Spectro-Holographic Instrument for Dynamic Sensing of Cancer Progression

项目摘要 在本申请中，我们建议开发一种本地对比光学光谱传感方法 识别和表征亚细胞结构，并量化它们的性质 癌症改变，通过使用光散射光谱作为自然光学生物标记物。这样一种技术 可以同时标记大量的亚细胞和亚核结构，而不需要 对早期癌症进展的研究将具有重要价值。没有污渍 也使得这种方法更容易在时间进程癌症进展研究中实施，并将 在人体上进行活体观察。 尽管细胞器和核结构的细胞变化很容易观察和研究 在癌症方面，现有的成像技术有根本的局限性，阻碍了对非常 早期癌前病变。与核等发育不良的细胞变化相反 肿大和组织，癌变的早期阶段有更细微的变化 用标准的显微镜技术很难辨别。可能是最常用的 观察细胞结构的成像工具是荧光显微镜。它可以达到有针对性的对比 然而，对于特定的细胞器或蛋白质，活细胞中的成像仍然仅限于几种类型的 荧光团，因此结构类型。尽管最近发展起来的光遗传方法和新的 活细胞荧光探针显著提高了荧光在生命系统中的效用， 方法本质上局限于在相对较短的时间尺度上观察几种类型的结构。一个偶数 传统光学成像的更本质的限制是它受到衍射极限的限制，并且 无法辨别比波长小得多的细胞结构的特性。在……上面 另一方面，电子显微镜成像方法是破坏性的，涉及广泛的操作 与样本一起，不能在生命系统中使用。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。这种方法 应该在理想的情况下识别活细胞中的所有重要细胞结构，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.