SYSTEM AND METHOD FOR CHANGING A CONCENTRATION OF PARTICLES IN A FLUID

Abstract

A system for changing a concentration of at least one group of particles in a fluid, which includes a first container and a second container; a first transfer device and a second transfer device, and an element for keeping the volume of fluid in the first and second containers constant. The first container is fluidly connected to an inlet of the first transfer device, and the second container is fluidly connected to an inlet of the second transfer device. The first and second transfer devices include a chamber associated with at least one acoustic wave generator for generating acoustic waves within the chamber; at least two outlets including a first outlet for fluid enriched with the particles and a second outlet for fluid depleted of the particles; the first outlet being fluidly connected to the first container and the second outlet being fluidly connected to the second container.

Claims

1-16. (canceled)

17. A system for changing a concentration of at least one given group of particles comprised in a fluid, the system comprising: a first container and a second container; a first transfer device and a second transfer device, wherein the first container is fluidly connected to an inlet of the first transfer device and the second container is fluidly connected to an inlet of the second transfer device, each of the first and second transfer devices comprising: a chamber configured to be associated with at least one acoustic wave generator for generating acoustic waves within the chamber; at least two outlets comprising a first outlet for fluid enriched with said given group of particles and a second outlet for fluid depleted of said given group of particles; the first outlet being fluidly connected to the first container and the second outlet being fluidly connected to the second container; and means for keeping the volume of fluid in each of the first and second containers constant.

18. The system according to claim 17, comprising at least three outlets comprising a first central outlet for fluid enriched with said given group of particles and two second peripheral outlets for fluid depleted of said given group of particles.

19. The system according to claim 17, comprising a control unit configured to keep the volume of fluid in each of the first and second containers constant by regulating the flow rate of the fluid circulating in the system as a function of measurements representative of the volume of fluid in the first container and/or the second container.

20. The system according to claim 19, wherein the measurements representative of the volume of fluid in the first container and/or the second container are measurements of the volume of fluid in the first container and/or the second container, and/or measurements of the weight of the first container and/or the second container, and/or measurements of the fluid flow rate in the first container and/or the second container.

21. The system according to claim 17, comprising measurement means for measuring the volume of fluid in the first container and/or the second container, and/or the weight of the first container and/or the second container, and/or the fluid flow rate in the first container and/or the second container.

22. The system according to claim 17, wherein the chamber of each transfer device extends along a longitudinal axis, has a cross section with a width measured along a first transverse axis and a thickness measured along a second transverse axis perpendicular to the first transverse axis, the width being greater than or equal to the thickness, the chamber having first and second walls along the second transverse axis.

23. The system according to claim 17, wherein the width/thickness ratio of the chamber is greater than 1.

24. The system according to claim 17, wherein the acoustic waves have a wavelength λ and the thickness of the chamber is substantially equal to a multiple of λ/4.

25. The system according to claim 17, wherein the flow rate at the inlet of each of the first and second transfer devices is between 0.1 mL/min and 50 mL/min.

26. The system according to claim 17, wherein the fluid is a biological fluid selected in the group comprising human and/or non-human cell suspension, cell cluster suspension, blood, whole blood, surgical blood, platelet rich plasma, buffy coat, urine, serum, lymph, fluidified feces, adipose tissue, bone marrow, cerebrospinal fluid, sperm, cord blood, milk, saliva, tissue, egg albumen, seashell mix, or a mixture thereof.

27. The system according to claim 17, wherein the particles are selected in the group comprising biological cells, dispersed cells in a dispersion medium, monodisperse or polydisperse cells, blood cells, platelets, red blood cells, white blood cells, cancer cells, bacteria, proteins, liposomes, organelles, cell clusters, viruses, vesicles, microparticles, nanoparticles, microbubbles, microbeads, microorganisms, parasites, algae, sand, sediment, dust, antibodies, powders, gametes, parasite eggs, plankton, tissue, fat, pollen, spores, metal particles, or a mixture thereof.

