportunities to the entire field of life and health sciences.