An Integrated Cross-Correlative Environmental Specimen Holder Platform for Optica

描述（由申请人提供）：为了获得对细胞结构和功能的全面理解，相关光学和电子显微镜已被广泛用于促进整个生物体的宏观视图和单分子或大分子复合物的定位的获取。然而，目前的相关显微镜解决方案需要大量的样品制备之前，可以使用EM，这可能会改变相关的结构，并诱导图像伪影的生物标本进行调查。因此，该第二阶段项目的目的是开发并推向市场的通用连续流动环境样品保持器系统的光学显微镜（LM）和扫描和透射电子显微镜（SEM和TEM）与生物功能的样品基板和微米精度的定位参考系统的相关显微镜，以利用多种成像方式跨越一系列的空间尺度和频率。这款功能强大、用途广泛的相关显微镜标本保持器平台将实现环境LM和EM，允许精确的温度调节（从0 ℃以下到100 ℃），并支持在室内进行的微流体实验，生物样品在室内进行孵育或药物处理。它还允许在LM、SEM和TEM平台上对样品进行成像，而无需额外的操作。在第一阶段，我们成功地创建了系统的工作原型。所提出的交叉相关的标本保持器平台提供了将任何现有的单个显微镜转换成完整的相关显微镜系统的能力，该系统允许整个显微镜领域联合起来，不仅发现许多见解和研究现代医学中最紧迫的进展，而且更重要的是打开新的大门，为整个生命和健康科学领域提供机会。

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.