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

Jakobs, S., Subramaniam, V., Schönle, A., Jovin, T. M., & Hell, S. W. (2000). EGFP and DsRed expressing cultures of Escherichia coli imaged by confocal, two-photon and fluorescence lifetime microscopy. FEBS Letters, 479, 131-135.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-F893-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-9BD7-4
Genre: Journal Article

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600257.pdf (Publisher version), 312KB
 
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 Creators:
Jakobs, S.1, Author              
Subramaniam, V.2, Author              
Schönle, A.1, Author              
Jovin, T. M.2, Author              
Hell, S. W.1, Author              
Affiliations:
1Research Group of High Resolution Optical Microscopy, MPI for Biophysical Chemistry, Max Planck Society, ou_578626              
2Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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Free keywords: 2-photon excitation; Imaging microscopy; Protein; Gfp; Resolution; Mutants; Cells; green fluorescent protein; red fluorescent protein; confocal; Two-photon; fluorescence lifetime microscopy; Escherichia coli; E. coli
 Abstract: 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.

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Language(s): eng - English
 Dates: 2000
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: eDoc: 226673
Other: 23125
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Title: FEBS Letters
Source Genre: Journal
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Pages: - Volume / Issue: 479 Sequence Number: - Start / End Page: 131 - 135 Identifier: -