Viability states of bacteria—Specific mechanisms of selected probes

Single cell techniques like flow cytometry combined with viability staining can help to obtain information on viability states of bacteria. Many fluorescent dyes are available for this purpose and can be chosen according to the available excitation source, the species used, and the background of scientific questions and relevant specifications. Within this short overview, we focus on two diverse groups of bacteria: the gram− Escherichia coli and representatives of the gram+ Mycobacterium to demonstrate differences and similarities in dye uptake principles, processing and binding. We call for attention to possible diverse responses of different species to various viability assays. The cell surface structure of bacteria and the chemical properties of fluorescent probes considerably determine the success of a certain staining practice. Particular focus was drawn on analysis of membrane integrity, uptake of substrates and transformation of fluorogenic substrates. © 2010 International Society for Advancement of Cytometry

[1]  S. Müller,et al.  Functional single-cell analyses: flow cytometry and cell sorting of microbial populations and communities. , 2010, FEMS microbiology reviews.

[2]  Thomas Bley,et al.  Origin and analysis of microbial population heterogeneity in bioprocesses. , 2010, Current opinion in biotechnology.

[3]  N. Dimitrijević,et al.  Interfacial charge transfer between CdTe quantum dots and gram negative vs gram positive bacteria. , 2010, Environmental science & technology.

[4]  I. Matic,et al.  Reliable Detection of Dead Microbial Cells by Using Fluorescent Hydrazides , 2009, Applied and Environmental Microbiology.

[5]  J. Collins,et al.  Hydroxyurea induces hydroxyl radical-mediated cell death in Escherichia coli. , 2009, Molecular cell.

[6]  Daniel Ahmed,et al.  Acoustic tweezers: patterning cells and microparticles using standing surface acoustic waves (SSAW). , 2009, Lab on a chip.

[7]  T. Hübschmann,et al.  NBDT (3‐(N‐(7‐nitrobenz‐2‐oxa‐1,3‐diazol‐4‐yl)amino)‐3‐toluene)—A novel fluorescent dye for studying mechanisms of toluene uptake into vital bacteria , 2009, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[8]  Y. Urano,et al.  A simple and effective strategy to increase the sensitivity of fluorescence probes in living cells. , 2009, Journal of the American Chemical Society.

[9]  S. Yoshida,et al.  Discrimination of live, anti-tuberculosis agent-injured, and dead Mycobacterium tuberculosis using flow cytometry. , 2009, FEMS microbiology letters.

[10]  J. Pagés,et al.  Mechanisms of drug efflux and strategies to combat them: challenging the efflux pump of Gram-negative bacteria. , 2009, Biochimica et biophysica acta.

[11]  A. Mulyukin,et al.  Dormant forms of Mycobacterium smegmatis with distinct morphology. , 2009, Microbiology.

[12]  S. Nuding,et al.  Antibacterial activity of human defensins on anaerobic intestinal bacterial species: a major role of HBD-3. , 2009, Microbes and infection.

[13]  Kinam Park,et al.  Detection of pathogenic E. coli O157:H7 by a hybrid microfluidic SPR and molecular imaging cytometry device , 2009, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[14]  Bo Kara,et al.  Studies related to antibody fragment (Fab) production in Escherichia coli W3110 fed‐batch fermentation processes using multiparameter flow cytometry , 2009, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[15]  M. Voget,et al.  Synergistic effects of Miconazole and Polymyxin B on microbial pathogens , 2008, Veterinary Research Communications.

[16]  J. Collins,et al.  Mistranslation of Membrane Proteins and Two-Component System Activation Trigger Antibiotic-Mediated Cell Death , 2008, Cell.

[17]  Pedro J J Alvarez,et al.  Fullerene water suspension (nC60) exerts antibacterial effects via ROS-independent protein oxidation. , 2008, Environmental science & technology.

[18]  R. Jerala,et al.  Alexidine and chlorhexidine bind to lipopolysaccharide and lipoteichoic acid and prevent cell activation by antibiotics. , 2008, The Journal of antimicrobial chemotherapy.

