Photonic Crystal Fiber Probe Fluorescence Biosensing

光子晶体光纤探针荧光生物传感

• 批准号: 7123836
• 负责人: JAMES R. BAKER
• 金额: $ 37.98万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7123836
• 关键词: antineoplastics; bioengineering /biomedical engineering; biomarker; biomedical equipment development; biosensor device; fiber optics; fluorescence; fluorescent dye /probe; molecular oncology; nanotechnology; neoplasm /cancer; neoplasm /cancer pharmacology; photonics; technology /technique development; tissue /cell culture

DESCRIPTION (provided by applicant): Methods using fluorescent probes to identify cancer signatures and biological activities of cancer cells hold great promise. Probes based on fluorescence resonance energy transfer (FRET) and techniques such as fluorescence correlation spectroscopy (FCS) and other similar technologies offer the ability to identify specific RNA or protein molecules that can identify a cancer and provide information on oncogenic pathways used by the tumor cells. Other probes can give insight into drug response by measuring apoptosis induction by chemotherapies and radiation. However, fluorescent analysis has several limitations. Most ex vivo analyses use a flow cytometer or complex, confocal microscope to perform analyses, and this requires that tissue be removed from the body and often disrupted into cells, then fixed and analyzed in a static manner. The problems with in vivo fluorescent analysis are even greater since background fluorescence and tissue scattering, even in the near-infrared range, limit signal acquisition to the skin. Two-photon excitation has been a critical advance in optics, facilitating FRET, FCS and CARS techniques in vitro. However, these applications are limited by the complex technology (confocal microscopy) necessary to employ these techniques. We have demonstrated the use of two-photon fluorescence analysis through optical fibers for analysis of cancer cells in vitro and human tumors in vivo in SCID mice. This prior work constitutes the equivalent of an R21 proposal, as we achieved our major objectives: to develop sensing system optics and electronics and to document the ability of this system to obtain and analyze fluorescence signals in vitro and in vivo. The primary goal of this R33 application is to develop a more sensitive prototype device based on a novel dual-clad photonic crystal fiber (DCPCF) that we hypothesize will provide the sensitivity and redundancy necessary for the clinical evaluation of fluorescence signals in vivo using several fluorescence techniques. We plan to carry out our studies in three Specific Aims: 1: Develop DCPCF for use in a two-photon optical fiber fluorescence probe (D-TPOFF). 2: Utilize the D-TPOFF to quantify cancer signatures in vitro and monitor drug effects in tumor cells using targeted nanoparticles ex vivo and in vivo. 3: Utilize D-TPOFF to adapt other fluorescent techniques to examine events in tumors in vivo. At the end of these studies, this technology will be at a point where it is ready for commercialization.

描述（由申请人提供）：使用荧光探针鉴定癌症特征和癌细胞生物学活性的方法具有很大的希望。 基于荧光共振能传递（FRET）以及诸如荧光相关光谱（FCS）和其他类似技术等技术的探针具有鉴定可以鉴定癌症并提供有关肿瘤细胞使用的致癌途径信息的特定RNA或蛋白质分子的能力。 其他探针可以通过测量化学疗法和辐射诱导凋亡诱导来深入了解药物反应。 但是，荧光分析有几个局限性。 大多数离体分析都使用流式细胞仪或复杂的共聚焦显微镜进行分析，这要求将组织从体内去除并经常被破坏到细胞中，然后固定并以静态方式进行分析。 体内荧光分析的问题甚至更大，因为背景荧光和组织散射，即使在近红外范围内，也将信号限制为皮肤。 在体外，两光子激发在光学，促进FCS和汽车技术方面一直是重要的进步。 但是，这些应用受到采用这些技术所需的复杂技术（共聚焦显微镜）的限制。 我们已经证明了通过光纤分析SCID小鼠体内的癌细胞和人类肿瘤的两光子荧光分析。 这项先前的工作构成了R21提案的等效，因为我们实现了主要目标：开发传感系统光学和电子设备，并记录该系统在体外和体内获得和分析荧光信号的能力。 该R33应用的主要目标是基于我们假设的新型双胶光子晶体纤维（DCPCF）开发一种更灵敏的原型设备，它将提供使用多种荧光技术在体内对荧光信号进行临床评估所需的敏感性和冗余。 我们计划以三个具体目标进行研究： 1：开发DCPCF用于两光子光纤荧光探针（D-TPOFF）。 2：利用D-TPOFF在体外量化癌症特征，并使用靶向纳米颗粒在体内和体内监测肿瘤细胞中的药物作用。 3：利用D-TPOFF适应其他荧光技术来检查体内肿瘤中的事件。 在这些研究结束时，这项技术将正准备进行商业化。