WO2019014692A1 - Procédé de mesure par spectroscopie électrique de l'impédance d'un fluide contenant des cellules vivantes - Google Patents

Procédé de mesure par spectroscopie électrique de l'impédance d'un fluide contenant des cellules vivantes Download PDF

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Publication number
WO2019014692A1
WO2019014692A1 PCT/AT2018/050017 AT2018050017W WO2019014692A1 WO 2019014692 A1 WO2019014692 A1 WO 2019014692A1 AT 2018050017 W AT2018050017 W AT 2018050017W WO 2019014692 A1 WO2019014692 A1 WO 2019014692A1
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Prior art keywords
electrodes
double
measurement
measuring method
cell
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PCT/AT2018/050017
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German (de)
English (en)
Inventor
Georg Brunauer
Christoph SLOUKA
Christoph Herwig
Alfred Gruber
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Novapecc Gmbh
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Publication of WO2019014692A1 publication Critical patent/WO2019014692A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
    • G01N27/026—Dielectric impedance spectroscopy
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
    • G01N27/22—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating capacitance
    • G01N27/221—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating capacitance by investigating the dielectric properties
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • G01N33/48707—Physical analysis of biological material of liquid biological material by electrical means
    • G01N33/48735—Investigating suspensions of cells, e.g. measuring microbe concentration
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
    • G01N27/22—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating capacitance
    • G01N27/226—Construction of measuring vessels; Electrodes therefor

Definitions

  • the invention relates to a measuring method for electrical spectroscopic impedance measurement of a fluid containing living cells, wherein at least two electrodes are arranged in the fluid, and a plurality of measuring signals in the form of alternating voltages are applied in succession between the electrodes and the current and the voltage are measured as measured values between the electrodes in which case the frequencies of the alternating voltages are at least partially different.
  • Such methods are particularly used for cell number determination of biotechnological samples application.
  • Such measuring methods for determining the cell count of any microbiological culture by means of an alternating voltage signal by driving a defined frequency range are already known.
  • the fluid is usually the medium in which the cells live. This may be a nutrient medium in the laboratory or in the industrial sector, for example, a malt extract with a wort to provide the minerals for beer production.
  • live cell determination is performed outside the process using fluorescence-labeled cells using flow cytometry or confocal microscopy as described in Microbiological Reviews, 1996, 60 (4), 641-696 and Journal of Immunological Methods , 2000, 243 (1), 191-210.
  • impedance spectroscopy is known from the literature as a method of investigating heterogeneous mixtures, so-called "dispersion.”
  • an alternating current / voltage signal at high frequencies (AC) is generally used, since large deflections of the electric field (amplitudes) occur , is also referred to here as a so-called ⁇ -dispersion.
  • AC alternating current / voltage signal at high frequencies
  • VCC living cell concentration
  • US Pat. No. 6,596,507 B2 describes a biomass sensor that can be used for such a measurement.
  • To determine the living cell density English: Viable Cell Density
  • probes based on the permittivity physical unit: As / Vm
  • Such a measuring system is of the Applicant known under the trade name "Incyte" sensor sold. In this case, the entire volume in which the living cells are located is alternately polarized and depolarized. The electric field is measured.
  • WO2017149005A1 describes a method for calibrating impedance-spectroscopic biomass sensors for detecting information about the amount or / and size of living cells in a biomass by means of an electric field.
  • the measurement systems described above are not mature or impractical, in particular the measurement accuracy and the reliability required for the operation is low, this relates in particular to the influence of the living cell volume by the extremely high electric fields.
  • the object of the invention is therefore to provide a method of the type described with which the life cell concentration can be determined more accurately.
  • the spectroscopic impedance measurement comprises a frequency range of 0.1 Hz to 10 6 Hz.
  • the values are surprisingly much better correlated with the cell concentration. This can be explained, at least in part, by the fact that the effect of the ⁇ dispersion occurs in the region of particularly low frequencies.