[19]  M. Bouix,et al.  Dynamic analysis of Lactobacillus delbrueckii subsp. bulgaricus CFL1 physiological characteristics during fermentation , 2008, Applied Microbiology and Biotechnology.

[20]  Thomas Dick,et al.  The protonmotive force is required for maintaining ATP homeostasis and viability of hypoxic, nonreplicating Mycobacterium tuberculosis , 2008, Proceedings of the National Academy of Sciences.

[21]  T. Egli,et al.  Rapid, cultivation-independent assessment of microbial viability in drinking water. , 2008, Water research.

[22]  S. Jayaraman A novel method for the detection of viable human pancreatic beta cells by flow cytometry using fluorophores that selectively detect labile zinc, mitochondrial membrane potential and protein thiols , 2008, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[23]  A. Chorny,et al.  Ghrelin Protects against Experimental Sepsis by Inhibiting High-Mobility Group Box 1 Release and by Killing Bacteria12 , 2008, The Journal of Immunology.

[24]  A. Driessen,et al.  Protein translocation across the bacterial cytoplasmic membrane. , 2008, Annual review of biochemistry.

[25]  Andrew Leis,et al.  Disclosure of the mycobacterial outer membrane: Cryo-electron tomography and vitreous sections reveal the lipid bilayer structure , 2008, Proceedings of the National Academy of Sciences.

[26]  M. Niederweis,et al.  Nutrient acquisition by mycobacteria. , 2008, Microbiology.

[27]  H. Moser,et al.  Physicochemical properties of antibacterial compounds: implications for drug discovery. , 2008, Journal of medicinal chemistry.

[28]  Y. Park,et al.  Antibacterial Activity and Mechanism of Action of the Silver Ion in Staphylococcus aureus and Escherichia coli , 2008, Applied and Environmental Microbiology.

[29]  Y. Shai,et al.  Analysis of in vitro activities and modes of action of synthetic antimicrobial peptides derived from an alpha-helical 'sequence template'. , 2008, The Journal of antimicrobial chemotherapy.

[30]  C. Bleck,et al.  On the killing of mycobacteria by macrophages , 2007, Cellular microbiology.

[31]  J. Trevors,et al.  Fluorescence polarization in studies of bacterial cytoplasmic membrane fluidity under environmental stress. , 2007, Progress in biophysics and molecular biology.

[32]  S. Günther,et al.  Fluorogenic surrogate substrates for toluene-degrading bacteria--are they useful for activity analysis? , 2007, Journal of microbiological methods.

[33]  S. Günther,et al.  Limits of propidium iodide as a cell viability indicator for environmental bacteria , 2007, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[34]  Bernard Faller,et al.  Physicochemical Parameters as Tools in Drug Discovery and Lead Optimization , 2007 .

[35]  R. Cox,et al.  Growth regulation in the mycobacterial cell. , 2007, Current molecular medicine.

[36]  Alvin W Nienow,et al.  A comparison of high cell density fed‐batch fermentations involving both induced and non‐induced recombinant Escherichia coli under well‐mixed small‐scale and simulated poorly mixed large‐scale conditions , 2007, Biotechnology and bioengineering.

[37]  Natalie Leys,et al.  Physiological changes induced in bacteria following pH stress as a model for space research , 2007 .

[38]  D. Kletsas,et al.  Acid Tolerance of Streptococcus macedonicus as Assessed by Flow Cytometry and Single-Cell Sorting , 2006, Applied and Environmental Microbiology.

[39]  A. Van Deun,et al.  Early and rapid microscopy-based diagnosis of true treatment failure and MDR-TB. , 2006, The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease.

[40]  D. Kletsas,et al.  Rapid assessment of the physiological status of Streptococcus macedonicus by flow cytometry and fluorescence probes. , 2006, International journal of food microbiology.

[41]  G. Robinson,et al.  Monitoring changes in nisin susceptibility of Listeria monocytogenes Scott A as an indicator of growth phase using FACS. , 2006, Journal of microbiological methods.

