EGFP and DsRed expressing cultures of Escherichia coli imaged by confocal, two-photon and fluorescence lifetime microscopy
EGFP and DsRed expressing cultures of Escherichia coli imaged by confocal, two-photon and fluorescence lifetime microscopy
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DOI：
10.1016/s0014-5793(00)01896-2
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发表时间：
2000-08-18
期刊：
影响因子：
3.5
通讯作者：
Hell, SW
中科院分区：
文献类型：
--
作者：
Jakobs, S;Subramaniam, V;Hell, SW
绿色荧光蛋白（GFP）已成为监测体内蛋白质定位和基因表达的宝贵标志物。最近，从热带珊瑚中分离出一种新的红色荧光蛋白（drFP583或DsRed） [Matz， M.V.等，（1999）Nature biotechnology, 17, 969-973]。EGFP和DsRed的最大发射峰分别为509 nm和583 nm，在同时激发下适合于几乎无交叉的双色标记。我们用单光子共聚焦和双光子显微镜对表达EGFP或DsRed的混合大肠杆菌群体进行了成像，两种激发模式都证明适合于表达任何一种荧光蛋白的细胞成像。DsRed成熟时间较长，表达该荧光蛋白的大肠杆菌明显小于表达EGFP的大肠杆菌。在老化的细菌培养物中，DsRed在细胞内聚集，荧光寿命成像显微镜检测到其荧光寿命明显缩短，(C) 2000年欧洲生化学会联合会。Elsevier Science B.V.版权所有。
The green fluorescent protein (GFP) has become an invaluable marker for monitoring protein localization and gene expression in vivo. Recently a new red fluorescent protein (drFP583 or DsRed), isolated from tropical corals, has been described [Matz, M.V. et al, (1999) Nature Biotech, 17, 969-973]. With emission maxima at 509 and 583 nm respectively, EGFP and DsRed are suited for almost crossover free dual color labeling upon simultaneous excitation, We imaged mixed populations of Escherichia coli expressing either EGFP or DsRed by one-photon confocal and by two-photon microscopy, Both excitation modes proved to be suitable for imaging cells expressing either of the fluorescent proteins. DsRed had an extended maturation time and E, coli expressing this fluorescent protein were significantly smaller than those expressing EGFP, In aging bacterial cultures DsRed appeared to aggregate within the cells, accompanied by a strong reduction in its fluorescence lifetime as determined by fluorescence lifetime imaging microscopy, (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.