The invention relates to a method for generating motion in a thin liquid film and a device for carrying out the method.
In chemical, biological or microbiological analysis it is often necessary to generate motion in liquid films so as to mix it thoroughly. In this way reactions can for example be forwarded or the liquid can be homogenised.
Such liquid films can be applied for example in microarray experiments for examining macromolecules such as proteins, nucleic acids, antigens or antibodies. A fast method for analysing macromolecules uses microarrays, in which known first, if required different macromolecules are arranged at various points for example in a matrix form on a substrate. These macromolecules are also designated as probe molecules. A liquid with second macromolecules (sample molecules) is sluiced over the microarray, which with at least a type of probe molecules on the microarray a specific can enter into a bond (hybridising). If the liquid is then removed from the surface, the sample molecules to be examined remain only at the sites of specific binding. Locally resolved measuring, for example fluorescence measuring, can be used to ascertain at which sites sample molecules are present. It can therefore be determined from the known position of the individual probe molecules in the matrix form of the microarray, with which type of macromolecules the macromolecules to be examined have entered into a specific bond.
The duration of a corresponding analytical experiment is determined to a considerable extent by the diffusion of the sample molecules to the probe molecules and can therefore take a while. If for example the concentration of the macromolecules to be examined in the liquid is only minimal, then it can last a very long time until it has found its specific bonding partner on the array. Therefore a device, with which the liquid can be thoroughly mixed, is preferred in order to achieve homogeneous distribution of the macromolecules on the microarray at any given time.
Thorough mixing in liquid drops on a surface is described in DE-A-101 17 772. The piezoelectric sound transducer, by means of which surface acoustic waves are generated, comprises for example an interdigital transducer.
Such interdigital transducers are metallic electrodes designed in the manner of combs, whereof the double finger distance defines the wavelength of the surface acoustic wave and which can be produced by the optical photolithography process for example in the range around the 10 μm finger distance. Such interdigital transducers are provided for example on piezoelectric crystals for the purpose of exciting surface acoustic waves in a manner known per se on said crystals.
With this type of known method there is the danger of corrosion of the interdigital transducer by liquid, or the influence of the liquid and reactants possibly contained therein through contact with the interdigital transducer. To avoid such disadvantages the surface of the piezoelectric crystal, on which the interdigital transducer is located and on which the surface acoustic wave is generated, can be provided with a passivation layer. This coating must be removed again on the electrical contact surfaces of the interdigital transducer, necessitating a lithography and etching stage.
The generation of a flow in liquids by means of acoustic waves is described in Wesley Le Mars Nyborg “Acoustic Streaming” in Physical Acoustics 2B; ed. W. P. Mason; Academic Press 265 (1965).
The motion of liquids by means of surface acoustic waves is illustrated in S. Shiokawa et al., IEEE Proceedings of the Ultrasonics Symposium 1989, pages 643 ff.
Finally, it would be desirable if a mixing process could be made available not only for liquid drops, but also for liquid films and which can be used for example also for liquid films in a capillary gap.
The object of the present invention is to provide an improved method and an improved device for generating motion in a thin liquid film, in particular in a capillary gap, which are in addition easy and cost-effective to produce and utilise.
This task is solved by a method having the characteristics of Claim 1 and a device having the characteristics of Claim 15.
The liquid film is separated from the at least one ultrasound generating device by the substrate. A separate passivation or protective layer, which would keep the ultrasound-generating device away from the liquid film, is not required. The method is easy and cost-effective to carry out. The application for liquid films, which are limited by a capillary gap, is particularly advantageous.
With the inventive method therefore a liquid film having a thickness for example of a few micrometers to 5 millimetres can be separated by the substrate from the ultrasound device, for example a piezoelectric sound transducer, which generates acoustic waves in a frequency range of a few MHz to a few 100 MHz. The piezoelectric sound transducer can have a size of a few square millimetres to a few square centimetres and a thickness of a few 10 micrometers to a few millimetres.
