Imaging Molecular Level Details of Collagen Fibers by VSFG Microscopy

在这个拟议的项目中，我们计划填补两国之间关系的知识空白 微观自组装结构、胶原-分子相互作用与宏观纤维 I型胶原的形态，是大多数人类组织的主要成分，也是一种常见的 用于组织工程的生物材料。通过研究胶原蛋白-水和胶原蛋白 在模拟生理相关的三个基本方面的体外系统中的相互作用- 三维纤维组织结构，我们的目标是填补基础胶原蛋白方面的知识空白 研究。我们将通过开发一种高光谱成像技术--振动成像来实现这一目标 和频率产生(VSFG)显微镜-在高重复频率(400 KHz)下，并将其应用于 胶原蛋白。长远的愿景是开发新的生物物理学方法来揭示分子水平 用于细胞周围空间研究和其他复杂生物学的结构和相互作用 环境，并最终将其应用于研究与之相关的各种细胞周围环境 疾病。为了将光谱特征与微观和宏观结构相关联 I型胶原，我们计划应用机器学习技术来分析我们的数据并提取 胶原蛋白的微观/宏观结构的光谱特征。我们将把两个主要科学重点放在：(A) 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(计划1和计划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，以研究其形态特征。 无标签和非侵入性方式 2.建立自组装胶原原纤维结构的分子光谱特征 3.了解胶原纤维第一溶剂层中胶原蛋白与水的相互作用。 4.成像与胶原蛋白相互作用的化学物质和多肽的空间位置。 如果成功，意义在于一种无标签、特定于振动模式的成像技术 针对细胞周围空间的特定基因将可用，这可以揭示分子水平的洞察力 胶原结构及其与周围分子的相互作用，与纤维化和细胞有关- 与细胞周隙相互作用相关的疾病。这一拟议项目有助于扩大 NIGMS通过开发新技术来揭示基本的分子水平原理， I型胶原微观和宏观形态的机理及表征 鳞片和胶原-分子相互作用，为生物物理/生化奠定了基础 与胶原蛋白相关的未来生物医学应用原则。

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.