1,2-Dioxetanes for Quantitative Chemiluminescence Imaging

在化学学部化学结构、动力学和机制B （CSDM-B）项目的支持下，南卫理公会大学的Alexander R. Lippert教授正在研究用于定量化学发光成像技术的化学发光分子的基本结构和机制。化学发光是在萤火虫和荧光棒中观察到的光，有时被称为化学反应过程中产生的冷光。在细胞和哺乳动物中使用化学发光成像具有显著的优势，包括低成本，高灵敏度，低光毒性，以及在深层组织中成像的能力。然而，由于缺乏对分析物进行精确定量成像的方法和最佳显微镜仪器，这种新兴的分子成像技术受到限制。提出的研究将推进化学发光传感领域的发展，从粗略的相对测量到精确的定量成像的各种分析物。该项目还将有助于提高弱势学生对STEM的参与。PI将与当地HBCU Prairie View a&m大学合作，招募并共同指导代表性不足的学生参与这项研究，并获得长期指导。该项目的三个具体目标是：(1)开发用于定量成像的近红外（NIR）和短波红外（SWIR）比例化学发光探针；(2)研制基于动力学的化学发光探针，用于定量成像；(3)发展用于定量成像的化学发光显微镜。这项工作旨在获得对化学发光机制的理解，以开发创新的分子、仪器和方法，用于细胞和动物中O2、pH、金属、活性物质和酶的定量化学发光成像，从而推进对化学发光的基本理解，并广泛地使生物学研究成为可能。具有近红外和SWIR比例发射的化学发光分子将提供更好的定量和在深层组织中成像的能力。化学发光反应和探针与分析物反应的动力学将得到详细的解决，并将开发方法来促进化学发光显微镜的理解。新型化学发光显微镜的光学元件将最大限度地提高光的聚集能力和减少光子损失。最后，将新分子和机制转化为细胞和全动物成像方法，将开启一个高灵敏度、低光毒性和深层组织定量化学发光成像的新领域。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

With the support of the Chemical Structure, Dynamics, and Mechanisms B (CSDM-B) Program in the Division of Chemistry, Professor Alexander R. Lippert of Southern Methodist University is studying the fundamental structures and mechanisms of chemiluminescent molecules for use in quantitative chemiluminescence imaging techniques. Chemiluminescence is the light observed in fireflies and glowsticks and is sometimes referred to as cold light generated during a chemical reaction. Imaging in cells and mammals using chemiluminescence offers significant advantages including low-cost, high sensitivity, low phototoxicity, and the ability to image in deeper tissue. This emerging technique for molecular imaging is limited, however, by a lack of methods for precise quantitative imaging of analytes and optimal microscopy instrumentation. The proposed study will advance the field of chemiluminescence sensing by progressing from rough relative measurements to precise quantitative imaging of a diverse set of analytes. This project will also serve to enhance participation of under-represented students in STEM. In collaboration with a local HBCU Prairie View A&M University, the PI will recruit and co-mentor under-represented students to participate in this research and receive long-term mentorship.The three specific aims of the project are: (1) develop near-infrared (NIR) and short-wave infrared (SWIR) ratiometric chemiluminescent probes for quantitative Imaging; (2) develop kinetics-based chemiluminescence probes for quantitative imaging; (3) develop chemiluminescence microscopy for quantitative imaging. The proposed work aims to gain mechanistic understanding to develop innovative molecules, instrumentation, and methods for quantitative chemiluminescence imaging of O2, pH, metals, reactive species, and enzymes in cells and animals, thereby advancing fundamental understanding of chemiluminescence and broadly enabling biological studies. Chemiluminescent molecules with NIR and SWIR ratiometric emission will provide better quantification and the ability to image in deeper tissue. The kinetics of the chemiluminescent reactions and reactions of probes with analytes will be solved in detail and methods will be developed to advance this understanding in the context of chemiluminescence microscopy. New types of chemiluminescence microscopes will be developed with optical components that maximize light gathering ability and minimize photon loss. Finally, the translation of novel molecules and mechanistic understanding into methods for cellular and whole animal imaging will unlock a new field of high sensitivity, low phototoxicity, and deep tissue quantitative chemiluminescence imaging.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.