This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Olsen, K. N.
Right arrow Articles by Ingmer, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Olsen, K. N.
Right arrow Articles by Ingmer, H.
Agricola
Right arrow Articles by Olsen, K. N.
Right arrow Articles by Ingmer, H.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, August 2002, p. 4145-4147, Vol. 68, No. 8
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.8.4145-4147.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Noninvasive Measurement of Bacterial Intracellular pH on a Single-Cell Level with Green Fluorescent Protein and Fluorescence Ratio Imaging Microscopy

Katja N. Olsen,1,2 Birgitte B. Budde,2 Henrik Siegumfeldt,2 K. Björn Rechinger,2,{dagger} Mogens Jakobsen,2 and Hanne Ingmer1*

Department of Veterinary Microbiology,1 Department of Dairy and Food Science, The Royal Veterinary and Agricultural University, DK-1870 Frederiksberg C, Denmark2

Received 17 January 2002/ Accepted 14 May 2002

We show that a pH-sensitive derivative of the green fluorescent protein, designated ratiometric GFP, can be used to measure intracellular pH (pHi) in both gram-positive and gram-negative bacterial cells. In cells expressing ratiometric GFP, the excitation ratio (fluorescence intensity at 410 and 430 nm) is correlated to the pHi, allowing fast and noninvasive determination of pHi that is ideally suited for direct analysis of individual bacterial cells present in complex environments.

* Corresponding author. Mailing address: Department of Veterinary Microbiology, The Royal Veterinary and Agricultural University, Stigbøjlen 4, DK-1870 Frederiksberg C, Denmark. Phone: 45 35282773. Fax: 45 35282757. E-mail: ingmer{at}biobase.dk.

{dagger} Present address: Foss Electric A/S, DK-3400 Hillerød, Denmark.

Applied and Environmental Microbiology, August 2002, p. 4145-4147, Vol. 68, No. 8
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.8.4145-4147.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Rasmussen, M. B., Oddershede, L. B., Siegumfeldt, H. (2008). Optical Tweezers Cause Physiological Damage to Escherichia coli and Listeria Bacteria. Appl. Environ. Microbiol. 74: 2441-2446 [Abstract] [Full Text]  
  • Wilks, J. C., Slonczewski, J. L. (2007). pH of the Cytoplasm and Periplasm of Escherichia coli: Rapid Measurement by Green Fluorescent Protein Fluorimetry. J. Bacteriol. 189: 5601-5607 [Abstract] [Full Text]  
  • Kim, Y. S., Cha, H. J. (2006). High-Throughput and Facile Assay of Antimicrobial Peptides Using pH-Controlled Fluorescence Resonance Energy Transfer.. Antimicrob. Agents Chemother. 50: 3330-3335 [Abstract] [Full Text]  
  • Brehm-Stecher, B. F., Johnson, E. A. (2004). Single-Cell Microbiology: Tools, Technologies, and Applications. Microbiol. Mol. Biol. Rev. 68: 538-559 [Abstract] [Full Text]  
  • Courville, P., Chaloupka, R., Veyrier, F., Cellier, M. F. M. (2004). Determination of Transmembrane Topology of the Escherichia coli Natural Resistance-associated Macrophage Protein (Nramp) Ortholog. J. Biol. Chem. 279: 3318-3326 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»