Rapid non-invasive biomechanical imaging of neural crest cell migration in vivo

体内神经嵴细胞迁移的快速非侵入性生物力学成像

• 批准号: 10811154
• 负责人: Jitao Zhang
• 金额: $ 32.99万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-18 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10811154
• 关键词: 3-Dimensional; Address; Affect; Atomic Force Microscopy; Behavior; Biomechanics; Birds; Birth; Cell physiology; Cells; Cephalic; Chemicals; Chick Embryo; Complex; Congenital Disorders; Cues; Data; Data Analyses; Detection; Development; Disease; Ectoderm; Elasticity; Embryo; Embryonic Development; Environment; Epithelium; Event; Evolution; Failure; Fluorescence; Gel; Genetic; Goals; Hour; Image; Imaging Device; In Vitro; Incubators; Inherited; Knowledge; Label; Lasers; Lighting; Literature; Measurement; Mechanics; Mesenchymal; Mesoderm; Methodology; Microscope; Microscopy; Modulus; Molecular; Molecular Analysis; Morphogenesis; Morphologic artifacts; Multimodal Imaging; Mus; Names; Neural Crest Cell; Neural Tube Closure; Neurophysiology - biologic function; Optics; Organ; Organogenesis; Phototoxicity; Play; Population; Prevention; Process; Refractive Indices; Resolution; Role; Sampling; Scanning; Signal Transduction; Speed; System; Techniques; Technology; Testing; Time; Tissues; Xenopus; blastomere structure; cell behavior; cell motility; design; developmental disease; embryo cell; embryo culture; embryo tissue; epithelial to mesenchymal transition; genetic analysis; improved; in vivo; in vivo imaging; innovation; insight; instrument; malformation; mechanical properties; mechanical signal; meter; microscopic imaging; migration; neural plate; neuroimaging; new technology; novel; success; tool; tumor

Project Summary Neural crest cells (NCCs) are a highly migratory cell population that collectively migrates to various places and involves organogenesis during embryo development. The aberrant NCC development can lead to severe congenital and hereditary malformations and diseases. In vitro and recent in vivo evidence show the NCCs coordinate their behaviors upon mechanical changes of external environment, suggesting the crucial involvement of biomechanical cues. However, there are literature controversies regarding data interpretation for implementing contact-based tool in 3D embryonic tissue where involves complex mechanical crosstalk, and it is unknown if there exist a universal mechanical mechanism across species among which NCCs behave very differently. One major reason is the lack of non-contact and non-invasive tool that can access 3D biomechanics of embryonic cell and tissue with high resolution and high speed in vivo. This proposal addresses this unmet need based on a novel optical technology named Brillouin microscopy. The goal of this project is to develop and validate a coaxial line scanning Brillouin microscopy (c-LSBM) for rapidly acquiring mechanical images of NCCs and surrounding tissues in vivo. Specifically, we will focus on the biomechanics during the onset of epithelial- mesenchymal transition and the collective migration, which are crucial events for enabling the function of NCCs in morphogenesis. To achieve this goal, we will first develop c-LSBM into an instrument, which overcomes several technical limitations of existing Brillouin technology and allows distortion-free measurement. In addition, the c-LSBM will be equipped with fluorescence channels for multimodal imaging, and the mechanical relevance of acquired Brillouin data will be validated against gold-standard AFM technique (Aim 1). We will then use this new non-invasive tool to elucidate the role of tissue biomechanics in affecting the migration behavior of NCCs in chick embryo in vivo, which enables us to address the current literature controversies regarding how cells adapt their stiffness to the mechanical environment (Aim 2). In summary, the c-LSBM instrument can serve as a new tool for in-depth biomechanical studies of embryo development in vivo. The non-contact and non-invasive characters of this Brillouin technology can provide new data to advance our knowledge of the physical aspects of development. Together with existing tools as well as genetic & molecular analysis, this will provide a complete methodology for investigating the developmental disorders and the prevention of birth diseases.

项目摘要 神经rest细胞（NCC）是一个高度迁移的细胞种群，共同迁移到各个地方， 在胚胎发育过程中涉及器官发生。异常的NCC开发会导致严重 先天性和遗传畸形和疾病。体外和最近的体内证据表明NCCS 协调其行为在外部环境的机械变化上，表明至关重要 生物力学提示的参与。但是，关于数据解释的文献争议 在3D胚胎组织中实现基于接触的工具，其中涉及复杂的机械串扰，这是 未知是否存在跨物种的普遍机械机制，其中NCC的行为非常 对不同。一个主要原因是缺乏可以访问3D生物力学的非接触和非侵入性工具 具有高分辨率和高速体内的胚胎细胞和组织的。该提案解决了这个未满足的 基于一种名为Brillouin显微镜的新型光学技术的需求。该项目的目的是开发和 验证同轴线扫描布里鲁因显微镜（C-LSBM），以快速获取NCC的机械图像 和周围的组织在体内。具体而言，我们将重点关注上皮开始期间的生物力学 间充质转变和集体迁移，这是启用NCC功能的关键事件 在形态发生中。为了实现这一目标，我们将首先将C-LSBM开发为一种克服的工具 现有的布里鲁因技术的几个技术局限性，并允许无失真的测量。此外， C-LSBM将配备用于多模式成像的荧光通道和机械相关性 获得的Brillouin数据将通过金标准的AFM技术进行验证（AIM 1）。然后我们将使用此 新的非侵入性工具，以阐明组织生物力学在影响NCC迁移行为中的作用 小鸡胚胎在体内，这使我们能够解决有关细胞如何适应的当前文献争议 它们对机械环境的刚度（AIM 2）。总而言之，C-LSBM仪器可以用作新的 用于体内胚胎发育的深入生物力学研究的工具。非接触和非侵入性 这种布里鲁因技术的角色可以提供新的数据以促进我们对物理方面的了解 发展。与现有工具以及遗传和分子分析一起，这将提供完整的 研究发育障碍和预防出生疾病的方法。