By way of advantage the substrate is thinner than a few centimetres, though thicker than ¼ of the ultrasound wavelength. So-called “flexural plate wave modes” or Lamb modes forming in the substrate can therefore effectively be prevented. It can have a surface of a few square millimetres to a few 10 square centimetres.
No coating of a piezoelectric crystal surface with subsequent lithography or etching is necessary in particular for the inventive method. The inventive method enables applications with liquid films on metallised surfaces, which can lead to short circuits for example of the interdigital transducer with the known method.
The surface facing the liquid is a planar substrate surface in the case of the inventive method. Fluidic problems, which result due to various lateral surface characters of the substrate, can be absent. A smooth substrate surface is easier to clean than a heterogeneous surface.
The acoustic wave input in the liquid causes a flow along closed flow lines. The acoustic wave itself is strongly localised in the liquid stark around the input site. The range of the flow depends on the gap thickness and is all the greater the wider the capillary gap is. As a rule the flow rate drops exponentially with the distance from the input site. In the case of a gap height of ca. 200 μm the decrease in the flow rate is to be seen by a factor of 10 pro millimetres distance. In a capillary gap of 100 μm high the performance must be increased ca. By a factor of 10 on the other hand to increase the range by 1 mm, in which homogeneous thorough mixing is achieved.
Yet to ensure homogeneous mixing of a larger liquid film, for example several ultrasound wave-generating devices can be provided to generate several ultrasound waves at different places. However, it is particularly advantageous if ultrasound is input into the liquid film by means of an ultrasound wave-generating device, such that the liquid at least is set in motion at two poles of motion or respectively input sites. A lateral distance of a few 100 μm, preferably several millimetres, is possible for example. Depending on the desired requirement the poles of motion can be arranged such that their working fields overlap or lie further apart.
Two poles of motion or respectively input sites can be received for example by means of an ultrasound wave-generating device, which radiates bidirectionally.
In an embodiment of the invention the ultrasound wave is generated by means of a surface wave-generating device, preferably an interdigital transducer on the side of the substrate facing away from the liquid film.
Using such an interdigital transducer volume acoustic waves can be generated variously in the substrate, which pass through the latter obliquely. The interdigital transducer generates a bidirectionally radiating limit surface wave (LSAW) on the limit surface between the piezoelectric crystal and the substrate, to which it is applied. This limit surface leakage wave radiates energy as volume acoustic waves (BAW) into the substrate. The amplitude of the LSAW decreases exponentially, whereby typical decay lengths are approximately 100 μm. The radiation angle α of the volume acoustic waves in the substrate measured against the normal of the substrate results from the arcussinus of the ratio of the speed of sound Vs of the volume acoustic wave in the substrate and the speed of sound VLSAW of the limit surface acoustic wave (α=arcsin (Vs/VLSAW) created with the interdigital transducer. Radiating in the substrate is therefore only possible if the speed of sound in the substrate is less than the speed of sound of the limit surface leakage wave. As a rule therefore transversal waves are excited in the substrate, because the longitudinal speed of sound in the substrate is greater than the speed of the limit surface leakage wave. A typical value for the limit surface leakage wave speed is for example 3900 m/s.
The piezoelectrically caused deformations under the interdigital transducer fingers engaging in one another in the manner of combs radiate volume acoustic waves (BAW) directly into the substrate. In this case this produces an angle of radiation a measured against the normal of the substrate as arcussinus of the ratio on the one hand to the speed of sound in the substrate Vs and on the other hand to the product from the period of the interdigital transducer IIDT and the applied high frequency f (α=arcsin (Vs/(IIDT·f)). For this sound input mechanism the angle of incidence relative to the normal, the angle of levitation α, can therefore be preset by the frequency. Both effects can occur adjacently.
Both mechanisms (LSAW, BAW) enable oblique radiating of the substrate. The entire electrical contacting of the interdigital transducer takes place on the side of the substrate facing away from the liquid film, so that corrosion of the electrical contacting by aggressive liquids is excluded.
