PROBE FOR MEASURING THE BIOMASS CONTENT IN A MEDIUM

Abstract

The invention relates to a probe for measuring the biomass content in a medium having a suspending fluid and cells. The probe has at least three electrodes, wherein two of the electrodes are configured as excitation electrodes for transmitting an excitation signal through a medium. Two of the electrodes are configured as signal electrodes for receiving an excitation signal that has passed through the medium. The or each signal electrode is located between the two excitation electrodes at a position where a high current density is generated. The probe can have two excitation electrodes and four signal electrodes. The signal electrodes are configured substantially in parallel and arranged in couples adjacent each other, at positions between the excitation electrodes. The signal electrodes are configured such that (i) a first couple of signal electrodes are arranged between the closest end points of the excitation electrodes at one side of the probe, and (ii) a second couple of signal electrodes are arranged between the closest end points of the excitation electrodes at the other side of the probe.

Claims

1. A probe for measuring the biomass content in a medium having a suspending fluid and cells, the probe comprising: at least three electrodes, wherein two of the electrodes are configured as excitation electrodes for transmitting an excitation signal through a medium, and said excitation electrodes are configured such that the current density of an excitation signal passing between said excitation electrodes through said medium varies along the length of the excitation electrodes, wherein two of the electrodes are configured as signal electrodes for measuring a voltage therebetween that has been generated by an excitation signal passing between the excitation electrodes, and where only three electrodes are provided the signal couple of electrodes comprise one of the excitation electrodes and a single sensing electrode, wherein at least one of the signal electrodes or each signal electrode is located between the two excitation electrodes at a position where high or higher current density occurs such that the signal electrode or each signal electrode is positioned upon, or located on or adjacent, a current path which has substantially the same current phase as the total excitation current.

2. A probe according to claim 1, wherein one of the signal electrodes or each signal electrode, are a different length or shape from the excitation electrodes, are different lengths or shapes.

3. A probe according to claim 1, wherein one of the signal electrodes or each signal electrodes is shorter than the excitation electrodes.

4. A probe according to claim 1, wherein the surface area of one of the excitation electrodes is greater than the other excitation electrode.

5. A probe according to claim 1, wherein the conductive surface area of the excitation electrodes is greater than the surface area of one of the signal electrodes.

6. A probe according to claim 1 wherein one of the signal electrodes or each signal electrodes is positioned in the region of a mid-point between the excitation electrodes.

7. A probe according to claim 1, wherein one of the signal electrodes or each signal electrodes is positioned and substantially half-way between the excitation electrodes.

8. A probe according to claim 1, wherein the excitation electrodes are substantially the same length and substantially parallel.

9. A probe according to claim 7, wherein one of the signal electrodes or each signal electrodes is configured to be substantially parallel to the excitation electrodes.

10. A probe according to claim 1, wherein the excitation electrodes and a single signal electrode are configured substantially in parallel, and the signal electrode is positioned between the excitation electrodes in a region between the closest end points of the excitation electrodes.

11. A probe according to claim 1, wherein the excitation electrodes and two signal electrodes are configured substantially in parallel, wherein the two signal electrodes are positioned adjacent each other at a position between the excitation electrodes in a region between the closest end points of the excitation electrodes.

12. A probe according to claim 1, wherein the excitation electrodes and four signal electrodes are configured substantially in parallel, wherein the four signal electrodes, arranged in couples adjacent each other, at positions between the excitation electrodes, and configured such that (i) a first couple of signal electrodes are arranged between the closest end points of the excitation electrodes at one side of the probe, and (ii) a second couple of signal electrodes are arranged between the closest end points of the excitation electrodes at the other side of the probe.

13. A probe according to claim 1, wherein the excitation electrodes are configured substantially in parallel and are shorter than a single signal electrode, which is positioned between the excitation electrodes, wherein the excitation electrodes are positioned in a region between the end points of the signal electrodes.

14. A probe according to claim 1, wherein the excitation electrodes are configured substantially in parallel and are shorter than two signal electrodes, which are parallel and adjacent to each other in a position between the excitation electrodes and the excitation electrodes are positioned in a region between end points of the signal electrodes.

