Spectro-Holographic Instrument for Dynamic Sensing of Cancer Progression

项目摘要 在本申请中，我们提出开发一种天然对比度光学光谱感测方法， 识别和表征亚细胞结构，并在细胞经历预 癌症改变，通过使用光散射光谱作为天然光学生物标志物。这种技术 能够同时标记大量的亚细胞和亚核结构，而不需要 使用染色剂，并且对于研究早期癌症进展具有很大价值。没有污点 也使得这种方法易于在时间进程癌症进展研究中实施， 适用于人体内观察。 虽然细胞器和核结构的细胞改变很容易观察和研究 在癌症中，现有的成像技术存在根本性的局限性，阻碍了对癌症的研究。 早期癌前病变与发育异常的细胞改变，如核 在肿瘤的扩大和组织化过程中，癌发生的最早阶段有更微妙的变化， 这是用标准的显微镜技术不容易辨别的。也许最常用的 用于观察细胞结构的成像工具是荧光显微镜。它可以实现针对性的对比 对于特定的细胞器或蛋白质，然而，活细胞中的成像仍然局限于少数几种类型， 荧光团和因此的结构类型。虽然最近开发的光遗传学方法和新的 活细胞荧光探针显著地提高了荧光在生命系统中的应用， 该方法固有地局限于在相对短的时间尺度上观察几种类型的结构。更 传统光学成像的更实质性的限制在于其受到衍射极限的影响， 不能辨别明显小于波长的细胞结构的性质。对 另一方面，电子显微镜成像方法是破坏性，涉及大量的操作 与样品，不能在生活系统中使用。为了克服两者的局限性， 方法，需要一种基于完全不同的物理原理的技术。该方法 理想情况下，应当识别活细胞中所有重要的细胞结构，同时动态地 当细胞经历癌前改变时量化它们的性质。

Project Summary In this application we propose to develop a native contrast optical spectroscopic sensing approach that identifies and characterizes subcellular structures and quantifies their properties when cells undergo pre- cancerous alterations, by using light scattering spectra as native optical biomarkers. Such a technique would enable simultaneous labeling of large number of subcellular and subnuclear structures without the use of stains and would be of great value for studying early cancer progression. The absence of stains also makes such methods easy to implement in time-course cancer progression studies and would be amenable for in vivo observations in humans. Although cellular alterations in organelle and nuclear structure are readily observed and studied in cancer, there are fundamental limitations in existing imaging techniques that prevent the study of very early stage pre-cancerous alterations. In contrast to dysplastic cellular alterations such as nuclear enlargement and organization, the earliest stages of carcinogenesis have much more subtle alterations that are not easily discernible with standard microscopy techniques. Perhaps the most often used imaging tool for observing cellular structure is fluorescence microscopy. It can achieve targeted contrast for specific organelles or proteins, however imaging in live cells remains limited to just a few types of fluorophores and therefore structure types. Although recently developed optogenetic methods and new live cell fluorescent probes have significantly improved the utility of fluorescence in living systems, the method is inherently limited to observing a few types of structures at relatively short time scales. An even more substantial limitation of conventional optical imaging is that it is subject to the diffraction limit and cannot discern the properties of cellular structures that are significantly smaller than a wavelength. On the other hand, electron microscopy imaging methods are destructive, involve extensive manipulations with the sample, and cannot be utilized in living systems. In order to overcome the limitations of both methods, a technique that is based on an entirely different physical principle is required. This method should ideally identify all important cellular structures in live cells, while simultaneously dynamically quantifying their properties when cells undergo pre-cancerous alterations.