[42]  H. Kress,et al.  Dynamic life and death interactions between Mycobacterium smegmatis and J774 macrophages , 2006, Cellular microbiology.

[43]  Thomas Egli,et al.  Flow-cytometric study of vital cellular functions in Escherichia coli during solar disinfection (SODIS). , 2006, Microbiology.

[44]  S. Nuding,et al.  A flow cytometric assay to monitor antimicrobial activity of defensins and cationic tissue extracts. , 2006, Journal of microbiological methods.

[45]  H. Harms,et al.  Analysis of living S. cerevisiae cell states—A three color approach , 2006, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[46]  Richard W. Horobin,et al.  Fluorescent cationic probes for nuclei of living cells: why are they selective? A quantitative structure–activity relations analysis , 2006, Histochemistry and Cell Biology.

[47]  S. Na,et al.  The survival response of Escherichia coli K12 in a natural environment , 2006, Applied Microbiology and Biotechnology.

[48]  M. G. Pinho,et al.  Bacterial Cell Wall Synthesis: New Insights from Localization Studies , 2005, Microbiology and Molecular Biology Reviews.

[49]  H. Engelhardt,et al.  The growth rate of Mycobacterium smegmatis depends on sufficient porin‐mediated influx of nutrients , 2005, Molecular microbiology.

[50]  S. Condón,et al.  Membrane Damage and Microbial Inactivation by Chlorine in the Absence and Presence of a Chlorine-Demanding Substrate , 2005, Applied and Environmental Microbiology.

[51]  S. Krämer,et al.  Permeation of aromatic carboxylic acids across lipid bilayers: the pH-partition hypothesis revisited. , 2005, Biophysical journal.

[52]  D. Knorr,et al.  Cellular injuries upon exposure of Escherichia coli and Lactobacillus rhamnosus to high‐intensity ultrasound , 2005, Journal of applied microbiology.

[53]  R. Lahiri,et al.  Application of a viability-staining method for Mycobacterium leprae derived from the athymic (nu/nu) mouse foot pad. , 2005, Journal of medical microbiology.

[54]  C. A. Kent,et al.  Monitoring population dynamics of the thermophilic Bacillus licheniformis CCMI 1034 in batch and continuous cultures using multi-parameter flow cytometry. , 2005, Journal of biotechnology.

[55]  R. Dapson Dye–tissue interactions: mechanisms, quantification and bonding parameters for dyes used in biological staining , 2005, Biotechnic & histochemistry : official publication of the Biological Stain Commission.

[56]  J. Rhee,et al.  Use of Pseudomonas putida EstA as an Anchoring Motif for Display of a Periplasmic Enzyme on the Surface of Escherichia coli , 2004, Applied and Environmental Microbiology.

[57]  P. Black,et al.  Bacterial Long Chain Fatty Acid Transport: Gateway to a Fatty Acid-responsive Signaling System* , 2004, Journal of Biological Chemistry.

[58]  N. Jiao,et al.  Membrane potential based characterization by flow cytometry of physiological states in an aerobic anoxygenic phototrophic bacterium , 2004 .

[59]  H. Nikaido,et al.  Efflux Pump-Mediated Intrinsic Drug Resistance in Mycobacterium smegmatis , 2004, Antimicrobial Agents and Chemotherapy.

[60]  M. Young,et al.  Formation of 'non-culturable' cells of Mycobacterium smegmatis in stationary phase in response to growth under suboptimal conditions and their Rpf-mediated resuscitation. , 2004, Microbiology.

[61]  H. Engelhardt,et al.  The MspA porin promotes growth and increases antibiotic susceptibility of both Mycobacterium bovis BCG and Mycobacterium tuberculosis. , 2004, Microbiology.

[62]  Hajime Unno,et al.  Application of glutaraldehyde for the staining of esterase-active cells with carboxyfluorescein diacetate , 2004, Biotechnology Letters.