15. A probe according to claim 1 wherein at least one electrode is a conductive exposed length of wire material.

16. A probe according to claim 1 wherein at least one electrode is a conductive exposed strip material.

17. A probe according to claim 1, wherein one of the signal electrodes or each signal electrodes is less than one third the length of an excitation electrode.

18. A probe according to claim 1, wherein one of the signal electrodes or each signal electrodes is less than one fourth the length of an excitation electrode.

19. A probe according to claim 1, wherein one of the signal electrodes or each signal electrodes is less than one fifth the length of an excitation electrode.

20. A probe according to claim 1, wherein one of the signal electrodes or each signal electrodes is less than one sixth the length of an excitation electrode.

21. An apparatus as recited in claim 1, further comprising: an output circuit configured to generate an excitation signal for transmission via the probe through a medium; an input circuit configured to receive the excitation signal from the probe after it has passed through a medium; and a means for generating and processing signals.

22. An apparatus as recited in claim 1, further comprising: an output circuit configured to generate an excitation signal for transmission via the probe through a medium, an input circuit configured to receive the excitation signal from the probe after it has passed through a medium; a processor and/or electronic circuit for generating and processing signals.

23.-24. (canceled)

Description

[0028] FIG. 1 illustrates a known probe and has been described above. In order that the invention may be more readily understood, reference will now be made, by way of example, to the drawings in which, FIGS. 2a to 6b are schematic views of an exposed face of a probe according to the invention;

[0029] FIG. 7 is a graph showing a frequency scan of a potassium chloride solution using the probe of FIG. 1; and

[0030] FIG. 8 is a graph showing a frequency scan of a potassium chloride solution using the probe of FIG. 5b.

[0031] FIGS. 2a to 6b show the exposed face of a probe 10, having an electrically non-conductive base 12 upon which electrodes are mounted. Excitation electrodes 14 are arranged in parallel across the face of the probe 10. One or more signal electrodes 16 are arranged between the excitation electrodes. Alternatively, excitation electrodes can be arranged between the signal electrodes.

[0032] Excitation electrodes 14 are alternatively referred to as drive electrodes or current electrodes—they introduce a signal to a medium to be measured. Signal electrodes 16 are alternatively referred to as sensing or voltage electrodes—they are used to measure a voltage therebetween.

[0033] The or each signal electrode 16 is substantially parallel to the excitation electrodes. The base 12 is substantially planar such that the electrodes lie on substantially the same plane. The invention (including but not limited to the embodiments of FIGS. 2a to 6b), however, can be implemented with non-parallel electrodes and/or electrodes configured in different planes.

[0034] End points A and B are indicated adjacent end points of the excitation electrodes, which are substantially aligned, as viewed, at one side of the probe. End points A′ and B′ are the end points of the excitation electrodes at the other side of the probe.

[0035] An outer end of the or each signal electrode 16 can be substantially aligned with the ends A, B, A′, B′ of the excitation electrodes.

[0036] In use, an excitation signal is passed through a medium between the excitation electrodes 14. The signal electrode(s) 16 are used to detect the potential drop across the medium therebetween. The potential can be detected with a high impedance voltmeter, such that there is virtually no current flowing across the signal electrodes 16 electrode/solution interfaces.

[0037] In FIGS. 2a, 3a, 4a, 5a and 6a a three electrode system is shown. The excitation electrodes 14 (the two outer electrodes, as viewed) form a first electrode couple or pair. The signal couple of electrodes 16, for receiving the excitation signal that has passed through the medium, comprise one of the excitation electrodes 14 and a single sensing electrode 16.

[0038] In FIGS. 2b, 3b, 4b, 5b and 6b separate couples or pairs of electrodes for excitation and signal detection are provided. To be clear, the signal electrodes 16 comprise first and second electrodes, neither being an excitation electrode.

