Multicolor whole-cell bacterial sensing using a synchronous fluorescence spectroscopy-based approach.
The wide collection of currently available fluorescent proteins (FPs) offers new possibilities for multicolor reporter gene-based studies of bacterial functions. However, the simultaneous use of multiple FPs is often limited by the bleed-through of their emission spectra. Here we introduce an origin...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4379052?pdf=render |
| _version_ | 1828355037325164544 |
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| author | Damien Parrello Christian Mustin David Brie Sebastian Miron Patrick Billard |
| author_facet | Damien Parrello Christian Mustin David Brie Sebastian Miron Patrick Billard |
| author_sort | Damien Parrello |
| collection | DOAJ |
| description | The wide collection of currently available fluorescent proteins (FPs) offers new possibilities for multicolor reporter gene-based studies of bacterial functions. However, the simultaneous use of multiple FPs is often limited by the bleed-through of their emission spectra. Here we introduce an original approach for detection and separation of multiple overlapping fluorescent signals from mixtures of bioreporters strains. The proposed method relies on the coupling of synchronous fluorescent spectroscopy (SFS) with blind spectral decomposition achieved by the Canonical Polyadic (CP) decomposition (also known as Candecomp/Parafac) of three-dimensional data arrays. Due to the substantial narrowing of FP emission spectra and sensitive detection of multiple FPs in a one-step scan, SFS reduced spectral overlap and improved the selectivity of the CP unmixing procedure. When tested on mixtures of labeled E. coli strains, the SFS/CP approach could easily extract the contribution of at least four overlapping FPs. Furthermore, it allowed to simultaneously monitor the expression of three iron responsive genes and pyoverdine production in P. aeruginosa. Implemented in a convenient microplate format, this multiplex fluorescent reporter method provides a useful tool to study complex processes with different variables in bacterial systems. |
| first_indexed | 2024-04-14T02:34:45Z |
| format | Article |
| id | doaj.art-ba304a638a46498e82eecd85a2941190 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-14T02:34:45Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-ba304a638a46498e82eecd85a29411902022-12-22T02:17:31ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01103e012284810.1371/journal.pone.0122848Multicolor whole-cell bacterial sensing using a synchronous fluorescence spectroscopy-based approach.Damien ParrelloChristian MustinDavid BrieSebastian MironPatrick BillardThe wide collection of currently available fluorescent proteins (FPs) offers new possibilities for multicolor reporter gene-based studies of bacterial functions. However, the simultaneous use of multiple FPs is often limited by the bleed-through of their emission spectra. Here we introduce an original approach for detection and separation of multiple overlapping fluorescent signals from mixtures of bioreporters strains. The proposed method relies on the coupling of synchronous fluorescent spectroscopy (SFS) with blind spectral decomposition achieved by the Canonical Polyadic (CP) decomposition (also known as Candecomp/Parafac) of three-dimensional data arrays. Due to the substantial narrowing of FP emission spectra and sensitive detection of multiple FPs in a one-step scan, SFS reduced spectral overlap and improved the selectivity of the CP unmixing procedure. When tested on mixtures of labeled E. coli strains, the SFS/CP approach could easily extract the contribution of at least four overlapping FPs. Furthermore, it allowed to simultaneously monitor the expression of three iron responsive genes and pyoverdine production in P. aeruginosa. Implemented in a convenient microplate format, this multiplex fluorescent reporter method provides a useful tool to study complex processes with different variables in bacterial systems.http://europepmc.org/articles/PMC4379052?pdf=render |
| spellingShingle | Damien Parrello Christian Mustin David Brie Sebastian Miron Patrick Billard Multicolor whole-cell bacterial sensing using a synchronous fluorescence spectroscopy-based approach. PLoS ONE |
| title | Multicolor whole-cell bacterial sensing using a synchronous fluorescence spectroscopy-based approach. |
| title_full | Multicolor whole-cell bacterial sensing using a synchronous fluorescence spectroscopy-based approach. |
| title_fullStr | Multicolor whole-cell bacterial sensing using a synchronous fluorescence spectroscopy-based approach. |
| title_full_unstemmed | Multicolor whole-cell bacterial sensing using a synchronous fluorescence spectroscopy-based approach. |
| title_short | Multicolor whole-cell bacterial sensing using a synchronous fluorescence spectroscopy-based approach. |
| title_sort | multicolor whole cell bacterial sensing using a synchronous fluorescence spectroscopy based approach |
| url | http://europepmc.org/articles/PMC4379052?pdf=render |
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