Spectroscopic approach for monitoring two-photon excited fluorescence resonance energy transfer from homodimers at the subcellular level
Spectroscopic approach for monitoring two-photon excited fluorescence resonance energy transfer from homodimers at the subcellular level
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DOI：
10.1117/1.1584052
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发表时间：
2003-07-01
影响因子：
3.5
通讯作者：
Tromberg, BJ
中科院分区：
文献类型：
--
作者：
LaMorte, VJ;Zoumi, A;Tromberg, BJ
我们采用了光谱方法监测荧光共振能量转移（FRET）在活细胞。该方法使用双光子激发荧光成像和光谱学（TPIS-FRET）在亚细胞成像体积内提供了绿色荧光蛋白（GFP）突变体信号的优异光谱分离。与目前基于FRET的方法相比，TPIS-FRET不依赖于滤光片的选择、比率图像分析或渗漏校正算法。利用TPIS-FRET的内在光学切片能力，我们已经确定了离散的亚细胞结构域内的蛋白质-蛋白质相互作用。为了说明这种技术的适用性，以检测同型二聚体的形成，我们证明了在体内协会的早幼粒细胞（PML）同型二聚体内相应的核体。(C)2003年，由光学仪器工程师学会（Society of Photo-Optical Instrumentation Engineers）主办。
We have employed a spectroscopic approach for monitoring fluorescence resonance energy transfer (FRET) in living cells. This method provides excellent spectral separation of green fluorescent protein (GFP) mutant signals within a subcellular imaging volume using two-photon excited fluorescence imaging and spectroscopy (TPIS-FRET). In contrast to current FRET-based methodologies, TPIS-FRET does not rely on the selection of optical filters, ratiometric image analysis, or bleedthrough correction algorithms. Utilizing the intrinsic optical sectioning capabilities of TPIS-FRET we have identified, protein-protein interactions within discrete subcellular domains. To illustrate the applicability of this technique to the detection of homodimer formation, we demonstrated the in vivo association of promyleocyte (PML) homodimers within their corresponding nuclear body. (C) 2003 Society of Photo-Optical Instrumentation Engineers.