MULTI-DIMENSIONAL MICROSCOPY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 光学成像的最新发展彻底改变了我们在体内检查细胞和组织的方式。现代光学成像技术的数量正在不断增加，每一种技术都对正在研究的系统中的重要生物化合物提供了独特的对比机制和灵敏度。例如，组织形态可以用光学相干断层扫描(OCT)进行非侵入性检查。在过去的十年里，双光子激发荧光(TPEF)、二次谐波产生(SHG)和相干反斯托克斯拉曼散射(CARS)显微镜等非线性方法的发展为生物医学研究人员提供了不需要标记就可以选择性可视化内源结构的便利工具。显微镜的多维方法集成了各种成像技术和对比机制，以更好地评估生物标本。光学显微镜以前已经实现了TPEF、SHG、CARS和OCT技术的组合，然而，这些成像模式的完全集成还没有实现。在这项提议中，我们以以下方式推动多维成像的极限： A)将TPEF、SHG、CARS和OCT合并为单一成像平台。 B)通过控制光谱相位来优化图像对比度和穿透深度。 建议的多维成像平台独一无二地允许绘制关键组织和细胞成分的三维图谱，如黄素代谢物、弹性蛋白网络、胶原丝和脂类以及详细的组织形态。这种对生物标本的光学评估将提供目前可能的最详细的样本显微图像之一。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Recent developments in optical imaging have revolutionized the way in which we can examine cells and tissue in vivo. The number of modern optical imaging technique is continuously expanding, each offering a unique contrast mechanism and sensitivity to important bio-compounds in the system under study. For instance, tissue morphology can be non-invasively examined with optical coherence tomography (OCT). Over the past decade, the development of nonlinear methods such as two photon excited fluorescence (TPEF), second harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) microscopy has given the biomedical researcher convenient tools for selectively visualizing endogenous structures without the need of labeling. The multi-dimensional approach to microscopy integrates various imaging techniques and contrast mechanisms to better assess the biological specimen. Combinations of the TPEF, SHG, CARS and OCT techniques have previously been realized into optical microscopes, however, a full integration of these imaging modalities has not been accomplished. In this proposal we push the envelope of multi-dimensional imaging in the following way: a)Combining the TPEF, SHG, CARS and OCT into a single imaging platform. b)Optimizing the image contrast and penetration depth by controlling the spectral phase. The proposed multi-dimensional imaging platform uniquely permits the three-dimensional mapping of key tissue and cell components such as flavin metabolites, elastin networks, collagen filaments and lipid pools along with the detailed tissue morphology. Such an optical assessment of the biological specimen would provide one of the most detailed microscopic views of the sample currently possible.