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）：使用荧光探针来识别癌症特征和癌细胞的生物活性的方法具有很大的前景。 基于荧光共振能量转移 (FRET) 和荧光相关光谱 (FCS) 等技术以及其他类似技术的探针能够识别特定的 RNA 或蛋白质分子，从而识别癌症并提供有关肿瘤细胞使用的致癌途径的信息。 其他探针可以通过测量化疗和放疗诱导的细胞凋亡来深入了解药物反应。 然而，荧光分析有一些局限性。 大多数离体分析使用流式细胞仪或复杂的共聚焦显微镜来进行分析，这需要从体内取出组织并通常破碎成细胞，然后以静态方式固定和分析。 体内荧光分析的问题甚至更大，因为即使在近红外范围内，背景荧光和组织散射也会限制皮肤的信号采集。 双光子激发是光学领域的一项重要进步，促进了体外 FRET、FCS 和 CARS 技术。 然而，这些应用受到采用这些技术所需的复杂技术（共焦显微镜）的限制。 我们已经展示了通过光纤进行双光子荧光分析的用途，用于在 SCID 小鼠中分析体外癌细胞和体内人类肿瘤。 这项先前的工作相当于 R21 提案，因为我们实现了我们的主要目标：开发传感系统光学和电子学，并记录该系统获取和分析体外和体内荧光信号的能力。 该 R33 应用的主要目标是开发一种基于新型双包层光子晶体光纤 (DCPCF) 的更灵敏的原型设备，我们假设该设备将提供使用多种荧光对体内荧光信号进行临床评估所需的灵敏度和冗余度。技术。 我们计划围绕三个具体目标开展研究： 1：开发用于双光子光纤荧光探针（D-TPOFF）的DCPCF。 图 2：利用 D-TPOFF 量化体外癌症特征，并使用靶向纳米粒子离体和体内监测肿瘤细胞中的药物作用。 图 3：利用 D-TPOFF 调整其他荧光技术来检查体内肿瘤事件。 这些研究结束后，这项技术将达到商业化的阶段。