Rapid estimation of bacterial antibiotic susceptibility with flow cytometry

Mason, David; Allman, Richard; Stark, J. M.; Lloyd, David

doi:10.1111/j.1365-2818.1994.tb03494.x

Cited by 92 publications

(79 citation statements)

References 16 publications

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“…Recently, new viability assays were developed for bacteria (Keer and Birch 2003). These tests were essentially based on respiratory activity (Bhupathiraju et al 1999;Rodriguez et al 1992), external membrane integrity (Nebe-von-Caron et al 2000), intracellular pH (Chitara et al 2000), and membrane potential (Deere et al 1995;Manson et al 1993Manson et al , 1994Manson et al , 1995. The major advantage of these new technologies is the rapidity of response and the multi-parametric nature of the collected information.…”

Section: Introductionmentioning

confidence: 99%

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Flow cytometry analysis of germinating Bacillus spores, using membrane potential dye

Laflamme

Veillette

et al. 2004

Arch Microbiol

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Section: Introductionmentioning

confidence: 99%

“…The measurement of membrane potential has been used as an indicator of viability in numerous studies to test the susceptibility to antimicrobial agents (Kaprelyants and Kell 1992;Manson et al 1994Manson et al , 1995Shapiro 1995). Cells with high membrane potential are often referred as energized or hyperpolarized cells.…”

Section: Introductionmentioning

confidence: 99%

Flow cytometry analysis of germinating Bacillus spores, using membrane potential dye

Laflamme

Veillette

et al. 2004

Arch Microbiol

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“…Oxonol is only taken up by A. castellanii cells with depolarized plasma membranes, i.e. nonviable cells [20][21][22]. The results are presented as dual-parameter histograms, with forward angle light scatter (an indicator of cell size) against oxonol fluorescence : the results in Figure 2 show clear differences in cell-associated fluorescence between dead ( Figure 2A) and live (control) (Figure 2B) cells.…”

Section: Oxygen Levels Correlate With Increased Unsaturationmentioning

confidence: 98%

“…The viability of cultures was measured by flow cytometry [20]. Samples (1 ml) were taken from cultures, oxonol was added at a concentration of 2 µg\ml and incubations were carried out for 5 min.…”

Section: Viability Of Culturesmentioning

confidence: 99%

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Oxygen induction of a novel fatty acid n−6 desaturase in the soil protozoon, Acanthamoeba castellanii

Rutter

Thomas

Herbert

et al. 2002

Biochemical Journal

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Induction of fatty acid desaturation is very important for the temperature adaptation of poikilotherms. However, in oxygen-limited late-exponential-phase Acanthamoebacastellanii cultures, oxygen alone was able to induce increased activity of a fatty acid desaturase that converts oleate into linoleate and which has been implicated in the temperature adaptation of this organism. Experiments with Δ10-nonadecenoate showed that the enzyme is an n-6 desaturase rather than a Δ12-desaturase. It also used preferentially 1-acyl-2-oleoyl-phosphatidylcholine as substrate and NAD(P)H as electron donor. The involvement of cytochrome b5 as an intermediate electron carrier was shown by difference spectra measurements and anti-(cytochrome b5) antibody experiments. Of the three protein components of the desaturase complex, oxygen only increased the activity of the terminal (cyanide-sensitive) protein during n-6 desaturase induction. The induction of this terminal protein paralleled well the increase in overall oleate n-6 desaturation. The ability of oxygen to induce oleate desaturase independently of temperature in this lower eukaryotic animal model is of novel intrinsic interest, as well as being important for the design of future experiments to determine the molecular mechanism of temperature adaptation in poikilotherms.

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Rapid assessment of ceftazidime, ciprofloxacin, and gentamicin susceptibility in exponentially-growingE. coli cells by means of flow cytometry

Walberg

Gaustad²,

Steen

1997

Cytometry

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Exponentially growing E. coli cells were cultivated in the presence of ceftazidime, ciprofloxacin, and gentamicin in concentrations ranging from 0.5–8 minimal inhibitory concentration (MIC), permeabilized by means of cold shock in EDTA/Na‐azide, and stained with the DNA‐specific dye combination of ethidium bromide and mithramycin before the fluorescence, light scattering, and cell number were measured flow‐cytometrically. In order to evaluate the applicability of the cold‐shock procedure, cells were also permeabilized by 70% ethanol. Permeabilization by cold shock, which eliminates washing of the cells, reduced the preparation time to <5 min. A statistically significant increase in light scattering and fluorescence, i.e., cell size and DNA content, could be detected already after 30 min of ceftazidime and ciprofloxacin exposure, even at sub‐MIC concentrations. The results obtained with these drugs with cold‐shock permeabilization were similar to those seen with ethanol fixation. For gentamicin‐treated cells, however, a majority of the cells lost their fluorescence after cold shock. In gentamicin‐treated cells fixed in ethanol, there was no consistent effect on either light scattering or fluorescence; however, we observed a substantial fragmentation and leakage of DNA in such cells. The cell proliferation was completely inhibited within 30 min of gentamicin incubation. For all three drugs, loss of light scattering and DNA were associated with cellular disintegration, i.e., reduced viability. The present results demonstrate that effects of ceftazidime, ciprofloxacin, and gentamicin on E. coli can be detected by flow cytometry within 1 h from the beginning of drug exposure to the completed measurement. Cytometry 27:169–178, 1997. © 1997 Wiley‐Liss, Inc.

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Rapid estimation of bacterial antibiotic susceptibility with flow cytometry

Cited by 92 publications

References 16 publications

Flow cytometry analysis of germinating Bacillus spores, using membrane potential dye

Flow cytometry analysis of germinating Bacillus spores, using membrane potential dye

Oxygen induction of a novel fatty acid n−6 desaturase in the soil protozoon, Acanthamoeba castellanii

Rapid assessment of ceftazidime, ciprofloxacin, and gentamicin susceptibility in exponentially-growingE. coli cells by means of flow cytometry

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