[0039] In FIGS. 5a and 5b two signal electrode couples 16 are provided, and represent the preferred configuration of the invention. In this configuration the field generated by the excitation signal is substantially symmetrical such that the potential in a fluid being measured is the same at each end and, therefore, senses the same voltage. In this configuration doubling the surface area of the signal electrodes, with respect to the configuration in FIGS. 4a and 4b, for example, halves the impedance between the electrode and the fluid or medium.

[0040] Referring back to FIG. 1, the probe 2 is used to measure a phase difference between the current passed between the outer wires 4 and the voltage measured between the inner wires 6, after having passed through a medium. The inventor has realised that the voltage of the outer wire is the same along its length but the current density varies according to the path the current takes between the wires. The local current density at the end points is higher than in the centre or mid-regions. By taking different paths the phase of the current along different paths will vary because the polarisation impedance (being current dependent) also varies along the excitation electrode's length. Measurement of bulk fluid impedance requires the total electrode current to be measured and a voltage which is a representative weighted mean of that voltage found along all current paths. Because current density is high or higher towards the end of the excitation electrodes the density varies along the length of the excitation electrodes. Known probes (see FIG. 1) provide a spatially averaged voltage which is not representative of that along all the current paths. Further, the phase of the voltage detected by known probes is a weighted average of the voltages on all current paths measurable along the whole length of the or each signal electrode.

[0041] In FIG. 2a, the excitation electrodes 14 are substantially the same length and are arranged to be substantially parallel to each other. The signal electrode is arranged between corresponding end-points of the excitation electrodes 14 i.e. between the closest end-points A and B of the excitation electrodes 14.

[0042] The polarisation impedance depends on current density and with excitation signals up to around 1 MHz the series-equivalent resistance decreases with current density while the capacitance increases. This varying polarisation impedance can introduce errors in the voltage measurements. The current density varies along the length of the excitation electrodes 16. The inventor has realised that the most representative voltage to be measured is proximal, or between, the regions of the end-points A and B, where current density is greatest. The regions of high current density carry the bulk of the total current and consequently errors in the voltage measurement are reduced such that the voltage to be measured is representative of the whole current flow in a medium between the excitation electrodes. In FIG. 2b a signal electrode couple 16 is provided between corresponding end-points of the excitation electrodes 14 i.e. between the closest end-points A and B of the excitation electrodes 14.

[0043] In FIG. 3a, the excitation electrodes 14 are different lengths, but are arranged to be substantially parallel to each other. To be clear, the length A-A′ of the uppermost electrode as viewed is shorter than the length B-B′ of the lowermost electrode. At one side of the probe 10 the ends of the excitation electrodes A, B are vertically aligned, as viewed. The distance between the corresponding end-points of the excitation electrodes 14 i.e. between the closest end-points A and B of the excitation electrodes 14, is comparable to the probe shown in FIGS. 2a and 2b.

[0044] The shorter excitation electrode results in the current density being biased towards the closest end-points A and B. It is in the region of the closest end-points that current density increases, such that there is a lower resistance due to polarisation and the voltage to be measured is more representative of the whole current flow in a medium between the excitation electrodes. In FIG. 3b a signal electrode couple is provided between corresponding end-points of the excitation electrodes 14 i.e. between the closest end-points A and B of the excitation electrodes 14. FIGS. 4a and 4b correspond, respectively, to FIGS. 2a and 2b, but the probes 10 are configured with one of the excitation electrodes 14 (the lowermost electrode, as viewed) having a surface area significantly larger than the other of the excitation electrode couple. The larger surface is at least twice (2×) and preferably at least 5×, and most preferably at least 10× the surface area of the other excitation electrode 14. Said larger electrode can, for example, form a part of the container for the medium being measured. The impedance of the larger electrode (including the contribution due to surface polarisation) is reduced compared to that of the other excitation electrode, and that of the test medium. As a result the voltage will be extremely close to that of the test medium adjacent the large electrode 44.

