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Use of Ratiometric Probes with a Spectrofluorometer for Bacterial Viability Measurement

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초록

Assessment of microorganism viability is useful in many industrial fields. A large number of methods associated with the use of fluorescent probes have been developed, including fluorimetry, fluorescence microscopy, and cytometry. In this study, a microvolume spectrofluorometer was used to measure the membrane potential variations of Escherichia coli. In order to estimate the sensitivity of the device, the membrane potential of E. coli was artificially disrupted using an ionophore agent: carbonyl cyanide 3-chlorophenylhydrazone. The membrane potential was evaluated using two ratiometric methods: a Rhodamine 123/ 4’,6-diamidino-2-phenylindole combination and a JC-10 ratiometric probe. These methods were used to study the impact of freezing on E. coli, and were compared with the conventional enumeration method. The results showed that it was beneficial to use this compact, easy-touse, and inexpensive spectrofluorometer to assess the viability of bacterial cells via their membrane potential.

Abstract

Assessment of microorganism viability is useful in many industrial fields. A large number of methods associated with the use of fluorescent probes have been developed, including fluorimetry, fluorescence microscopy, and cytometry. In this study, a microvolume spectrofluorometer was used to measure the membrane potential variations of Escherichia coli. In order to estimate the sensitivity of the device, the membrane potential of E. coli was artificially disrupted using an ionophore agent: carbonyl cyanide 3-chlorophenylhydrazone. The membrane potential was evaluated using two ratiometric methods: a Rhodamine 123/ 4’,6-diamidino-2-phenylindole combination and a JC-10 ratiometric probe. These methods were used to study the impact of freezing on E. coli, and were compared with the conventional enumeration method. The results showed that it was beneficial to use this compact, easy-touse, and inexpensive spectrofluorometer to assess the viability of bacterial cells via their membrane potential.

참고문헌 (40)

