Enzyme-Activatable Substrate Probes for Fluorescence Imaging and Quantification in Cells

在这个由化学部门化学测量和成像项目资助的项目中，路易斯安那州立大学的麦卡利教授正在开发一种高效和选择性地检测生物细胞中特定蛋白质的光学方法。为了应对这一挑战，麦卡利小组正在设计、制造并在人类细胞研究中实施一系列新的智能染料分子，这些分子对细胞内特定蛋白质分析物的存在非常敏感。高度跨学科的方法为团队提供了丰富的学习环境，他们的训练和多样性使得能够解决跨越物理和生物科学的测量科学挑战成为可能。本项目的重点是开发和评价新的染料分子，其荧光信号可以被打开，或者由于目标分析物作用于染料分子而使现有的荧光转变为另一种能量。这个项目中的反应性染料被设计成选择性地对特定酶的存在做出反应，例如只在人类癌细胞中大量存在的与癌症相关的酶。利用显微镜和细胞计数方法，利用荧光染料对癌细胞进行高完整性检测和从正常细胞中分化。该项目更广泛的影响包括为癌细胞的高保真检测和成像提供可靠的途径；在多样化、丰富的教育环境中对科学家进行跨学科培训，使他们能够应对日益复杂的科学挑战；通过专业学会会议和公开演讲向科学同行和公众传播研究成果；并有机会通过高影响力的外展项目影响pK-12女学生。

In this project, funded by the Chemical Measurement and Imaging Program of the Chemistry Division, Professor McCarley of Louisiana State University is developing optical methods for detecting specific proteins in biological cells in a highly efficient and selective manner. To meet this challenge, the McCarley group is designing and making, and implementing in human cell studies, a new series of smart dye molecules that are sensitive to the presence of specific protein analytes within the cells. The highly interdisciplinary approach provides an enriched learning environment for a team whose training and diversity make it possible to capably address measurement science challenges that span the physical and biological sciences. The focus of this project is development and evaluation of new dye molecules whose fluorescent signal can be switched on, or the existing fluorescence can be changed to another energy, as a result of target analyte action on the dye molecules. The responsive dyes in this project are designed to selectively respond to the presence of particular enzymes, for example cancer-linked enzymes that are highly populated only within human cancer cells. High-integrity detection and differentiation of cancer cells from normal cells with the fluorescent dyes is being demonstrated by use of microscopy and cell counting methods. The broader impacts of this project include proven routes for the high-fidelity detection and imaging of cancer cells; the interdisciplinary training of scientists in a diverse, rich educational environment, that will enable them to address the ever-growing complexity of scientific challenges; dissemination of the work to scientific peers and the public through professional society meetings and public presentations; and the opportunity to influence female pK-12 students through high-impact outreach programs.