Ultrasound-enabled two-photon FRET microscopy

超声双光子 FRET 显微镜

基本信息

批准号：
8249835
负责人：
Jerome Mertz
金额：
$ 8.18万
依托单位：
BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-01 至 2014-03-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Fluorescence resonance energy transfer (FRET) can serve as a nanoscale molecular ruler. When used in imaging applications, it is a highly sensitive reporter of donor-acceptor molecular configuration. In most cases, FRET is utilized with standard one-photon excitation. Extensions of FRET to two-photon excitation are generally hampered by the problem of donor and acceptor bleed through, which imposes the requirement of technically complicated spectral unmixing algorithms or fluorescence lifetime measurements. We propose to considerably simplify the detection of FRET with two-photon excitation. Our goal is to establish the feasibility of modulating FRET signal using ultrasonic waves. Since FRET is sensitive to distance between donor-acceptor pairs at the nanoscale, mechanical compression and tension produced by sound waves are likely to modulate the FRET signal, serving as a useful signature to distinguish FRET from donor and acceptor bleed-through, which is a particular problem in multiphoton FRET. In particular, we propose to conduct preliminary experiments to establish the premise that ultrasound modulates FRET. We will investigate two types of samples: genetically encoded FRET probes in tissue (or tissue-like environments), and labeled microbubbles in solution. In the former case, we will investigate the application of our technique to in-vivo imaging in C.Elegans. FRET is a powerful tool for functional imaging. We believe that our hybrid ultrasound-optical technique will significantly improve the isolation of FRET signals and therefore be of benefit to any FRET-based functional imaging application. Moreover, when coupled with two-photon excitation, as proposed here, our technique should be well adapted to thick tissue imaging, thereby benefiting the in-vivo imaging community. PUBLIC HEALTH RELEVANCE: Fluorescence resonance energy transfer (FRET) is a powerful research technique to image cellular function in tissue. We propose to improve FRET signal generation with the use of ultrasound. This will benefit any FRET-based imaging applications, and will be specifically adapted to in-vivo microscopy in thick tissue.

描述（申请人提供）：荧光共振能量转移（FRET）可以充当纳米级分子尺。当用于成像应用时，它是供体-受体分子构型的高度灵敏的报告基因。在大多数情况下，FRET 与标准单光子激发一起使用。 FRET 扩展到双光子激发通常受到供体和受体渗漏问题的阻碍，这提出了技术上复杂的光谱解混合算法或荧光寿命测量的要求。我们建议通过双光子激发大大简化 FRET 的检测。我们的目标是建立使用超声波调制 FRET 信号的可行性。由于 FRET 对纳米级供体-受体对之间的距离敏感，声波产生的机械压缩和张力可能会调制 FRET 信号，作为区分 FRET 与供体和受体渗漏的有用特征，这是一种有用的特征。多光子 FRET 中的特殊问题。特别是，我们建议进行初步实验来建立超声调制 FRET 的前提。我们将研究两种类型的样品：组织（或类组织环境）中的基因编码 FRET 探针，以及溶液中的标记微泡。在前一种情况下，我们将研究我们的技术在线虫体内成像中的应用。 FRET 是功能成像的强大工具。我们相信，我们的混合超声光学技术将显着改善 FRET 信号的隔离，因此有利于任何基于 FRET 的功能成像应用。此外，当与双光子激发相结合时，正如这里所提出的，我们的技术应该很好地适应厚组织成像，从而使体内成像界受益。公共健康相关性：荧光共振能量转移 (FRET) 是一种强大的研究技术，可对组织中的细胞功能进行成像。我们建议使用超声波来改善 FRET 信号的生成。这将有利于任何基于 FRET 的成像应用，并将特别适用于厚组织的体内显微镜检查。