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活性的方法具有很大的前景。 基于荧光共振能量转移（FRET）的探针和诸如荧光相关光谱（FCS）和其他类似技术的技术提供了鉴定特定RNA或蛋白质分子的能力，所述特定RNA或蛋白质分子可以鉴定癌症并提供关于肿瘤细胞所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了解药物反应。 然而，荧光分析有几个局限性。 大多数离体分析使用流式细胞仪或复杂的共聚焦显微镜进行分析，这需要将组织从体内取出并通常破碎成细胞，然后固定并以静态方式分析。 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限制了对皮肤的信号采集。 双光子激发是光学领域的一个重要进展，促进了FRET，FCS和汽车技术在体外的应用。 然而，这些应用受到采用这些技术所必需的复杂技术（共焦显微镜）的限制。 我们已经证明了使用双光子荧光分析，通过光纤分析癌细胞在体外和人体肿瘤在体内的SCID小鼠。 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器件，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，我们假设该新型双包层光子晶体光纤将提供使用几种荧光技术进行体内荧光信号临床评价所需的灵敏度和冗余度。 我们计划在三个具体目标下开展研究： 1：开发用于双光子光纤荧光探针（D-TPOFF）的DCPCF。 第二章：利用D-TPOFF在体外定量癌症特征，并使用靶向纳米颗粒在体外和体内监测肿瘤细胞中的药物作用。 3：利用D-TPOFF来调整其他荧光技术以检查体内肿瘤中的事件。 在这些研究结束时，这项技术将处于商业化的准备阶段。