The piezoelectric crystal supporting the interdigital transducer can be stuck, pressed or bonded on the substrate or can be stuck, pressed or bonded on the substrate via a coupling medium (for example electrostatically or via a gel film). Likewise the piezoelectric crystal can constitute the substrate itself.
The use of a substrate material, which has a slight acoustic damping at operating frequency, is also particularly advantageous. The volume acoustic wave in the substrate is partly reflected on the limit surface to the gap, and only a fraction of the sound energy enters the liquid. With weaker damping in the substrate the reflected beam can be again input into the gap following further reflection on another substrate surface at another place. In this way the substrate is utilised like a fibre optic to conduct and distribute the volume acoustic wave in the substrate to several places of the gap. At the same time the substrate is selected advantageously such that on the limit surface between substrate and liquid part of the ultrasound energy is decoupled, which serves as motion for the liquid film. On the other limit surface of the substrate the greatest possible reflection should enter. Quartz glass for example has proven particularly suitable at a frequency of 10 MHz to 250 MHz, preferably 100 MHz to 250 MHz, which has almost complete reflection on a limit surface to air, and has approximately 10% to 20% decoupling on the limit surface between substrate and liquid.
Located on the piezoelectric substrate in an embodiment are one or more interdigital transducers for generating the ultrasound waves, which are contacted either separately or are contacted together in series or in parallel to one another, and which can be controlled separately by choice of frequency with varying finger electrode distance.
It is particularly easy to produce homogeneous thorough mixing also by frequency-controlled variation of the input site. This raises the limit of the mixing range, in that the input site and thus active mixing area is pushed around the latter over the surface to be mixed.
The use of interdigital transducers with non-constant finger distance (“tapered interdigital transducer”), as described for another application for example in WO 01/20781 A1, enables selection of the radiation site of the interdigital transducer by means of the applied frequency. In this way it can be ascertained precisely at which place the ultrasound wave is input into the liquid.
By using a tapered interdigital transducer, which additionally has finger electrodes not designed straight, for example finger electrodes engaging in one another in particular curved, the direction of radiation, therefore the azimuthal angle θ in the limit surface, can be regulated by varying the operating frequency. And on the other hand also the angle of levitation a can be changed with the frequency by direct BAW generation on the interdigital transducer.
It is possible to very precisely determine those parts of the liquid film, which are to be moved by the ultrasound wave, by the frequency dependencies described. Mixing patterns, which are better suited to mixing liquids than stationary flows, can thus be created temporally non-stationary for example through continuous variation of the frequency.
An advantage with the inventive method according to these configurations using interdigital transducers on the side of the substrate facing away from the liquid is the independence of the electrical impedance of the interdigital transducer electrode from the electrical conductivity of the sample and the possibility of being able to use this method on metallicised slide supports. Since the interdigital transducer electrode required to create the acoustic wave is removed through the thickness of the substrate from the metallicised surface and the possibly electrical conductive liquid, no capacitive short circuit can occur, as can happen in the method described in DE-A-101 17 722, and also the impedance of the interdigital transducer electrode is not influenced by the conductivity of the sample solution. Therefore more stable electrical impedance matching to the high-frequency generator electronics is possible than with the previously known method.
In an alternative embodiment a limit surface acoustic wave is generated by means of a surface wave-generating device, preferably an interdigital transducer, on a front surface of the substrate. This produces oblique radiation of a volume acoustic wave in the substrate in the manner described. This volume acoustic wave is likewise input obliquely into the liquid film in contact with a main surface possibly by reflection on the main surfaces.
With the described embodiments ultrasound energy can be input at different places into the liquid film. By suitable selection of geometry, for example the thickness of the substrate, these input sites can be ascertained locally exactly. With such a method several input sites are produced, without a large number of surface wave-generating devices being necessary. Problems, which might arise with wiring or a plurality of surface wave-generating devices, are thus avoided.
