Imaging Molecular Level Details of Collagen Fibers by VSFG Microscopy

在这个拟议的项目中，我们计划填补知识差距的关系， 微观自组装结构、胶原-分子相互作用和宏观纤维 I型胶原蛋白的形态，大多数人体组织的主要成分， 用于组织工程的生物材料。通过对胶原-水和胶原-蛋白的研究， 体外系统中的相互作用，模拟生理学相关的三个基本方面- 三维纤维组织结构，我们的目标是填补知识空白，在基本胶原蛋白 research.我们将通过开发一种高光谱成像技术来实现这一目标-振动 和频发生（VSFG）显微镜-在高重复率（400 kHz），并将其应用于 胶原长期的愿景是发展新的生物物理学方法，以揭示分子水平的 结构和相互作用的研究和其他复杂的生物 环境，并最终将其应用于研究各种细胞周围环境相关 疾病为了将光谱特征与晶体的微观和宏观结构相关联， I型胶原蛋白，我们计划应用机器学习技术来分析我们的数据， 胶原蛋白微观/宏观结构的光谱特征。我们将重点关注两大科学问题：（一） 理解微观自组装纤维结构的分子特征及其 与宏观形态的关系（方案1和2）;和（B）研究分子水平 胶原蛋白-分子相互作用（计划3和4）。具体计划包括： 1.获取胶原组织的高光谱VSFG图像，以研究其形态学， 无标记和非侵入性方式 2.建立自组装胶原纤维结构的分子光谱特征 3.理解胶原纤维第一溶剂化层中胶原-水的相互作用。 4.与胶原蛋白相互作用的化学物质和肽的成像空间位置。 如果成功，其意义在于，一种无标记的、振动模式特异性成像技术， 特异性的细胞周间隙，这可以揭示分子水平的见解， 胶原蛋白结构及其与周围分子的相互作用，与纤维化和细胞- 与细胞周围空间相互作用有关的疾病。该项目将有助于扩大 NIGMS通过开发新技术来揭示基本的分子水平原理， 在微观和宏观两个方面与胶原I形态相关的机制和特征 规模和胶原蛋白分子相互作用，奠定了生物物理/生物化学 与胶原蛋白相关的未来生物医学应用的原则。

In this proposed project, we plan to fill the knowledge gap of the relationships between microscopic self-assembled structures, collagen-molecule interactions and macroscopic fiber morphologies of type-I collagen, the primary component of most human tissues and a commonly used biomaterial for tissue engineering. By investigating collagen-water and collagen-protein interactions in in vitro systems that mimic basic aspects of physiologically relevant three- dimensional fibrillar tissue architectures, we aim to fill knowledge gaps in fundamental collagen research. We will achieve this goal by developing a hyperspectral imaging technique – vibrational sum frequency generation (VSFG) microscopy – at high repetition rates (400 kHz) and apply it to collagen. The long-term vision is to develop new biophysics methods to reveal molecular-level structures and interactions for pericellular space research and other complex biological environments, and eventually applying it to study various pericellular environment related diseases. In order to correlate spectral features to microscopic and macroscopic structures of type I collagen, we plan to apply machine-learning techniques to analyze our data and extract spectral signatures of collagen’s micro/macrostructures. We will two major scientific focuses: (A) understanding molecular signatures of microscopic self-assembly fibrils structures and its relationship to the macroscopic morphology (plan 1 and 2); and (B) investigating molecular level collagen-molecule interactions (plan 3 and 4). Specific plans include: 1. Obtaining hyperspectral VSFG images of collagen tissues to study their morphology in a label free and non-invasive manner 2. Establishing molecular spectral signatures of self-assembled collagen fibril structures 3. Understanding collagen-water interaction in first solvation layer of collagen fibers. 4. Imaging spatial locations of chemicals and peptides that interact with collagens. If successful, the significance is that a label free, vibrational mode specific imaging technique specific for pericellular space will be available, which can reveal molecular level insights of collagen structures and its interactions with surrounding molecules, pertinent to fibrosis and cell— pericellular space interaction related diseases. This proposed project contributes to the scope of NIGMS by developing new technology to reveal fundamental molecular-level principle, mechanism and signatures related to morphology of collagen I at both micro- and macroscopic scales, and collagen-molecule interactions, laying foundations for biophysical/biochemical principles for future biomedical applications related to collagens.