Acoustic Concentration Of Particles In Fluid Flow

Abstract

An apparatus for acoustic concentration of particles in a fluid flow includes a substantially acoustically transparent membrane and a vibration generator that define a fluid flow path therebetween. The fluid flow path is in fluid communication with a fluid source and a fluid outlet and the vibration generator is disposed adjacent the fluid flow path and is capable of producing an acoustic field in the fluid flow path. The acoustic field produces at least one pressure minima in the fluid flow path at a predetermined location within the fluid flow path and forces predetermined particles in the fluid flow path to the at least one pressure minima.

Claims

1. A method, comprising: applying an acoustic radiation pressure to a population of particles disposed in a medium within a fluid flow channel, wherein the fluid flow channel is at least partially defined by a surface of a material configured to act as a pressure release surface, and wherein the acoustic radiation pressure is applied with a vibration generator disposed opposite the material so as to give rise to a pressure node, and the acoustic radiation pressure being applied as to give rise to one or both of: (a) an accumulation of one or more particles with positive acoustic contrast, if present, at the surface of the material, or (b) an accumulation of one or more particles with negative acoustic contrast, if present, in the direction of the vibration generator relative to the surface of the material.

2. The method of claim 1, wherein the material is characterized as acoustically transparent.

3. The method of claim 1, further comprising optically inspecting one or more of the accumulation of positive acoustic contrast particles at the surface of the material.

4. The method of claim 1, further comprising optically inspecting one or more of the accumulation of negative acoustic contrast particles in the direction of the vibration generator relative to the surface of the material.

5. The method of claim 1, further comprising separating particles outside the accumulation of positive acoustic contrast particles at the surface of the material.

6. The method of claim 1, further comprising separating particles outside the accumulation of negative acoustic contrast particles in the direction of the vibration generator relative to the surface of the material.

7. The method of claim 1, further comprising washing one or more of the accumulation of positive acoustic contrast particles at the surface of the material, exposing one or more of the accumulation of positive acoustic contrast particles at the surface of the material to one or more reagents, or both.

8. The method of claim 7, wherein the one or more reagents are diffused through the material.

9. The method of claim 1, further comprising washing one or more of the accumulation of negative acoustic contrast particles in the direction of the vibration generator relative to the surface of the material, exposing one or more of the accumulation of negative acoustic contrast particles in the direction of the vibration generator relative to the surface of the material to one or more reagents, or both.

10. The method of claim 9, wherein the one or more reagents are diffused through the material.

11. The method of claim 1, further comprising changing a frequency of the vibration generator so as to change a location of the pressure node, to give rise to a pressure node, or both.

12. The method of claim 11, further comprising changing a frequency of the vibration generator so as to change a location of the pressure node.

13. A system, comprising: a fluid flow channel having an inlet and an outlet, the fluid flow channel being at least partially defined by a surface of a material configured to act as a pressure release surface; a vibration generator configured to, during operation, effect an acoustic radiation pressure that produces an acoustic displacement field within a medium disposed within the fluid flow channel, the acoustic displacement field being applied to as to give rise to one or both of: (a) an accumulation of one or more positive acoustic contrast particles, if present in the medium, at the surface of the material, or (b) an accumulation of one or more negative acoustic contrast particles, if present in the medium, in the direction of the vibration generator relative to the surface of the material.

14. The system of claim 10, further comprising an imager configured to image one or more particles accumulated at the surface of the material.

15. The system of claim 10, further comprising an imager configured to image one or more particles accumulated in the direction of the vibration generator relative to the surface of the material.

16. The system of claim 10, wherein the material is optically transparent.

17. The system of claim 10, wherein the material is permeable.

18. The system of claim 17, wherein the material is selectively permeable.

19. The system of claim 10, further comprising a channel configured to receive particles outside an accumulation of one or more positive acoustic contrast particles at the surface of the material.

20. The system of claim 10, further comprising a channel configured to receive particles outside an accumulation of one or more negative acoustic contrast particles, if present in the medium, in the direction of the vibration generator relative to the surface of the material.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The accompanying drawings, which are incorporated into and form a part of the specification, illustrate one or more embodiments of the present invention and, together with the description, serve to explain the principles of the invention. The drawings are only for the purpose of illustrating one or more preferred embodiments of the invention and are not to be construed as limiting the invention. In the drawings:

[0013] FIG. 1 is a schematic view of a separator according to the prior art;

[0014] FIG. 2 is a schematic view of an embodiment of an apparatus in accordance with the present invention;

[0015] FIG. 3 is a schematic graph showing the location of pressure nodes and antinodes in the apparatus of FIG. 2;

[0016] FIG. 4 is a schematic view of particles being separated by the apparatus of FIG. 2; and

