CN108136283A - Large-scale acoustically separated device - Google Patents

Abstract

Disclose the device (100) for secondary fluid or particle to be detached with main fluid.Described device includes sound chamber (107), the fluid outlet (114) positioned at the top of sound chamber, the concentrated solution outlet (116) positioned at the bottom end of sound chamber and the entrance (112) positioned at first side (122) of sound chamber (107).It ultrasonic transducer (106) on the side wall of sound chamber and generates multidimensional standing acoustic waves in sound chamber (107) in the reflector of opposite side-walls, multidimensional standing acoustic waves capture particle (such as cell) and detaches particle (such as cell) with main fluid.Main fluid is collected by fluid outlet (114), and particle exports (116) by concentrate and collects.Described device is a large-scale plant that can handle several litres of liquid per hour, and with larger inner space.

Description

Large-scale acoustically separated device

Cross reference to related applications

This application claims the preferential of the U.S. Provisional Patent Application Serial No. 62/211,142 submitted for 28th in August in 2015 Power；And the it is required that priority of U.S. Provisional Patent Application Serial No. 62/252,068 submitted on November 6th, 2015.These The disclosure of application is incorporated herein by reference in their entirety.

Background technology

Sound swimming is particle and secondary fluid to be detached from secondary working fluid or main fluid, and not using high intensity standing acoustic waves Use film or physical size exclusion filter.It is well known that when density and/or compressibility have difference, high intensity sound is stayed Wave can be to the particle applied force in fluid, further referred to as acoustics reduced factor.Pressure distribution in standing wave includes its node The region of local maximum at the Local Minimum pressure amplitude at place and its antinodal points.According to the density and compressibility of particle, They will be captured at the node or antinodal points of standing wave.In general, the frequency of standing wave is higher, due to the pressure of standing wave, can catch The particle obtained is smaller.

Have been proven that (density and sound pass through with different acoustics reduced factors on MEMS (MEMS) scale The combination of the speed of material) material separation (for example, from the acoustically separated secondary fluid of secondary working fluid or from secondary working fluid stream sound Credit is from particle).On the mems scale, traditional sound swimming system is depended on using half-wave or quarter-wave sound chamber, this Sound chamber is usually less than a millimeters thick under several megahertzs of frequency, and is worked with low-down flow velocity (for example, μ L/ minute).This The system of sample does not have scalability, because they benefit from extremely low Reynolds number, laminar flow operation, and needs minimum fluid Dynamics Optimization.

On a macroscopic scale, plane standing acoustic waves has been used for completing this separation process.However, single plane wave is usual Particle or secondary fluid are captured in this way, i.e., can only be detached them with secondary working fluid by closing plane standing wave.This Do not allow to operate continuously.Moreover, the energy generated needed for acoustics plane standing wave often passes through wasted energy amount heating primary fluid.

Therefore, traditional sound swimsuit is put due to many factors and has effects that limited, these factors include fever, use Plane standing wave, limitation fluid flow and cannot capture different types of material.Therefore it needs to provide the cluster of particle for generating optimization Roll into a ball the system and method to improve Gravity Separation and collection efficiency.The sound improved swimsuit is needed to put, is moved using improved fluid Mechanics, so that sound swimming becomes a continuous process.

Invention content

It in various embodiments, can this disclosure relates to be put with improved fluid dynamic macro-scale sound swimsuit For improving the separation of particle (such as cell) and particle/fluid mixture.More specifically, described device is included containing ultrasound The sound chamber of energy converter and reflector, the ultrasonic transducer and reflector generate multidimensional standing acoustic waves.

Disclosed herein is put for the sound swimsuit that detaches secondary fluid or particle with primary/main fluid.For example, particle can To be the cells such as Chinese hamster ovary (CHO) cell, NS0 hybridomas, baby hamster kidney (BHK) cell or people's cell； Lymphocyte, such as T cell (such as regulatory T cells (Treg), Jurkat T cells), B cell or NK cells；Before them Body, such as peripheral blood mononuclear cells (PBMC)；Algae or other plant cell, bacterium, virus or microcarrier.

Sound swimsuit is disclosed in various embodiments to put, including：Sound chamber, including at least one at its first end Entrance；In at least one fluid outlet for the top end that sound swimsuit is put；In at least one concentrate for the bottom end that sound swimsuit is put Outlet；At least one ultrasonic transducer is coupled to the sound chamber, and at least one ultrasonic transducer includes piezoelectric material, The piezoelectric material is configured as being driven to generate multidimensional standing acoustic waves in the sound chamber by voltage signal；And relative to institute State the reflector that at least one ultrasonic transducer is located at sound chamber's opposite side；Wherein, the sound chamber is included by length and width The planar cross-sectional area of restriction and by the width and the side section area of High definition, wherein the length is more than or waits In the width, and wherein described planar cross-sectional area is more than side section area.

At least one entrance can be a part for dump diffuser.At least one entrance can be included across piezoelectricity About 60% height of the height of material.The base portion of at least one entrance can be positioned along the base portion of piezoelectric material.Dump Diffuser can be included at least one entrance flowing ports at the upper end of pumping chamber and the stream at the lower end of pumping chamber Dynamic outlet, the flowing outlet have such shape, which is provided perpendicular to being generated by least one ultrasonic transducer The flow direction of the axial direction of multidimensional standing acoustic waves.

In general, dump diffuser is used to make the flowing of entrance more by reducing in sound chamber the inhomogeneities caused by gravity Uniformly, so as to which the efficiency for putting sound swimsuit maximizes.At least one entrance can be configured to allow fluid at least 800 millis Liter/min flow velocity enter sound chamber, and fluid collector can be configured to allow for fluid with the stream of at least 25 ml/mins Speed leaves sound chamber.

In some embodiments, at least one entrance enters including first at the first end of the sound chamber Mouthful and the second entrance at the second end opposite with its first end of the sound chamber so that fluid stream enter sound chamber be it is uniform and Symmetrically.

Some embodiments that the sound swimsuit is put are additionally included in below at least one entrance and lead to described at least one First skew wall of a concentrate outlet, wherein first skew wall is included relative to first level face into about 11 ° to about 60 ° of angle Degree.

At least one energy converter can be across multiple energy converters of the length of sound chamber.Multiple energy converters can be with arranged in series Into single.In some embodiments, multiple energy converters include first row, which includes at least two energy converters, positioned at packet The top of second row containing at least two energy converters.At least one concentrate outlet can include multiple concentrates and export.

The sound chamber can include at least 40 cubic inches of volume.

In the various embodiments put in the sound swimsuit, what oblique top wall, the roof of parabolic curved or hypocycloid were bent Roof leads at least one fluid outlet from the first end and second end of sound chamber.In other embodiments, at least one stream Body exports the middle section for being connected to sound chamber.

The multidimensional standing acoustic waves can include axial force component and transverse force component with same order.

The ultrasonic transducer can include：Housing with top, bottom end and inner space；And positioned at housing bottom end Crystal, the crystal have exposure outer surface and inner surface, the crystal sound wave can be generated when being driven by voltage signal. In some embodiments, back sheet contacts the inner surface of crystal, and back sheet is made of substantially acoustic window material.Substantially acoustic window material Can be cork wood, cork or foam.Substantially acoustic window material can have up to 1 inch of thickness.Substantially acoustic window material can To be grid pattern.In other embodiments, the outer surface of crystal is covered by thickness for half-wavelength or smaller wearing face material Lid, wearing face material are polyurethane, epoxy resin or silicone coatings.The outer surface of crystal can also have wearing face, institute Wearing face is stated to be formed by the matching layer or wearing plate for adhering to the material of crystal outer surface.Matching layer or wearing plate can be by oxygen Change aluminium to form.In other embodiments, crystal does not have back sheet or wearing layer, that is, crystal is without back sheet or wearing layer.

The multidimensional standing acoustic waves can be three-dimensional standing wave.

A kind of sound swimsuit is also disclosed in various embodiments to put, including：Sound chamber is included at its first end at least One entrance；In at least one fluid outlet for the top end that sound swimsuit is put；In at least one dense of the bottom end that sound swimsuit is put Contracting object exports；At least one ultrasonic transducer, is coupled to the sound chamber, and at least one ultrasonic transducer includes piezoresistive material Material, the piezoelectric material are configured as being driven to generate multidimensional standing acoustic waves in the sound chamber by voltage signal；It is and opposite It is located at the reflector of sound chamber's opposite side at least one ultrasonic transducer；Wherein, at least one entrance is in turn The form of diffuser is stored up, the dump diffuser is included in the flowing outlet at the preceding lower end of pumping chamber, in the pumping chamber First entrance flowing ports at upside and the second entrance flowing ports at the rear upper end of the booster.

It can be from 1 milliliter about per minute to 800 milliliters about per minute by the flowing velocity of sound chamber.The dress of the disclosure It puts for cell concentration from every milliliter of fluid down to 50,000 cells to 80,000,000 cell of every milliliter of fluid, can have There is more than 90% separative efficiency.

In a particular embodiment, the multi-dimensional standing-wave generates acoustic radiation power, which has same order Axial force component and transverse force component.In a particular embodiment, standing acoustic waves can be multidimensional standing acoustic waves (for example, three-dimensional Standing acoustic waves).The example of this multidimensional standing acoustic waves can be found in jointly owned U.S. Patent number 9,228,183, Full content is entirely incorporated by reference.

These and other unrestricted features are described in further detail below.

