TIME OF FLIGHT MASS SPECTROMETER

Abstract

A time of flight mass spectrometer that includes a first electrode; and a second electrode that is spaced apart from the first electrode. The ion source is configured to apply voltages to the first and second electrodes to produce an electric field in a region between the first and second electrodes so as to influence ions present in the region between the first and second electrodes when the mass spectrometer is in use. A shield is formed on the first electrode and/or second electrode. The shield is configured to inhibit an electric field formed between edges of the first and second electrodes from penetrating into the region between the first and second electrodes when the mass spectrometer is in use

Claims

1. A time of flight mass spectrometer including: a first electrode; and a second electrode that is spaced apart from the first electrode; wherein the ion source is configured to apply voltages to the first and second electrodes to produce an electric field in a region between the first and second electrodes so as to influence ions present in the region between the first and second electrodes when the mass spectrometer is in use; wherein a shield is formed on the first electrode and/or second electrode, wherein the shield is configured to inhibit an electric field formed between edges of the first and second electrodes from penetrating into the region between the first and second electrodes when the mass spectrometer is in use.

2. A time of flight mass spectrometer according to claim 1, wherein the shield is a raised element formed on a surface of one of the first and second electrodes that faces the other of the first and second electrodes so that the shield extends towards the other of the first and second electrodes.

3. A time of flight mass spectrometer according to claim 1, wherein the shield surrounds an axis extending between the first and second electrodes.

4. A time of flight mass spectrometer according to claim 1, wherein the first and/or second electrode include an aperture formed therein.

5. A time of flight mass spectrometer according to claim 1, wherein an inwardly facing surface of the shield is outwardly spaced apart from an axis extending between the first and second electrodes by a distance that is at least the width of an aperture formed in the first and/or second electrode.

6. A time of flight mass spectrometer according to claim 1, wherein an outwardly facing surface of the shield is outwardly spaced apart from an axis extending between the first and second electrodes by a distance that is at least 1.5 times the width of an aperture formed in the first and/or second electrode.

7. A time of flight mass spectrometer according to claim 1, wherein a secondary shield is formed on the electrode on which the shield is formed, wherein the secondary shield is configured to inhibit an electric field from penetrating into the region between the first and second electrodes through an aperture formed in the electrode on which the shield is formed, wherein the secondary shield surrounds the aperture.

8. A time of flight mass spectrometer according to claim 7, wherein the height of the shield is larger than the height of the secondary shield.

9. A time of flight mass spectrometer according to claim 1, wherein: the first and second electrodes are included in an ion source of the mass spectrometer; the ion source is configured to apply voltages to the first and second electrodes to produce an extraction electric field in an extraction region between the first and second electrodes so as to extract ions from the extraction region through an aperture in the second electrode when the mass spectrometer is in use.

10. A time of flight mass spectrometer according to claim 9, wherein the first electrode is a sample plate for carrying a sample.

11. A time of flight mass spectrometer according to claim 9, wherein the sample plate may be mounted on a sample plate carrier and the mass spectrometer includes a mechanism configured to translate the sample plate carrier laterally with respect to an ion optic axis so as to laterally offset the sample plate carrier with respect to the ion optic axis.

12. A time of flight mass spectrometer according to claim 11, wherein the shield is formed on the second electrode and an inwardly facing surface of the shield is outwardly spaced from a boundary between the second electrode and the aperture formed in the second electrode by a distance that is adequately small so that the inwardly facing surface of the shield remains within the outer boundary of the first electrode when viewed along an ion optic axis, when a centre of the sample plate carrier is at a maximum permitted lateral offset with respect to the ion optic axis.

13. A time of flight mass spectrometer according to claim 9, wherein the ion source includes a laser for ionising a sample carried on the sample plate by firing light at the sample.

14. A time of flight mass spectrometer according to claim 9, wherein the ion source is a MALDI ion source.

15. A time of flight mass spectrometer or ion source substantially as any one embodiment herein described with reference to and as shown in the accompanying drawings.

Description

[0084] Examples of the present proposals are discussed below, with reference to the accompanying drawings in which:

[0085] FIG. 1 shows an example TOF mass spectrometer that is not an embodiment of the present invention, but is useful for understanding the present invention.

[0086] FIG. 2 shows another example TOF mass spectrometer that is not an embodiment of the present invention, but is useful for understanding the present invention.