Since an interdigital transducer electrode generally sends out two volume acoustic waves in the substrate, two unidirectionally driven surface elements of the same size separated lateral from one another on the limit surface between substrate and liquid film result as flow source with the inventive method, instead of a locally concentrated bidirectionally driven surface element of the size of the interdigital transducer electrode. Accordingly the fluidic effective cross-section, therefore the surface via which an interdigital transducer electrode is fluidically active and which can intermix an interdigital transducer electrode, is clearly enlarged. An additional outcome of this is greater flexibility in the arrangement of the flow sources. For example, the distance of both unidirectionally driven flow sources from one another grows with the thickness of the substrate. Two liquids separated from one another can thus be mixed, for example using an interdigital transducer element.
To prevent reflections occurring in unwanted places the ultrasound wave can be randomly dispersed by appropriate selection of a randomly dispersive front surface of the substrate. To this end at least one surface of the substrate is roughened, for example. This effect can also be utilised for targeted enlargement.
In order to guide the spread of the ultrasound wave in the substrate in preferred directions, correspondingly and angularly arranged reflection surfaces can be provided preferably on the front surfaces not corresponding to the main surfaces. The ultrasound wave can be guided in a predetermined manner with such reflection surfaces.
An inventive device for carrying out an inventive method has a substrate with an ultrasound wave-generating element. A main surface of the substrate is provided for contact with the liquid film. The ultrasound wave-generating device is designed such that the ultrasound wave is input obliquely into the substrate. It is particularly advantageous if the ultrasound wave-generating device is arranged on a main surface of the substrate, which is arranged opposite the liquid film. However it is also conceivable that the ultrasound wave-generating device is arranged on another surface and the ultrasound wave is guided to the liquid film by reflection inside the substrate.
By using a substrate material, which has minimal acoustic damping and corresponding reflection coefficients on the limit surfaces, a device can be provided, in which a greater range of the acoustic beam is achieved as described by reflection on the limit surfaces.
Another inventive device for carrying out an inventive method has a substrate with an ultrasound wave-generating element on a front surface, which is not a main surface. The angle of radiation of the volume acoustic wave generated in the substrate is generated by the speed of sounds inside and outside the substrate. In this way the volume acoustic wave is generated obliquely in the substrate and the part propagating away from the liquid film is reflected at least in part by reflection on the main surface facing away from the liquid film in the direction of the limit surface between substrate and liquid film.
Devices with interdigital transducers of the already abovedescribed geometry can be employed to create the abovementioned effects.
The invention will now be explained in detail by means of the accompanying schematic figures, in which:
a: is a schematic lateral sectional view through an arrangement for carrying out a first design of the inventive method,
b: is a schematic lateral sectional view through an arrangement for carrying out a first embodiment of the inventive method, whereby the direction of radiation is indicated for different frequencies,
a to 6c: are schematic sectional views of different embodiments of the electrical contacting of a device for carrying out an inventive method,
a: is a lateral schematic sectional view through an arrangement for carrying out a sixth embodiment of the inventive method,
b: a schematic sectional view in the direction of viewing A of
a: is a lateral schematic sectional view through an arrangement for carrying out a seventh embodiment of the inventive method,
b: is a sectional view in the direction of viewing B of
a: is a schematic plan view of a cross-section of an arrangement for carrying out an eighth embodiment of the inventive method,
b: is a schematic plan view of a cross-section of an arrangement for carrying out a ninth embodiment of the inventive method,
In
The substrate 1 is mounted by way of spacers 13 on a further substrate 11, for example likewise a slide support made of glass. The spacers can be separate elements or can be formed integrally with one of the substrates 1, 11.
In between the substrates 1 and 11 is a liquid film 7, to be mixed. The capillary gap, in which the liquid 7 is located, is a few micrometers, for example 30 to a few 100 micrometers. Positioned on the substrate 11, for example a slide support, can be for example a microarray, which has spots in a regular arrangement, to which different macromolecules are bound. In the liquid 7 for example other macromolecules are present, whereof the reaction properties are to be examined with the macromolecules of the microarray.