Description of the drawings

It is the brief description to attached drawing below, for illustrating exemplary embodiment disclosed herein rather than in order to limit The present invention.

Fig. 1 is the exterior perspective view put according to the first exemplary sound swimsuit of the disclosure.The device has level cross-sectionn Product is more than the sound chamber of its vertical cross sectional area.

Fig. 2 is the cross-sectional view that the sound swimsuit of Figure 12 is put.

Fig. 3 A-3D illustrate four exemplary embodiments of the roof to form fluid path, and the fluid path is swum from sound The sound chamber of device leads to the fluid outlet at the top of device.Fig. 3 A show that its flat outer surface has different angles from flat inner surface The roof of degree.Fig. 3 B show its flat outer surface with roof of the flat inner surface with equal angular (that is, with constant thickness Roof).Fig. 3 C show the outer surface being bent with hypocycloid and interior surface (that is, fluid path quickly becomes It is narrow) roof.Fig. 3 D show to form the roof of fluid path, and the fluid path is connected only to the middle section of sound chamber.

Fig. 4 A-4D show the exemplary arrangement that the sound swimsuit with one or more concentrate outlets is put.With more In the device of a concentrate outlet, outlet is evenly spaced from each other.Fig. 4 A show the base portion there are one concentrate outlet with tool Device.Fig. 4 B show the device there are one the base portion of concentrate outlet with tool.Fig. 4 C show that there are three dense with tool The device of the base portion of contracting object outlet.Fig. 4 D show device of the tool there are four the base portion of concentrate outlet.

Fig. 5 A-5C show the exemplary embodiment of transducer assemblies put according to the sound swimsuit of the disclosure.Fig. 5 A are shown Piezoelectric energy-conversion device assembly, the piezoelectric energy-conversion device assembly include total of six with the rectangle transducing of two rows of three energy converters arrangements Device.Fig. 5 B show piezoelectric energy-conversion device assembly, the piezoelectric energy-conversion device assembly include total of six with it is single be arranged side by side it is rectangular Energy converter.Fig. 5 C show piezoelectric energy-conversion device assembly, which is arranged to two rows of rectangles including total of five and changes Energy device, wherein upper row includes two energy converters, and lower row includes three energy converters.

Fig. 6 show to standing acoustic waves capture particle cluster simulation, the standing acoustic waves by Fig. 5 C transducer assemblies Energy converter generate.

Fig. 7 A and Fig. 7 B show the more exemplary implementations for the piezoelectric energy-conversion device assembly put according to the sound swimsuit of the disclosure Example.Fig. 7 A are shown including total of three with the piezoelectric energy-conversion device assembly of the single matrix transducer being arranged side by side.Fig. 7 B are shown Include total of eight with the piezoelectric energy-conversion device assembly of the single square transducer being arranged side by side.

Fig. 8 is the stereogram of exemplary dump diffuser.

Fig. 9 is the side view of the exemplary dump diffuser of Figure 10.

Figure 10 is the front cross sectional view put according to the second exemplary sound swimsuit of the disclosure.The device also has level cross-sectionn Product is more than the sound chamber of its vertical cross sectional area.

Figure 11 is the forepart exterior perspective view of the device of Figure 10.

Figure 12 is the rear exterior stereogram of the device of Figure 10.

Figure 13 is that 1.5% yeast mixt of display is passed through with 810 ml/mins (mL/minute) flowing according to the disclosure 9 inches multiply 3 inches of reduction/clarification percentages multiplied when 2 inches of (length by width multiply height) sound swimsuits are put at any time (above Line) and phase contrast microscopy (following line) curve graph, the sound swimsuit put without dump diffuser and 60 volts, 80 Volt and 100 volts under work.Shallower line with dot represents device using film, and represents that device does not use with the deeper line just put Film.

Figure 14 is to show 3% yeast mixt with the flowing of 810 ml/mins by multiplying 3 inches according to 9 inches of the disclosure Multiply reduction/clarification percentage (line above) and difference when 2 inches of (length by width multiplies height) sound swimsuits are put at any time to show The curve graph of micro- art (following line), the sound swimsuit are put without dump diffuser, tool there are five alternate tangential flow It (alternating tangential flow, ATF) film and works under 60 volts, 80 volts and 100 volts.

Figure 15 is to show 3% yeast mixt with the flowing of 810 ml/mins by multiplying 3 inches according to 9 inches of the disclosure Multiply the curve graph of reduction/clarification percentage when 2 inches of (length by width multiplies height) sound swimsuits are put at any time, the sound swimsuit Tool is put there are five alternate tangential flow (ATF) film and is worked under 80 volts and 100 volts.Shallower line represents that device uses root According to the dump diffuser with double-row hole role of the disclosure, and deeper line represents that device is used and is turned according to half plate of the disclosure Store up diffuser.

Figure 16 is to show 3% yeast mixt with the flowing of 810 ml/mins by multiplying 3 inches according to 9 inches of the disclosure Multiply reduction/clarification percentage (line above) and difference when 2 inches of (length by width multiplies height) sound swimsuits are put at any time to show The curve graph of micro- art (following line), the sound swimsuit is put using half plate dump diffuser, five alternating tangential flows (ATF) The transducer assemblies of film and use with two rows of energy converters, wherein top row is closed and bottom comes and works under 100 volts.

Figure 17 is the exterior perspective view put according to the exemplary sound swimsuit of third of the disclosure.The embodiment, which particularly uses, to be turned Store up diffuser, wherein fluid along two different axis rather than only one axis (such as in Fig. 1) into dump diffuser pumping chamber.

Figure 18 is the stereogram of the lateral cross-section of the device of Figure 17.

Figure 19 is the side sectional view that the sound swimsuit of Figure 19 is put, and another aspect is shown together with Figure 18.

Figure 20 is the front view that the sound swimsuit of Figure 17 is put, and there is transparent wall to show supplementary features.

Figure 21 is the enlarged drawing of the flow chamber of the device of Figure 17.

Figure 22 is the amplification cross-sectional view for the transducer assemblies that the sound swimsuit of Figure 17 is put.

Figure 23 is the cross-sectional view of conventional ultrasound transducer.

Figure 24 is the cross-sectional view of the ultrasonic transducer of the disclosure.There are air gap in energy converter, and there is no back sheets Or wearing plate.

Figure 25 is the cross-sectional view of the ultrasonic transducer of the disclosure.There are air gap in energy converter, and there are back sheet and Wearing plate.

Figure 26 is the figure for showing the relationship between acoustic radiation power, gravity-buoyancy and Stokes resistance and particle size. Horizontal axis is with micron (μm) for unit, and the longitudinal axis is with newton (N) for unit.

Figure 27 is the electrical impedance magnitude of the square transducer driven with different frequency and the relational graph of frequency.

The captured line configuration of seven peak amplitudes of the ultrasonic transducer of Figure 28 A diagram disclosure.Figure 28 B are to show this public affairs The stereogram for the separator opened.Show fluid flow direction and captured line.Figure 28 C are the fluid flow directions along Figure 28 B (arrow 814) sees view during fluid inlet, shows the capture node of the standing wave of capture particle.Figure 28 D are along as in Figure 28 B The view that shown arrow 816 is taken along capture cable architecture across energy converter planar.

Specific embodiment

By reference to below to desired embodiment and including exemplary detailed description, can be more easily understood The disclosure.Following specification and claims will refer to many terms, should be defined as following meanings.

Although for the sake of clarity having used specific term in the following description, these terms are intended to refer only to What is selected in attached drawing is used to illustrate the specific structure of embodiment, and is not intended to define or limit the disclosure.In following attached drawing and In following description, it shall be noted that identical numeral mark represents the component with identity function.

Unless the context is clearly stated, otherwise "one" and " described " of singulative including plural referents.

Term " comprising " is used to that the component to be required to exist and other components is allowed to exist herein.Term " comprising " should Be construed to include term " by ... form ", only allow the component and may be by manufacturing the component and what be generated appoint What impurity exists.

Numerical value be construed as including when the effective digital for being reduced to identical quantity the identical numerical value of numerical value and with it is described The difference of value is less than in the application to determine the numerical value of the experimental error of the conventional measurement technology of the value.

All ranges disclosed herein includes the endpoint being previously mentioned and can be independently combinable (for example, the model of " 2 grams to 10 grams " It encloses including 2 grams and 10 grams of endpoint and all medians).The endpoint and any value of range disclosed herein are not limited to accurate model It encloses or is worth；They can be inaccurate, to include the value close to these ranges and/or value.

With the modifier " about " that quantity is used together including described value and with as the meaning indicated by context.When with When range is used together, modifier " about " is also considered as disclosing the range defined by the absolute value of two endpoints.Example Such as, the range " from about 2 to about 10 " also discloses that the range of " from 2 to 10 ".Term " about " can represent signified number Positive and negative 10%.For example, " about 10% " can represent 9% to 11% range, " about 1 " can represent 0.9 to 1.1.

It should be noted that many terms used herein are relative terms.For example, term "up" and "down" is that in position This is opposite, that is, and in given directions, upper member is located at position more higher than lower member, but if device is overturn, These terms can change.For given structure, term " entrance " and " outlet " be relative to flowing through their fluid, such as Fluid flows into the structure by the entrance and flows out the structure by the outlet.Term " upstream " and " downstream " are phases Flow through the direction of all parts for fluid, i.e., streaming flow first flow flowed again by upstream components by components downstream it. It should be noted that in the loop, the first component can be described as be in the upstream and downstream of second component.

