Fluorescence lifetime-based optical molecular imaging.

Fluorescence lifetime-based optical molecular imaging.
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基于荧光寿命的光学分子成像。

DOI:
10.1007/978-1-60761-901-7_12
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发表时间:
2011
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Kumar,AnandTN
Kumar,AnandTN
中科院分区:
--
文献类型:
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作者:
Kumar,AnandTN

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荧光寿命是体内分子成像的一种强有力的对比机制。在本章中,我们描述了使用时间域荧光断层成像系统来优化利用寿命对比度的仪器和方法。该系统的主要特点是使用超短激光脉冲在自由空间使用点激励,并使用门控、增强型CCD相机进行非接触检测。成像体积的表面边界是使用与成像系统集成的摄影测量相机获取的,并在生物组织中光传播的理论模型中实现。使用基于寿命的层析方法来优化分析时间域数据,该方法基于从时间域数据的长时间衰减部分提取寿命和衰减幅度的层析集合。这种方法提高了定位体内目标的能力,其分辨率比传统的光学方法更好。以体模和荷瘤小鼠乳腺癌模型为例,说明了时间域寿命复用法和层析成像技术的应用。在后一种应用中,时间域方法允许在存在背景自体荧光的情况下改进对来自完整裸鼠的荧光蛋白信号的检测。这一特征在药物治疗反应的纵向临床前评估以及解决与肿瘤生理学和转移相关的基本问题方面具有潜在的应用价值。
Fluorescence lifetime is a powerful contrast mechanism for in vivo molecular imaging. In this chapter, we describe instrumentation and methods to optimally exploit lifetime contrast using a time domain fluorescence tomography system. The key features of the system are the use of point excitation in free-space using ultrashort laser pulses and non-contact detection using a gated, intensified CCD camera. The surface boundaries of the imaging volume are acquired using a photogrammetric camera integrated with the imaging system, and implemented in theoretical models of light propagation in biological tissue. The time domain data are optimally analyzed using a lifetime-based tomography approach, which is based on extracting a tomographic set of lifetimes and decay amplitudes from the long time decay portion of the time domain data. This approach improves the ability to locate in vivo targets with a resolution better than conventional optical methods. The application of time domain lifetime multiplexing and tomography are illustrated using phantoms and tumor bearing mouse model of breast adenocarcinoma. In the latter application, the time domain approach allows an improved detection of fluorescent protein signals from intact nude mice in the presence of background autofluorescence. This feature has potential applications for longitudinal pre-clinical evaluation of drug treatment response as well as to address fundamental questions related to tumor physiology and metastasis.
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