Multi-modal high-resolution technology for tissue diagnostics

描述（由申请人提供）：该项目的总体目标是开发一种多模态高分辨率自由标签近实时技术，使生物组织水平的分子和细胞转化研究成为可能。我们建议将三种技术结合成一个独特的系统：(i)双光子时间分辨荧光光谱（p- trfs），它提供了对细胞和组织组成的直接评估；（ii）超声后向散射显微镜（UBM），它允许对组织微观解剖进行视觉重建；（iii）光声显微镜（PAM），它允许对与组织的特定形态和生理相关的光吸收进行映射。这种结构将提供有关所研究生物系统的补充信息，从而促进对所讨论的生物系统（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、工程和验证仪器和方法，以实现上述三种模式的集成；并展示了从活生物系统中同时检测几个生物相关参数的综合方法的可行性。将实现三个目标。目标1：设计和设计一个集成的混合诊断系统，允许高分辨率并发检测/分析组织(a)自身荧光（通过2p-TRFS）， (b)微观解剖（通过UBM）和(c)光学吸光度（通过PAM）。目的2：通过结合光学和超声对比度的组织模体模型验证混合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、数据共配准）。目的3：以仓鼠口腔癌为模型，验证p- trfs - UBM - PAM杂交系统的性能；并演示该系统在肿瘤体积内同时检测和解析多种生物特征（如组织组成、微结构和新生血管生成）的可行性。

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.