[63]  Alvin W Nienow,et al.  Further studies related to the scale‐up of high cell density escherichia coli fed‐batch fermentations: , 2003, Biotechnology and bioengineering.

[64]  Alex Avdeef,et al.  In vitro trans-monolayer permeability calculations: often forgotten assumptions. , 2003, Drug discovery today.

[65]  S. Zivanovic,et al.  Influence of Inorganic Salts and Bases on the J-Band in the Absorption Spectra of Water Solutions of 1,1‘-Diethyl-2,2‘-cyanine Iodide , 2003 .

[66]  C. Saint,et al.  A comparative study of carboxyfluorescein diacetate and carboxyfluorescein diacetate succinimidyl ester as indicators of bacterial activity. , 2003, Journal of microbiological methods.

[67]  W. Lubitz,et al.  Online Monitoring of Escherichia coli Ghost Production , 2003, Applied and Environmental Microbiology.

[68]  W. D. de Vos,et al.  Multiparametric Flow Cytometry and Cell Sorting for the Assessment of Viable, Injured, and Dead Bifidobacterium Cells during Bile Salt Stress , 2002, Applied and Environmental Microbiology.

[69]  K. Oh,et al.  Effect of 2-NBDG, a Fluorescent Derivative of Glucose, on Microbial Cell Growth , 2002 .

[70]  D. Kell,et al.  Formation and resuscitation of "non-culturable" cells of Rhodococcus rhodochrous and Mycobacterium tuberculosis in prolonged stationary phase. , 2002, Microbiology.

[71]  Y. St-Pierre,et al.  Rapid antimicrobial susceptibility testing of urinary tract isolates and samples by flow cytometry. , 2002, Journal of medical microbiology.

[72]  A. Neyfakh,et al.  Efflux-mediated drug resistance in Gram-positive bacteria. , 2001, Current opinion in microbiology.

[73]  R. Benz,et al.  Study of the Properties of a Channel-forming Protein of the Cell Wall of the Gram-positive Bacterium Mycobacterium phlei , 2001, The Journal of Membrane Biology.

[74]  C. Hewitt,et al.  An industrial application of multiparameter flow cytometry: assessment of cell physiological state and its application to the study of microbial fermentations. , 2001, Cytometry.

[75]  E. Duque,et al.  Three Efflux Pumps Are Required To Provide Efficient Tolerance to Toluene in Pseudomonas putidaDOT-T1E , 2001, Journal of bacteriology.

[76]  E. Pauwels,et al.  Human Lactoferrin and Peptides Derived from Its N Terminus Are Highly Effective against Infections with Antibiotic-Resistant Bacteria , 2001, Infection and Immunity.

[77]  C. Hewitt,et al.  Studies related to the scale-up of high-cell-density E. coli fed-batch fermentations using multiparameter flow cytometry: effect of a changing microenvironment with respect to glucose and dissolved oxygen concentration. , 2000, Biotechnology and bioengineering.

[78]  F. Srienc,et al.  Glucose uptake rates of single E. coli cells grown in glucose-limited chemostat cultures. , 2000, Journal of microbiological methods.

[79]  M. Labra,et al.  Two and three-color fluorescence flow cytometric analysis of immunoidentified viable bacteria. , 2000, Cytometry.

[80]  C. Böttcher,et al.  Supramolecular Structures of J-Aggregates of Carbocyanine Dyes in Solution , 2000 .

[81]  D. J. Mason,et al.  Flow Cytometric Investigation of Filamentation, Membrane Patency, and Membrane Potential in Escherichia coli following Ciprofloxacin Exposure , 2000, Antimicrobial Agents and Chemotherapy.

[82]  S. Müller,et al.  Flow cytometric techniques to characterise physiological states of Acinetobacter calcoaceticus. , 2000, Journal of microbiological methods.

[83]  N. Yamaguchi,et al.  Viability of Escherichia coli O157:H7 in natural river water determined by the use of flow cytometry , 2000, Journal of applied microbiology.

