Optical photothermal IR (O-PTIR) microscopy for chemical imaging of living cells at sub-micron resolution

用于活细胞化学成像的光学光热红外(OPTIR)显微镜 亚微米分辨率 项目摘要/摘要 光热光谱公司(PSC)和波士顿大学的郑继新教授与Rohith Reddy教授合作 (休斯顿大学)提议开发、验证一种名为光学光热的新技术并将其商业化 红外(OPTIR)光谱。这种基于光学显微镜的仪器将提供微观化学分析 以及用于生命科学的亚微米空间分辨率的化学成像。新的OPTIR技术基于 红外(IR)光谱分析是化学分析中最常用的分析技术之一。红外光谱分析 已经被应用于生物医学中的各种研究问题，但还没有被广泛应用于 活细胞研究有两个关键限制。首先，空间分辨率(~3-30微米)的基本限制 阻碍了红外显微镜在单细胞和亚细胞研究中的广泛应用。第二，强红外吸收 水的存在极大地阻碍了红外光谱技术在活细胞和其他水合样品上的应用。利用 成功的第一阶段研究，这项建议旨在克服传统红外光谱的关键障碍，同时还 提供约10倍更高的空间分辨率。 该项目将涉及两个主要方面：(1)开发一种商用OPTIR显微镜，该显微镜旨在 专门针对生命科学研究界；(2)OPTIR技术在特定领域的示范和验证 癌症研究中的问题。商业仪器的开发将在加利福尼亚州圣巴巴拉的PSC设施进行 与波士顿大学郑继新教授团队合作。将进行与癌症相关的研究 在波士顿大学和休斯顿大学。仪器开发活动将包括开发 生命科学界完全工程化的OTPIR原型，重点是提高空间分辨率， 测量速度、易用性以及与活细胞和其他水合样品的测量兼容性 生理环境。与癌症相关的研究将集中在使用OPTIR技术测量和绘制关键字 光谱标记物是癌细胞和癌症侵袭性的指示器。进一步的研究将调查 红外光谱标记物与肿瘤抗癌疗效的相关性。 项目团队是唯一有资格执行这项工作的人。郑教授是一位获奖的研究先驱 生命科学新的光谱和成像技术的发展和应用 PSC的商业团队拥有数十年开发和商业化高分辨率成像的成功经验 和分析仪器。项目合作者雷迪教授是一位具有广泛专业知识的获奖研究员 红外光谱在癌症研究中的应用。该项目的成功完成将导致 下一代单细胞分析工具，将通过提供 有关细胞状态变化的功能信息，包括膜顺序、蛋白质折叠和DNA损伤。 这些变化可以在活细胞中测量，并随着时间的推移提供对健康和疾病过程的新见解。

Optical photothermal IR (OPTIR) microscopy for chemical imaging of living cells at sub-micron resolution Project Summary/Abstract Photothermal Spectroscopy Corp (PSC) and Prof. Ji-Xin Cheng of Boston University in collaboration with Prof. Rohith Reddy (University of Houston) propose to develop, validate, and commercialize a novel technical called Optical Photothermal Infrared (OPTIR) spectroscopy. This optical microscope-based instrument that will provide microscopic chemical analysis and chemical imaging for the life sciences with sub-micron spatial resolution. The new OPTIR technique is based on infrared (IR) spectroscopy, one on the most commonly used analytical techniques for chemical analysis. IR spectroscopy has already been applied to diverse research problems in the biomedical sciences, but it has not been widely applied for live cell research because of two key limitations. First, fundamental limits on spatial resolution (~3 – 30 µm) have prevented broad application of IR microscopy to single-cell and sub-cellular investigations. Second, strong IR absorption by water has dramatically hindered application of IR spectroscopy to live cells and other hydrated samples. Leveraging successful Phase I research, this proposal aims to overcome the key barriers of conventional IR spectroscopy, while also providing ~10X better spatial resolution. This project will involve two major thrusts: (1) the development of a commercial OPTIR microscope that is designed specifically for the life sciences research community; (2) demonstration and validation of the OPTIR technology on specific problems in cancer research. Commercial instrument development will be performed at PSC facilities in Santa Barbara CA in collaboration with the group of Prof. Ji-Xin Cheng at Boston University. The cancer related research will be performed at both Boston University and the University of Houston. Instrument development activities will include the development of a fully engineering OTPIR prototype for the life sciences community, focus on improvements in spatial resolution, measurement speed, ease of use, and compatibility with measurements of live cells and other hydrated samples in physiological environments. Cancer related research will focus on using the OPTIR technique to measure and map key spectroscopic markers that are indicators of cancerous cells and cancer aggressiveness. Further studies will investigate correlations between IR spectroscopic markers and tumor fighting efficacy. The project team is uniquely qualified to perform this work. PI Prof. Cheng is an award-winning research pioneer in development and application of novel spectroscopic and imaging techniques for the life sciences while the technical and commercial team at PSC has decades of successful experience developing and commercializing high resolution imaging and analytical instrumentation. Project collaborator Prof. Reddy is an award winning researcher with extensive expertise in the application of infrared spectroscopy to issues in cancer research. Successful completion of this project will lead to a next-generation single-cell analysis tool that will profoundly impact cellular and sub-cellular biology by providing functional information about changes in cellular state, including membrane order, protein folding, and DNA damage. These changes can be measured in live cells and over time to provide new insights into processes in health and disease.