Complementary ex vivo multimodal optical imaging and in vivo Raman spectroscopy to understand tissue dynamics

项目摘要 有限的方法存在检查纵向变化的结构和生物化学性质的组织 在各种生理条件下，如怀孕。光和基于光的技术提供了 用于响应于组织生理学的纵向移位的非侵入性、实时、体内监测， 发育、激素或环境影响。拟议研究的目标是寻求一个 利用互补拉曼散射方法研究组织纵向变化的多管齐下平台 生理学在体内以及离体和比较结果与常规测定，以验证我们的方法。 此外，我们将结合联合收割机各种形式的体内拉曼成像，包括（i）双波长、双区域 拉曼，（ii）偏振敏感拉曼，和（iii）共振拉曼光谱，以定量分析 组织生理学的变化。由于对早产及其组织缺乏详细的了解， 生物化学，我们的小组已经报道了在体内RS检测和提取纵向生化的效用 小鼠和人类子宫颈的变化，子宫颈组织在整个过程中经历了广泛的重塑， 怀孕。我们建立了一个传统的在体RS探针识别显着的光谱变化的能力， 胶原蛋白，弹性蛋白，水和血液中。此外，我们可以将这些拉曼光谱与 小鼠宫颈的生物力学特性，包括刚度和扩张性，作为宫颈组织 经历正常的重塑/成熟以准备分娩。基于我们之前的研究，目前 一项提案将集中于多模态离体和体内拉曼方法，以获得深入的定量信息 在生理性宫颈成熟和过早重塑的小鼠模型中。我们假设 多模式拉曼方法将能够检测关键的生化变化，如细胞外基质 (ECM)组织、组织水合作用、脂质和蛋白质流入以及血管分布，这将允许分子和 子宫颈的结构表型作为生理组织重塑的原型。结果将相互关联 通过离体成像和生物力学测试获得空间信息。我们的目标是：1）实施 多模式非线性成像以表征小鼠子宫颈中结构蛋白的变化，2）使用双 波长，双区域拉曼光谱，以跟踪组织水合作用和脂质动力学的变化，以及3） 评价在给药过程中小鼠宫颈中血液和血管化的空间和时间变化。 怀孕最终，该项目将整合这些目标的结果，以提供一个更完整的画面， 分子和结构的变化，可以用来了解正常的，以及受损的， 怀孕。由此产生的深入生物分子概况和空间组织图获得的正常术语和 采用创新的拉曼方法的早产将提供有关该机制的重要信息 早期宫颈重塑和监测纵向变化的结构和生化 其他组织与RS的特性。

PROJECT SUMMARY Limited approaches exist to examine longitudinal changes in the structural and biochemical properties of tissues under various physiological conditions such as pregnancy. Light and light based technologies offer the potential for non-invasive, real-time, in vivo monitoring of longitudinal shifts in tissue physiology in response to developmental, hormonal, or environmental influences. The objective of the proposed research is to pursue a multi-pronged platform with complementary Raman scattering approaches to study longitudinal changes in tissue physiology in vivo as well as ex vivo and compare the results with conventional assays to validate our approach. In addition, we will combine various forms of in vivo Raman imaging, including (i) dual wavelength, dual region Raman (ii) polarization-sensitive Raman, and (iii) resonance Raman spectroscopy to quantitatively analyze changes in tissue physiology. In response to a lack of detailed understanding of preterm birth and its tissue biochemistry, our group has reported the utility of in vivo RS to detect and extract longitudinal biochemical changes in the mouse as well as human cervix, a tissue that undergoes extensive remodeling over the course of pregnancy. We established the ability of a conventional in vivo RS probe to identify significant spectral changes in collagen, elastin, water, and blood. Moreover, we can correlate these Raman spectra with changes in biomechanical properties of the mouse cervix, including stiffness and distensibility, as the cervical tissue undergoes normal remodeling/maturation in preparation for labor. Building on our prior research, the present proposal will focus multimodal ex vivo and in vivo Raman approaches to gain in-depth, quantitative information during physiologic cervical maturation and in mouse models of premature remodeling. We hypothesize that multimodal Raman approaches will enable detection of key biochemical changes, such as extracellular matrix (ECM) organization, tissue hydration, lipid and protein influx and vascularity that will allow molecular and structural phenotyping of the cervix as a prototype of physiologic tissue remodeling. Results will be correlated with spatial information obtained via ex vivo imaging and biomechanical testing. Our Aims are to: 1) Implement multimodal non-linear imaging to characterize changes in structural proteins in the mouse cervix, 2) Use dual wavelength, dual region Raman Spectroscopy to track changes in tissue hydration and lipid dynamics, and 3) Evaluate spatial and temporal changes in blood and vascularization in the mouse cervix over the course of pregnancy. Ultimately, this project will integrate the results of these aims to provide a more complete picture of the molecular and structural changes that can be used to understand normal as well as compromised pregnancies. The resultant in-depth biomolecular profiles and spatial tissue maps obtained for normal term and preterm pregnancies with the innovative Raman approaches will provide vital information about the mechanism of premature cervical remodeling and for monitoring longitudinal changes in the structural and biochemical properties of other tissues with RS.