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

点击翻译按钮获取中文摘要

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.