  • the measured values are preferably distributed over the entire frequency range, particularly preferably substantially uniformly. It can also be provided that particularly interesting or characteristic frequency ranges with more measured values are examined in more detail.
  • the method according to the invention differs from the prior art in that the living cell determination takes place very close to the electrodes, and Very small polarization voltages are used in order not to influence the microbiological cell culture to be determined via the voltage measurement signal.
  • the entire measuring volume around the electrodes is polarized there with a voltage signal, which results in the fact that partly large electric fields act on the microbiological culture to be determined.
  • the method has a higher measurement accuracy compared to the known systems.
  • metallic electrodes are provided in / or outside a housing, which dive into a medium of a microbiological fluid to be examined, such as a cell culture.
  • the measured bilayer capacity can be correlated as a function of cell culture vitality or living cell concentration and cell number.
  • the measured signal increases with cell vitality.
  • the measurement signal changes very much with the temperature.
  • living cell determination is understood to mean the determination of the vitality of a microbiological cell culture, the cell count of the living cells in a defined volume and thus the cell concentration.
  • the temperature must be kept constant.
  • the measurement usually takes place at a temperature at which a microbiological cell culture can be usefully fermented or cultured.
  • the biomass increase increases with increasing temperature. In the present case, a temperature of 28 ° C was chosen.
  • “Online” and “inline” measuring system is understood in the context of this disclosure, the embodiments of the measuring system. If a measuring system or a probe is operated in the bypass of a main vessel for storage or processing of the fluid such as a fermenter, it is called an online probe, while the electrodes are preferably encapsulated and sealed in a housing.
  • the electrodes can be arranged for measurement as online electrodes in a bypass channel of a main vessel of the fluid.
  • the fluid passed through the bypass channel can either be returned to the main vessel, discarded or otherwise used.
  • Bypass is a flow path away from the main flow path or the main flow path. understood understood over the small amounts of fluid, for example, for investigation.
  • the measuring probe is located directly in a fermenter, this is an inline measuring system, with the electrodes protruding into the environment to be analyzed and flowing around it.
  • the electrodes can be arranged for measurement as in-line electrodes in a main vessel of the fluid.
  • macroscopic structure is understood to mean the embodiment of the electrodes which have a larger areal structure.
  • microstructure is understood to mean the embodiment of the electrodes which have a microstructured structure.
  • the metallic electrodes are made of a high-alloy, stainless stainless steel. These electrodes are extremely suitable for measuring the "double-layer capacitance" of different microbiological cultures In a further embodiment of the invention, it is provided that these stainless steel electrodes have a coating of precious metals, preferably of gold or platinum.
  • a further embodiment provides that the electrodes have a microscopic structure consisting of very small punctiform stainless steel electrodes, particularly preferably finger electrodes, preferably with or without gold or platinum coating. This construction is extremely suitable for living cell determination of very low cell densities.
  • double-layer capacitance is understood to mean an electrochemical capacity which occurs as soon as a cell is polarized from a microbiological culture.
  • polarizing or “polarization” is understood to mean the application of a voltage signal between the electrodes.
  • the measurement of the double-layer capacitance and the polarization of the electrodes is preferably carried out by means of a spectroscopic impedance measuring device.
  • a spectroscopic impedance measuring device there for example, the complex resistance values consisting of a real part and an imaginary part are recorded in a frequency range of 10 6 to 10 1 Hertz (Hz) and an amplitude of preferably 100 to 250 mV.
  • the double-layer capacitance is determined from the measured current and voltage values. From this, finally, the number of living cells are derived.
  • the method can be used in different cell cultures for the determination of a wide variety of cell types, but especially in the food industry such as the brewery for Saccharomyces cerevisiae or in the laboratory field for Escherichia coli or tumor cells.
  • the inventive method of the type mentioned above for living cell determination is suitable for all cell cultures.
  • the method is performed with a suitable measuring device having a high-power impedance device.