[0087] FIG. 3 shows a view along the ion optic axis of the extraction electrode and sample plate of the ion source of the TOF mass spectrometer of FIG. 2: (a) with the ion optic axis aligned with the centre of the sample plate and (b) with the ion optic axis aligned with an extreme analysis point on sample plate.

[0088] FIG. 4 shows a cross-section view of electric field contours obtained from an electrostatic model of region around the sample plate and extraction electrode of the ion source of the TOF mass spectrometer of FIG. 2: (a) with the ion optic axis aligned with the centre of the sample plate and (b) with the ion optic axis aligned with an extreme analysis point on sample plate.

[0089] FIG. 5 shows an example TOF mass spectrometer whose ion source includes an extraction electrode having an annular shield formed thereon: (a) where the extraction electrode has a plane aperture (no secondary shield) and (b) where the extraction electrode has an aperture extending through the extraction electrode to form a through channel (with secondary shield).

[0090] FIG. 6 shows preferred limits and values for parameters defining the shield of FIG. 5.

[0091] FIG. 7 shows a cross-section view of electric field contours obtained from an electrostatic model of region around the sample plate and extraction electrode of the ion source of FIG. 5(b): (a) with the ion optic axis aligned with the centre of the sample plate and (b) with the ion optic axis aligned with an extreme analysis point on sample plate.

[0092] FIG. 8 shows a mass spectrum (normalised to maximum signal) of peptides in CHCA matrix obtained using a mass spectrometer having the ion source of FIG. 2: (a) with the ion optic axis aligned with the centre of the sample plate and (b) with the ion optic axis aligned with an extreme analysis point on sample plate.

[0093] FIG. 9 shows a mass spectrum (normalised to maximum signal) of peptides in CHCA matrix obtained using a mass spectrometer having the ion source of FIG. 5(b): (a) with the ion optic axis aligned with the centre of the sample plate and (b) with the ion optic axis aligned with an extreme analysis point on sample plate.

[0094] In general, the following discussion describes examples of the present proposals in the context of a time of flight (“TOF”) mass spectrometer including an ion source which has an extraction electrode located above a sample plate. In the example depicted, the extraction electrode is a plate-shaped element with an aperture formed therein, through which ions are extracted. The extraction electrode also has a shield formed thereon that extends towards a sample plate. The form of the shield is preferably optimised for the particular geometry of the extraction electrode to control side field penetration, preferably to ensure that pre- and post-extraction electric fields are axially symmetrical and invariant with sample plate carrier position (relative to the extraction electrode).

[0095] The present invention may be viewed as relating to an ion optic system for a time of flight (TOF) mass spectrometer.

[0096] As shown in FIG. 1, a TOF mass spectrometer typically comprises an extraction region 1, an acceleration region 2, a field free region 6 and associated TOF mass analyser (not shown). The mass analyser may be linear or reflectron, for example.

[0097] The extraction region is typically formed between a first electrode 3 and a second electrode 4. The acceleration region is typically formed between the second electrode 4 and a third electrode 5.

[0098] In a simple form, the second electrode 4 and third electrode 5 are planar parallel plates with appropriate size central apertures to enable the ions to pass through.

[0099] In a MALDI ion source, the first electrode 3 may be a sample plate. The MALDI process is often used to facilitate the vaporization and ionization of biomolecules and large organic molecules.

[0100] In a typical MALDI ion source, the molecules are embedded in a matrix which absorbs UV light. When a UV laser is fired on a sample, located on the sample plate 3, to initiate the MALDI process, a plume of ionised and neutral analyte and matrix molecules is ejected from the sample plate 3.

[0101] The ionised molecules are subsequently extracted from extraction region 1 through the aperture in the second electrode 4, often referred to as the extraction electrode, by applying appropriate voltages to the first and second electrodes 3, 4 to produce an extraction electric field in the extraction region 1. The ions are further accelerated by a field formed in the acceleration region 2 between the extraction electrode 4 and the third electrode 5. The third electrode may be at a ground potential with the ions passing through it into the field free region 6 of the mass spectrometer, e.g. to an associated linear or reflection mass analyser. For this reason, the third electrode 5 is often referred to as the ground electrode.

