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

在该项目中，由化学部的化学测量和成像计划资助，路易斯安那州立大学的McCarley教授正在开发以高效和选择性的方式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，McCarley团队正在设计、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敏感。 高度跨学科的方法为团队提供了丰富的学习环境，其培训和多样性使其能够有效地解决跨越物理和生物科学的测量科学挑战。 该项目的重点是开发和评估新的染料分子，其荧光信号可以被打开，或者现有的荧光可以被改变为另一种能量，作为目标分析物作用于染料分子的结果。 该项目中的响应染料被设计为选择性地响应特定酶的存在，例如仅在人类癌细胞中高度聚集的癌症相关酶。 通过使用显微镜和细胞计数方法，证明了用荧光染料从正常细胞中检测和区分癌细胞的高完整性。 该项目的更广泛影响包括：癌细胞高保真检测和成像的经验证的途径;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;通过专业协会会议和公开演讲向科学同行和公众传播工作成果;并有机会通过高影响力的外展计划影响女性pK-12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.