[0045] The probe of FIG. 5a is comparable to the probe of FIG. 2a with an additional signal electrode couple 18. To be clear, a first signal electrode 16 is arranged between corresponding end-points of the excitation electrodes 14 i.e. between the closest end-points A and B of the excitation electrodes 14. A second signal electrode 18 is arranged between corresponding end-points A′ and B′ of the excitation electrodes 14. In FIG. 5b a first signal electrode couple 16 and a second signal electrode couple 18 are provided between corresponding end-points A and B, and A′ and B′, respectively i.e. between the closest end-points at each end of the excitation electrodes 14. In FIGS. 5a and 5b the signal electrodes are substantially the same length.

[0046] The signal electrodes of FIGS. 5a and 5b are located in the two regions where current density is greatest. By measuring the voltage from the excitation current in the region between each of end-points the signal can be (i) more relevant or stronger, because the signal electrode is provided in two locations thus receiving a voltage, compared to FIGS. 2a and 2b, that has a lower source impedance (twice the contact area with the substance being measured) and/or (ii) more representative of the whole current flow in a medium and/or (iii) the excitation signal is received at two spatially separated positions on the face of the probe 10, for providing a more representative indication of biomass in a medium in which biomass distribution is not evenly distributed.

[0047] In FIGS. 5a the signal electrodes are connected together. In FIG. 5b the upper-most signal electrodes, as viewed, are connected together, while the lower-most signal electrodes, as viewed, are also connected together but isolated from the upper-most electrodes.

[0048] Each signal electrode 16,18 shown in FIGS. 5a and 5b, or any of the other Figures of the invention, can be sensed individually or in groups.

[0049] In light of the teaching herein it will be appreciated that the optimum position for a probe configuration can be evaluated. Further, in light of the teaching herein it would appear that the theoretical position of a signal electrode would be right between the very ends of the excitation electrodes.

[0050] The inventor has realised through practical tests, however, that positioning the outer ends of the signal electrodes substantially in line with the outer ends of the excitation electrodes can provide improved results. This is illustrated by the embodiments of FIGS. 2a to 5b.

[0051] The inventor has also realised through practical tests that the length of the signal electrode influences the results. If the signal electrodes are too short (i.e. below 2 mm in the configuration of FIG. 5b) the polarisation effect is over-compensated. The length of a signal electrode can be less than around one third the length of an excitation electrode i.e. the ratio is 1:3 (e.g. if the excitation electrode was 15 mm long then the signal electrode(s) would be no longer than 5 mm). The ration may be less that 1:4, may be preferably less than 1:5. An optimum ratio can be 1:6.

[0052] In FIG. 6a, the excitation electrodes 14 are substantially the same length and are arranged to be substantially parallel to each other. The signal electrode 16 is arranged between the excitation electrodes 14 and extend in a parallel direction with respect to the excitation electrodes. In this probe configuration the excitation electrodes are shorter than the signal electrodes and are positioned approximately midway between the end points of signal electrode, as viewed. The current flow between the excitation electrodes 14 is concentrated between points A to B and points A′ to B′ such that the current density is greater along the path between said points. The current density will vary less along the length of the excitation electrodes 14 and, therefore, the most representative voltage to be measured is proximal, or between, the closest end-points A and B, and A′ and B′, where current density is greatest. The signal electrode 16 spans a path between points A to B and points A′ to B′ such that errors in the voltage measurement are reduced because the voltage to be measured is more representative of the whole current flow in a medium between the excitation electrodes. In FIG. 6b a signal electrode couple i.e. two signal electrodes 16 is provided between points A to B and points A′ to B′ of the excitation electrodes 14.

[0053] In FIGS. 4a and 4b one of the electrodes has a greater surface area, which can be implemented using a larger diameter wire or strip, such as an exposed strip material of conductive material. One or more of the aforementioned electrodes 14, 16, 18 can be implemented with such a strip.

[0054] The electrodes in FIGS. 2a to 6b are generally formed by an exposed conductive wire. The invention, however, is not so limited and the or each electrode can be formed, by way of example, by: conductive film; deposited conductive material; conductive strips; printed conductive film; conductive strips; or a combination thereof. The shape or footprint of the or each electrode can be configured to optimise the performance in light of the teaching herein. By way of example, the or each electrode can be disc-shaped, having a circular or elliptical footprint, 4-sided footprint or hexagonal footprint.

