MULTI-DIMENSIONAL MICROSCOPY

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 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不断扩大，每种技术都提供了独特的对比机制和对所研究系统中重要生物化合物的灵敏度。例如，可以利用光学相干断层扫描（OCT）非侵入性地检查组织形态。在过去的十年中，非线性方法，如双光子激发荧光（TPEF），二次谐波产生（SHG）和相干反斯托克斯拉曼散射（汽车）显微镜的发展给生物医学研究人员提供了方便的工具，选择性地可视化内源性结构，而不需要标记。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。TPEF，SHG，汽车和OCT技术的组合以前已经实现到光学显微镜，然而，这些成像方式的完全集成尚未完成。在该提案中，我们以以下方式推动多维成像的包络： a）将TPEF、SHG、汽车和OCT组合到单个成像平台中。 B）通过控制光谱相位来优化图像对比度和穿透深度。 所提出的多维成像平台独特地允许关键组织和细胞组分（例如黄素代谢物、弹性蛋白网络、胶原蛋白丝和脂质池）的三维映射沿着详细的组织形态。对生物样本的这种光学评估将提供目前可能的样本的最详细的显微镜视图之一。

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.