28. A method for changing a concentration of at least one given group of particles comprised in a fluid, by means of a system comprising: a first container and a second container; a first transfer device and a second transfer device each comprising: a chamber configured to be associated with at least one acoustic wave generator for generating acoustic waves within the chamber; an inlet and at least two outlets comprising a first outlet for fluid enriched with said given group of particles and a second outlet for fluid depleted of said given group of particles; the method comprising preliminary steps of: (a) introducing a volume of fluid in the first container and a volume of fluid in the second container; (b) applying an acoustic field by generating acoustic waves inside the chamber of each of the first and second transfer devices; followed by steps of: (c) simultaneously transferring fluid contained in the first container into the chamber of the first transfer device, and transferring fluid contained in the second container into the chamber of the second transfer device; (d) simultaneously transferring fluid enriched with said given group of particles, collected from the first outlet of the first and second transfer devices, into the first container, and transferring fluid depleted of said given group of particles, collected from the second outlet of the first and second transfer devices, into the second container; wherein the respective volumes of fluid in the first and second containers are kept constant during steps (c) and (d).

29. The method according to claim 28, wherein the fluid is circulated continuously from one of the first and second containers, through one of the first and second transfer devices, and to one of the first and second containers.

30. The method according to claim 28, comprising the measurement of the volume of fluid in the first container and/or the second container, and/or the weight of the first container and/or the second container, and/or the fluid flow rate in the first container and/or the second container.

31. The method according to claim 30, comprising the regulation of the respective flow rates at the inlets of the first and second containers and the first and second transfer devices as a function of measurements representative of the volume of fluid in the first container and/or the second container.

32. The method according to claim 28, wherein the steps (c) and (d) are repeated until the concentration of the given group of particles in the fluid contained in the second container reaches a predefined level.

Description

DESCRIPTION OF THE DRAWINGS

[0233] The following detailed description will be better understood when read in conjunction with the drawings. For the purpose of illustrating, the system is shown in the preferred embodiments. It should be understood, however that the application is not limited to the precise arrangements, structures, features, embodiments, and aspect shown. The drawings are not drawn to scale and are not intended to limit the scope of the claims to the embodiments depicted. Accordingly, it should be understood that where features mentioned in the appended claims are followed by reference signs, such signs are included solely for the purpose of enhancing the intelligibility of the claims and are in no way limiting on the scope of the claims.

[0234] Features and advantages of the invention will become apparent from the following description of embodiments of a system, this description being given merely by way of example and with reference to the appended drawings in which:

[0235] FIG. 1 is a schematic representation of the application of an acoustic field on particles within a chamber.

[0236] FIG. 2 is a schematic representation of a system according to a first embodiment of the invention.

[0237] FIG. 3 is a schematic representation of a transfer device of the system of FIG. 2.

[0238] FIG. 4 is a schematic representation of a system according to a second embodiment of the invention.

[0239] FIG. 5 is a flow chart showing steps of a method according to one embodiment of the invention.

[0240] While various embodiments have been described and illustrated, the detailed description is not to be construed as being limited hereto. Various modifications can be made to the embodiments by those skilled in the art without departing from the true spirit and scope of the disclosure as defined by the claims.

ILLUSTRATIVE EMBODIMENTS OF THE INVENTION

[0241] As shown in FIG. 1, the application of an acoustic field on particles 2 within a chamber between an acoustic wave generator 16 and a reflector 4 induces a movement of said particles 2, allowing to gather said particles 2 at a node of said acoustic field.

[0242] In this example, ultrasonic waves are generated in a chamber between a reflector 4 and a wall 5 associated with an acoustic wave generator 16 coupled with a transmitter layer. This enables the creation of an acoustic pressure node in the center of the chamber (depending on the chosen frequency at which the acoustic wave generator 16 operates) and therefore of acoustic radiation forces (ARF). The ARF push the particles 2 towards the pressure node with a force of up to a hundred times gravity equivalent. The particles 2 suspended in the fluid will then migrate to the sound pressure node and can then remain trapped in this position.

