Upconverting Chelates for Cancer Detection and Diagnosis

这项工作的长期目标是通过强大的新分子上转换标记来增强诊断，预测和治疗癌症的工具。这些标记物具有许多潜在用途，包括体外诊断、患病组织的体内检测和分界以及掺入微流体系统中用于高通量筛选。这项工作的近期目标是通过应用于组织样品的体外免疫诊断，更好地理解和发展我们的分子上转换器的能力。这种新的和革命性的类型的报告是基于上转换镧系元素（稀土）螯合物。这些材料没有自发荧光背景，不光漂白，使用红外激发（其光毒性较小，提供良好的组织穿透，并且可以使用二极管激光器提供），并且具有窄的发射带（其有利于多路测定）。上转换螯合物比上转换磷光体（也在SRI开发）小得多;上转换螯合物具有几百的分子量。我们将上转换螯合物应用于组织样品的免疫诊断，其中多路复用。光漂白的能力和不存在将具有很大的价值。在第一阶段的工作（R21），我们的具体目标包括生产一种类型的上转换螯合物和测量其光学性质，组装一个上转换显微镜，其使用，缀合的螯合物的抗体，并测试螯合物-抗体探针在组织细胞系的测定。在第二阶段（R33），我们将扩展螯合物的多重检测，具体目标是制备两个额外的上转换螯合物，扩展上转换显微镜检测所有三种螯合物，并同时检测淋巴瘤组织样本中的三种抗原。这项工作将结合联合收割机的努力，光学物理学家，化学家，免疫学家，癌症生物学家和病理学家。

The long term objectives of this proposed work are to enhance the tools for diagnosing, predicting, and treating cancer through powerful new molecular upconverting labels. These labels have a number of potential uses, including in vitro diagnostics, in vivo detection and demarkation of diseased tissue, and incorporation in microfluidic systems for high throughput screening. The near term objective of this proposed work is to better understand and develop the capabilities of our molecular upconverters through application to in vitro immunodiagnosis of tissue samples. This new and revolutionary type of reporter is based on upconversion in lanthanide (rare earth) chelates. These materials have no autofluorescent background, do not photobleach, use infrared excitation (which is less phototoxic, offers good tissue penetration, and can be provided using diode lasers), and have narrow emission bands (which is advantageous for multiplexed assays). The upconverting chelates are much smaller than the upconverting phosphors (also developed at SRI); the upconverting chelates have molecular weights of a few hundred. We will apply upconverting chelates to immunodiagnosis of tissue samples, where the multiplexing. capabilities and absence of photobleaching will be of great value. In the first phase of work (R21), our specific aims include producing one type of upconverting chelate and measuring its optical properties, assembling an upconverting microscope for its use, conjugating the chelate to antibodies, and testing the chelate-antibody probe in assays on tissue cell lines. In the second phase (R33), we will extend the chelates to multiplexed assays, with specific aims of preparing two additional upconverting chelates, extending the upconverting microscope to detect all three chelates, and simultaneously detecting three antigens in lymphoma tissue samples. This work will combine the efforts of optical physicists, chemists, immunologists, a cancer biologist, and a pathologist.