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首页> 外文会议>Conference on Multiphoton Microscopy in the Biomedical Sciences IV; 20040125-20040127; San Jose,CA; US >Long-Wavelength Long-Lifetime Luminophores for Cellular and Tissue Imaging
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Long-Wavelength Long-Lifetime Luminophores for Cellular and Tissue Imaging

机译：用于细胞和组织成像的长波长长寿命发光体

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摘要

Fluorescence detection is the dominant technology for cellular imaging, clinical diagnostics, DNA analysis and drug discovery. In fluorescence microscopy, the fluorophores are subject to photobliching and detectability is limited by cellular autofluorescence. We describe new approach to develop fluorescent probes that display long emission wavelength, long decay times, and high quantum yield and high fluorescence brightness. These luminophores are covalently linked pairs of long-lifetime fluorophores (like metal-ligand complexes) and a short-fluorescence-lifetime and high quantum yield dyes. Using resonance energy transfer (RET) it is possible of obtaining desirable spectral properties and long fluorescence lifetime in covalently linked pairs. The long-lifetime donor results in a long-lived fluorescence component in the acceptor decay. Importantly the emission spectrum of the luminophore is that of the acceptor and quantum yield of the luminophore approaches that of the higher quantum yield acceptor. Such luminophores are suitable for fluorescence measurements in biological samples with the use of real time background suppression to eliminate autofluorescence. We discuss experimental examples based on long lived metal-ligand-complexes and long wavelength acceptors like Texas Red. The emission maxima (spectra) and decay time of such RET tandems can be readily adjusted by selection of the donor, acceptor and distance between them. Such luminophores with long-wavelength emission and adjustable long lifetime can have numerous applications in one-photon and multi-photon cellular and tissue imaging with the use of off-gating the excitation pulse and sample autofluorescence.
机译：荧光检测是细胞成像，临床诊断，DNA分析和药物发现的主要技术。在荧光显微镜下，荧光团易受光致变色，并且可检测性受到细胞自发荧光的限制。我们描述了开发显示长发射波长，长衰变时间，高量子产率和高荧光亮度的荧光探针的新方法。这些荧光团是长寿命荧光团（如金属-配体配合物）和短荧光寿命和高量子产率染料的共价连接对。使用共振能量转移（RET），可以在共价连接的对中获得所需的光谱特性和较长的荧光寿命。寿命长的供体在受体衰变中导致了寿命长的荧光成分。重要的是，发光体的发射光谱是受体的发射光谱，并且发光体的量子产率接近较高量子产率的受体的发射光谱。此类荧光团适合使用实时背景抑制来消除自发荧光的生物样品中的荧光测量。我们讨论了基于长寿命金属配体配合物和长波长受体（例如Texas Red）的实验示例。通过选择供体，受体和它们之间的距离，可以很容易地调节此类RET串联的发射最大值（光谱）和衰减时间。这种具有长波长发射和可调节的长寿命的发光体通过使用非门控激发脉冲和样品自发荧光，可以在单光子和多光子细胞和组织成像中有许多应用。

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