Term " level " and " vertical " are used to indicate the direction relative to absolute reference, i.e. ground level.However, these terms It is not necessarily to be construed as requiring structure absolute parallel each other or absolute upright.For example, the first vertical stratification and the second vertical stratification are not It is certain parallel to each other.For referring to relative to absolute reference (i.e. earth surface), top is always high for term " top " and " bottom " In the surface of bottom.Term " upward " and it is " downward " be also for absolute reference；Up often fight the earth Gravity.

Term " parallel " should be interpreted in stereovision to keep approximately constant distance rather than tight between two surfaces These surfaces in the mathematical meaning of lattice will not intersect when extending to infinity.

This application involves " same orders ".If plurality divided by the quotient compared with decimal are at least 1 and the value less than 10, Two numbers have the identical order of magnitude.

Term " virus " refers to the infectious agent that can be only replicated in another living cells, and in addition with virion Form exists, and capsid of the virion by surrounding and containing DNA or RNA is formed, and in some cases, there is encirclement clothing The lipid envelope of shell.

Term " crystal " refers to monocrystalline or polycrystalline material as piezoelectric material.

Sound swimming be for from fluid dispersion remove particle low-power, without pressure drop, clog-free solid-state approach：That is, its It is used to realize the separation more generally carried out with porous filter, but it does not have the shortcomings that filter.Particularly, the disclosure Sound swimsuit, which is put, to be suitble to point in high flow rate running system is used together and operated on a macroscopic scale with bioreactor From.Sound swimsuit, which is put, to be designed to generate high intensity multi-dimensional ultrasound standing wave, and high intensity multi-dimensional ultrasound standing wave causes to be more than fluid resistance With the acoustic radiation power of buoyancy or the combined effect of gravity, and (i.e. remains stationary) suspended phase therefore can be captured (that is, thin Born of the same parents) sound wave is made there are more times to increase the concentration of particle, cohesion and/or coalescence.This has critically important with pervious method Difference, in pervious method, particle trajectories are only changed by the effect of acoustic radiation power.Therefore, in the present apparatus, radiation Power is used as the filter for preventing target particles (such as biological cell) from traversing standing wave plane.The capture ability of standing wave can root According to needing to change, such as the shape by changing the flow velocity of fluid, acoustic radiation power harmony swimsuit is put, with by capturing and settling Maximize cell retention.The technology provides a kind of green and sustainable alternative solution to detach secondary phase, and significantly reduces Energy cost.For as low as 1 micron of particle size, it has been demonstrated that outstanding particle separative efficiency.The sound swimming of the present invention Device has the ability for generating ultrasonic wave stationary field, which can capture flow velocity in flow field and be more than 1 ml/min (mL/minute) particle.

The sound field scattered out from particle forms three-dimensional acoustics radiant force, plays the role of 3 D captured field.When particle phase For wavelength it is smaller when, acoustic radiation power is proportional to particle volume (such as cube of radius).It is compared with frequency and acoustics The factor is directly proportional.It changes also with acoustic energy (such as square of acoustic pressure amplitude).For harmonic excitation, the sine space of power becomes Change be by particle driving to settling position in standing wave the reason of.When the acoustic radiation power being applied on particle is better than fluid resistance During with the combined effect of buoyancy/gravity, particle is trapped in standing acoustic waves field.Acoustic force (i.e. laterally and axially acoustic force) is right The concentration for acting through particle, reunion, aggregation, cohesion and/or the coalescence of captured particle form closelypacked cluster, when up to During to critical dimension, for the particle heavier than main fluid by the gravity continuous sedimentation of enhancing or for than main fluid more Light particle is risen by the buoyancy of enhancing.In addition, power between secondary paticle, such as Bjerkness power, contributes to particle aggregation.

Higher density and lower compressibility is presented than their culture medium that suspends in most of biological cell types, makes The acoustics reduced factor obtained between cell and culture medium has positive value.Therefore, axial acoustic radiation power (acoustic Radiation force, ARF) cell is driven towards standing pressure node.The axial component of acoustic radiation power will have face The cellular driven of specific factor is to pressure node plane, and it is anti-to be driven to pressure with the cell or other particles for bearing reduced factor Nodal plane.The radially or laterally component of acoustic radiation power is by the power of cell capture.The radially or laterally component of ARF is more than stream The combined effect of body resistance and gravity.For minicell or lotion, resistance FD can be expressed as：

Wherein U_fAnd U_pIt is fluid and cell speed, R_pIt is particle radius, μ_fAnd μ_pIt is the dynamic viscosity of fluid and cell, andIt is the ratio of dynamic viscosity.Buoyancy F_BIt is expressed as：

For the cell to be captured in multi-dimensional ultrasound standing wave, it can be assumed that dynamic balance or force vector on cell and be Zero, therefore can be given by the hope of the expression formula of lateral acoustic radiation power FLRF：

F_LRF=F_D+F_B。

Cell for known dimensions and material properties and for giving flow velocity, can estimate transverse direction using the equation The size of acoustic radiation power.

It is based on the formula developed by Gor'kov for calculating acoustic radiation power theoretical model.Primary acoustics spoke Penetrate the function that power FA is defined as field potential U, F_A=-▽ (U),

Wherein field potential U is defined as

And f₁And f₂It is the monopole and bipolar contribution being defined by the formula

Wherein p is acoustic pressure, and u is fluid particle speed, and Λ is cell density ρ_pWith fluid density ρ_fRatio, σ is cell velocity of sound c_p With fluid sound speed c_fRatio, V_oIt is cell volume, and<>Represent the time averaged with the period of wave.

The model of Gork'ov is the individual particle being directed in standing wave, and is limited to relative to the sound field in fluid and particle The small particle size of wavelength.It does not account for influence of the viscosity of fluid and particle to radiant force yet.Thus, which cannot For the macroscopical ultrasonic separation device being discussed herein, because particle cluster may increase quite big.Therefore will use not by The more complicated and complete model of the acoustic radiation power of particle size limitation.The model implemented be based on Yurii Ilinskii and The theoretical work of Evgenia Zabolotskaya such as AIP minutes (Conference Proceedings), rolls up 1474- 1, described in the 255-258 pages (2012).These models further include the influence of fluid and particle viscosity, and therefore more accurate Ground calculates acoustic radiation power.Additional internal model for calculating the sound force trapping of cylindrical object has been developed that , such as in standing wave be captured particle " hockey disk ", be very similar to cylinder.

The cross force of total acoustic radiation power (ARF) generated by the ultrasonic transducer of the disclosure is significant and is enough gram Take fluid resistance.Therefore, this transverse direction ARF can be used for staying in cell in standing acoustic waves when fluid flows through standing wave.In addition, such as It is upper described, this concentration that acts through particle, cohesion of the acoustic force (that is, laterally and axially acoustic force) to the particle that is captured And/or coalesce and result in closelypacked cluster, the cluster is due to the gravity (particle heavier than main fluid) of enhancing Or buoyancy (particle lighter than main fluid) and settle.It therefore, can be by the Gravity Separation of enhancing, by a kind of the opposite of material Larger solid is detached with the smaller particle of different materials, identical material and/or main fluid.

Multi-dimensional standing-wave is in the axial direction (that is, on the direction of standing wave, between energy converter and reflector, perpendicular to stream Dynamic direction) and horizontal direction (that is, in flow direction) generation acoustic radiation power.When mixture flows through sound chamber, suspended particulate exists By strong axial force component on standing wave direction.Since this acoustic force is perpendicular to flow direction and resistance, it can root Particle is quickly moved to pressure node plane or antinodal points plane according to the reduced factor of particle.Then, lateral acoustic radiation power Play a part of moving the particle of concentration towards the center of each plane node, so as to cause coalescing or assembling.Lateral acoustics Radiation force component overcomes fluid resistance, this allows particle agglomerate constantly to increase, and is then settled from mixture due to gravity.Often The resistance of a particle declines with the size increase of particle cluster and the acoustic radiation power of each particle is with particle cluster Size increases and declines, can together consider in the operation of acoustically separated device device or individually consider.In the disclosure extremely In some few examples, the transverse force component and axial force component of multidimensional standing acoustic waves have the identical order of magnitude.With regard to this point Speech, it is noted that in multidimensional standing acoustic waves, axial force can have the value different from cross force, for example, it is weaker or relatively strong or Can be equal or equivalent, but the cross force of multidimensional acoustic standing wave is more than the cross force of plane standing wave, sometimes big two orders of magnitude or More.

Sound swimming filter device can be used in a manner of at least two different.First, standing wave can be used for capture specific raw Object molecule (such as phytochemicals, recombinant protein or monoclonal antibody), and by these required products and cell, cell fragment It is detached with culture medium.Then specific biomolecule can be transferred and collect to be further processed.Secondly, standing wave can be used for Capture cell and cell fragment present in cell culture medium.Cell and cell fragment with positive reduced factor are moved to standing wave Node (rather than antinodal points).Since cell and cell fragment agglomerate at the node of standing wave, it also occur that cell training The physics for supporting base is cleaned, and thus more many cells are captured when contact with the cell having been held in standing wave.This usually will be thin Born of the same parents and cell fragment are separated from cell culture medium.When the cell condensation in standing wave to quality is no longer able to be kept by sound wave Degree when, the cell of captured aggregation and cell fragment can be settled by gravity from fluid stream, and can be with It is collected separately.In order to help this gravitational settling of cell and cell fragment, standing wave can be interrupted to allow all cells It is settled from the fluid stream filtered.This process is very useful for being dehydrated.Specific biomolecule may be removed in advance, Or it is retained in fluid stream (that is, cell culture medium).