[0017] FIGS. 5a and 5b are microscopic photographs showing latex particles acoustically trapped on a membrane surface of the apparatus; and

[0018] FIG. 6 is a schematic view of an embodiment of an apparatus in accordance with the present invention showing profiles of various pressure minima.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0019] Successful meso- to microfluidic sample preparation is dependent upon efficient sorting, concentration, and washing of targets. Numerous successful analytical lab-on-a-chip micro-devices capable of a wide range of detection techniques from spectroscopy to gene detection have been demonstrated in both clinical and homeland security arenas. In the present evolution of these devices, however, their increased application to real world problems of interest has been severely limited by inadequate provisions for handling samples. The heart of this problem lies in concentrating and purifying a large dilute sample that contains interferents. These microfabricated devices generally require a clean sample with a representative population of target species that can be analyzed only in microliter and nanoliter volumes. In applications where the sample volume is measured in milliliters to liters, the sample preparation is a daunting task that has not been adequately addressed.

[0020] Several field-based methods for sample processing have been applied to this problem including immunomagnetic separation, electrophoresis, dielectrophoresis and ultrasonic separation. Ultrasonic separation is particularly attractive for many applications as it typically does not require reagents and can be performed in complex media with little regard for sample conductivity or pH.

[0021] Ultrasonic separation is typically achieved in resonant chambers in which standing waves are established using a vibration generator, such as a piezoelectric transducer or the like. The force on a particle is given by the following equation derived by Gor'kov:

[00001]$F=-\left(\frac{2}{3}.Math..Math..Math.R^3\left[\frac{Z_0}{{}_f.Math.c_f^2}.Math.\stackrel{\_}{p^2}-\frac{3.Math.Z_1.Math.{}_f}{2}.Math.\stackrel{\_}{v^2}\right]\right)$

[0022] Where R is particle radius, .sub.f is fluid density c.sub.f is fluid sound speed p.sup.2 is mean square fluctuations of pressure at the particle, v.sup.2 is mean square fluctuations of velocity at the particle and Z.sub.0 and Z.sub.1 are functions of particle and fluid properties called acoustic contrast factors. Most particles and cells of interest have positive acoustic contrast in water or buffers and therefore they typically migrate to positions of lowest pressure (pressure nodes or pressure minima). Materials such as fat globules and gas bubbles have negative acoustic contrast and tend to move toward positions of highest pressure (pressure antinodes or pressure maxima).

[0023] Referring to FIG. 1, an ultrasonic separator in accordance with the prior art is indicated generally at 10. Separator 10 includes fluid channel 12, wavelength glass acoustic reflector top 14 and wavelength matching layer resonator bottom 16 coupled to transducer 18. Typically, separator 10 operates at a resonant frequency approximately or wavelength of fluid layer 12. The thickness and composition of the material of top reflector 14 and bottom matching layer 16 are chosen such that the phase relationship of incident and reflected waves results in a pressure node or pressure minima either at the center of fluid channel 12 or at the surface of the top reflector 14. Separator 10 uses acoustic standing waves in channel 12 to force particles with positive acoustic contrast to move towards one wall of the channel. Device 10 is tuned such that a standing wave can be established for which a pressure node or minima forces particles with positive acoustic contrast to migrate toward top of channel 12.

[0024] Referring to FIG. 2, an embodiment of apparatus in accordance with the present invention is indicated generally at 20. Apparatus 20 includes fluid flow path or channel 22 preferably in fluid communication with a fluid source (not shown) and a fluid outlet (not shown) having membrane 24 as a top surface coupled to vibration generator 26 disposed adjacent flow channel 22. Flow channel 22 is preferably defined by an upper surface of vibration generator 26 and by membrane 24. The fluid source may supply water, or any suitable liquid to flow path or channel 22, as will be appreciated by those skilled in the art. Fluid flow path or channel 22 preferably has a predetermined dimension that is a function of the resonance of the fluid source. Preferably, vibration generator 26 is a piezoelectric transducer. Alternatively, vibration generator 26 is a line-drive element, a displacement generator, or any other type of vibration generator capable of producing an acoustic or displacement field within fluid channel 22. When vibration generator 26 is driven, plane waves incident on the boundary of membrane 24 are reflected back out of phase. Membrane 24 functions as a pressure release surface with a reflection coefficient of near 1. Therefore, the reflected wave is 180 degrees out of phase with the incident wave and the pressure wave is 90 degrees out of phase with the displacement wave. This results in a pressure node or minima at the surface of membrane 24, best seen in FIG. 3 and discussed in more detail below. Membrane 24 can be made of any suitable material but it should be thin enough to be substantially acoustically transparent to the acoustic wave generated by vibration generator 26 such as, but not limited to, thin Mylar, glass, mica or similar suitable materials.