[0243] A plurality of acoustic nodes can be created in the chamber, said acoustic nodes can be located at the center of the chamber, or off the center of the chamber.

[0244] This is particularly advantageous as it allows the isolation of particles 2 within a fluid without any mechanical force, filtration, or centrifugation steps that could damage said particles 2, especially if said particles 2 are fragile like cells.

[0245] In the first embodiment shown in FIG. 2 and FIG. 3, the system 1 according to the invention comprises: [0246] a first container 11 comprising an inlet 111 and an outlet 112; [0247] a second container 12 comprising an inlet 121 and an outlet 122; [0248] a first transfer device 13 and a second transfer device 14, wherein the first container 11 is fluidly connected to an inlet 133 of the first transfer device 13 and the second container 12 is fluidly connected to an inlet 143 of the second transfer device 14, each of the first and second transfer devices 13, 14 comprising: [0249] a chamber 131, 141; [0250] three outlets 134, 135, 144, 145 comprising a first central outlet 134, 144 for fluid enriched with said given group of particles and two second peripheral outlets 135, 145 for fluid depleted of said given group of particles; [0251] pumps 15 configured to regulate flow rates at the inlets 111, 121, 133, 143.

[0252] FIG. 3 shows a schematic larger-scale view of the chamber 131 or 141 of the transfer devices 13, 14 comprised in the system represented in FIG. 2.

[0253] In this embodiment, the first and second containers 11, 12 are each configured to comprise a fluid, the fluid comprised in the first container 11 being enriched with at least one group of particles and the fluid comprised in the second container 12 being depleted of said group of particles.

[0254] In this embodiment, operating the system 1 comprises the following: a volume of fluid is introduced in the first container 11 and a volume of fluid is introduced in the second container 12; an acoustic field is applied to said fluid by generating acoustic waves inside the chamber 131, 141 of each of the first and second transfer devices 13, 14; then the fluid contained in the first container 11 is transferred into the chamber 131 of the first transfer device 141, and the fluid contained in the second container 12 is simultaneously transferred into the chamber 141 of the second transfer device 14, whereby a given group of particles migrates to a sound pressure node in the chamber 13, 14 created by the generation of an acoustic field in said chamber 131, 141 and are delivered at the central outlet 134, 144 whereas other components of the fluid are delivered on the sides of the chamber 131, 141 at the peripheral outlets 135, 145; the fluid enriched with said given group of particles, collected from the central outlet 134, 144 of the first and second transfer devices 13, 14, is transferred into the first container 11, and the fluid depleted of said given group of particles, collected from the peripheral outlets 135, 145 of the first and second transfer devices 13, 14, is simultaneously transferred into the second container. The respective volumes of fluid in the first and second containers are kept constant during operating through the regulation of the flow rate of the fluid circulating in the system 1, obtained by means of the pumps 15, as a function of measurements representative of the volume of fluid in the first container 11 and/or the second container 12.

[0255] This embodiment is particularly advantageous as the given group of particles is separated from other components of the fluid without using any mechanical force, thus preventing any damage to said given group of particles.

[0256] This embodiment is particularly advantageous as the flow rates at the inlets 111, 121, 133, 143 of the containers and chambers are regulated to ensure that the volume of fluid in each of said containers 11, 12 stays constant at all times, preventing an inopportune emptying of one of said containers 11, 12. By keeping these volumes constant, the volumes and particle concentrations of the final products are perfectly controlled.

[0257] As shown in FIG. 3, in this embodiment, each transfer device 13, 14 comprises: [0258] a chamber 131, 141 configured to be associated with an acoustic wave generator 16 for generating acoustic waves; [0259] a reflector located opposite said at least one acoustic wave generator 16, the reflector being the air 3 surrounding the chamber 131, 141 in this example; and [0260] three outlets 134, 135, 144, 145 comprising a first central outlet 134, 144 for fluid enriched with said given group of particles and two second peripheral outlets 135, 145 for fluid depleted of said given group of particles.