  • a suitable measuring device having a high-power impedance device.
  • measuring electronics consisting of a frequency generator including software are used. According to the invention, it is provided that this measuring electronics takes over the evaluation of the measurement signal with the algorithm of the underlying biochemical-physical model.
  • a process-integrated measuring system which is inexpensive to produce, and with very small polarization voltages determines the cell count of a cell culture, and has improved measurement accuracy and reliability.
  • a first preferred embodiment of the invention provides that the subject measuring method for living cell determination of Saccharomyces cerevisiae, in short S. cerevisiae, is used in aerobic and anaerobic processes.
  • a further preferred embodiment of the invention provides that the subject measuring method for living cell determination and for the diagnostic characterization of tumor cells is used.
  • the above-described measuring system for living cell determination with a macroscopic or microscopic structure as electrodes is designed as an online or inline measuring system and has a high accuracy of measurement in all microbiological cultures with very low polarization voltages. In a further development, this measuring system is combined with additional analysis and evaluation electronics.
  • the sensor part hereby has an online or inline housing, particularly preferably a high-alloy stainless steel, preferably coated with gold or platinum. These housings are in particular able to keep the electrodes at a defined distance.
  • the electrodes are electrically insulated, gas- and liquid-tight introduced into an online housing.
  • the housing in this case particularly preferably consists of a jacket for temperature control. This probe has proven to be particularly suitable due to the laminar flow.
  • the temperature is measured in the liquid-flow-through cooling jacket and regulated to a specified value.
  • the electrodes are electrically insulated, gas- and liquid-tight out and immersed in the measuring medium.
  • the opposite part of this inline measuring probe carries the electrical contacts to the measurement evaluation. This probe is particularly suitable for live cell determination inside containers.
  • total cell number is understood to mean the total number of live and dead cells in the measurement volume of a microbiological culture. In order to keep the live cell determination as exact as possible even under changing conditions, it is favorable if the total number of cells is made available to the measuring system from the input variable.
  • an optical probe for total cell number is preferably arranged close to the electrodes, which detects the instantaneous total number of cells and transmits them as an input quantity to the measuring and evaluation system for living cell determination.
  • C represents the capacity in Farad [F]
  • the distance d corresponds to the distance between the plates
  • ⁇ is the dielectric constant (ER * £ O)
  • A is the area of the electrodes.
  • the capacitance reading should decrease by a factor of four. The stirring in a container or in a fermenter has an influence on the electrode surface. This reduces the measurement signal of the capacitance.
  • the applied physical-mathematical model hereinafter referred to as mathematical algorithm, consists of an electrical resistance RL, which is connected in parallel to a non-ideal capacity CPEDL, with the parameters Q, n.
  • This equivalent circuit diagram depicts the natural processes of formation of the double layer region close to the electrodes, and can thus be described to a good approximation as follows:
  • ⁇ DL l,,. ⁇ _ ( 2 )
  • KDL ⁇ is the angular frequency and i is the imaginary part
  • n and Q are determined as parameters in the physical-mathematical model and compared with the experimental data. With these parameters, the double-layer capacitance CDL can subsequently be calculated with the following equation:
  • the alternating voltages are in the range of 100 mV to 250 mV. This voltage is sufficient to obtain measurements of sufficient accuracy without allowing excessive currents to flow. Furthermore, it can be provided that initially an ohmic double-layer resistance RDL and a charge QDL from a measured double-layer impedance ZDL are calculated from the measured values according to the formula:
  • ZDL ViRöl + QDL (4) is calculated and then a double-layer capacitance CDL is calculated by multiplying the ohmic double-layer resistance RDL with the charge QDL.
  • the following formula is used:
  • the calculated double-layer capacity CDL is very well correlated with the living cell concentration. Over a wide range, the double-layer CDL capacity may even be proportional to the living cell concentration. This measures the complex resistance (ie the impedance).
  • the imaginary part is applied over the real part. From this, the double-layer capacity CDL is determined.