In the disclosure, filling type bioreactor, which can also be used for generating, is subsequently used in various applications (including cell treatment Method) cell.In such method, the biological cell used in cell therapy is cultivated simultaneously in the bioreactor Amplification (that is, increasing the quantity of cell in bioreactor by cell proliferation).These cells can be lymphocyte, as T is thin Born of the same parents' (such as regulatory T cells (Tregs), Jurkat T cells), B cell or NK cells；Their precursor, such as peripheral blood mononuclear Cell (PBMC)；Etc..In filling type bioreactor, cell culture medium containing some cells (also known as main fluid) is from life Object reactor is sent to the filter device for generating standing acoustic waves.Most cells are captured and are maintained in standing acoustic waves, and remaining Main fluid and main fluid in other cells return to bioreactor.With the increase of the cell quantity of capture, their shapes Into the cluster of bigger, these clusters can be settled under critical dimension due to gravity from standing acoustic waves.Cluster can fall into sound Learn outside standing wave region such as less than standing acoustic waves, concentrate outlet, recycling cell can be exported from this concentrate for cell treatment Method.Only sub-fraction cell is captured and is removed from bioreactor by concentrate outlet, and rest part continues in life It is bred in object reactor, allows cell needed for continuous production and recycling.

In such applications, the sound swimsuit of the disclosure, which is put, can serve as cell holding meanss.System described herein is one Determine to operate under the cell recirculation rate of range, effectively cell is maintained in perfusion (or culture medium removal) speed range, and And it can be adjusted to be fully retained or optionally through certain percentage by the manipulation of fluid flow rate, transducer power or frequency The cell of ratio.Power and flow velocity can be monitored and used as the feedback of automatic control system.

Target cell can also be maintained at by using standing acoustic waves in the flow chamber of external filter devices so that other Part can be in close proximity to and in order to changing the purpose of target cell and be introduced into.Such operation will include capture T cell and It is subsequently introduced the slow virus material of the modification with specific gene montage so that the slow virus with specific gene montage will transfect T cell simultaneously generates the Chimeric antigen receptor T cell for being also referred to as CAR-T cells.

The acoustics filter device of the disclosure is designed to keep high-strength three-dimensional standing acoustic waves.The device is by function generator It is driven with amplifier (not shown).Device performance is by computer monitoring and control.It there may come a time when to need to adjust since acoustic streaming acts on The frequency or voltage amplitude of standing wave processed.The modulation can be modulated by amplitude and/or be completed by frequency modulation(PFM).Standing wave is propagated Duty ratio can be used for realize material capture certain results.In other words, it can be opened and closed with different frequencies Acoustic beam is to obtain desired result.

Compared with conventional equipment, the sound swimsuit of the disclosure puts the flow that can handle higher flow velocity and bigger.For inciting somebody to action The first exemplary embodiment that the sound swimsuit that secondary fluid or particle are detached with primary/main fluid puts 100 is shown in fig. 1 and 2 Go out.Fig. 1 is exterior perspective view, and Fig. 2 is the front cross sectional view of the device.The design of the device provides vertical plane or flowing Line of symmetry so that flowing evenly occurs in fluid when passing through the device.

Referring initially to Fig. 1, sound swimsuit is put 100 and is formed by side wall 110.As shown here, side wall 110 has rectangular shape, So that the device has the first side 122；It is spaced apart with the first side 122 and second side opposite with the first side 122 124；Front side 126；It is spaced apart with front side 126 and the rear side 128 opposite with front side 126；Top 130；And between top 130 It separates and the bottom end 132 opposite with top 130.Here there is also shown the braced frames 160 for the device.Supporting leg 162 is shown It is shown as extending from braced frame.Braced frame can be with the integral structure either detached with device 100 of device 100.

Referring now to Figure 2, roof 140 is located at the top of side wall 110, and base portion 150 is located at 110 lower section of side wall.Side wall 110th, roof 140 and base portion 150 closed interior space 107 together.At least one concentrate outlet 116 is located at the bottom of device 100 End.As will be further explained in this article, the particle of concentration will be exported by concentrate and leave inner space 107.Base portion is illustrated as For tool there are two skew wall 152, wall 152 slopes downwardly into concentrate outlet 116.It is noted that due to sectional view, these walls 152 are presented For straight line, and in three dimensions, wall is conical.

There are at least one fluid inlets 112 at the first side 122, and fluid is allowed to enter from the external of device 100 Inner space 107.As shown here, there are two fluid inlets 112, each fluid in each in side 122,124 Entrance.In the top end of device 100, there are at least one fluid outlets 114.As will be further explained in this article, fluid will pass through Fluid outlet leaves inner space 107.Fluid inlet 112 and fluid outlet 114 are also visible in Fig. 1.

Now referring to Fig. 1 and Fig. 2, at least one ultrasonic transducer 106 and at least one reflector 108 are internally positioned On the opposite side in space, and there are sound chambers 120 between them.As shown here, three ultrasonic transducers 106 are located at device Rear side 128 on, and reflector 108 is located on the front side 126 of device.

It should be noted that the volume of sound chamber 120 and inner space 107 are not coextensive.The volume of sound chamber is by side Wall 110 limits.On the contrary, inner space 107 further includes the space from roof 140 and base portion 150.It should also be noted that skew wall 152 have the interior angle A that with respect to the horizontal plane (being limited herein by the bottom of acoustics 121) measures, and wherein angle A is in embodiment About 10 ° to about 60 °, including about 30 ° to about 45 °.

Still referring to Fig. 1 and Fig. 2, sound chamber 120 has the length 101 between the first side and the second side； Width 103 between front side and rear side；And the height 105 of the High definition by ultrasonic transducer.Therefore, length 101 and width Degree 103 defines planar cross-sectional area (i.e. horizontal cross-sectional area), and width 103 and height 105 define side section area (i.e. vertical cross sectional area).As seen here, planar cross-sectional area is more than side section area.

In a particular embodiment, sound chamber 120 can have at least 40 cubic inches of volume so that can be handled in sound chamber big The fluid of volume.In this regard, the fluid inlet 112 of device may be configured to allow fluid per minute at least 800 milliliters (mL/min) flow velocity enters sound chamber.

Fig. 3 A-3D are the front views of the different roofs 140 of can be used in sound swimsuit is put four.Roof is formed swims from sound The fluid path of the fluid outlet 114 at the top of device is led in the sound chamber 120 of device.In these figures, each roof 140 has interior Surface 142 and outer surface 144.It should be noted that other roof shapes and construction can also be used, will such as see later herein 's.In a particular embodiment, fluid outlet, which can be configured as, allows fluid to flow out sound chamber with the flow velocity of at least 25mL/min.

Fig. 3 A show the roof 140 with flat outer surface 144, have the angle different from flat inner surface 142, So that roof is thicker near fluid outlet 114.Inner surface 142 from fluid outlet 114 extend to about with the sound chamber of Fig. 2 107 The identical length 141 of length 101.

Fig. 3 B show that flat outer surface 144 has the roof 140 with 142 equal angular of flat inner surface, i.e. roof has There is constant thickness.Equally, inner surface 142 extends to roughly the same with the length of the sound chamber of Fig. 2 107 101 from fluid outlet 114 Length 141.

Fig. 3 C show the inner surface 142 being bent with hypocycloid and the roof 140 of outer surface 144.Equally, inner surface 142 extend to the length 141 roughly the same with the length of the sound chamber of Fig. 2 107 101 from fluid outlet 114.The hypocycloid of inner surface Shape causes fluid path quickly to narrow up until fluid outlet 114.

Fig. 3 D show the roof 140 with inner surface 142, and inner surface 142 extends only to short length 141.In the reality It applies in example, length 141 is than 101 much shorter of length of sound chamber so that fluid is only discharged to fluid outlet from the middle section of sound chamber 114。

Fig. 4 A-4D are the front views of the different base portions 150 of can be used in sound swimsuit is put four.Base portion is formed swims from sound The fluid path of the concentrate outlet 116 of bottom of device is led in the sound chamber 120 of device.It should be noted that other shapes and construction Base portion is can be used for, as that will see later herein.Supporting leg 162 can also be seen herein, although they are also not required to It to be integrally formed with base portion.

Fig. 4 A show base portion of the tool there are one concentrate outlet 116.Two skew walls 152 lead to concentration from the side of sound chamber Object outlet 116.

Fig. 4 B also show base portion of the tool there are one concentrate outlet 116.Here skew wall 152 is than the skew wall in Fig. 4 A more It is shallow.

Fig. 4 C show base portion of the tool there are three concentrate outlet 116.It is evenly spaced from each other to export 116.Skew wall 152 is logical It is exported to each concentrate.

Fig. 4 D show base portion of the tool there are four concentrate outlet.It is evenly spaced from each other to export 116.Skew wall 152 leads to Each concentrate outlet.

Fig. 5 A-5C show three different embodiments of the transducer assemblies formed by multiple ultrasonic transducers, energy converter Component can be used for the sound swimsuit of the disclosure to put.Multiple energy converters allow the particle capture efficiency of bigger, especially when transducing utensil When having different resonant frequencies, so as to capture larger range of particle (such as cell) size.These transducer assemblies 170 along The length orientation of sound chamber, and side 122,124 and top 130 have been marked in every width figure, for the orientation of component.

