IRaman: Breakthrough biomedical microscope with simultaneous infrared and Raman spectroscopy at sub-micron spatial resolution

该第二阶段提案旨在开发和商业化IR+拉曼，这是一种突破性的仪器， 在同一台仪器上同时进行红外（IR）和拉曼光谱分析，空间分辨率达到亚微米。 该项目与NIH的目标保持一致，因为它包含了国家生物医学研究所的几个关键目标。 成像和生物工程，包括光学成像和光谱学，红外成像，共聚焦显微镜， 多模态成像红外光谱和拉曼光谱在研究有机硅的组成和分子结构方面已引起人们的兴趣。 生物材料由于其无标记地操作并且对大分子组合物，例如蛋白质， 脂质、核酸和碳水化合物，以及检测这些大分子的同位素标记， 较小的代谢物。红外光谱和拉曼光谱都广泛用于分析实验室中，并且经常被用于分析。 称为“互补技术”，因为它们都探测不同类型的分子振动。例如ir 光谱学对蛋白质二级结构非常敏感，而拉曼对脂质以及 某些氨基酸在制药应用中，拉曼对药物更敏感，而IR对药物更敏感。 赋形剂（添加剂）通常具有弱的拉曼信号和/或具有大的荧光背景。拉曼可以实现 亚微米空间分辨率，但IR受限于较长的激发波长，空间分辨率为~10 μ m。这 该项目旨在通过提供亚微米空间分辨率的IR和拉曼光谱来克服这一限制。 最近一个引人注目的例子表明，在健康科学中，红外和拉曼的多峰组合具有强大的功能， 分析疟原虫感染的红细胞（D. Perez-Guaita等人光谱91，46-58（2017））。研究 表明“两种技术的结合提供了互补的信息不明显”使用的技术 单独地然而，这项研究是通过一个艰苦的过程进行的， 在两个不同的仪器中的完全相同的细胞位置，需要大量的额外时间和成本。这项建议 旨在开发一种仪器，使同时进行红外和拉曼测量简单、可靠和常规。 该项目将利用成功的第一阶段研究开发和商业化一种新的光学显微镜为基础的 该平台可以在同一台仪器上同时进行IR和拉曼。该项目将涉及一个合作 光热光谱公司与Ji-Xin Cheng博士（波士顿大学）和Lynne Taylor博士之间的对话 （Purdue）。光热团队将设计和建造下一代IR+拉曼仪器，以克服关键的 限制，并扩大了在第一阶段开发的原型的能力。这个为期两年的项目将开发alpha 和beta原型单位的应用程序测试在光热的应用实验室在圣巴巴拉，加利福尼亚州，并将安装一个 在普渡大学Lynne Taylor教授实验室的beta装置上，重点展示IR+拉曼的适用性 制药科学中的关键问题。光热科学家也将与郑教授的小组密切合作 在波士顿大学的细胞生物学，特别是在单个细菌水平上研究抗体敏感性。 β IR+拉曼也将用于研究细胞/组织和微塑料表征中的其他应用。

This Phase II proposal aims to develop and commercialize IR+Raman a breakthrough instrument that will for the first time enable simultaneous infrared (IR) and Raman spectroscopy on the same instrument with sub-micron spatial resolution. This project is well aligned with NIH goals as it incorporates several key thrusts of the National Institute of Biomedical Imaging and Bioengineering, including optical imaging and spectroscopy, infrared imaging, confocal microscopy, and multimodal imaging. IR and Raman have gained interest in investigating the composition and molecular structure of biological materials as they operate label free and are sensitive towards macromolecular composition, such as proteins, lipids, nucleic acids and carbohydrates, as well as the detection of isotopic labelling of these macromolecules and even smaller metabolites. Both Infrared and Raman spectroscopies are widely used in analytical laboratories and are often referred to as “complementary techniques” as they both probe different types of molecular vibrations. For example, IR spectroscopy is very sensitive to protein secondary structure, whereas Raman is particularly sensitive to lipids as well as certain amino acids. And in pharma applications Raman is more sensitive to drugs, whereas IR is more sensitive to excipients (additives) that often have weak Raman signals and/or have large fluorescent backgrounds. Raman can achieve sub-micron spatial resolution, but IR is limited by the longer excitation wavelengths to spatial resolution ~10 um. This project aims to overcome this limitation by providing IR and Raman spectroscopy, both at sub-micron spatial resolution. A compelling recent example of the power of the multimodal combination of IR and Raman in health sciences involved analysis of malaria parasite infected red blood cells (D. Perez-Guaita et al Vib. Spectrosc. 91, 46-58 (2017)). The research showed “that the combination of both techniques provides complementary information not evident” using the techniques individually. This research was performed however using a painstaking process of separately and sequentially measuring the exact same cell locations in two different instruments, requiring substantial additional time and cost. This proposal aims to develop an instrument that makes simultaneous IR and Raman measurements simple, robust, and routine. This project will leverage successful Phase I research to develop and commercialize a new optical microscope-based platform that can perform simultaneous IR and Raman on the same instrument. The project will involve a collaboration between proposer Photothermal Spectroscopy Corp and Dr. Ji-Xin Cheng (Boston University) and Dr. Lynne Taylor (Purdue). The team at photothermal will design and build a next generation IR+Raman instrument to overcome key limitations and expand the capabilities over the prototype developed in Phase I. The two year project will develop alpha and beta prototype units for applications testing at Photothermal’s applications lab in Santa Barbara, CA, and will install a beta unit in the labs of Prof. Lynne Taylor at Purdue University, with a focus on demonstrating applicability of IR+Raman to key problems in pharmaceutical sciences. Photothermal scientists will also collaborate closely with Prof. Cheng’s group at Boston University in cell biology, specifically related to investigate antibody susceptibility at the single bacterium level. The beta IR+Raman will also be used to investigate other applications in cells/tissue and microplastics characterization.