Compact all-fiber based CARS microscope for biomedical imaging

描述（由申请人提供）：荧光成像是目前用于细胞和组织分析的主要光学方法。它依赖于荧光分子探针的使用，所述荧光分子探针由已经被修饰的基因表达，或者被标记到感兴趣的分子上。这种修改可能会干扰正常功能并妨碍对实验结果的解释。此外，荧光团本身会受到光漂白的影响，这导致成像时间有限，并妨碍了长期研究。相干反斯托克斯拉曼散射（汽车）显微镜是一种非线性光学成像技术，允许分子的高速振动成像。汽车显微镜提供了几个独特的优势：（a）汽车成像是无标记和非破坏性的。(b)对激发强度的非线性依赖性确保汽车信号仅在焦点中心产生，从而提供固有的亚微米3D空间分辨率。(c)汽车成像提供了内在的化学选择性，并允许键特异性对比。因此，汽车成像有可能成为一种重要的方法，用于对细胞和组织进行微创和定量监测，这对于生物研究和医学诊断至关重要。生物医学研究人员广泛采用这项技术的关键障碍是现有激光系统在产生所需的可调谐、高功率和短脉冲激光束方面的技术和操作限制。这些系统体积庞大、价格昂贵并且需要定期维护。领先的便携式拉曼仪器生产商Alumentron与普渡大学合作，提出通过开发一种紧凑、坚固且价格实惠的基于光纤激光器的汽车显微镜，将汽车研究级实验室仪器推向市场。在第一阶段，我们将证明这一努力的可行性，开发和原型几个光纤组件，目前还没有商业化，然后构建一个功能的全光纤汽车系统。我们将展示细胞和组织的化学选择性，高对比度和空间分辨率成像。在第二阶段，我们将致力于设计优化，以极低的成本制造相关组件，并提高性能，目标是将这种强大的技术商业化。我们也将设计和展示一种小型化的光纤汽车成像内窥镜，以促进临床环境中的微创原位诊断。所提出的研究的成功实施将导致稳健和可靠的化学成像显微镜和内窥镜，其提供关于体内和体外细胞的真实的时间、定量和局部结构和分子信息， 组织中 公共卫生相关性：对细胞和组织进行非侵入性、实时、成分和亚细胞测量的能力将极大地促进我们对肥胖和相关疾病的理解，提供对动脉粥样硬化的快速准确诊断，并增强对乳腺、肺、结肠和咽喉肿瘤的检测和诊断。环境和其他研究领域。

DESCRIPTION (provided by applicant): Fluorescence imaging is the current major optical method for cell and tissue analysis. It relies on the use of fluorescent molecular probes which are either expressed by genes which have been modified, or are tagged on to molecules of interest. The modification can interfere with normal function and hamper the interpretation of experimental results. Furthermore, the fluorophores themselves are subject to photobleaching which results in limited imaging time and prevents long term studies. Coherent anti-Stokes Raman scattering (CARS) microscopy is a nonlinear optical imaging technique that allows high-speed vibrational imaging of molecules. CARS microscopy offers several unique advantages: (a) CARS imaging is label-free and non-destructive. (b) The nonlinear dependence on excitation intensity ensures that the CARS signal is only generated in the focal center, providing an inherent sub-micron 3D spatial resolution. (c) CARS imaging offers intrinsic chemical selectivity and allows bond-specific contrast. Thus CARS imaging has the potential to become a vital method for minimal invasive and quantitative monitoring of cell and tissue which is critical for biological research and medical diagnostics. The key hurdles for this enabling technology to be widely adopted by biomedical researchers are the technical and operational limitations of the existing laser systems to generate the required tunable, high-power, and short-pulsed laser beams. These systems are bulky, expensive and require regular maintenance. Agiltron, a leading portable Raman instrument producer, in collaboration with Purdue University, proposes to bring the CARS research level laboratory instrument into the market by developing a compact, robust and affordable fiber laser based CARS microscope. In Phase I we will demonstrate the feasibility of this effort by developing and prototyping several fiberoptic components that are currently not commercially available and then constructing a functional all fiberoptic CARS system. We will demonstrate chemical selective, high contrast and spatially resolution imaging of cells and tissues. In Phase II, we will address design optimization, fabrication of associated components at drastically lower cost, and performance improvement with the goal of commercializing this powerful technique. We will also design and demonstrate a miniaturized fiber CARS imaging endoscope to facilitate minimal invasive in-situ diagnostics in clinical environment. The successful implementation of the proposed research will result in robust and reliable chemical imaging microscopes and endoscopes that provide real time, quantitative and localized structural and molecular information about in-vivo and in-vitro cell and tissues. PUBLIC HEALTH RELEVANCE: The capability to perform non-invasive, real-time, compositional, and sub-cellular measurement of cell and tissue will greatly facilitate our understanding of obesity and related diseases, provide rapid and accurate diagnostics of atherosclerosis, and enhance the detection and diagnosis of tumor in breast, lung, colon, and throat .The all fiber microscope will also allow high-speed spectral analysis and chemical imaging of samples in pharmaceutical, environmental, and other research fields.