[0025] There is shown in FIG. 3 a pressure profile in fluid flow path or channel 22 indicating pressure node or minima 28 adjacent membrane 24 and pressure antinode or maxima 30 adjacent vibration generator 26. The thickness of channel 22 thickness is wavelength (2) of the resonance. Particles and/or cells with positive acoustic contrast are driven to the surface of membrane 24 surface or pressure node 28. Pressure nodes 28 can also be created within the fluid by tuning fluid layer 22 to alternate frequencies e.g. or 5/4. For example, there is shown in FIG. 6, various locations of pressure minima 28a, 28b, and 28c in flow path or channel 22 of apparatus 20, based on the resonance of medium disposed in the fluid layer in flow channel 22. Pressure minima 28a for a 5/4 wavelength is shown in three locations within channel 22. Pressure minima 28b for a wavelength is shown at a pair of locations within channel 22 and pressure minima 28c for a wavelength is shown at a single location adjacent membrane 24. Those skilled in the art will appreciate that pressure minima, such as pressure minima 28a, 28b, and 28c may be located at any predetermined location within channel 22 between vibration generator 26 and membrane 24 and that the predetermined location is a function of the resonance and frequency of the fluid source and the predetermined dimension of flow channel 22 between vibration generator 26 and membrane 24. Alternatively, apparatus 20 includes a wavelength matching layer 25 on an upper surface of vibration generator 26 opposite membrane 24. Matching layer 25 is preferably a wavelength matching layer and is operable to isolate vibration generator 26 from the fluid within channel 22 and/or to better match the acoustic impedance of the fluid within channel 22.

[0026] Apparatus 20 can be applied to separate and or concentrate target particles and cells. When device 20 is embodied as a channel 22 with laminar flow, indicated by arrow 34, particles or cells 32 are forced into slower streamlines where they become concentrated, best seen in FIG. 4. For particles 32 of different sizes or with different acoustic contrasts, device 20 can perform field flow fractionation (FFF), as will be appreciated by those skilled in the art. In FIG. 4, particles or cells 32 with larger volumes or greater acoustic contrast are forced to surface of membrane 24 more quickly.

[0027] Alternatively, when the flow of the fluid in device 20 is slowed sufficiently or stopped altogether, particles or cells 32 are trapped at surface of membrane 24. There, particles or cells 32 are washed or exposed to other reagents. This is preferably done by replacing the sample fluid in channel 22 or, if membrane 24 is made permeable, reagents are preferably added to the opposite side of membrane 24 where the reagents can diffuse through membrane 24 to the trapped targets 32.

[0028] Thin membrane 24 advantageously allows optical observation with high numerical aperture close working distance lenses (not shown). This is useful in applications in oncology or microbiology. In addition, cells or particles 32 can be observed in an imaging plane in flow away from the membrane if an alternate tuning that provides for pressure nodes or minima in the fluid is used. In FIGS. 5a and 5b there is shown microscopic photographs of test results for apparatus 20 using 3 micron red latex particles 32. Particles 32 are trapped on the surface of membrane 24.

[0029] For apparatus 20, it is only necessary to tune to the resonance of the fluid layer (, , 5/4, etc wavelength). It is therefore simpler to accommodate fluid property or temperature changes that may affect the tuning of apparatus 20. Added advantages to the membrane configuration of apparatus 20 include possible viewing of trapped or moving plane focused species with close working distance objectives and possible incorporation of particular membrane properties, such as selective permeability.

[0030] Acoustic separations utilizing apparatus 20 can advantageously be accomplished without the use of reagents and without regard for fluid pH or conductivity, making apparatus 20 well suited for use in complex media such as blood or sewer water. Apparatus 20 uses membrane top 24 that can be fabricated inexpensively from polymers. Membrane top 24 is thin enough to accommodate high power microscopic observation of trapped species 32. Membrane 24 can also advantageously be made selectively permeable such that reagents or analytes could diffuse across membrane 24.

[0031] The primary commercial applications for apparatus 20 are contemplated to be sample preparation (concentration/separation/washing) and imaging for medical, industrial, and environmental samples. Apparatus 20 of the present invention pushes positive acoustic contrast particles 32 to channel wall 24 opposite vibration generator 26 that comprises a thin membrane top 24, which advantageously eliminates the need for precise tuning of paired matching layer 16 and reflector 14 as in the prior art device 10 shown in FIG. 1.

[0032] Although the invention has been described in detail with particular reference to these preferred embodiments, other embodiments can achieve the same results. Variations and modifications of the present invention will be obvious to those skilled in the art and it is intended to cover in the appended claims all such modifications and equivalents. The entire disclosures of all references, applications, patents, and publications cited above and/or in the attachments, and of the corresponding application(s), are hereby incorporated by reference.