Concurrent volumetric imaging with multimodal optical systems

项目摘要 快速采用转基因动物和荧光标记的生物相容性纳米颗粒， 生物医学科学中的药物递送增加了对成像技术的需求， 体积成像，使纵向调查的细胞动力学在其自然环境。的 理解组织对损伤和治疗的反应所需的信息的复杂性也产生了 对组合了联合收割机互补性能优势的多模态成像系统的兴趣增加。 在这方面，我们开发了一种集成的双模态成像系统， 相干显微镜（OCM）和双通道共焦荧光显微镜（CFM），使 同时测量深层组织层的荧光和反射率。组合系统 提供了一个独特的机会，通过提供互补的信息， 单独使用任何一个系统都无法获得信息。CFM检测荧光标记的 细胞中的分子。OCM是一种无标记的episcopic方法，可提供有关细胞边界的信息， 细胞外基质，以及正常和致病组织层的厚度变化。 尽管这些技术具有潜力，但如果没有并发的能力，它们的集成是不完整的。 体积成像允许在同一样本中逐步跟踪细胞动力学。至于扫描- 基于系统，OCM和CFM中的快速体积成像需要在个体水平上动态聚焦， 扫描点，这是具有挑战性的，由于有限的时间分辨率的动态聚焦设备。 该项目旨在建立可调声梯度透镜的能力， 迄今为止，OCM和CFM中的聚焦技术能够实现快速体积和并发成像， 深度分辨反射率和荧光从体内深层组织层。这种整合将有一个 对生物医学研究产生巨大影响，因为OCM和CFM技术仍然是最经济实惠，灵活， 并且后者在许多研究机构中容易获得。我们提出以下两个具体目标： 目标1：在共聚焦荧光显微镜中实现快速体积成像。 目标2：在光学相干显微镜中实现扩展景深。 意义 能够同时快速获取体积信息，如反射率和荧光， 深层组织层将为纵向研究发育和 病理生理学机制，并促进在相同的体内细胞跟踪 同时通过减少受试者间的变异性来提高研究的严谨性和重现性， 会影响细胞过程的结果

PROJECT SUMMARY The rapid adoption of genetically modified animals and fluorescently labeled biocompatible nanoparticles for drug delivery in biomedical science have increased demand for imaging technologies capable of fast volumetric imaging to enable longitudinal investigations of cell dynamics in their natural environment. The complexity of information required to understand tissue response to injury and treatment has also generated increased interest in multimodal imaging systems that combine complementary performance strengths. In this regard, we have developed an integrated dual-modality imaging system that combines optical coherence microscopy (OCM) and dual-channel confocal fluorescence microscopy (CFM) to enable the simultaneous measurements of fluorescence and reflectance from deep tissue layers. The combined system provides a unique opportunity to inform cells’ behavior in their natural environment by offering complementary information not available with either system alone. CFM interogates the distribution of fluorescently labeled molecules in cells. OCM is a label-free, episcopic method providing information about the cell boundaries, extracellular matrix, and thickness changes in layers of normal and pathogenic tissues. Despite its potential, the integration of these technologies is incomplete without the capability of concurrent volumetric imaging allowing the tracking of cell dynamics progressively in the same specimen. As for scanning- based systems, fast volumetric imaging in OCM and CFM requires dynamic focusing at the level of individual scanning points, which is challenging due to the limited temporal resolution of dynamic focusing devices. This project aims to establish the capability of the tunable acoustic gradient lens, the fastest dynamic focusing technology to date, in OCM and CFM to enable a fast volumetric and concurrent imaging of depth-resolved reflectance and fluorescence from deep tissue layers in vivo. This integration will have a tremendous impact on biomedical research as OCM and CFM technologies remain the most affordable, flexible, and the latter is readily available in many research facilities. We propose the following two specific aims: Aim 1: Enable fast volumetric imaging in confocal fluorescence microscopy. Aim 2: Enable extended depth-of-field in optical coherence microscopy. Significance The ability to concurrently acquire volumetric information such as reflectance and fluorescence rapidly from deep tissue layers will provide a powerful tool for longitudinal investigations into developmental and pathophysiological mechanisms in various research fields and facilitate in vivo cell tracking in the same specimen while increasing the rigor and reproducibility of studies by decreasing the inter-subject variability that can affect the outcomes of cellular processes.