[0261] In this embodiment, the chamber 131, 141 of each transfer device 13, 14 extends along a longitudinal axis (x), has a cross section with a width measured along a first transverse axis (y) and a thickness measured along a second transverse axis (z) perpendicular to the first transverse axis, the width being greater than or equal to the thickness, the chamber 131, 141 having first and second walls 132, 136, 142, 146 along the second transverse axis (z). The chamber 131, 141 has a thickness between 350 and 450 μm, a width between 0.7 and 2.1 cm and a length between 1 and 6 cm and the walls 132, 136, 142, 146 are made of PMMA.

[0262] In this embodiment, the fluid is introduced at the inlet 133, 143 of the chamber 131, 141. The flow rate at the inlet 133, 143 of the chamber 131, 141 ranges between 0.4 and 0.6 mL/min The acoustic generator 16 associated with said chamber 131, 141 generates acoustic waves having a frequency ranging between 1.8 and 2 MHz in the chamber 131, 141 that are reflected by the reflector being the air 3 located at the outside of the chamber 131, 141. This creates at least one pressure node in the chamber 131, 141 allowing a selective migration of a given group of particles towards the central outlet 134, 144 while the other components of the fluid are evacuated at the peripheral outlets 135, 145. The acoustic wave generator 16 can be coupled with a transmitter layer (not represented in FIG. 3).

[0263] This embodiment is particularly advantageous as an acoustic field is generated within the chamber 131 by the acoustic wave generator 16. As explained in FIG. 1, this simple arrangement allows the isolation of particles within a fluid without any mechanical force, filtration, or centrifugation steps that could damage said particles, especially if said particles are fragile like cells.

[0264] In the second embodiment shown in FIG. 4, the elements similar to those of the first embodiment bear identical references. The system 1 of the second embodiment differs from the first embodiment in that each transfer device 13, 14 comprises only two outlets. More precisely, the system 1 according to the second embodiment of the invention comprises: [0265] a first container 11 comprising an inlet 111 and an outlet 112; [0266] a second container 12 comprising an inlet 121 and an outlet 122; [0267] a first transfer device 13 and second transfer device 14, wherein the first container 11 is fluidly connected to an inlet 133 of the first transfer device 13 and the second container 12 is fluidly connected to an inlet 143 of the second transfer device 14, each of the first and second transfer devices 13, 14 comprising: [0268] a chamber 131, 141; [0269] two outlets 134, 135, 144, 145 comprising a first outlet 134, 144 for fluid enriched with said given group of particles and a second outlet 135, 145 for fluid depleted of said given group of particles; [0270] pumps 15 configured to regulate flow rates at the inlets of the first and second containers and the chambers 111, 121, 133, 143.

[0271] In this embodiment, the first and second containers 11, 12 are each configured to comprise a fluid, the fluid comprised in the first container 11 being enriched with at least one group of particles and the fluid comprised in the second container 12 being depleted of said group of particles.

[0272] In this embodiment, the respective volumes of fluid in the first and second containers 11, 12 are kept constant at all times.

[0273] This embodiment is particularly advantageous as the flow rates at the inlets of the containers and chambers 111, 121, 133, 143 are regulated to ensure that the volume of fluid in each of said containers 11, 12 stays constant at all times, preventing an inopportune emptying of one of said containers 11, 12.

[0274] As shown in FIG. 5, the method of the invention comprises the following steps: [0275] providing a system 1 of the invention; [0276] applying an acoustic field by generating acoustic waves inside each chamber 131, 141 of the first and second transfer devices 13, 14, by means of each acoustic wave generator 16; [0277] simultaneously transferring the fluid contained in the first container 11 to the first transfer device 13, and transferring the fluid contained in the second container 12 to the second transfer device 14; [0278] simultaneously transferring the fluid enriched with at least one group of particles, collected from the first outlet(s), to the first container 11 and the fluid depleted of said group of particles, collected from the second outlet(s), to the second container 12.