  • the formula corresponds to a serial circuit consisting of ohmic and capacitive resistance.
  • a limit capacitance Cthreshoid is subtracted from the double-layer capacitance CDL.
  • the conductivity is dependent on the medium and is preferably determined with a probe - particularly preferably with the same probe with which the measurement takes place - as the reciprocal of the resistance in Siemens per unit length [S / cm]. This can be distortions and displacements by impurities in the fluid such as various sugars or alcohols are prevented.
  • the threshold capacity Cthreshoid may be made dependent on previously measured parameters of the fluid such as the maltose or glucose concentration or the like.
  • the accuracy of the estimation of the living cell concentration can be improved if the double-layer capacitance CDL is smoothed, preferably via a Fast Fourier Transformation smoothing (FFT smoothing), particularly preferably via a 5-point FFT smoothing.
  • FFT smoothing a Fast Fourier Transformation smoothing
  • double-layer capacitances CDL from previous times can be included in the calculation and thus an improved statement can be made.
  • a probe for living cell determination of a microbiological cell culture preferably of Saccharomyces cerevisiae or tumor cells, is used, with a supply for the microbiological cell culture, with electrodes, via electrical connection points with a measurement and evaluation system, for generating a high and low frequency Measuring signals are connected, wherein two opposite electrodes are provided for the assignment of the analysis voltage and the detection of the measurement signal.
  • the electrodes are made of a metallic material, in particular of a stainless steel, preferably high-alloy chromium-nickel stainless steels.
  • the metallic electrodes are coated, in particular coated with precious metals or non-noble metals, preferably gold.
  • the electrodes are arranged at a distance of about 2 mm. As a result, only a small part of the fluid, which is located very close to the electrodes, is measured. This improves the correlability of the measured values. In addition, the risk of temperature fluctuations is reduced.
  • the cell biology double-layer capacitance near the electrode can be determined by means of impedance measurement.
  • the temperature of the fluid in the region of the electrodes is adjusted during the measurements. This can be done, for example, by a temperature control device in the region of the electrodes, for example in a jacket. Thereby, that part of the fluid which is most essential for the measurement, namely that which is directly at the electrodes and between the electrodes, is brought to a certain, known temperature and thus the correlation of the measured values to the living cell concentration can be improved. This can be done by heating and / or cooling systems.
  • CDL (CPE) - (RDL 1 * QDL) 1 / U a non-idealized double-layer CDL (CPE) capacity is calculated, which is characteristic of the physiological state of the cells. It has become more surprising Shown manner that the nephew-idealized double layer capacitance CDL (CPE), called in wei ⁇ more excellent result and double layer capacitance CPE-Q, well to the physiological condition, and thus can be correlated with the cell viability.
  • the double-layer CDL (CPE) capacity is directly proportional to the metabolic activity or growth of the cells.
  • the vitality can be determined in particular via the observation of the double-layer capacitance CDL (CPE) over a longer period of time. If the calculated values are plotted over time and a curve is generated for these values, then the vitality can be determined from the slope of the curve.
  • FIG. 1 shows a schematic view of an online probe for living cell determination
  • FIG. 2 is a schematic view of an inline probe for live cell determination
  • Figure 3A, 3B is a schematic representation of a microstructured electrode system for living cell determination of a cell culture with a very low cell count;
  • FIGS. 5A, 5B measured values for biomass determination of optical density
  • FIG. 6 Diagram for anaerobic cell cultivation at various levels
  • Figures 7A, 7B, 7C show time course in biomass growth in aerobic and anaerobic cell cultivation of S. cerevisiae, impedance signal and maltose / ethanol concentrations over time, impedance signal and glucose / ethanol concentrations over time;
  • FIGS. 8A, 8B show raw data from the impedance measurement in the Nyquist representation in aerobic cell cultivation in comparison of online probe to inline probe;
  • FIG. 9 shows a comparison of the calculated cell weight or the biomass from the raw data of the impedance measurement with the measured cell weight;
  • FIG. 10 shows a schematic block diagram of the design of the measuring and evaluation system for living cell determination.