[0055] The shape of the or each electrode can be configured to optimize the contact impedance with a substance in which the probe is immersed. By way of example, minimizing the size of an electrode to lie precisely on a selected current path would result in an increased contact impedance that would detriment performance. In light of the teaching herein a skilled person would be able to optimise the performance

[0056] Further, the cross-sectional profile can, by way of example, be: substantially circular or elliptical, as it would be for a wire; shallow substantially rectangular; having a flat base with an arcuate upper section, such as a hemispherical profile; or a combination thereof.

[0057] The electrodes in FIGS. 2a to 6b have been illustrated, by way of example, in a substantially planar configuration. The invention can alternatively be configured in three-dimensional configurations, such as a cylindrical form. The invention can alternatively be configured to increase the length of the current path between excitation electrodes (with respect to those shown in FIGS. 2a to 6b such that the influence of polarisation impedance is minimised.

[0058] A number of embodiments have been described above and in order that the performance improvement can be appreciated a comparison will be made between the performance of the probe of FIG. 1 and a representative embodiment of the invention, as shown in FIG. 5b.

[0059] The component 2 of FIG. 1 is 25 mm in diameter. All four electrodes are substantially centrally positioned on a flat insulated mount, are 0.5 mm in diameter and have an exposed length of 12 mm. They lie on substantially the same plane. The outer wires 4 are 14.5 mm apart. The inner wires 6 are each positioned 3.5 mm from the outer wires, such that there is 7.5 mm between the inner wires. The outer ends of the electrodes are substantially aligned.

[0060] As described above, in order to measure a fluid's specific impedance at a particular frequency it is necessary to measure the voltage across and the current flowing through a representative part of the fluid. The current we can most easily actually measure is the current flowing in the excitation electrodes. This current passes through the metallic electrode, through the metal/liquid interface and into the bulk liquid where it can take many different paths before returning via the other excitation electrode. Although the relationship between the voltage gradient and the current density in the fluid is constant (at a fixed frequency and in a homogeneous liquid) the current density will vary strongly between different current paths. The current density crossing the metal liquid interface layer will also vary significantly over the surface of the electrode (for most electrode configurations, and particularly if the electrodes are to be in one plane). This can be shown using finite element analysis.

[0061] The polarisation impedance of this metal liquid interface layer is itself current density dependent, so the layer will introduce series impedance into all the current paths, but this impedance will not be the same for all paths. The currents flowing along the many different paths will therefore not all be in phase with each other. The total excitation current measured will be the sum of all these currents, and will have a phase angle somewhere between the phase extremes of the currents which make it up.

[0062] The probe of the invention has one or more signal electrodes, each configured to measure a voltage, which is an average (or weighted average) of the voltages adjacent to it in the fluid, on each current path extending between the excitation electrodes through that region.

[0063] In order to measure the impedance of the fluid without interference from the polarisation impedance it is necessary that the phase relationship between the excitation current and the measured voltage is the same as that between the voltage gradient and the current in the bulk fluid.

[0064] During a measurement using the probe of FIG. 1 the current paths spread out in 3 dimensions in the adjacent fluid. A consideration of the expected current density distribution on the excitation electrodes suggests that the high and low extremes will be at the ends and centers of the strips. A current path most representative of the total current phase is expected to meet the electrodes away from both of these extreme zones. Using a KCl solution as a test fluid, the impedance and parallel—equivalent capacitance and conductance were calculated from the measured current and voltage using the probe of FIG. 1. For this test solution, the capacitance and the conductance should be frequency independent (within a given frequency range).

[0065] The frequency scan results are shown in FIG. 7. Note the large capacitance error due to electrode polarisation increasing at low frequencies. The configuration of the electrodes in FIG. 1 are such that the errors are large because the voltage electrodes are not sensing the voltage along a representative current path (they will be averaging the voltage over a lot of different paths).