Fig. 5 A show piezoelectric energy-conversion device assembly 170, and including total of six rectangular transducer 106, they are arranged as two rows 172,174, often arrange three energy converters.It is contemplated that in this arrangement, energy converter crosses over the entire width and height of sound chamber jointly Degree.

Fig. 5 B show piezoelectric energy-conversion device assembly 170, the piezoelectric energy-conversion device assembly 170 include total of six with single 172 simultaneously The square transducer 106 that arrangement is put.Energy converter crosses over the entire width of transducer assemblies jointly, but is not the whole of transducer assemblies A height.

Fig. 5 C show piezoelectric energy-conversion device assembly 170, which includes being arranged to two rows of total of five Rectangular transducer 106, wherein upper row 174 includes two energy converters, and lower row 172 includes three energy converters.It should be noted that Energy converter in upper row 174 interlocks/deviates relative to the energy converter in lower row 172.One benefit of this arrangement is shown in figure 6 Go out, indicate the position of multidimensional standing acoustic waves 176 that will be generated by energy converter.It can be seen, interlocking for energy converter makes It is also staggeredly, so as to be interlocked by the standing wave that upper row 174 generates and the standing wave in lower row 172 to obtain standing acoustic waves 176.Such as preceding institute It states, the particle/cell being trapped in multidimensional standing acoustic waves will agglomerate and form cluster, and the cluster is finally settled from standing wave And it is downwardly towards concentrate outlet.It is this staggeredly to avoid generating across in lower row 172 from upper row 174 to the cluster of whereabouts Standing acoustic waves so that the cluster formed in lower row 172 is not disturbed or broken.

It is also contemplated that multiple energy converters can be with arranged in series into single, such as in Fig. 7 A and Fig. 7 B like that.Figure 7A is shown including with the transducer assemblies 170 of the single total of three rectangular transducer 106 being arranged side by side.Fig. 7 B show tool Have with the transducer assemblies 170 of the single total of eight square transducer 106 being arranged side by side.

Turning now to Fig. 1 and Fig. 2, device 100 has the asymmetrical fluid entrance 112 being arranged on the opposite side of sound chamber. In specific embodiment, these entrances are the forms of dump diffuser (dump diffusers), and dump diffuser provides mainstream The mixture of body and particle more uniformly flows into sound chamber.

In brief, each dump diffuser includes entrance, and the mixture of main fluid/secondary fluid or particle is entered by this Mouth flows into hollow chamber.The mixture is filled with the chamber in dump diffuser, this reduce/eliminate gravity influence account for it is leading Flowing pulsation and flow irregularity as caused by pump, hose and horizontal inlet flow in the case of status.Then, mix level Ground flows out into sound chamber 107 from dump diffuser.Sound chamber's (black arrow) that dump diffuser brings heavier mixture into, position On cavity bottom and under ultrasonic transducer 106 and the node cluster formed in standing acoustic waves.This can be maximum It reduces to limit and flows into any interference that material generates cluster.

Dump diffuser is shown in Fig. 8 and Fig. 9 structurally and operationally.Fig. 8 is the dump diffuser 530 for removing foreboard Stereogram shows the inside and outside of dump diffuser.Fig. 9 is the stereogram of the foreboard of dump diffuser.

Since Fig. 9, dump diffuser 530 is included with upper end 532, opposite lower end 534, two side 538, preceding table Face 536 and the housing 531 of rear surface 539.Hollow chamber 540 is present in housing 531.Dump diffuser further includes reception mixing Object and the ingress port 542 for leading to chamber 540.Ingress port 542 is present on the upper end and side 538 of housing；Here have two A ingress port is visible.Figure 11 is mounted in the picture of the foreboard 546 in the front surface 536 of housing.As shown here, diffuser Outlet 544 is located on lower end 534 and in the form of two rounds, although these can also be the form of slit.

Referring now to Fig. 2 and Fig. 8, in use, the mixture of main fluid/secondary fluid or particle passes through ingress port 542 enter dump diffuser 530 and filled chamber 540.Then, pressure equably pushes away mixture by diffuser exit 544 Go out.These diffuser exits 544 also pass through the side wall 110 of device 100, and can also be considered as to enter inner space 107 Fluid inlet 112.Diffuser exit is placed on 121 top of bottom of sound chamber.In embodiment, diffuser exit is located at chamber Between the 0% and 100% of height 105 above bottom 121, in sound chamber, and 5% of the height specifically in sound chamber Between 25%, height 515.Diffuser exit 544 provides the axial direction for being parallel to the standing acoustic waves generated by ultrasonic transducer The flow direction in direction.Multiple diffuser exits are also disposed to so that they are in opposite position so that the level speed of fluid It spends the centre in sound chamber and will be reduced to zero.

It is desirably symmetrical by the flowing streamline of sound chamber because this minimize drop to concentrate outlet 116 with Inhomogeneities, vortex disturbance, cycle and the disturbance for the cluster being collected.Symmetry can also make inlet flow rate distribution and particle Gravity during collection maximizes.Because it is heavier than the penetrant discharged at the top of device, (opposite) weight enters mixing Object enters near sound chamber bottom.Axisymmetric inlet also assures the mixture of entrance due to gravity and in cavity bottom diffusion, and And it is provided from bottom to top close to uniform VELOCITY DISTRIBUTION.Due to two opposite inlet flow rates, the horizontal velocity of mixture With its center close to sound chamber, direction zero reduces and can be equal to zero.In this illustration, uniform speed contributes to Separation and collection result.Uniform speed avoids peak velocity, and peak velocity may prevent standing acoustic waves that particle is overcome to hinder Power, particle resistance may hinder cluster to increase and by gravity or buoyancy continuous leave standing acoustic waves.

As particle cluster settles, axial direction acoustic force associated with standing wave keeps cluster complete.This effect ensures Cluster with high terminal velocity hurtles down, the order of magnitude of about 1 cm/s.Compared with chamber flow velocity, the rate is very Soon, chamber flow velocity is the order of magnitude of 0.1 cm/s to 0.3 cm/s.The shallow wall angle of base portion means cylindrical particle cluster The distance declined before sound chamber is left can be very short, so as to which the dispersion of cluster hardly occur.Ideally, system with Energy converter carrys out work using 3 to 12 captured lines per square inch.Symmetry, the minimal flow in centeral acquisition zone domain disturbance and it is shallow Collector wall provide good powder collection.

Show that sound swimsuit puts 600 the second exemplary embodiment in figs. 10-12.Figure 10 is front cross sectional view.Figure 11 It is the positive exterior perspective view of the device.Figure 12 is the exterior perspective view at device rear portion.In the apparatus, the planar cross-sectional of sound chamber Face area is also greater than the lateral cross-section area of sound chamber.

Since Figure 10, the device 100 that sound swimsuit puts 600 and Fig. 1 has many similarities.Device 600 has first Side 122 and the second opposite side 124.There are side wall 110, roof 140 and base portions 150 to limit inner space 107.Dump Diffuser 530 is present on each side 122,124, serves as the fluid inlet 112 of the inner space 107 of device.Here, it pushes up Wall 140 includes the parabola inner surface 142 for leading to the fluid outlet 114 at top 130.Two concentrate outlets 116 are present in In base portion 152, skew wall 152 leads to each outlet at bottom end 132.Five ultrasonic transducers 106 are shown, rectangle indicates to produce The piezoelectric material of raw multidimensional standing acoustic waves.

One noticeable aspect (becoming apparent from Fig. 10) of the device is 530/ fluid inlet 112 of dump diffuser With respect to the placement relationship of ultrasonic transducer 106.As seen here, it is about piezoelectric material height 176 that fluid inlet 112, which has, 60% height 113.Moreover, the base portion 111 of fluid inlet 112 is positioned along the base portion 177 of piezoelectric material, i.e., it is aligned. In embodiment, the height of fluid inlet can be about the 5% to about 75% of piezoelectric material height.

Referring now to Figure 11, reflector 108 is visible on the front side of device 600 126.In addition, it could be seen that fluid outlet 114 and the rear sides 128 that lead to device from top/bottom end of inner space of concentrate outlet 116.

In fig. 11 it also can be seen that the alternative constructions of dump diffuser 530.There are two positions for dump diffuser tool in Fig. 8 In the entrance flowing ports 542 on side 538.Contrastingly, there are three entrance flowing ports for dump diffuser tool shown in Fig. 8 542.Two entrances flowing ports 542 are located on side 538.Third entrance flowing ports 542 are located on the upper end 532 of diffuser Rear surface 539 on.

Referring now to Figure 12, see ultrasound transducer assembly 170.Five connectors 171 are visible, a connector A visible energy converter 106 in 171 corresponding diagrams 10.

It is put and is tested using the sound swimsuit of Fig. 1.The size of sound chamber multiplies 3 inches for 9 inches and multiplies 2 inches of (length by width Multiply height).As shown in Figure 5A, which, which has, is arranged to six two rows of energy converters.Experiment measure water inlet/yeast mixt with The reduction of time/clarification percentage and compacting cell quality (packed cell mass, PCM).