  • FIG. 1 schematically shows a first embodiment of a measuring probe 1, which is intended for on-line use, with metallic electrodes 2 as measuring elements, wherein the electrodes 2 in this embodiment are made of stainless high-grade nickel-chromium stainless steel.
  • the active surfaces of the electrodes 2 protrude into the measuring channel 3, through which a microbiological culture 14 flows as a fluid, which can have a noble metal coating 4, preferably gold, on the measuring surface.
  • the metallic electrodes 2 are sealed, for example with O-rings 5 in a cylindrical housing 6 from the flowed through measuring channel 3.
  • the sensor housing 6 is for example made of glass, but can of course also be made of other materials, such as stainless steel, which meet the food requirements, or the biotechnological requirements.
  • a jacket 9 which acts as a temperature control.
  • the exact measurement temperature can be adjusted so that the temperature in the cell culture to be analyzed can be kept constant during the measurement.
  • a medium for temperature control for example, water can be used.
  • other suitable media for tempering can be used.
  • the temperature control medium is fed via an inlet and outlet into an externally guided circuit.
  • the live cell determination is carried out by means of a 4-point impedance measurement, in which an amplitude of 100 to 500 mV is applied to the electrodes 2 in a frequency range from 10 6 to 10 1 Hz.
  • an electrical line pair 11, 1 is electrically isolated and gas and liquid-tight out to the outside.
  • FIGS. 2 and 3 each schematically show the application of the measuring probe 1 shown in FIG. 1 for living cell determination.
  • FIG. 2 schematically shows a second embodiment of the measuring probe 1 according to the invention, which is intended for in-line use.
  • the measuring probe 1 consists, as described in FIG. 1, of two stainless-steel high-grade nickel-chromium stainless steel electrodes 2 arranged opposite one another.
  • the electrodes can, for example, have a precious metal coating 4, preferably of gold, on the side facing one another. Of course, other noble metal coating can be applied.
  • the rod-shaped or tubular housing 12 is also made of a stainless high-alloy nickel-chromium stainless steel, which in the interior of the electrical leads 8 for the electrodes 2 electrically insulated, and sealed, leads to the electrical connection points 11, 1.
  • a union fitting 13 functions for gas- and liquid-tight screwing in a container or vessel.
  • FIG. 3 shows a further embodiment variant of a measuring cell 1.
  • the two electrodes 2 are designed microstructured.
  • the electrodes 2 are designed in this embodiment of the invention, without touching each other, interlocking as finger electrodes.
  • it is intended to use any microstructured electrode geometries in order to increase the specific surface area.
  • the microstructured electrodes are coated with precious metals, preferably gold.
  • the distance between the electrodes 2 is between 100-500 ⁇ .
  • the measuring probe 1 can be actively flowed through with a medium of a microbiological cell culture 14 by means of a pump.
  • This variant embodiment of the measuring cell 1 according to the invention is particularly preferably suitable for microbiological cell cultures with a lower cell density or smaller numbers of cells.
  • FIGS. 4A and 4B depict the complex resistors T and Z "from the impedance measurement in the so-called" Nyquist representation ".
  • FIG. 4A shows the impedance measurement data of a cell culture at different dilutions.
  • FIG. 4A shows the results from the impedance measurement at different cell densities.
  • FIG. 4B shows the mathematically modeled functions for the cell densities are depicted as so-called “data fit”.
  • FIGS. 5A and 5B depict the measurement results of aerobic cell cultivation on glucose and maltose.
  • the biomass determination is summarized by means of two measurement methods depending on the ideal capacity Cideai. In FIG. 5A, the optical density measurement OD.sub.10 IO is used and in FIG. 5B the dry cell weight DCW has been determined.
  • FIG. 6 shows graphically the results of different anaerobic cell cultivations at different sugars and sugar concentrations.
