A Magnetic Microrobotic System with Raman Spectroscopy and Confocal Fluorescence Microscopy for Molecular Diagnostics and Cell Biology

磁控微型机器人为细胞和生物分子的操纵和表征提供了一种很有前途的平台技术。目前的磁性微型机器人系统通常使用明场/荧光显微镜或超声成像作为视觉反馈，无法检索操作下的生物样品的任何生化信息。这一应用旨在建立第一个无缝集成拉曼光谱和共聚焦荧光显微镜的磁性微机器人系统，用于同时操纵细胞内细胞器/生物分子并表征其生物物理和生化性质。拉曼和共聚焦荧光成像模式的独特组合为微型机器人系统提供了生物物理和生化反馈，并允许在分子诊断和细胞生物学方面进行新的应用。 提出的微型机器人系统包括：(1)基于倒置显微镜的拉曼成像系统，用于生物样品的生化表征；(2)定制的基于线圈的电磁控制模块，用于定位亚微米分辨率的磁珠基微型机器人；以及(3)激光扫描共聚焦荧光成像模块，其可以容易地集成到拉曼成像系统的倒置显微镜上，用于磁珠基微型机器人和生物样本的3D成像。在CFI和NSERC Discovery Grant的支持下，申请者购买了带有倒置显微镜的拉曼成像系统，并开发了定制的电磁控制模块。此RTI应用程序要求激光扫描共聚焦荧光模块与拉曼成像系统集成。 利用这个独特的系统，我们将开展以下研究项目：(1)新型冠状病毒(严重急性呼吸综合征冠状病毒2 SARS-CoV-2)的显微机器人捕获、浓缩和无扩增检测；(2)脑癌细胞核机制和相关分子机制的微流体表征；以及(3)微流控细胞共培养过程中信号分子的无标记检测和空间分布。所要求的共聚焦荧光成像模块将用于：(1)垂直扫描来自磁珠的荧光信号，用于无放大检测SARS-CoV-2；(2)脑癌细胞内基于磁珠的微型机器人的3D成像以及在磁珠压入过程中细胞的核变形；以及(3)在微流控设备上共同培养的骨细胞(骨细胞)和骨吸收细胞(破骨细胞)的荧光成像。该系统将用于培训来自8个研究实验室的15名HQP，并将成为一个强大的多功能设施，使机器人、诊断和细胞研究等领域的新协作成为可能。

Magnetically controlled microrobots represent a promising platform technology for manipulation and characterization of cells and biomolecules. Current magnetic microrobotic systems typically involve brightfield/fluorescence microscopy or ultrasound imaging as visual feedback, which cannot retrieve any biochemical information of the bio-sample under manipulation. This application aims to establish the first magnetic microrobotic system seamlessly integrating both Raman spectroscopy and confocal fluorescence microscopy, for simultaneous manipulation of intracellular organelles/biomolecules and characterization their biophysical and biochemical properties. The unique combination of the Raman and confocal fluorescence imaging modalities provides the microrobotic system with both biophysical and biochemical feedback and allows novel applications in molecular diagnostics and cell biology. The proposed microrobotic system includes: (1) a Raman imaging system based on an inverted microscope for biochemical characterization of bio-samples; (2) a custom-made, coil-based electromagnetic control module for positioning of magnetic bead-based microrobots with sub-micrometer resolution; and (3) a laser scanning confocal fluorescence imaging module, which can be readily integrated onto the inverted microscope of the Raman imaging system, for 3D imaging of the magnetic bead-based microrobot and the bio-sample. With CFI and NSERC Discovery Grant support, the applicants have purchased a Raman imaging system with an inverted microscope, and developed a custom-made electromagnetic control module. This RTI application is to request the laser scanning confocal fluorescence module for integration with the Raman imaging system. Using this unique system, we will pursue the following research projects: (1) microrobotic capture, enrichment, and amplification-free detection of the novel coronavirus (severe acute respiratory syndrome coronavirus 2 SARS-CoV-2); (2) microfluidic characterization of nucleus mechanics and associated molecular mechanisms of brain cancer cells; and (3) label-free detection and spatial profiling of signaling molecules during microfluidic cell co-culture. The requested confocal fluorescence imaging module will be used for: (1) vertical scanning of fluorecence signals from a magnetic bead for amplification-free detection of SARS-CoV-2; (2) 3D imaging of a magnetic bead-based microrobot inside a brain cancer cell and the cell's nuclear deformation during magentic bead indentation; and (3) fluorecence imaging of bone cells (osteocytes) and bone resorption cells (osteoclasts) co-cultured on a microfluidic device. This system be used for training 15 HQPs from 8 research labs, and will serve as a powerful multifunctional facility to enable new collaborations at the interface of robotics, diagnostics and cell research.