title: Non-linear dielectric spectroscopy of microbiological suspensions.
abstract: BACKGROUND
  Non-linear dielectric spectroscopy (NLDS) of microorganism was characterized by the generation of harmonics in the polarization current when a microorganism suspension was exposed to a sinusoidal electric field. The biological nonlinear response initially described was not well verified by other authors and the results were susceptible to ambiguous interpretation. In this paper NLDS was performed to yeast suspension in tripolar and tetrapolar configuration with a recently developed analyzer.
  
  
  METHODS
  Tripolar analysis was carried out by applying sinusoidal voltages up to 1 V at the electrode interface. Tetrapolar analysis was carried on with sinusoidal field strengths from 0.1 V cm(-1) to 70 V cm(-1). Both analyses were performed within a frequency range from 1 Hz through 100 Hz. The harmonic amplitudes were Fourier-analyzed and expressed in dB. The third harmonic, as reported previously, was investigated. Statistical analysis (ANOVA) was used to test the effect of inhibitor an activator of the plasma membrane enzyme in the measured response.
  
  
  RESULTS
  No significant non-linearities were observed in tetrapolar analysis, and no observable changes occurred when inhibitor and activator were added to the suspension. Statistical analysis confirmed these results.When a pure sinus voltage was applied to an electrode-yeast suspension interface, variations higher than 25 dB for the 3rd harmonic were observed. Variation higher than 20 dB in the 3rd harmonics has also been found when adding an inhibitor or activator of the membrane-bounded enzymes. These variations did not occur when the suspension was boiled.
  
  
  CONCLUSIONS
  The lack of result in tetrapolar cells suggest that there is no, if any, harmonic generation in microbiological bulk suspension. The non-linear response observed was originated in the electrode-electrolyte interface. The frequency and voltage windows observed in previous tetrapolar analysis were repeated in the tripolar measurements, but maximum were not observed at the same values.
  
  
  CONCLUSIONS
  Contrary to previous assertions, no repeatable dielectric non-linearity was exhibited in the bulk suspensions tested under the field and frequency condition reported with this recently designed analyzer. Indeed, interface related harmonics were observed and monitored during biochemical stimuli. The changes were coherent with the expected biological response.
authors.affiliation: Laboratorio de Medios e Interfases, Departamento de Bioingeniería, Facultad de Ciencias Exactas y Tecnología (FACET), Universidad Nacional de Tucumán (UNT), CC327, Correo Central, CP4000, San Miguel de Tucumán, Tucumán, Argentina. etreo@herrera.unt.edu.ar
authors.forename: Ernesto F
authors.initials: EF
authors.lastname: Treo
authors.forename: Carmelo J
authors.initials: CJ
authors.lastname: Felice
mesh_terms: D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D017076:Computer-Aided Design; D004553:Electric Conductivity; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017711:Nonlinear Dynamics; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D012680:Sensitivity and Specificity; D013057:Spectrum Analysis / Q000295:instrumentation* / Q000379:methods; D013535:Suspensions
chemical_list: D013535:Suspensions