In curve graph in fig. 13, yeast mixt is for 1.5% yeast and with 810 ml/mins (mL/minute) Flow velocity flows through device.The entrance of device is not a part (that is, without foreboard as shown in Figure 9) for dump diffuser.Device Ultrasonic transducer is run under 60 volts, 80 volts and 100 volts.Device uses sound passing membrane (acoustically transparent Films, ATF) it is operated, also operated in the situation without using any this ATF.As can be seen from Figure 28, having And without ATF in the case of, the PCM (following line) measured under 60 volts is about 20%-28%, under 80 volts about 28%-35% and the about 35%-38% under 100 volts.Have and without ATF in the case of, reduction/clarification hundred of mixture It is being about 75%-80% under 60 volts point than (line above), under 80 volts is being about 80%-90% and is about under 100 volts 85%-90%, although these devices seem there is slightly better separation/clarifying efficiency in the case of no ATF.Reduce by hundred Ratio and PCM values is divided all to show that operation causes water and yeast preferably to detach at higher voltages.

In the curve graph of Figure 14, yeast mixt is 3.0% yeast, and flows through dress with the flow velocity of 810 ml/mins It puts.Equally, the entrance of device is not a part for dump diffuser.The ultrasonic transducer of device is under 60 volts, 80 volts and 100 volts Operation.These devices are operated using five sound passing membranes (ATF).Here, the PCM (following line) of measurement is about under 60 volts 18%-25%, it under about 25%-29% and 100 volt is about 29%-30% under 80 volts to be.The reduction of mixture/clarification percentage It is about 55%-75% under 60 volts than (line above), is about 75%-82% under 80 volts, is about 75%- under 100 volts 82%.

In the curve graph of Figure 15, yeast mixt is 3.0% yeast, and flows through dress with the flow velocity of 810 ml/mins It puts.The entrance of the device is a part for the dump diffuser with foreboard.Deeper line represents dump diffuser, wherein foreboard It is configured as half plate (i.e. a large trough of foreboard bottom) and shallower line represents dump of the foreboard with two rounds and spreads Device.The ultrasonic transducer of device is run under 80 volts and 100 volts.These devices are operated using five ATF.For two rounds Foreboard, PCM (following line) are measured as about 20%-30% under 80 volts, about 30%-35% are measured as under 100 volts.With It is about 20%-30% under 80 volts in the PCM of half plate foreboard, is about 30%-35% under 100 volts.It is worth noting that, Once the top row of sonac is closed, the PCM for half plate foreboard does not have significant changes.For the mixture of two round foreboards Reduction/clarification percentage (line above) be about 68%-80% under 80 volts, and be about under 100 volts when top row is opened 85%-90%.It is about 68%-80% at 80 volts for reduction/clarification percentage of the mixture of half plate foreboard, is opened in top row Under 100 volts be when opening about 85%-90% then in the case where top row is closed, be about 65%-75% under 100 volts.

In curve graph in figure 16, yeast mixt is 3.0% yeast, and flows through dress with the flow velocity of 810 ml/mins It puts.The entrance of the device is a part for dump diffuser, uses half plate foreboard.Using only bottom row's energy converter, and The ultrasonic transducer of device works under 100 volts.The PCM (following line) measured is about 18%-32%, the reduction of mixture/ It is about 60%-78% to clarify percentage (line above).

Show that sound swimsuit puts 700 third exemplary embodiment in Figure 17-22.Figure 17 is exterior perspective view.Figure 18 is The perspective side sectional view of the device.Figure 19 is the side sectional view of the device.Figure 20 is the front cross sectional view of the device.Figure 21 be the magnification fluoroscopy cross-sectional side view of sound chamber.Figure 22 is the magnification fluoroscopy cross-sectional side view of ultrasonic transducer.This is special Determine embodiment also to be built in modular fashion by multiple components.

Since Figure 17, sound swimsuit puts 700 has some similarities with Fig. 1 and device shown in Fig. 10.Device 700 has There are the first side 122 and the second opposite side 124.There are side wall 110, roof 140 and base portions 150 to limit inner space 107.Dump diffuser 530 is present on each side 122,124, serves as the fluid inlet of the inner space 107 of device 112.Entrance flowing ports 542 that there are three dump diffuser tools herein, as described in the device similar to Figure 10.Roof 140 include leading to the conical inner surface 142 of the fluid outlet 114 at top 130.Concentrate outlet 116 is present in base portion 150 In, conical surface leads to the concentrate outlet at bottom end 132.Ultrasonic transducer 106 is present in rear side, and reflector 108 is present in On the front side opposite with energy converter.

Referring now to Figure 18, as can be seen that fluid outlet 114 and concentrate outlet 116 from the top of inner space 107/ A side 124 of device, the i.e. public side there are fluid inlet 112 are led in bottom end.As described above, fluid outlet 112 is usual Clear fluid is recycled in permission from inner space 107.Concentrate outlet 116 allows generally for recycling or collects particle, cell.

Referring now to Figure 19, it can be seen that leading to the inner surface 142 of the roof 140 of fluid outlet 114, skew wall 152 leads to Concentrate outlet 116 is also such.The bottom dotted line 121 of sound chamber represents that the top dotted line 119 of sound chamber represents.Inner surface 142 have the interior angle B measured relative to dotted line 119, and wherein angle B is about 11 ° to about 60 ° in embodiment, including about 30 ° to about 45°.Similarly, skew wall 152 has angle A relative to dotted line 121, and wherein angle A is about 11 ° to about 60 ° in embodiment, including about 30 ° to about 45 °.O-ring 180 can be arranged in roof/between base portion and sound chamber, to provide Fluid Sealing therebetween.

Referring now to Figure 20, see again, fluid outlet 114 and concentrate outlet 116 from the top of inner space 107/ One of side 124 of device, the i.e. public side there are fluid inlet 112 are led in bottom end.

It can be seen that the piezoelectric material 178 of ultrasonic transducer and the fluid from dump diffuser 530 into sound chamber 107 enter Mouth 112.It can also be seen that hollow chamber 540.Piezoelectric material 178 has height 176.Fluid inlet 112 also has height 113. The height 113 of fluid inlet 112 is about the 60% of the height 176 of piezoelectric material 178.In embodiment, the height of fluid inlet It can be about the 5% to about 75% of piezoelectric material height.Again, the bottom of the bottom margin 111 of fluid inlet 112 and piezoelectric material Portion edge 177 is aligned.

Turning now to Figure 21, it can be seen that the enlarged drawing of the sound chamber 107 of device 700, sound chamber 107 are sandwiched in ultrasonic transducer Between 106 and reflector 108.There is very small gap between fluid inlet 112 and energy converter 106 (for example, 0.010 English It is very little).The gap can fill such as O-ring, as shown in figure 19.In the bottom margin 177 and base portion of the piezoelectric material of energy converter Skew wall 152 between there is also very short gap (for example,<0.025 inch).

Figure 22 provides the amplification cross-sectional view of ultrasonic transducer 106.As shown in Figure 18 and Figure 19, ultrasonic transducer 106 are usually located in the side wall of device.As shown here, ultrasonic transducer includes housing 190.There are gas in the housing of energy converter Gap 194.Connector 191 exists and is spaced apart with the piezoelectric material 178 of crystal form.Use the embedding material of such as epoxy resin Piezoelectric material 178 is attached to housing by material 192.It then, will such as adhesive-backed film made of polyether-ether-ketone (PEEK) 193 are attached to the outer surface of piezoelectric material 178 and housing.The film can serve as wearing layer.Wearing layer usually have half-wavelength or The thickness of smaller (for example, 0.050 inch).Here the additional spy of the ultrasonic transducer in the present apparatus will be explained in further detail Sign.

The concrete application disclosed herein put for sound swimsuit is processing bioreactor material.Into these devices Fluid stream be master/secondary working fluid (such as water, cell culture medium) and secondary paticle mixture.Secondary paticle can include thin Born of the same parents and certain material, such as biomolecule (such as recombinant protein or monoclonal antibody or virus).These devices can be used for assembling Larger particles in mixture, such as cell, so as to which there are two types of different opening apparatus of wandering about as a refugee.First, the cell of aggregation and some The stream of fluid can be exported by concentrate and be discharged.Secondly, the clear fluid stream containing predetermined substance (such as biomolecule) can lead to Cross fluid outlet discharge.Depending on the substance for wishing to recycle, any one in the two streams of separating device can recycle Into bioreactor.

The sound swimsuit using three-dimensional acoustics standing wave of the disclosure puts standard mistake that can also as needed with upstream or downstream Filtering technology couple, such as using diatomaceous depth-type filtration, tangential flow filtration (tangential flow filtration, ) or other physical filtering processes TFF.

It is desirable that the flow velocity for the device for passing through the disclosure can be minimum 1 ml/min (mL/min) or it is minimum about 800mL/min, and more preferably, higher flow velocity can also be realized.In the unit of replacement, these flow velocitys can be sound The about 0.005mL/min or about 4.5mL/min/cm2 every square centimeter of the cross-sectional area of room.Batch-type reactor is mended in batches Material formula bioreactor and filling type bioreactor are not always the case.

