Quantifying the Dynamics of Bacterial Secondary Metabolites by Spectral Multiphoton Microscopy
Abstract
Phenazines, a group of fluorescent small molecules produced by the bacterium Pseudomonas aeruginosa, play a role in maintaining cellular redox homeostasis. Phenazines have been challenging to study in vivo due to their redox activity, presence both intra- and extracellularly, and their diverse chemical properties. Here, we describe a noninvasive in vivo optical technique to monitor phenazine concentrations within bacterial cells using time-lapsed spectral multiphoton fluorescence microscopy. This technique enables simultaneous monitoring of multiple weakly fluorescent molecules (phenazines, siderophores, NAD(P)H) expressed by bacteria in culture. This work provides the first in vivo measurements of reduced phenazine concentration as well as the first description of the temporal dynamics of the phenazine-NAD(P)H redox system in Pseudomonas aeruginosa, illuminating an unanticipated role for 1-hydroxyphenazine. Similar approaches could be used to study the abundance and redox dynamics of a wide range of small molecules within bacteria, both as single cells and in communities.
Additional Information
© 2011 American Chemical Society. Received: March 21, 2011. Accepted: June 14, 2011. Published: June 14, 2011. Published In Issue September 16, 2011. We thank L. Dietrich for the gift of the ΔphzM strain, and A. Price-Whelan and L. Dietrich for helpful advice and discussions. D.K.N. is a Howard Hughes Medical Institute Investigator, and this work was supported by the HHMI. D.S.T. and P.T.C.S. were supported by NIH, NSF, SMA2, and SMART. Author Contributions: These authors contributed equally to this work.Attached Files
Accepted Version - nihms-312850.pdf
Supplemental Material - cb200094w_si_003.pdf
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Additional details
- PMCID
- PMC3212935
- Eprint ID
- 26601
- Resolver ID
- CaltechAUTHORS:20111005-113650941
- NIH
- NSF
- SMA2
- SMART
- Created
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2011-10-05Created from EPrint's datestamp field
- Updated
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2021-11-09Created from EPrint's last_modified field
- Caltech groups
- Division of Geological and Planetary Sciences