[0102] In a simple MALDI ion source, the ions may be promptly extracted by a static extraction electric field, of typically 2 to 5 kV, formed between the sample plate 3 and the extraction electrode 4 (for avoidance of any doubt, this field may be achieved by lowering an existing voltage applied to the extraction plate). The extracted ions pass may then pass through the aperture in the extraction electrode 4 and may then be further accelerated by a field formed in the accelerating region between the extraction electrode 4 and the ground electrode 5 before passing through the aperture in the ground electrode 5 into the field free region 6 and an associated mass analyser.

[0103] However, in many MALDI ion sources in TOF mass spectrometers, the ion source implements a technique known as pulsed extraction to improve the instruments mass resolution by focusing the kinetic energy spread of the ions. In such a technique, the resolution can be improved by holding the potential of the extraction electrode 4 at the same potential as the sample plate 3, creating a field free region, whilst ions are formed. Then, after a short predetermined delay, pulsing the extraction electrode 4, e.g. by between 2 kV and 5 kV, to produce the extraction electric field. The short delay may be chosen to be a period of time optimum for focusing the kinetic energy spread of the ions of interest. Essentially, with an appropriate delay, typically 10 ns to a few μs, ions with lower velocity are able to receive enough extra potential energy to catch ions with higher velocity after flying some distance from the ion source, usually the detector.

[0104] The electrodes 4, 5 used in the extraction and acceleration regions in a simple form may be plane parallel plates with a central aperture (the central aperture may be gridded or ungridded). The aperture in the extraction electrode 4 is usually fairly small, e.g. 2 mm to 20 mm, because once the size is increased beyond a few mm the electric field created by the potential difference between the extraction electrode 4 and the ground plate 5 extends through the aperture in the extraction electrode into the portion of the extraction region 1 immediately in front of the sample plate 3. This effect, which may be referred to as axial field penetration, may compromise the field free region in front of the sample plate 3 (prior to producing the extraction electric field, for pulsed extraction) and can therefore result in ions being extracted at an undesired time and/or having an undesired trajectory, which can significantly decrease both mass analyser resolution and mass analyser sensitivity. Therefore, it is usually desirable to maintain a small aperture.

[0105] However, there are advantages to having a larger aperture in the extraction electrode 4. For example, it may be desirable to be able to both direct the laser light beam into the ion source close to the ion optic axis and also view the sample plate 3 at an angle close to the ion optic axis, which both require a larger aperture diameter. Further, along with the charged analyte that is extracted through the ion lens, there is a great deal of neutral analyte and matrix ejected from sample that can rapidly contaminate elements of the extraction electrode 4 and may adversely affects the ion source performance. The rate at which this contamination builds up can be reduced with larger apertures.

[0106] It has been reported in U.S. Pat. No. 6,888,129 that axial field penetration can be controlled to an acceptable level with a larger aperture if the aperture in the extracting electrode is extended in the form of a through channel. FIG. 2 shows the TOF mass spectrometer of FIG. 1 modified to have a second electrode 7 whose aperture is extended in the form of a tube 11 that extends in the direction of the sample plate 3, such that the ions may pass through from one side of the second electrode 7 to the opposite side by passing through said channel. As taught in U.S. Pat. No. 6,888,129, the channel length may be slightly less than, equal to, or greater than the diameter of the aperture. As discussed in U.S. Pat. No. 6,888,129, the tube 11 helps to reduce the field penetration from the acceleration region 2 into the extraction region 1 through the aperture in the second electrode 7 to an acceptable level, without compromising the effectiveness of the pulsed extraction. In practice there will always be some residual field penetration through the extraction electrode 7 and a compromise must be achieved between the benefits of the larger aperture in the extraction electrode 7 and the detrimental effects on ion source performance. The tube 11 provided by the extended aperture proposed by U.S. Pat. No. 6,888,129 may be referred to as a secondary shield herein.

[0107] In a typical MALDI ion source, ions are produced from a small area on the sample plate 3, which area is typically no larger than the size of the beam waist of the irradiating laser light, typically 5 μm to 500 μm in diameter. In most practical applications it is required to analyse ions from several points on the same sample plate that may extend over several cm, or from several smaller samples arranged over an area of several cm. Typically the samples are arranged on a sample plate of rectangular form that may have a width that is in the range 20 mm to 150 mm (though other widths and forms are possible). It is possible to scan the laser beam (which may be UV light) over a stationary sample plate or move the sample plate relative to a fixed laser position. For most applications, it is more practical to translate the sample plate in a plane perpendicular to an ion optic axis. This is usually achieved by mounting the sample plate on a sample plate carrier, using a mechanism configured to translate the sample plate carrier laterally (e.g. in two orthogonal directions within a plane perpendicular to the ion optic axis).