[84]  J. Warmington,et al.  The mode of antimicrobial action of the essential oil of Melaleuca alternifolia (tea tree oil) , 2000, Journal of applied microbiology.

[85]  S. Müller,et al.  Adaptive responses of Ralstonia eutropha to feast and famine conditions analysed by flow cytometry. , 1999, Journal of biotechnology.

[86]  F. Srienc,et al.  Dynamics of glucose uptake by single Escherichia coli cells. , 1999, Metabolic engineering.

[87]  R. Gross,et al.  Phospholipid-subclass-specific partitioning of lipophilic ions in membrane-water systems. , 1999, The Biochemical journal.

[88]  D. Gibson,et al.  Distinguishing between living and nonliving bacteria: Evaluation of the vital stain propidium iodide and its combined use with molecular probes in aquatic samples , 1998 .

[89]  I. Paulsen,et al.  Evolutionary origins of multidrug and drug-specific efflux pumps in bacteria. , 1998, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[90]  H. Sahl,et al.  The Lantibiotic Mersacidin Inhibits Peptidoglycan Synthesis by Targeting Lipid II , 1998, Antimicrobial Agents and Chemotherapy.

[91]  J. Vives-Rego,et al.  Assessment of the effects of gramicidin, formaldehyde, and surfactants on Escherichia coli by flow cytometry using nucleic acid and membrane potential dyes. , 1997, Cytometry.

[92]  H. Yoneyama,et al.  Use of Fluorescence Probes to Monitor Function of the Subunit Proteins of the MexA-MexB-OprM Drug Extrusion Machinery inPseudomonas aeruginosa * , 1997, The Journal of Biological Chemistry.

[93]  Cindy Lee,et al.  Hydrolysis of peptides in seawater and sediment , 1997 .

[94]  H. Abé,et al.  Intracellular fate of 2-NBDG, a fluorescent probe for glucose uptake activity, in Escherichia coli cells. , 1996, Bioscience, biotechnology, and biochemistry.

[95]  J. Liu,et al.  Mycolic Acid Structure Determines the Fluidity of the Mycobacterial Cell Wall* , 1996, The Journal of Biological Chemistry.

[96]  R. W. Horobin,et al.  Phloem mobility of fluorescent xenobiotics in Arabidopsis in relation to their physicochemical properties , 1996 .

[97]  G L Amidon,et al.  Transport approaches to the biopharmaceutical design of oral drug delivery systems: prediction of intestinal absorption. , 1996, Advanced drug delivery reviews.

[98]  S. Müller,et al.  Membrane-potential-related fluorescence intensity indicates bacterial injury , 1996 .

[99]  K. Sigler,et al.  Slow fluorescent indicators of membrane potential: a survey of different approaches to probe response analysis. , 1996, Journal of photochemistry and photobiology. B, Biology.

[100]  P. Monfort,et al.  Cell cycle characteristics and changes in membrane potential during growth of Escherichia coli as determined by a cyanine fluorescent dye and flow cytometry , 1996 .

[101]  K. Oh,et al.  A novel fluorescent derivative of glucose applicable to the assessment of glucose uptake activity of Escherichia coli. , 1996, Biochimica et biophysica acta.

[102]  C. Edwards,et al.  Rapid assessment of physiological status in Escherichia coli using fluorescent probes. , 1995, The Journal of applied bacteriology.

[103]  T. Tsuji,et al.  A new fluorescence staining assay for visualizing living microorganisms in soil , 1995, Applied and environmental microbiology.

[104]  D. Veal,et al.  Rapid screening of the antimicrobial activity of extracts and natural products. , 1994, The Journal of antibiotics.

[105]  C. Edwards,et al.  Rapid assessment of bacterial viability by flow cytometry , 1992, Applied Microbiology and Biotechnology.

[106]  M. Vaara,et al.  Agents that increase the permeability of the outer membrane. , 1992, Microbiological reviews.