  1. Joux F, "Use of fluorescent probes to assess physiological functions of bacteria at single-cell level", Microbes Infect, 2, pp.1523~1535 (2000)
  2. Xu L, "Mechanisms of ultraviolet disinfection and chlorination of Escherichia coli:culturability, membrane permeability, metabolism, and genetic damage", J. Environ. Sci. (China), 65, pp.356~366 (2017)
  3. Strahl H, "Membrane potential is important for bacterial cell division", Proc. Natl. Acad. Sci. USA, 107, pp.12281~12286 (2010)
  4. McFeters GA, "Physiological assessment of bacteria using fluorochromes", J. Microbiol. Methods, 21, pp.1~13 (1995)
  5. Breeuwer P, "Assessment of viability of microorganisms employing fluorescence techniques", Int. J. Food Microbiol, 55, pp.193~200 (2000)
  6. Sträuber H, "Viability states of bacteria-specific mechanisms of selected probes", Cytometry A, 77, pp.623~634 (2010)
  7. Kaprelyants A, "Rapid assessment of bacterial viability and vitality by rhodamine 123 and flow cytometry", J. Appl. Microbiol, 72, pp.410~422 (1992)
  8. Diaper J, "Rapid assessment of bacterial viability by flow cytometry", Appl. Microbiol. Biotechnol, 38, pp.268~272 (1992)
  9. Novo D J, "Multiparameter flow cytometric analysis of antibiotic effects on membrane potential, membrane permeability, and bacterial counts of Staphylococcus aureus and Micrococcus luteus", Antimicrob. Agents Chemother, 44, pp.827~834 (2000)
  10. Zhao J-Y, "Labeling efficiency and toxicity evaluation of CdSe", ZnS quantum dots on Escherichia coli, 2014, pp.16, 2424~
  11. Chung J, "Fast and continuous microorganism detection using aptamer-conjugated fluorescent nanoparticles on an optofluidic platform", Biosens. Bioelectron, 67, pp.303~308 (2015)
  12. Emaus RK, "Rhodamine 123as a probe of transmembrane potential in isolated rat-liver mitochondria: spectral and metabolic properties", Biochim. Biophys. Acta, 850, pp.436~448 (1986)
  13. Porter J, "Membrane hyperpolarisation by valinomycin and its limitations for bacterial viability assessment using rhodamine 123 and flow cytometry", FEMS Microbiol. Lett, 132, pp.259~262 (1995)
  14. Williams S, "Distinguishing between living and nonliving bacteria: evaluation of the vital stain propidium iodide and its combined use with molecular probes in aquatic samples", J. Microbiol. Methods, 32, pp.225~236 (1998)
  15. Boulos L, "LIVE", DEAD BacLight: application of a new rapid staining method for direct enumeration of viable and total bacteria in drinking water, 1999, pp.37, 77~86
  16. Nescerecka A, "A pipeline for developing and testing staining protocols for flow cytometry, demonstrated with SYBR Green I and propidium iodide viability staining", J. Microbiol. Methods, 131, pp.172~180 (2016)
  17. Bunthof CJ, "Fluorescent method for monitoring cheese starter permeabilization and lysis", Appl. Environ. Microbiol, 67, pp.4264~4271 (2001)
  18. Kumar A, "Engineered ZnO and TiO2 nanoparticles induce oxidative stress and DNA damage leading to reduced viability of Escherichia coli", Free Radic. Biol. Med, 51, pp.1872~1881 (2011)
  19. Kralj JM, "Electrical spiking in Escherichia coli probed with a fluorescent voltage-indicating protein", Science, 333, pp.345~348 (2011)
  20. Matsuyama T., "Staining of living bacteria with rhodamine 123", FEMS Microbiol. Lett, 21, pp.153~157 (1984)
  21. Monfort P, "Cell cycle characteristics and changes in membrane potential during growth of Escherichia coli as determined by a cyanine fluorescent dye and flow cytometry", J. Microbiol. Methods, 25, pp.79~86 (1996)
  22. Comas J, "Assessment of the effects of gramicidin, formaldehyde, and surfactants on Escherichia coli by flow cytometry using nucleic acid and membrane potential dyes", Cytometry, 29, pp.58~64 (1997)
  23. Jiao N, "Membrane potential based characterization by flow cytometry of physiological states in an aerobic anoxygenic phototrophic bacterium", Aquat. Microb. Ecol, 37, pp.149~158 (2004)
  24. Rezaeinejad S, "Heterogeneity of Escherichia coli population by respiratory activity and membrane potential of cells during growth and long-term starvation", Microbiol. Res, 166, pp.129~135 (2011)
  25. Gao W, "Freezing inactivation of Escherichia coli and Enterococcus faecalis in water: response of different strains", Water Environ. Res, 81, pp.824~830 (2009)
  26. Sherr B, "Enumeration of total and highly active bacteria", Methods Microbiol, 30, pp.129~159 (2001)
  27. Muthukrishnan T, "Evaluating the reliability of counting bacteria using epifluorescence microscopy", J. Mar. Sci. Eng, 5, pp.4~ (2017)
  28. Silva F, "Bacteriostatic versus bactericidal activity of ciprofloxacin in Escherichia coli assessed by flow cytometry using a novel farred dye", J. Antibiot. (Tokyo), 64, pp.321~325 (2011)
  29. Xu C, "Antibacterial activity and a membrane damage mechanism of Lachnum YM30melanin against Vibrio parahaemolyticus and Staphylococcus aureus", Food Control, 73, pp.1445~1451 (2017)
  30. Ma J, "Self-defense of Escherichia coli against damages caused by nanoalumina", Environ. Toxicol. Pharmacol, 55, pp.110~117 (2017)
  31. Biggerstaff J, "New methodology for viability testing in environmental samples", Mol. Cell. Probes, 20, pp.141~146 (2006)
  32. Breeuwer P, "Molecular Microbial Ecology Manual", pp.1563~1580 (2004), Springer
  33. Sekyere JO, "Carbonyl cyanide mchlorophenylhydrazine (CCCP) reverses resistance to colistin, but not to carbapenems and tigecycline in multidrugresistant Enterobacteriaceae", Front. Microbiol, 8, pp.228~ (2017)
  34. Chimerel C, "Indole prevents Escherichia coli cell division by modulating membrane potential", Biochim. Biophys. Acta, 1818, pp.1590~1594 (2012)
  35. Kennedy D, "Responses of Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus to simulated food processing treatments, determined using fluorescence-activated cell sorting and plate counting", Appl. Environ. Microbiol, 77, pp.4657~4668 (2011)
  36. Guo N, "The preservative potential of Amomum tsaoko essential oil against E. coli, its antibacterial property and mode of action", Food Control, 75, pp.236~245 (2017)
  37. Novo D, "Accurate flow cytometric membrane potential measurement in bacteria using diethyloxacarbocyanine and a ratiometric technique", Cytometry A, 35, pp.55~63 (1999)
  38. Berney M, "Assessment and interpretation of bacterial viability by using the LIVE", DEAD BacLight Kit in combination with flow cytometry, 2007, pp.73, 3283~3290
  39. Miyanaga K, "Optimization of distinction between viable and dead cells by fluorescent staining method and its application to bacterial consortia", Biochem. Eng. J, 37, pp.56~61 (2007)
  40. Lopez-Amoros R, "Flow cytometric assessment of Escherichia coli and Salmonella typhimurium starvation-survival in seawater using rhodamine 123, propidium iodide, and oxonol", Appl. Environ. Microbiol, 61, pp.2521~2526 (1995)

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