EGFP and DsRed expressing cultures of Escherichia coli imaged by confocal, two‐photon and fluorescence lifetime microscopy

FEBS Letters

Hell, Stefan W.¹  Jovin, Thomas M.²  Schönle, Andreas¹  Subramaniam, Vinod²  Jakobs, Stefan¹ 

[1] Max Planck Institute for Biophysical Chemistry, High Resolution Optical Microscopy Group, 37070 Göttingen, Germany;Max Planck Institute for Biophysical Chemistry, Department of Molecular Biology, 37070 Göttingen, Germany

关键词: Green fluorescent protein;    Red fluorescent protein;    Confocal;    Two-photon;    Fluorescence lifetime microscopy;    Escherichia coli;   

DOI  :  10.1016/S0014-5793(00)01896-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown