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

磁控微型机器人代表了一种很有前途的平台技术，用于操纵和表征细胞和生物分子。目前的磁性微型机器人系统通常采用明场/荧光显微镜或超声成像作为视觉反馈，无法检索操作下生物样品的任何生化信息。该应用程序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型机器人系统，用于同时操作细胞内细胞器/生物分子并表征其生物物理和生化特性。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的新应用。

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.