An embodiment is shown, in which the ultrasound wave-generating device is arranged on the side of the substrate 1 opposite the liquid film. It is understood that in both this and in the following embodiments the ultrasound wave-generating device can also be arranged on the side of the other substrate 11 opposite the liquid film.
By means of the interdigital transducer ultrasound waves 9 can be generated in the given direction, which pass through the substrate 1 at an angle α to the normal of the substrate 1 as volume acoustic wave, as described hereinabove. Reference numeral 15 schematically indicates those areas of the limit surface between liquid 7 and substrate 1, which are essentially affected by the volume acoustic wave 9.
If float glass of for example 2 mm in thickness is used as substrate material the exit points 15 of the acoustic wave in the liquid are at a distance of ca. 8 mm and are arranged symmetrically to the sound source. If the interdigital transducer is operated at a high-frequency output of 500 mW, the range is then approximately 5 mm, in this case adequate for thorough mixing a liquid in a capillary gap over a microarray on the substrate 11 of a surface of 0.8 to 1.25 cm2.
b assists in explaining to point out, how different input angles can be set with an embodiment of
It is also possible, by applying a high-frequency field of suitable frequency to the interdigital transducer 3 on the piezoelectric crystal 5, to generate a surface acoustic wave, which is converted into a volume acoustic wave in the piezoelectric crystal 5 after brief propagation by means of structures (etched in periodic trenches, damped periodic metal strips) produced on the surface.
The electrical contacting of the interdigital transducer electrode in the embodiments of
In the embodiment of electrical contacting, as is illustrated in
In the embodiment of electrical contacting, as is illustrated in
A further possibility for supplying the electrical power to the piezoelectric sound transducer is inductive input. At the same time the electrical supply lines to the interdigital transducer electrodes are configured such that they act as an antenna for contactless control of the high-frequency signal. In the simplest case this is an annular electrode on the piezoelectric sound transducer, which acts as a secondary circuit of a high-frequency transformer, whereof the primary circuit is connected to the high-frequency generator electronics. This is held externally and is arranged directly adjacent to the piezoelectric sound transducer.
In the extreme case of such an arrangement the angle β can also be 90°. Then the acoustic modulator 300 is arranged on a front surface of the substrate 1. This arrangement of the acoustic modulator 300 is indicated in
Not illustrated in
In
The described inventive devices can be used with an inventive method as follows.
For carrying out an inventive method it suffices for the piezoelectric element 5 to be pressed firmly onto the substrate 1. The substrate 11 can be provided with a microarray, for example.
A substrate 1 with a surface acoustic wave-generating device is set thereon by way of spacers 13, as is shown in
In an embodiment of
In an embodiment of
Further inventive arrangements are described hereinafter.
Through appropriate selection of the geometry, for example the thickness of the substrate, in this way the points 75, at which a part of the ultrasound wave is input from the substrate 71 into the liquid 72, can be determined locally precisely, and in this way a desired pattern of motion can be generated in the liquid 72.
By using suitable modulation of the ultrasound output, for example by switching the interdigital transducer 73 off and on, a current can be generated in this way, which is suited to move the fluid in one direction. With the arrangement of
a shows a plan view of a cross-section of an arrangement, approximately at the level of the limit surface between liquid 72 and substrate 71, which enables particular deflection of the acoustic beam in the substrate 71. Acoustic beams 74, which encounter the limit surface between the liquid and the substrate 71 at points 75, go out from the interdigital transducer 73 such as described with reference to
In
The described reflection effect via selection of a suitable substrate material can likewise be generated by means of a volume oscillator 83, as shown in
A similar effect is achievable with an embodiment of
In the embodiment of
Individual embodiments of the method or respectively the features of the described embodiments can also be combined in suitable form also so that the targeted effects can be achieved at the same time.
Number | Date | Country | Kind |
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10308622.6 | Feb 2003 | DE | national |
10309183.1 | Mar 2003 | DE | national |
10325313.0 | Jun 2003 | DE | national |
Number | Date | Country | |
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Parent | 10547263 | Jul 2006 | US |
Child | 12870033 | US |