It turns now to be helpful on how to generate multidimensional standing acoustic waves (particularly three-dimensional acoustics standing wave). The multidimensional standing acoustic waves needed for powder collection is obtained by driving ultrasonic transducer with a frequency, the frequency generates acoustics and stays Wave and the basic 3D vibration modes (vibration mode) for exciting transducer crystal.To in ultrasonic transducer in a manner of multi-mode Piezo-electric crystal carry out disturbance allow generate multidimensional standing acoustic waves.Piezo-electric crystal can be specifically designed to be under design frequency with Multi-mode mode deforms, so as to allow to generate multidimensional standing acoustic waves.Multidimensional standing acoustic waves can be produced by the different mode of piezo-electric crystal 3 × 3 patterns that are raw, will such as generating multidimensional standing acoustic waves.By allowing piezo-electric crystal with many different vibration shape (mode Shapes it) vibrates, multiple multidimensional standing acoustic waves can also be generated.Therefore, crystal can excite various modes, as 0x0 patterns (that is, Piston mode) to 1x1,2x2,1x3,3x1,3x3 and other higher order patterns, then returned by the relatively low cycle of modes of crystal (not necessarily by directly sequence).Crystal this switching between modes or shake allow within a specified time to generate various more Tie up waveform and the single piston vibration shape.

Some being explained further for ultrasonic transducer used in devices, systems, and methods to the disclosure can also be had It helps.In this respect, energy converter uses piezo-electric crystal, is usually made of PZT-8 (lead zirconate titanate).This crystal can have 1 inch of diameter and the nominal resonance frequency of 2MHz, and can also have the size of bigger.Each ultrasonic transducer module can Only to have there are one crystal or can have multiple crystal, each crystal is used as independent ultrasonic transducer and can be by One or more amplifier controls.Crystal can be square, rectangle, irregular polygon or usual arbitrary shape.It changes Energy device is for generating pressure field, and the pressure field is in the direction of (transverse direction) orthogonal with standing wave direction and on standing wave direction (axial direction) Generate the power of same order.

Figure 23 is the cross-sectional view of conventional ultrasonic transducer.The energy converter has wearing plate 50, the asphalt mixtures modified by epoxy resin in bottom end Lipid layer 52, ceramic crystal 54 (being made of such as PZT), epoxy resin layer 56 and back sheet 58.On the either side of ceramic crystal There is electrode：Positive electrode 61 and negative electrode 63.Back sheet 58 is attached to crystal 54 by epoxy resin layer 56.Entire component is received In housing 60, housing 60 can be made of such as aluminium.Electrical adaptor with dummy earth pin 62 provides connection to pass through housing for conducting wire and connects To the lead (not shown) for being attached to crystal 54.It damps and generates in wide frequency ranges in general, back sheet is designed to increase The wide-band transducer of uniform displacement, and be designed to inhibit the excitation under certain vibration eigen mode.Wearing plate is commonly designed For impedance transformer, preferably to match the characteristic impedance that energy converter radiates the culture medium of entrance.

Figure 24 is the cross-sectional view of the ultrasonic transducer 81 of the disclosure.Energy converter 81 is shaped as disk or plate, and has Aluminum enclosure 82.Piezo-electric crystal is perovskite ceramics crystal block, and each perovskite ceramics crystal is (logical by larger bivalent metal ion Often lead or barium) and the lattice of O2- ions in small quadrivalent metallic ion (be usually titanium or zirconium) form.As an example, PZT (lead zirconate titanate) crystal 86 defines the bottom end of energy converter, and exposes from the exterior of housing.The crystal is on its periphery by being located at crystalline substance Small elastic layer 98 (for example, silicones or similar material) support between body and housing.In other words, there is no wearing layers. In a particular embodiment, crystal is irregular polygon, and is asymmetric irregular polygon in a further embodiment Shape.

The aluminium top plate 82a of housing is attached to the main body 82b of housing by screw thread by screw 88.Top plate includes being used for as transducing The connector 84 of device power supply.The top surface of PZT crystal 86 is connected to the positive electrode 90 separated by insulating materials 94 and negative electrode 92. Electrode can be made of any conductive material (such as silver or nickel).Electric power is supplied to PZT crystal 86 by the electrode on crystal.Please Note that crystal 86 does not have back sheet or epoxy resin layer.In other words, it is deposited between aluminium top plate 82a and crystal 86 in energy converter Air gap 87 (that is, air gap is complete empty).As shown in figure 5, can set in some embodiments minimum backing 58 and/or Wearing plate 50.

Transducer designs can influence the performance of system.One typical energy converter is a kind of layered structure, wherein ceramics Crystal is bonded on back sheet and wearing plate.Since energy converter has the high mechanical impedance presented by standing wave, traditional wearing plate Design criteria (such as half-wavelength thickness is used for radiation application for standing wave application or quarter-wave thickness) and manufacturer Method is possible and improper.On the contrary, in a transducer embodiment of the disclosure, there is no wearing plate or backings, allow crystal It is vibrated under one eigen mode (that is, near eigenfrequency) with high Q factor.Vibration ceramic crystal/disk is directly exposed to flow The dynamic fluid by flow chamber.

Removal backing (for example, the crystal back side is made to ventilate) also allows ceramic crystal to be vibrated with higher order vibration mode, and several Undamped (such as higher order modal displacement).In the energy converter comprising the crystal with backing, crystal as piston with Displacement vibration evenly.Removal backing can make crystal with non-uniform displacement mode vibration.The exponent number of the vibration shape of crystal is higher, The nodal line that crystal may have is more.The high order mode displacement of crystal can generate more captured lines, although captured line and section Point correlation be not necessarily it is one-to-one, but with higher frequency driving crystal can not necessarily generate more captured lines.

In some embodiments, crystal can have the backing of the Q factor (being, for example, less than 5%) of minimum influence crystal.The back of the body Lining can be made, such as cork wood, foam or cork of the material of substantially entrant sound, and crystal is allowed to be vibrated simultaneously with higher first order mode And high Q factor is kept, while remain as crystal and some mechanical supports are provided.Back sheet can be solid or can be tool There is the lattice across the hole of layer so that lattice follows the node of vibration crystal with specific higher order vibration mode, in node position It puts place and provides support, while allow the rest part free vibration of crystal.Lattice works or the target of acoustic window material is to provide branch Support, the excitation of Q factor or the specific vibration shape of interference without reducing crystal.

By avoiding the damping of epoxy resin layer and wearing plate and energy absorption effect, crystal is made to be in direct contact with fluid Contribute to high Q factor.Other embodiment can have wearing plate or wearing face to prevent the PZT comprising lead from contacting main fluid.Example Such as, in the biologic applications such as separation blood, this may be needed.This application may use wearing layer, as chromium, electrolytic nickel or Electroless nickel plating.Chemical vapor deposition can also be used for applying poly- (to penylene dimethyl) (such as Parylene) or other polymers Or the layer of polymer film.Organic and biocompatible coating such as silicones or polyurethane also are used as wearing face.

Figure 26 is to show the log-log plot (logarithm of acoustic radiation power, fluid resistance and buoyancy with the variation of particle radius Y-axis, logarithm x-axis), and provide the explanation using acoustic radiation power separation particle.Buoyancy is a kind of dependent on particle volume Power, therefore micron-sized particle size can be ignored, but can increase, and for of hundreds of microns magnitude Become for particle size notable.Fluid resistance (Stokes resistance) is micro- with fluid velocity linear change, therefore usually more than The buoyancy of scale particles, but can be neglected for the particle of the large-size of hundreds of microns grade.Acoustic radiation power scales Ratio is different.When the particle size becomes smaller, Gor'kov equations are accurate, and sound force trapping is proportional to the volume of particle. Finally, when particle size increases, acoustic radiation power no longer with particle radius cube and increase, and will be critical at some It rapidly disappears under particle size.For further increasing particle size, the size of radiant force increases again, but opposite in phase (figure In do not show).For increasing particle size, this pattern repeats.

First, when suspended matter mainly flows through system with the particle of small micron-scale, acoustic radiation power needs balanced fluid The combined effect of resistance and buoyancy, so that particle is trapped in standing wave.In fig. 26, this is happened at the particle labeled as Rc1 Size.The chart is bright, and all larger particles also will be captured.Therefore, when capturing little particle in standing wave, particle occurs and gathers Knot/agglomerating/aggregation/cohesion, leads to the continuous increase of effective particle size.When particle aggregation is into cluster, total resistance in cluster Power is far below the resistance summation on each particle.Substantially, as particle aggregation is into cluster, they are each other from fluid stream Move and reduce the overall resistance of cluster.As particle cluster size increases, acoustic radiation power is reflect off cluster so that per unit The net acoustic radiation power of volume reduces.Acoustics cross force on particle may be different from resistance so that cluster remains stationary and Size increases.For example, sound cross force can be more than resistance, with particle is allowed to be captured, be gathered into cluster and size increase.

Continue particle size increase until buoyancy becomes leading, this is represented by the second critical particle size Rc2.Cluster is per single The buoyancy of position volume is kept constant with cluster size, because it is the function of grain density, cluster concentration and gravity constant.Cause This, as cluster size increases, the buoyancy in cluster increases faster than acoustic radiation power.In size Rc2, particle will rise Or avale, this depends on their relative densities relative to main fluid.Under this size, acoustic force is secondary, gravity/floating Power becomes leading, and particle is settled or risen naturally from main fluid.And not all particle can all be deviate from, remaining Grain and the new particle of entrance sound chamber will move on three-dimensional nodes position, repeat increase and infall process.This phenomenon is explained In the rapid decrease of acoustic radiation power and rising after size Rc2.In this way, Fig. 6 explain little particle how It is continuously captured in standing wave, grows up to the particle or agglomerate of bigger, then finally risen or settle due to buoyancy increase.

