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 神经嵴细胞（NCC）是一种高度迁移的细胞群，其集体迁移到各个地方， 涉及胚胎发育期间的器官形成。异常的NCC发展可能导致严重的 先天性和遗传性畸形和疾病。体外和最近的体内证据表明， 协调他们的行为对外部环境的机械变化，这表明， 生物力学线索的参与。然而，关于数据解释存在文献争议， 在涉及复杂机械串扰的3D胚胎组织中实施基于接触的工具，并且 目前尚不清楚是否存在一种跨物种的普遍机械机制，其中NCC的行为非常 不同.一个主要原因是缺乏非接触和非侵入性的工具，可以访问3D生物力学 高分辨率、高速度的胚胎细胞和组织活体成像。该提案解决了这一未得到满足的问题 需要基于一种名为布里渊显微镜的新型光学技术。该项目的目标是开发和 验证同轴线扫描布里渊显微镜（c-LSBM）用于快速获取NCC的机械图像 和周围组织中。具体来说，我们将重点关注上皮细胞发病期间的生物力学- 间充质转化和集体迁移，这是使NCC发挥功能的关键事件 在形态发生中。为了实现这一目标，我们将首先将c-LSBM开发成一种仪器，它克服了 该技术克服了现有布里渊技术的几个技术限制，并允许无失真测量。此外，本发明还提供了一种方法， c-LSBM将配备用于多模态成像的荧光通道， 获得的布里渊数据将验证金标准AFM技术（目标1）。然后我们会用这个 一种新的非侵入性工具，用于阐明组织生物力学在影响NCC迁移行为中的作用， 鸡胚在体内，这使我们能够解决目前的文献争议，关于细胞如何适应 它们对机械环境的刚度（目标2）。总之，c-LSBM仪器可以作为一种新的 用于深入研究体内胚胎发育的生物力学工具。非接触式和非侵入式 这种布里渊技术的特性可以提供新的数据，以推进我们对物理方面的认识 发展质量和结合现有的工具以及遗传和分子分析，这将提供一个完整的 调查发育障碍和预防出生疾病的方法。