[107]  R. Hancock,et al.  Interaction of aminoglycosides with the outer membranes and purified lipopolysaccharide and OmpF porin of Escherichia coli , 1991, Antimicrobial Agents and Chemotherapy.

[108]  T. Smith,et al.  J-aggregate formation of a carbocyanine as a quantitative fluorescent indicator of membrane potential. , 1991, Biochemistry.

[109]  G. Steele,et al.  Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate-forming lipophilic cation JC-1. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[110]  H. Labischinski,et al.  Direct proof of a "more-than-single-layered" peptidoglycan architecture of Escherichia coli W7: a neutron small-angle scattering study , 1991, Journal of bacteriology.

[111]  H. Tapiero,et al.  Relevance of the chemical charge of rhodamine dyes to multiple drug resistance. , 1989, Biochemical pharmacology.

[112]  B. Erni Glucose transport in Escherichia coli. , 1989, FEMS microbiology reviews.

[113]  H. Nikaido,et al.  Molecular basis of bacterial outer membrane permeability. , 1985, Microbiological reviews.

[114]  P. Garland,et al.  Synthesis of cytoplasmic membrane during growth and division of Escherichia coli. Dispersive behaviour of respiratory nitrate reductase. , 1979, The Biochemical journal.

[115]  E. Racker,et al.  Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ. , 1979, Biochemistry.

[116]  A. Waggoner Optical probes of membrane potential , 1976, The Journal of Membrane Biology.

[117]  V. Braun,et al.  Covalent lipoprotein from the outer membrane of Escherichia coli. , 1975, Biochimica et biophysica acta.

[118]  C H Wang,et al.  Studies on the mechanism by which cyanine dyes measure membrane potential in red blood cells and phosphatidylcholine vesicles. , 1974, Biochemistry.

[119]  L. Leive THE BARRIER FUNCTION OF THE GRAM‐NEGATIVE ENVELOPE , 1974, Annals of the New York Academy of Sciences.

[120]  A. Leo,et al.  Partition coefficients and their uses , 1971 .

[121]  R. Hughes,et al.  Cross-linking of bacterial cell walls with glutaraldehyde. , 1970, The Biochemical journal.

[122]  M. Whitehouse,et al.  7-chloro-4-nitrobenzo-2-oxa-1,3-diazole: a new fluorigenic reagent for amino acids and other amines. , 1968, The Biochemical journal.

[123]  E. Jelley Molecular, Nematic and Crystal States of I : I′-Diethyl-Ψ-Cyanine Chloride , 1937, Nature.

[124]  D. Radzioch,et al.  Role of TLR2- and TLR4-mediated signaling in Mycobacterium tuberculosis-induced macrophage death. , 2010, Cellular immunology.

[125]  H. Shapiro Flow cytometry of bacterial membrane potential and permeability. , 2008, Methods in molecular medicine.

[126]  K. Miyanaga,et al.  A Recombinant Bacteriophage‐Based Assay for the Discriminative Detection of Culturable and Viable but Nonculturable Escherichia coli O157:H7 , 2006, Biotechnology progress.

[127]  Andrew B. Mahon,et al.  Optimisation of the fluorescein diacetate antibacterial assay. , 2005, Journal of microbiological methods.

[128]  Christopher J Hewitt,et al.  The application of multi-parameter flow cytometry to monitor individual microbial cell physiological state. , 2004, Advances in biochemical engineering/biotechnology.

[129]  H. Sahm,et al.  L-glutamate efflux with Corynebacterium glutamicum: why is penicillin treatment or Tween addition doing the same? , 2001, Journal of molecular microbiology and biotechnology.

[130]  H. Shapiro,et al.  Accurate flow cytometric membrane potential measurement in bacteria using diethyloxacarbocyanine and a ratiometric technique. , 1999, Cytometry.

[131]  J. van Heijenoort Assembly of the monomer unit of bacterial peptidoglycan , 1998, Cellular and molecular life sciences : CMLS.

[132]  G. Mie Beiträge zur Optik trüber Medien, speziell kolloidaler Metallösungen , 1908 .