Multi-modal high-resolution technology for tissue diagnostics

描述（由申请人提供）：本项目的总体目标是开发一种多模式高分辨率自由标记近实时技术，该技术能够在活生物体的组织水平上研究分子和细胞转化。我们建议将三种技术联合收割机组合成一个独特的系统：（i）双光子时间分辨荧光光谱（2 p-TRFS），其提供细胞和组织组成的直接评估，（ii）超声背散射显微镜（UBM），其允许组织显微解剖的视觉重建，以及（iii）光声显微术（PAM），其允许映射与组织的特定形态和生理学相关的光学吸收。这样的配置将提供关于所研究的生物系统的补充信息，并且因此促进所讨论的生物系统（例如，肿瘤、动脉粥样硬化斑块、生物工程组织、伤口愈合）的更完整的表征/研究。具体而言，该项目将侧重于设计、工程和验证能够整合上述三种模式的仪器和方法;并侧重于证明同时检测活生物系统中若干生物相关参数的综合方法的可行性。将讨论三个目标。目标1：设计和设计一个集成的混合诊断系统，允许对组织进行高分辨率的同时检测/分析：（a）自体荧光（通过2 p-TRFS），（B）显微解剖（通过UBM），和（c）光吸收（通过PAM）。目标二：旨在确认混合2 p-TRFS- UBM - PAM系统的性能（空间分辨率、穿透深度、成像对比度、灵敏度、信噪比、动态范围、采集速度、数据配准），该系统采用组织体模模型，该模型包含光学和超声对比度。目标3：使用体内仓鼠口腔癌作为模型系统，验证杂交2 p-TRFS- UBM - PAM系统的性能;并证明该系统同时检测和分辨肿瘤体积内多种生物学特征（例如，组织组成、微观结构和新血管生成）的可行性。 公共卫生相关性：这里提出的技术将有可能同时检测和监测感兴趣的生物组织的组成，形态和功能特征。潜在的应用包括在体内研究人类疾病的生物模型，如癌症和动脉粥样硬化性心血管疾病，生物工程组织/结构的非破坏性评估，在开发和伤口愈合机制的纵向研究。

DESCRIPTION (provided by applicant): The overall objective of this project is to develop a multi-modal high-resolution free-label near- real-time technology that enables research of molecular and cellular transformations at tissue level in living organisms. We propose to combine into one unique system three techniques: (i) two-photons time-resolved fluorescence spectroscopy (2p-TRFS) which provides a direct evaluation of cells and tissues composition, (ii) ultrasound backscatter microscopy (UBM) which allows a visual reconstruction of tissue microanatomy, and (iii) photoacoustic microscopy (PAM) which allows for mapping of optical absorption associated to specific morphology and physiology of tissue. Such configuration will provide complementary information about the investigated biological system and thus facilitate a more complete characterization/study of the bio-systems in question (e.g. tumor, atherosclerotic plaque, bioengineered tissue, wound healing). Specifically, this project will focus on design, engineering, and validation of instrumentation and methodologies which allow for integration of the three modalities noted above; and on demonstrating the feasibility of the integrated approach for concurrent detection of several biological-relevant parameters from living biological systems. Three aims will be addressed. Aim 1: To design and engineer an integrated hybrid diagnostic system that allows for high-resolution concurrent detection/analysis of tissue (a) autofluorescence (via 2p-TRFS), (b) micro-anatomy (via UBM), and (c) optical absorbance (via PAM). Aim 2: To validate the performance (spatial resolution, penetration depth, imaging contrast, sensitivity, signal-to-noise ratio, dynamic range, acquisition speed, data co-registration) of the hybrid 2p-TRFS - UBM - PAM system with a tissue phantom model which incorporates optical and ultrasound contrast. Aim 3: To validate the performance of the hybrid 2p-TRFS - UBM - PAM system using an in vivo hamster oral carcinoma as a model system; and to demonstrate the system feasibility for detecting and resolving concurrently multiple biological features (e.g. tissue composition, microstructures, and neo-angiogenesis) within the tumor volume. PUBLIC HEALTH RELEVANCE: The technology proposed here will make possible simultaneous detection and monitoring of compositional, morphological and functional features of biological tissues of interest. Potential applications include research in vivo of biological models of human diseases such as cancer and atherosclerotic cardiovascular disease, non-destructive evaluation of bioengineered tissues/constructs, longitudinal studies during development and wound healing mechanisms.