[0279] In addition, the flow rates at the inlets 111, 121, 133, 143 of the first and second containers 11, 12 and the chambers 131, 141 are regulated so that the respective volumes of fluid in the first and second containers 11, 12 are kept constant during the steps of the method.

[0280] As illustrated above, the method of the invention is a simple and fast method for separating at least one group of particles from a fluid without any dilution required. Furthermore, no steps of filtration, centrifugation or any steps requiring mechanical forces are needed during said method. This prevents damage to the group of particles to be separated.

EXAMPLES

[0281] The present invention is further illustrated by the following examples. The following examples are implemented, in particular, using the system of FIGS. 2 and 3.

Example 1

Platelet Enrichment

[0282] Materials and Methods

[0283] Platelet rich plasma is injected in equal amount in the first and second container of the present invention, hence each of them holds 50% of the total amount of platelets in the system. The platelet concentration may vary from high concentration to diluted samples. The platelets have an average diameter of 2 μm.

[0284] A flow is induced through the transfer devices by the flowing means. The flow control means are activated so each container holds a constant volume of fluid through the process with the flow rate controlled accordingly. The flow in the transfer devices inlets is kept within 0.4 to 0.6 mL/min. The transfer devices chambers have a thickness between 350 and 450 μm, a width between 0.7 and 2.1 cm and a length between 1 and 6 cm.

[0285] An acoustic force field is induced in the transfer device by means of the acoustic wave generator. The frequency of the acoustic wave is set between 1.8 and 2 MHz with a sinusoidal waveform.

[0286] Results

[0287] The first container is enriched with platelets while the second container is depleted of platelets until a predefined platelet level is reached in the second container. After 2.5 hours of processing, the first container holds between 60 and 80% of the platelets while the second container holds between 20 and 40% of the platelets.

Example 2

Blood Fractionation

[0288] Materials and Methods

[0289] Diluted whole blood is injected in equal amount in the first and second container of the present invention, hence each of them holds 50% of the total amount of blood cells in the system. The concentration of blood cell may vary from high concentration to diluted samples. Red blood cells have an average diameter of 6 μm while the platelets have an average diameter of 2 μm.

[0290] A flow is induced through the transfer devices by the flowing means. The flow control means are activated so that each container holds a constant volume of fluid through the process with the flow rate controlled accordingly. The flow in the transfer devices inlets is kept within 0.6 to 1 mL/min. The transfer devices chambers have a thickness between 350 and 450 μm, a width between 0.7 and 2.1 cm and a length between 1 and 6 cm.

[0291] An acoustic force field is induced in the transfer device by means of the acoustic wave generator. The frequency of the acoustic wave is set between 1.8 and 2 MHz with a sinusoidal waveform. This acoustic force field induce the migration of red blood cells toward the central outlet while the platelets tend to stay in the lateral outlets.

[0292] Results

[0293] The first container is enriched with red blood cells while the second container is depleted of red blood cells until a predefined red blood cell level is reached in the second container. After 2.5 hours of processing, the first container holds between 95 and 99% of the red blood cells while the second container holds between 1 and 5% of the red blood cells.

REFERENCES

[0294] 1—System

[0295] 11—First container

[0296] 111—Inlet

[0297] 112—Outlet

[0298] 12—Second container

[0299] 121—Inlet

[0300] 122—Outlet

[0301] 13—First Transfer device

[0302] 131—Chamber

[0303] 132—First wall of the chamber

[0304] 133—Inlet

[0305] 134—First outlet

[0306] 135—Second outlet

[0307] 136—Second wall of the chamber

[0308] 14—Second Transfer device

[0309] 141—Chamber

[0310] 142—First wall of the chamber

[0311] 143—Inlet

[0312] 144—First outlet

[0313] 145—Second outlet

[0314] 146—Second wall of the chamber

[0315] 15—Flowing means

[0316] 16—Acoustic wave generator

[0317] 2—Particle

[0318] 3—Air

[0319] 4—Reflector

[0320] 5—Wall

[0321] x—Longitudinal axis

[0322] y—First transverse axis

[0323] z—Second transverse axis