Use of fluorescent probes to assess physiological functions of bacteriaat single-cell level

Joux, Fabien; Lebaron, Philippe

doi:10.1016/s1286-4579(00)01307-1

Cited by 333 publications

(301 citation statements)

References 74 publications

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“…None of these PI positive bacteria were able to recover and grow on appropriate media after cell sorting, which agrees with others [22,43,44]. PI was therefore considered testimony of dead cells.…”

Section: Discussionsupporting

confidence: 84%

“…After sorting and plating on agar plates, 82% of these cells were able to proliferate, thereby confirming their viability. The discrepancy between SYTO 9 positive (intact) and culturable bacteria may relate to the presence of VBNC individuals, correctly identified as viable by FCM analysis [22]. Also, the fact that some bacteria did not resist the stress applied during cell sorting cannot be ruled out.…”

Section: Discussionmentioning

confidence: 99%

“…Although this technique is still considered as the gold-standard for bacterial viability assessment, it fails to identify subpopulations of viable but not cultivable (VBNC) bacteria exposed to antimicrobial agents [21,22]. Unlike culture plating, flow cytometry (FCM) in combination with fluorescent labeling of cells can distinguish different physiological states of single-cell microorganisms [23,24].…”

Section: Introductionmentioning

confidence: 99%

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Rose bengal uptake by E. faecalis and F. nucleatum and light-mediated antibacterial activity measured by flow cytometry

Manoil

Filieri

Schrenzel

et al. 2016

Journal of Photochemistry and Photobiology B: Biology

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Rose bengal uptake by E. faecalis and F. nucleatum and light-mediated antibacterial activity measured by flow cytometry Antibacterial photodynamic therapy (aPDT) using rose bengal (RB) and blue-light kills bacteria through the production of reactive oxygen derivates. However, the interaction mechanism of RB with bacterial cells remains unclear. This study investigated the uptake efficiency and the antibacterial activity of blue light-activated RB against Enterococcus faecalis and Fusobacterium nucleatum. Spectrophotometry and epifluorescence microscopy were used to evaluate binding of RB to bacteria. The antibacterial activity of RB after various irradiation times was assessed by flow cytometry in combination with cell sorting. Uptake of RB increased in a concentration dependent manner in both strains although E. faecalis displayed higher uptake values. RB appeared to bind specific sites located at the cellular poles of E. faecalis and at regular intervals along F. nucleatum. Blue-light irradiation of samples incubated with RB significantly reduced bacterial viability. After incubation with 10 μM RB and 240 s irradiation, only 0.01% (± 0.01%) of E. faecalis cells and 0.03% (±0.03%) of F. nucleatum survived after treatment. This study indicated that RB can bind to E. faecalis and F. nucleatum in a sufficient amount to elicit effective aPDT. Epifluorescence microscopy showed a yet-unreported property of RB binding to bacterial membranes. Flow cytometry allowed the detection of bacteria with damaged membranes that were unable to form colonies on agars after cell sorting.

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Section: Discussionsupporting

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Rose bengal uptake by E. faecalis and F. nucleatum and light-mediated antibacterial activity measured by flow cytometry

Manoil

Filieri

Schrenzel

et al. 2016

Journal of Photochemistry and Photobiology B: Biology

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“…Polybrominated biphenyls (PBB) and polybrominated diphenyl ethers (PBDE) have similar applications: in flame-retardant PCBs, connectors, and plastic housings. Through a literature review, possible substitutes for these materials have been identified to meet the same functional demands [21][22][23][24][25][26][27][28][29][30] (see Table 1). In relative design practice, designers can select substitutes according to their functions and costs.…”

Section: Materials Selectionmentioning

confidence: 99%

Design Strategies for Developing Sustainable Consumer Electronics

You¹,

Wang²,

Ma³

et al. 2016

JRST

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consumer electronics produce a huge amount of waste and harmful residuals every year. This study investigated how to avoid or reduce environmental pollution through the design of consumer electronic products. Through a literature review, analyses, discussions, and case studies, we defined design strategies for developing sustainable consumer electronics. First, material selection should be considered from the perspectives of users, usages, and use environment in accordance with environmental protection requirements. The application of recyclable and remanufacture-able materials is the inexorable trend. Second, electronic products should be designed with simply disassembled structures that employ as many recyclable functional components as possible. Third, the information on the packaging or labels of consumer electronic products should be designed with graphics that are as easy to understand as possible and within an appropriate amount. Moreover, ingenious design solutions that can motivate users positively and actively to alter their environmental protection behaviors are expected.

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“…The reliability and significance of CTC staining have been evaluated in comparison to other viability assays with single-cell resolution, such as direct viability counting 47) and fluorescence microscopy using 5-carboxyfluorescein diacetate (CFDA) 22) , 5-(4,6-dichlorotriazinyl) aminofluoroscein (DTAF) 6) , ChemChrome B 32) and a LIVE/DEAD Baclight TM Bacterial Viability kit 8,31) . Recently, the CTC staining method was also compared to microautoradiography and fluorescence in situ hybridization (FISH) to detect viable bacteria in activated sludge 27) .…”

mentioning

confidence: 99%

An Improved Redox Dye-Staining Method Using 5-Cyano-2,3-Ditoryl Tetrazolium Chloride for Detection of Metabolically Active Bacteria in Activated Sludge

Yoshida

Hiraishi

2004

Microb. Environ.

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A fluorochrome-staining method using 5-cyano-2,3-ditoryl tetrazolium chloride (CTC) was improved to enhance the sensitivity of detection of metabolically active bacteria in activated sludge. The highest efficiency of CTC staining was obtained with 6 mM CTC in MOPS buffer at pH 6.5. CTC staining efficiency was also increased by adding a substrate mixture (0.05% peptone, 0.05% yeast extract and 1 mM glucose), Meldola's Blue as an electron transfer mediator and KCN. The bacterial counts detected by the CTC staining method thus improved accounted for 41-76% of the total counts in activated sludge with the average value being 54%. This value was approximately two-fold higher than those obtained by the traditional CTC staining assay which has been performed in phosphate buffer or phosphate-buffered saline without any addition of external substrate or electron transfer mediator. There was a high positive correlation between CTC-stained direct counts and the plate counts of aerobic chemoorganotrophic bacteria or viable cell counts measured using a LIVE/DEAD BacLight Bacterial Viability kit. The viable population in the activated sludges as measured by BacLight staining and plate counting accounted for 73±7.6% and 8.7±4.7% of the total population, respectively. These data suggested that most of the "viable but nonculturable" bacteria (i.e., BacLight-positive but unculturable population) potentially existing in the sludge system were so metabolically active as to be detectable by the improved CTC staining method.

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Use of fluorescent probes to assess physiological functions of bacteriaat single-cell level

Cited by 333 publications

References 74 publications

Rose bengal uptake by E. faecalis and F. nucleatum and light-mediated antibacterial activity measured by flow cytometry

Rose bengal uptake by E. faecalis and F. nucleatum and light-mediated antibacterial activity measured by flow cytometry

Design Strategies for Developing Sustainable Consumer Electronics

An Improved Redox Dye-Staining Method Using 5-Cyano-2,3-Ditoryl Tetrazolium Chloride for Detection of Metabolically Active Bacteria in Activated Sludge

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