[0108] FIG. 3(a) shows the extraction electrode 7 of FIG. 2 viewed along the ion optic axis with the aperture in the extraction electrode 7 aligned with the centre of the sample plate 3, i.e. with zero lateral offset.

[0109] FIG. 3(b) shows the extraction electrode 7 of FIG. 2 viewed along the ion optic axis when the sample plate carrier (and therefore the sample plate 3) is at a maximum permitted lateral offset with respect to the second electrode 7. Thus, ion optic axis which extends through the aperture in the extraction electrode 7 is aligned with an extreme measurement position in one corner of the sample plate 3.

[0110] With reference to FIG. 3, the field formed between the sample plate 3 and extraction electrode 7 can be disturbed due to side field penetration between the sample plate 3 and the extraction electrode 4, as the sample plate carrier is translated from its zero offset (central alignment) position to a maximum lateral offset. This effect is more significant when there is not complete overlap between the extraction electrode 7 and sample plate 3.

[0111] This effect is illustrated by FIG. 4, which shows field contours, on electrostatic model of the ion source of FIG. 2, in a region around the sample plate 3 and the extraction electrode 7. As can be seen from FIG. 4, the field is symmetrical when the sample plate 3 is centred on the axis of the extraction electrode 7 (zero lateral offset), but asymmetrical when the sample plate 3 is laterally offset with respect to the extraction electrode 7.

[0112] The side field penetration between the sample plate 3 and extraction electrode 7 is potentially more problematic than axial field penetration, described above, due to its asymmetry and variation with sample plate carrier position.

[0113] The effect of the side field penetration can be significant before and during production of the pulsed extraction field between the sample plate 3 and the extraction electrode 7. Ideally, the region of initial ion formation in the extraction region 1 would be completely free of any side field penetration effects formed between edges of the extraction electrode 7 and the sample plate 3 (/sample plate carrier). Ideally, this field free region would extend to the distance traveled by the fastest moving (e.g. lowest mass) ions of interest during the period of time prior to the extraction field being formed (i.e. the pre-extraction period). Otherwise the effects of non-axisymmetric electric penetration could cause axial spreading of the ions, leading to loss of resolution, and divergence and deviation of the ions leading to loss of sensitivity.

[0114] During pulsed extraction, side field penetration may distort the lens formed by the electric field between the sample plate 3 and the extraction electrode 7. This could adversely influence a focusing effect, which could in turn cause undesirable aberrations, again leading to loss of resolution and loss of sensitivity. Similar problems would also occur for an ion source configured to produce a static extraction electric field that is present during both the formation and extraction of ions.

[0115] Uncontrolled and varying side field penetration as the sample plate 3 is translated, can therefore distort potentials between the sample plate 3 and extraction electrode 7 during pre-extraction and the pulsed extraction periods. Such uncontrolled and distorted potentials in the ion beam path may give rise to significant differences in both mass analyser resolution and mass analyser sensitivity as the sample plate carrier position varies.

[0116] The following examples aim to reduce the penetration of side fields to areas traversed by ions so as to reduce variation (preferably such that that there is no significant change in) instrument performance as the sample plate is laterally offset with respect to second electrode 7 (/ion optic axis), as well as to improve the quality of the mass spectra obtained (preferably so that the quality of mass spectra obtained is invariant with sample position on the sample plate 3).

[0117] Accordingly, with reference to FIG. 5(a), there is provided a TOF mass spectrometer in which an ion source has an extraction electrode 9. The extraction electrode 9 is a plate-shaped element having an aperture formed therein. A shield 10, which is a raised element, is formed on a surface of the extraction electrode 9 that faces the sample plate 3 so that the shield 10 extends towards the sample plate 3. The shield 10 surrounds an ion optic axis that extends through the aperture. The shield 10 is outwardly spaced apart from a boundary 10a between the extraction electrode 9 and the aperture. In this way, the shield 10 helps to inhibit an electric field formed between edges of the sample plate 3 and extraction electrode 4 from penetrating into the extraction region 1. In turn, this helps to shield the extraction region 1 (specifically the portion of the extraction region 1 where ions are present when the mass spectrometer is in use) from changes in the side penetration fields as the sample plate carrier (and therefore the sample plate 3) is translated with respect to the second electrode 9 (/ion optic axis).