  • ANA1 32.7 g / L glucose
  • ANA2 44.4 g / L maltose and 54.2 g / L glucose
  • ANA3 22.5 g / L glucose have been added.
  • the detection limit is at a threshold value of about 0.3 g / L of the cell dry weight.
  • a measuring probe 1 according to FIG. 3 with a microstructured electrode geometry is suitable.
  • FIGS. 7A and 7B show the time course of over twenty hours of aerobic and anaerobic microbial cell cultivation using the example of S. cerevisiae.
  • the growth course in particular the increase and decrease, can be observed in the microbiological culture.
  • FIG. 7B shows the time profile of the impedance signal as the calculated double-layer capacitance CPE-Q (the non-ideal double-layer capacitance CCL (CPE)) and the maltose / ethanol concentration during the aerobic cultivation.
  • CPE-Q the non-ideal double-layer capacitance CCL (CPE)
  • FIG. 7C shows the time profile of the impedance signal as calculated double-layer capacitance CPE-Q and the glucose / ethanol concentration during anaerobic cultivation.
  • FIG. 8A shows the raw data from the impedance measurement during an aerobic cultivation by means of the measuring probe 1 according to FIG.
  • the capacitance is smaller by an order of magnitude than that of measuring probe 1 according to FIG. 1. This is due to the fact that smaller electrode surfaces are present in measuring probe 1 according to FIG.
  • FIG. 8B shows the course of the impedance signal as well as the glucose consumption and the ethanol production during an aerobic cultivation using the measuring probe 1 according to FIG.
  • FIG. 9 shows the calculated biomass of the cell culture from the impedance signal of FIG. 6, including the measured cell dry weight DCW.
  • the flow cytometry measurement confirmed that no dead population was present.
  • the cell dry weight DCW can thus be adjusted with live cell concentration.
  • FIG. 10 shows, in a schematic block diagram, the structure 15 of the measuring and evaluation system 25 for living cell determination.
  • the measurement and evaluation system 25 includes a mathematical algorithm 16 for determining the cell biological double-layer capacitance, a microcontroller 17 for performing arithmetic operations, a current and voltage source 26 for generating a frequency-dependent measurement signal, a visualization and operation unit 18 for the representation and handling of the measuring process, and a measuring process control 19.
  • the measurement and evaluation system 25 has a connection or data interface 22 to a process control system in order to transmit the measurement data.
  • a measuring process controller 19 serves to monitor the analysis temperature 20; the temperature is controlled by a circulation pump 23.
  • the pump 24 here ensures a gentle promotion of the microbiological culture 14.
  • the structure 15 has a calibration of the cell number 21, which is requested by the measuring process control 19 and is used by the mathematical algorithm 16 for determining living cell determination.

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Abstract

L'invention concerne un procédé de mesure permettant de mesurer par spectroscopie électrique l'impédance d'un fluide contenant des cellules vivantes, au moins deux électrodes (2) étant agencées dans le fluide, plusieurs signaux de mesure étant appliqués successivement entre les électrodes (2) sous la forme de tensions alternatives, et le courant et la tension étant mesurés entre les électrodes (2) en tant que valeurs de mesure, les fréquences des tensions alternatives étant au moins en partie différentes. L'invention vise à proposer un procédé du type ci-dessus qui permette de déterminer plus précisément le nombre de cellules. Cet objectif est atteint en ce que la mesure spectroscopique de l'impédance comprend une plage de fréquence de 0,1 Hz à 106 Hz.
PCT/AT2018/050017 2017-07-21 2018-07-20 Procédé de mesure par spectroscopie électrique de l'impédance d'un fluide contenant des cellules vivantes WO2019014692A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022033395A1 (fr) * 2020-08-14 2022-02-17 南京原码科技合伙企业(有限合伙) Appareil et procédé de concentration rapide pour micro-organismes pathogènes

Citations (3)

* Cited by examiner, † Cited by third party
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