[0066] The probe configuration of FIG. 5b was tested. The dimensions of the probe are substantially the same as those in FIG. 1 except that the signal electrodes 16, 18 are shortened with respect to the excitation electrodes 14. Measurements were made with three different variations of the probe shown in FIG. 5b. The length of each signal electrode was the same, but in each variation the signal electrodes were a different length. Three configurations of the probe of FIG. 5b were tested, with each configuration having a different lengths of signal electrode—the configurations tested had signal electrode lengths of 2 mm, 2.5 mm and 3 mm respectively.

[0067] For accurate voltage sensing the measuring circuit connected to the sensing electrodes should draw as little current as possible. While this is achievable at low frequencies, measurements at higher frequencies can result in errors in measuring the voltage due to capacitive loading of the electrodes, which exhibit a fairly high source impedance (which increases as electrode area is reduced). This source impedance can be reduced by connecting the sensing electrodes in parallel with an identical pair (or pairs) of electrodes which are sensing an identical voltage—as per the embodiment of FIG. 5b. It is also to be noted that the invention is not limited to connecting sensing electrodes in parallel (as per FIG. 5b) and that the invention provides improvements in performance when only a single electrode couple is used (as per FIG. 5a). In the case of FIG. 5b the field is symmetrical, so a second pair can be placed at the other side of the axis of symmetry.

[0068] The frequency scan results using the probe of FIG. 5b is shown in FIG. 8. The capacitance values for each variation (i.e. 3 mm, 2.5 mm and 2 mm probes) are shown and labelled on the left-hand-side of the graph. It can be seen that the capacitance measurement of a probe having the 3 mm long electrodes 16, 18 is almost halved compared to that of the known probe (see FIG. 1). As the length is reduced further the capacitance measurements become more linear across the lower end of the frequency range, such that a probe having sensing electrodes 2 mm in length is able to measure capacitance substantially linearly across the frequency range. The inventor has also realised that in the embodiment of FIG. 5b reducing the length of the sensing electrodes to be less than 2 mm resulted in a lower capacitance reading at low frequencies (sub 1 MHz) resulting in a deviation from a correct value or linear measurement (as viewed). In practice, the error goes the other way and the measurement results using a signal electrode shorter than 2 mm in the given example is such that the measurement would start low, close to the origin, and track upwards before straightening out in a straight-line fashion as the frequency increased. This is because, if too short, over emphasis on the high current flow at the ends becomes unrepresentative of the current passing through the fluid or medium.

[0069] Measurements were compared using a known probe (FIG. 1) and an improved probe (FIG. 5b) for illustrative purposes only, and the inventive concept herein can be applied to probes having different configurations such that positioning the sensing electrodes in a position where the current path has substantially the same current phase as the total excitation current. The embodiments of the invention herein are configured such that voltage sensing electrodes, or signal electrodes, are placed in a position along a current path which has substantially the same current phase as the total excitation current, which is passed in to a medium to be measured by the excitation electrodes. The inventor has realised that this can be achieved by the or each signal electrode being located between the two excitation electrodes at a position where high current density occurs i.e. the or each signal electrode is positioned such that the average of the phases of all current paths measurable along the length of the or each signal electrode is substantially the same of the phase of the total excitation current. In the embodiments of the invention herein this occurs in the region between the end of the excitation electrodes. The probe is suitable for use with apparatus for measuring the biomass content in a medium, the apparatus utilising the probe to pass an excitation signal through a medium and subsequently receive the excitation signal from the probe after it has passed through the medium before processing and analyzing the signals.

[0070] The probe can be used in a system for performing a fermentation process utilising a culture comprising a suspending liquid and cells, the system having apparatus for measuring the biomass content in a medium, the system utilising the a probe to pass an excitation signal through a medium and subsequently receive the excitation signal from the probe after it has passed through the medium before processing and analyzing the signals.

[0071] The subject matter of EP1018025, EP0281602, EP0282532 and GB2481832 is incorporated herein by reference. The present invention has been described above purely by way of example, and modifications can be made within the spirit and scope of the invention, which extends to equivalents of the features described and combinations of one or more features described herein. The invention also consists in any individual features described or implicit herein.