[0118] The shield 10 of the extraction electrode 9 thereby helps to prevent significant changes in the pre-extraction and pulse extraction fields, thus helping to maintaining mass analyser resolution and mass analyser sensitivity as the sample plate carrier (and therefore the sample plate 3) position varies.

[0119] The shield 10 can be incorporated both as part of an extraction electrode 9 that incorporates a plane aperture (no secondary shield), not extending beyond the planar surfaces of the electrode as shown in FIG. 5(a), or as part of an extraction electrode 9′ that incorporates an aperture extending beyond the planar surfaces of the electrode to provide a through element in form of a tube 11′ as shown in FIG. 5 (b).

[0120] The tube 11′ of FIG. 5(b) may be referred to as a secondary shield herein. Here, the secondary shield is configured to inhibit an electric field from penetrating into the extraction region 1 through the aperture formed in the extraction electrode 9′.

[0121] The preferred form of the shield 10 is circular, being provided here as an annular ring, concentric with the aperture in the extraction electrode 9. However, in practice the shield does not have to be circular, but could be square, rectangular or have another form.

[0122] Parameters defining the shield may include its height (above the surface of the extraction electrode 9 that faces the sample plate 3) and, if the shield is circular, its inner radius and outer radius. These parameters are not completely independent of each other and are preferably within certain bounds for the shield to be effective. It is highly preferably for the shield 10 to be of such a form to prevent side field penetration into the portion of the extraction region 1 where ions are present when the mass spectrometer is in use up to a maximum lateral offset of the sample plate carrier (and therefore the sample plate 3), but without significantly distorting the shape of the extraction electric field used for focusing ions during pulsed extraction.

[0123] In general, the inner radius of the shield 10 may be determined by the size and shape of the sample plate 3/sample plate carrier and the height of the shield 10 and outer radius may be optimised to control the side field penetration within other instrument constraints.

[0124] The inner radius of the shield is preferably such that when the sample plate at a maximum permitted lateral offset, the inwardly facing surface of the shield 10 is within the boundary of the sample plate carrier (when viewed along the ion optic axis). This is because if the inwardly facing surface of the shield 10 is outside this boundary (when viewed along the ion optic axis), the side fields will readily penetrate into the extraction region. Lower values for the inner radius could be used and may be desirable to define the shape of the lens formed by the extraction electric field produced between the sample plate 3 and extraction electrode 9, the requirements for which depend greatly on the particular ion source geometry, for example, whether the extraction electrode 9 has a plane aperture as in FIG. 5(a) or tube 11′ as in FIG. 5(b).

[0125] In the above paragraph, it has been assumed that the sample plate carrier is conductive and therefore forms the first electrode together with the sample plate 3, and that the sample plate carrier provides the outer boundary of the first electrode. However, this need not be the case in other examples.

[0126] The outer radius and the height of the shield may be optimised to control the side field penetration. Generally, within limits, a wider low shield has similar effectiveness as a higher narrow shield. The limits for the outer radius and height of the shield 10, 10′ may therefore be determined by particular ion source geometry.

[0127] The height of the shield 10, 10′ is preferably such that there is a clearance between the shield 10, 10′ and the sample plate 3 of at least 2 mm, so as to avoid electrical breakdown between the sample plate 3 and extraction electrode 4, which may typically have a potential difference of up to between 2 kV and 5 kV across them. There may also be other practical considerations that impose a minimum clearance between the shield 10, 10′ and the sample plate 3 that relate to the mechanism used to translate the sample plate carrier (and therefore sample plate 3). Further, MALDI mass spectrometers often incorporate a viewing system to enable imaging of the sample, the illumination for which is preferably directed at a low angle of incidence with respect to the sample plate 3, which may requires a clearance of a few mm between the shield 10, 10′ and the sample plate 3, depending on the particular illumination system employed.

