The use of epi-fluorescence to determine the viability of Saccharomyces cerevisiae subjected to osmotic shifts

Raynal, Laurent; Barnwell, Philip; Gervais, Patrick

doi:10.1016/0168-1656(94)90048-5

Cited by 21 publications

(12 citation statements)

References 12 publications

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“…The nonpolar FDA enters the cell, where it is hydrolyzed by esterases to yield fluorescein, which is retained by the cell since it is a polar compound [5]. The degree of fluorescence depends on the physical and metabolic state of the cell and has been proven as a reliable indicator of the toxic effects of pollutants [6,7]. The application of this method to marine microalgae has been proposed to evaluate water quality [8].…”

Section: Introductionmentioning

confidence: 99%

Bioassay Technique Using Nonspecific Esterase Activities of Tetrahymena Pyriformis for Screening and Assessing Cytotoxicity of Xenobiotics

Bogaerts¹,

Sénaud²,

Bohatier³

1998

Environ Toxicol Chem

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Abstract-A simple and rapid test for screening and assessing the cytotoxicity of xenobiotics was developed with Tetrahymena pyriformis. The method estimates the activities of nonspecific esterases of a cell by concentrating within it a specific amount of fluorescence associated with fluorescein dye. The 1-h median effective concentration (EC50) values of 10 inorganic and eight organic substances are presented and compared to those of three other bioassays: the conventional T. pyriformis proliferation rate 9-h median inhibitory concentrations, the Microtox 30-min EC50s, and the Daphnia magna 4-methylumbelliferyl ␤-D galactoside 1-h EC50s. A highly significant correlation was found between the results obtained with the fluorescein diacetate test and those obtained with the growth inhibition and Microtox tests. This in vivo enzymatic test showed high sensitivity to all compounds tested except Cr 6ϩ and sodium dodecyl sulfate.

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Section: Introductionmentioning

confidence: 99%

Bioassay Technique Using Nonspecific Esterase Activities of Tetrahymena Pyriformis for Screening and Assessing Cytotoxicity of Xenobiotics

Bogaerts¹,

Sénaud²,

Bohatier³

1998

Environ Toxicol Chem

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“…Fluorescein Diacetate (FDA), a non-polar substance which crosses the membrane and is hydrolyzed by intracellular esterases in viable cells to produce fluorescein, exhibits yellow-green fluorescence when excited at 490 nm. Damaged or non-viable cells in general are unable to hydrolyze FDA or to retain fluorescein within the cell [172,173]. In combination with Ethidium Homodimer or Propidium Iodide, a similar esterase substrate, calcein acetoxy methyl ester (CAM) has been found to be reliable for viability assessment of protozoans, but not on Candida yeast, neither on bacteria such as Bacillus cereus and Escherichia coli [174].…”

Section: ) Pellet Convex Area -Core Convex Areamentioning

confidence: 99%

“…Binnerup et al [189] have proposed a protocol to determine rapidly the Gram characteristics of single cells or microcolonies: the polycarbonate membrane filters on which the bacteria have been deposited, immobilized with a thin layer of highly viscous silicone (which does not affect either cell growth or Acridine Orange subsequent staining), stained for Gram characterization and by Acridine Orange are first observed by light microscopy then by epifluoresence. The viability of Saccharomyces cerevisiae cells subjected to osmotic stresses and stained with Acridine Orange was assessed by visualization under white light, to detect all cells, followed by a visualization of the same field by epifluorescence, to detect the viable cells [172]. Multispectral imaging [190], by simultaneous or sequential capture of images using a range of excitation and emission wavelengths, has been tested by Lawrence et al [191] for the quantification of algal (by far red autofluorescence, l ex = 647 nm, l em = 680 -632 nm), bacterial (by a fluorescent nucleic acid stain SYTO 9, l ex = 488 nm, l em = 522 -532 nm) and exopolymer components (by a lectin probe, Triticium vulgaris-TRITC, l ex = 568 nm, l em = 605 -632 nm) of microbial biofilms observed by CLSM.…”

Section: ) Pellet Convex Area -Core Convex Areamentioning

confidence: 99%

Biomass Quantification by Image Analysis

Pons

Vivier

1999

Bioanalysis and Biosensors for Bioprocess Monitoring

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Microbiologists have always rely on microscopy to examine microorganisms. When microscopy, either optical or electron-based, is coupled to quantitative image analysis, the spectrum of potential applications is widened: counting, sizing, shape characterization, physiology assessment, analysis of visual texture, motility studies are now easily available for obtaining information on biomass. In this chapter the main tools used for cell visualization as well as the basic steps of image treatment are presented. General shape descriptors can be used to characterize the cell morphology, but special descriptors have been defined for filamentous microorganisms. Physiology assessment is often based on the use of fluorescent dyes. The quantitative analysis of visual texture is still limited in bioengineering but the characterization of the surface of microbial colonies may open new prospects, especially for cultures on solid substrates. In many occasions, the number of parameters extracted from images is so large that data-mining tools, such as Principal Components Analysis, are useful for summarizing the key pieces of information.

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Section: Introductionmentioning

confidence: 99%

Bioassay technique using nonspecific esterase activities of tetrahymena pyriformis for screening and assessing cytotoxicity of xenobiotics

Bogaerts

Sénaud

Bohatier

1998

Enviro Toxic and Chemistry

View full text Add to dashboard Cite

A simple and rapid test for screening and assessing the cytotoxicity of xenobiotics was developed with Tetrahymena pyriformis. The method estimates the activities of nonspecific esterases of a cell by concentrating within it a specific amount of fluorescence associated with fluorescein dye. The 1‐h median effective concentration (EC50) values of 10 inorganic and eight organic substances are presented and compared to those of three other bioassays: the conventional T. pyriformis proliferation rate 9‐h median inhibitory concentrations, the Microtox® 30‐min EC50s, and the Daphnia magna 4‐methylumbelliferyl β‐D galactoside 1‐h EC50s. A highly significant correlation was found between the results obtained with the fluorescein diacetate test and those obtained with the growth inhibition and Microtox tests. This in vivo enzymatic test showed high sensitivity to all compounds tested except Cr6+ and sodium dodecyl sulfate.

show abstract

The use of epi-fluorescence to determine the viability of Saccharomyces cerevisiae subjected to osmotic shifts

Cited by 21 publications

References 12 publications

Bioassay Technique Using Nonspecific Esterase Activities of Tetrahymena Pyriformis for Screening and Assessing Cytotoxicity of Xenobiotics

Bioassay Technique Using Nonspecific Esterase Activities of Tetrahymena Pyriformis for Screening and Assessing Cytotoxicity of Xenobiotics

Biomass Quantification by Image Analysis

Bioassay technique using nonspecific esterase activities of tetrahymena pyriformis for screening and assessing cytotoxicity of xenobiotics

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