Rienstra Nicholas M, Garvis Steve, Sanchez Juan C, Sibert Bryan, Wright Elizabeth R
Department of Biochemistry, University of Wisconsin, Madison, WI, USA.
Department of Mechanical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
bioRxiv. 2025 Jun 21:2025.06.17.660254. doi: 10.1101/2025.06.17.660254.
We present a novel microfluidic flow cell, or Flow-enabled Light and Ultrastructural Imaging Device for Correlative Electron and Light Localization (FLUID-CELL), that connects fluorescence light microscopy (FLM) and cryo-electron microscopy (cryo-EM) for advanced biological imaging and ultrastructural and structural analyses. The design of the FLUID-CELL features a precisely engineered microchannel that maintains native cell culturing conditions, supporting correlation and enabling real-time observation by FLM, as well as subsequent cryo-EM analysis. In this study, this device enabled practical FLM imaging over extended experimental periods, consistent sample handling, and the ability to perform correlative imaging. This capability connects dynamic cellular events imaged by fluorescence light microscopy with high-resolution ultrastructural data collected with cryo-EM. Our dual-modality approach streamlines the workflow and opens new possibilities for investigating the relationship between cellular function and molecular architecture at the nanoscale.
我们展示了一种新型微流控流动池，即用于相关电子和光定位的流动式光与超微结构成像装置（FLUID-CELL），它将荧光显微镜（FLM）和冷冻电子显微镜（cryo-EM）连接起来，用于先进的生物成像以及超微结构和结构分析。FLUID-CELL的设计特点是有一个精确设计的微通道，可维持天然细胞培养条件，支持相关性并能通过FLM进行实时观察，以及随后的冷冻电镜分析。在本研究中，该装置能够在延长的实验期间进行实际的FLM成像、实现一致的样品处理以及进行相关成像的能力。这种能力将通过荧光显微镜成像的动态细胞事件与用冷冻电镜收集的高分辨率超微结构数据联系起来。我们的双模态方法简化了工作流程，并为在纳米尺度上研究细胞功能与分子结构之间的关系开辟了新的可能性。