[0128] Some preferred limits and values for the parameters discussed above are shown in FIG. 6, in which the extraction electrode 9 is assumed to have a tube 11′ aperture. In this example: the height of the shield is bound by the height of the tube 11′ and a 2 mm clearance with sample plate 3; the maximum inner radius of the shield is defined by the preferred requirement (discussed above) for the inwardly facing surface of the shield to remain within the outer boundary of the sample plate carrier when the sample plate carrier is at a maximum permitted lateral offset with respect to the second electrode; the minimum inner radius of the shield can be optimised to control ion optic lensing in the extraction region; the maximum outer radius of shield is defined by the outer radius of the extraction electrode; the minimum outer radius of the shield can be optimised to control side field penetration for a given shield height. Thus, any combination of shield defining parameters within the hatched area of FIG. 6 may be preferred to control the side field penetration, but the most preferred values are defined by the solid line within hatched area of FIG. 6. This line defines the combinations of shield defining parameters, established by electrostatic modelling, that minimise the side field penetration into the region of ion formation and extraction within the extraction region. This line is of course specific to the ion source design shown here as example and similar analysis would be required to define parameters to achieve optimum field penetration control for other ion source geometries.

[0129] The plot of FIG. 6 plots minimum outer radius against height, and was produced by adjusting height and then calculating the optimum minimum outer radius for that height (having optimised for other parameters).

[0130] FIG. 7 shows field contours, on electrostatic model of an ion source incorporating the extraction electrode 9′ of FIG. 5(b) in a region around sample plate 3 and the extraction electrode 9′. As shown in this drafting, the field is symmetrical BOTH when the sample plate 3 is centred on the extraction electrode axis and when the sample plate 3 is laterally offset with respect to the extraction electrode 9′. This insensitivity to sample plate position is directly due to the shield preventing side field penetration into extraction region.

[0131] FIG. 8 shows a mass spectrum of peptides in the range 1-5 kDa, obtained experimentally using a mass spectrometer having an ion source that includes the extraction electrode 7 of FIG. 2 (with tube 11, but lacking shield 10), where the mass spectrometer was optimised/tuned with a sample located at centre of sample plate.

[0132] FIGS. 8 (a) and (b) show spectra obtained from centre and corner of the sample plate, respectively. As can be seen from FIG. 8, the amplitude of signal obtained at corner of sample plate has suffered a loss in intensity of approximately 90% with respect to amplitude at centre of sample plate.

[0133] FIG. 9 shows a mass spectrum of the same peptides obtained experimentally using a mass spectrometer having an ion source that includes the extraction electrode 9′ (with tube 11′, with shield 10′), where the mass spectrometer was again optimised/tuned with sample located at centre of sample plate. FIGS. 9 (a) and (b) show spectra obtained from centre and corner of plate, respectively. As can be seen from FIG. 9, the amplitude of signal obtained at corner of sample plate this time can be seen not to have suffered any significant loss in intensity or skewing of intensity distribution with respect to amplitude at centre of sample plate.

[0134] When used in this specification and claims, the terms “comprises” and “comprising”, “including” and variations thereof mean that the specified features, steps or integers are included. The terms are not to be interpreted to exclude the possibility of other features, steps or integers being present.

[0135] The features disclosed in the foregoing description, or in the following claims, or in the accompanying drawings, expressed in their specific forms or in terms of a means for performing the disclosed function, or a method or process for obtaining the disclosed results, as appropriate, may, separately, or in any combination of such features, be utilised for realising the invention in diverse forms thereof.

[0136] While the invention has been described in conjunction with the exemplary embodiments described above, many equivalent modifications and variations will be apparent to those skilled in the art when given this disclosure. Accordingly, the exemplary embodiments of the invention set forth above are considered to be illustrative and not limiting. Various changes to the described embodiments may be made without departing from the spirit and scope of the invention.

[0137] For example, although the examples depicted herein show the shield formed on the extraction electrode of an ion source, the shield could equally be applied to the sample plate of the ion source.

[0138] Moreover, although the examples depicted herein show the proposed shield as applied to the sample plate and extraction electrode of an ion source, it should be appreciated that the same principles could be applied to any electrode pair in a TOF mass spectrometer where voltages are applied to produce an electric field in a region between the first and second electrodes so as to influence (e.g. accelerate, decelerate, influence trajectory of, focus, defocus) ions present in the region between the first and second electrodes when the mass spectrometer is in use.

[0139] For the avoidance of any doubt, any theoretical explanations provided herein are provided for the purposes of improving the understanding of a reader. The inventors do not wish to be bound by any of these theoretical explanations.

[0140] All references referred to above are hereby incorporated by reference.