The size of energy converter, shape and thickness determine displacement of the energy converter under different stimulating frequencies, this is influenced again Grain separative efficiency.Higher order modal displacement generates the three-dimensional acoustics standing wave on all directions in sound field with strong gradient, by This generates similary strong acoustic radiation power in all directions, leads to the quantity and energy converter of multiple captured lines, wherein captured line The specific vibration shape it is related.

Figure 27 shows the electrical impedance magnitude of the measurement of energy converter, as the frequency in 2.2MHz energy converter near resonances Function.Minimum value in energy converter electrical impedance corresponds to the acoustic resonance of water column, and represents potential working frequency.Numerical value Modeling shows under these acoustic resonance frequencies, transducer displacement curve significant changes, so as to directly affect standing acoustic waves and thus The force trapping of generation.Since energy converter works near its thickness resonance, the substantially different phase of displacement of electrode surface.Energy converter The typical displacement of electrode is uneven and is based on stimulating frequency and changes.The transducer displacement pattern of higher order causes, higher The captured line of force trapping and multiple stabilizations is used for the particle of capture.

The influence of power and particle separative efficiency is captured to study transducer displacement curve to acoustics, experiment is repeated ten Secondary, other than stimulating frequency, all conditions are all identical.The ten continuous sound represented by circled numerals 1-9 in Figure 27 and letter A It learns resonant frequency and is used as stimulating frequency.Condition is the duration of experiment of 30 minutes, about 5 microns of SAE-30 oil droplets 1000ppm oil concentrations, the flow velocity of 500ml/min and the application power of 20W.

When lotion passes through energy converter, the captured line of observation and characterization oil droplet.Characterization be related to for identified in Figure 27 ten Seven in a resonant frequency, the quantity across the captured line of fluid channel is observed and patterns, as shown in Figure 28 A.

Figure 28 B show the isometric view of system, wherein capture line position is just determined.Figure 28 C be when along arrow 814 to Under the view of system that occurs when seeing entrance.Figure 28 D are the systems occurred when directly seeing energy converter planar along arrow 816 View.

The effect of stimulating frequency clearly determines the quantity of captured line, number the swashing from acoustic resonance 5 and 9 of captured line Single captured line at hair frequency changes to nine captured lines for acoustic resonant frequencies 4.It is observed under other stimulating frequencies To four or five captured lines.The different Displacements Distributions of energy converter can generate different (more) captured lines, position in standing wave It moves more gradients in distribution and generally produces higher force trapping and more captured lines.Although it should be noted that institute in Figure 28 A The different captured line distributions shown are what is obtained under the frequency shown in Figure 27, and the distribution of these captured lines can also be in different frequencies It is obtained under rate.

Figure 28 A are shown vibrates mould by the way that crystal is driven to be vibrated under different fundamental vibration frequencies and possible different crystal Formula.The 3D patterns of crystal vibration are passed through the one road direction reflector of fluid in chamber to propagate and are returned by standing acoustic waves.It is generated Multi-dimensional standing-wave can be considered including two components.First component is the flat motion outside plane component for the crystal for generating standing wave (uniform displacement on plane of crystal), second component are that the position of peak value and valley occur on two horizontal directions of plane of crystal Shifting amplitude changes.Three-dimensional force gradient is generated by standing wave.These three-dimensional force gradients lead to lateral radiation power, lateral radiation power relative to It flows through and overcomes viscosity resistance that particle is made to stop and capture particle.In addition, lateral radiation power is responsible for generating closelypacked Grain cluster.Therefore, particle separation and collection gravity driven be depending on multi-dimensional standing-wave is generated, and the multi-dimensional standing-wave of generation can be Mixture flowing during standing acoustic waves by overcoming particle resistance.As being schematically shown in Figure 28 A, along on standing wave axial direction Captured line forms multiple particle clusters.

The disclosure is described with reference to exemplary embodiment.It, can after the detailed description of front is read and understood Do corresponding modifications and changes.Disclosure permission falls with appended claims or modification and change in its equivalency range More.

Claims (21)

1. a kind of sound swimsuit is put, including：

Sound chamber, at least one entrance being included at its first end；

In at least one fluid outlet for the top end that the sound swimsuit is put；

It is exported at least one concentrate for the bottom end that the sound swimsuit is put；

At least one ultrasonic transducer, is coupled to the sound chamber, and at least one ultrasonic transducer includes piezoelectric material, institute Piezoelectric material is stated to be configured as being driven to generate multidimensional standing acoustic waves in the sound chamber by voltage signal；And

It is located at the reflector of sound chamber's opposite side relative at least one ultrasonic transducer；

Wherein, the sound chamber includes the planar cross-sectional area limited by length and width and by the width and High definition Side section area, wherein the length is greater than or equal to the width, and wherein described planar cross-sectional area is more than described Side section area.

2. sound swimsuit according to claim 1 is put, wherein at least one entrance is a part for dump diffuser.

3. sound swimsuit according to claim 2 is put, wherein at least one entrance is included across the piezoelectric material About 60% height of height.

4. sound swimsuit according to claim 2 is put, wherein the base portion of at least one entrance is along the piezoelectric material Base portion positioning.

5. sound swimsuit according to claim 2 is put, wherein, the dump diffuser is included at the upper end of pumping chamber extremely Few entrance flowing ports and the flowing outlet at the lower end of the pumping chamber, the flowing outlet are first-class with providing The shape in dynamic direction, the flow direction is perpendicular to the multidimensional standing acoustic waves generated by least one ultrasonic transducer Axial direction.

6. sound swimsuit according to claim 1 is put, wherein, at least one entrance is included in described the of the sound chamber The first entrance of at one end and the second entrance at the second end opposite with its first end of the sound chamber so that fluid flows into The sound chamber is uniform symmetrical.

7. sound swimsuit according to claim 1 is put, further include below at least one entrance and lead to described First skew wall of at least one concentrate outlet, wherein, first skew wall is included relative to first level face into about 11 ° to about 60 ° of angle.

8. sound swimsuit according to claim 1 is put, wherein, at least one energy converter is across the length of the sound chamber Multiple energy converters.

9. sound swimsuit according to claim 8 is put, wherein, the multiple energy converter arranged in series is into single.

10. sound swimsuit according to claim 8 is put, wherein, the multiple energy converter includes first row, the first row packet Containing at least two energy converters, above the second row comprising at least two energy converters.

11. sound swimsuit according to claim 1 is put, wherein, at least one concentrate outlet is that multiple concentrates go out Mouthful.

12. sound swimsuit according to claim 1 is put, wherein, the sound chamber includes at least 40 cubic inches of volume.

13. sound swimsuit according to claim 1 is put, wherein, sloping top walls, parabolic curved roof or hypocycloid bending Roof leads at least one fluid outlet from the first end and second end of the sound chamber.

14. sound swimsuit according to claim 1 is put, wherein, at least one fluid outlet is connected to the sound chamber Middle section.

15. sound swimsuit according to claim 1 is put, wherein, the multidimensional standing acoustic waves is included with same order Axial force component and transverse force component.

16. sound swimsuit according to claim 1 is put, wherein at least one ultrasonic transducer includes：

Housing, including top, bottom end and inner space；And

There is the outer surface of exposure and inner surface, the crystal to exist for crystal positioned at the bottom end of the housing, the crystal It can be vibrated when being driven by voltage signal.

17. sound swimsuit according to claim 16 is put, wherein back sheet contacts the inner surface of the crystal, the back of the body Lining is made of substantially acoustic window material.

18. sound swimsuit according to claim 1 is put, wherein, the multidimensional standing acoustic waves is three-dimensional standing wave.

19. a kind of sound swimsuit is put, including：

Sound chamber, at least one entrance being included at its first end；

In at least one fluid outlet for the top end that the sound swimsuit is put；

It is exported at least one concentrate for the bottom end that the sound swimsuit is put；

At least one ultrasonic transducer, is coupled to the sound chamber, and at least one ultrasonic transducer includes piezoelectric material, institute Piezoelectric material is stated to be configured as being driven to generate multidimensional standing acoustic waves in the sound chamber by voltage signal；And

It is located at the reflector of sound chamber's opposite side relative at least one ultrasonic transducer；

Wherein, at least one entrance in the form of dump diffuser, the dump diffuser is included in the front lower of pumping chamber Flowing ports at end, the first entrance flowing ports at the upside of the pumping chamber and the booster it is rear on Second entrance flowing ports at end side.

20. it is a kind of for detaching secondary fluid or the method for particle from mixture, including：

Secondary working fluid and the mixture of the secondary fluid or particle is made to flow through sound swimsuit with the rate of at least 25 ml/mins Put, the sound swimsuit put including：

Sound chamber, at least one entrance being included at its first end；

In at least one fluid outlet for the top end that the sound swimsuit is put；

It is exported at least one concentrate for the bottom end that the sound swimsuit is put；

At least one ultrasonic transducer, at least one ultrasonic transducer are coupled to the sound chamber, at least one ultrasound Energy converter includes piezoelectric material, and the piezoelectric material is driven to generate multidimensional standing acoustic waves in the sound chamber by voltage signal； And

It is located at the reflector of sound chamber's opposite side relative at least one ultrasonic transducer；

Wherein, the multidimensional standing acoustic waves captures the secondary fluid or particle, and by the secondary fluid or particle with it is described Secondary working fluid detaches.

21. according to the method for claim 20, wherein the sound chamber includes at least 40 cubic inches of volume.