WO2002072236A1 - Particle separation - Google Patents

Particle separation Download PDF

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Publication number
WO2002072236A1
WO2002072236A1 PCT/SE2002/000429 SE0200429W WO02072236A1 WO 2002072236 A1 WO2002072236 A1 WO 2002072236A1 SE 0200429 W SE0200429 W SE 0200429W WO 02072236 A1 WO02072236 A1 WO 02072236A1
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WO
WIPO (PCT)
Prior art keywords
particles
particle type
standing wave
wave pattern
ultrasonic standing
Prior art date
Application number
PCT/SE2002/000429
Other languages
French (fr)
Inventor
Henrik JÖNSSON
Thomas Laurell
Mats Allers
Hans W Persson
Original Assignee
Erysave Ab
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Erysave Ab filed Critical Erysave Ab
Publication of WO2002072236A1 publication Critical patent/WO2002072236A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/34—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration
    • A61M1/3496—Plasmapheresis; Leucopheresis; Lymphopheresis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/34—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration
    • A61M1/3472—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration with treatment of the filtrate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/34—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration
    • A61M1/3472—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration with treatment of the filtrate
    • A61M1/3479—Filtering material out of the blood by passing it through a membrane, i.e. hemofiltration or diafiltration with treatment of the filtrate by dialysing the filtrate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3678—Separation of cells using wave pressure; Manipulation of individual corpuscles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D21/00—Separation of suspended solid particles from liquids by sedimentation
    • B01D21/28—Mechanical auxiliary equipment for acceleration of sedimentation, e.g. by vibrators or the like
    • B01D21/283—Settling tanks provided with vibrators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2205/00—General characteristics of the apparatus
    • A61M2205/33—Controlling, regulating or measuring
    • A61M2205/3375—Acoustical, e.g. ultrasonic, measuring means

Definitions

  • the present invention deals with separation of particles in a fluid comprising particulate matter. Especially the present invention relates to a method and an apparatus for separating particles, and more specifically for separating particles in a blood solution by means of an acoustic filter using ultrasound.
  • particles or particulate matter comprised in a fluid subjected to an ultrasonic or acoustic standing wave field will be displaced to locations or positions at or in relation to the nodes or antinodes of the standing wave.
  • certain patterns of accumulations of particles will be formed. This is an advantage when separating out particles or particulate matter of a certain weight and/or size from a medium containing a spectrum of particles or particulate matter of different density or acoustic impedance.
  • particles having a density higher than the fluid without particles accumulate in or in the proximity of the nodes, and particles having a density lower than the fluid without particles, accumulate in or in the proximity of the antinodes.
  • particle separation is important when recycling blood in connection with major surgical operations, were the separation is carried out by means of so called blood washing.
  • This method of recycling blood is particularly advantageous in surgical operations when the patient looses large amounts of blood and the need of expensive blood transfusions decreases. Furthermore, the risk for infection and immunization is decreased since the patient's own blood is recycled.
  • blood washing is performed only in extreme cases, since the methods today suffers from low availability and a demand for comparatively expensive manual handling. Instead blood is often recycled unwashed, but more often expensive blood transfusions are used.
  • blood that is lost in connection with a heart operation comprises blood plasma with a number of harmful substances, such as fat emboli. If this lost blood is recycled to the patient without preceding blood washing, these fat emboli can cause thrombus formation, i.e. clots of fat, in the brain.
  • fat emboli can cause thrombus formation, i.e. clots of fat, in the brain.
  • activated complementary complex, deranged coagulation factors and massive fibrinolysis Increased levels of a number of protein usually used as serum markers for different types of organ damages have also been registered, entailing that these markers cannot be used with the previously expected specificity when diagnosing.
  • Another important field for particle separation is the separation of blood plasma from blood cells, were the separated blood plasma can be used for example in dialysis treatment.
  • Yet another important field for particle separation is the separation of blood plasma from blood cells in donated blood plasma. In the latter context there are generally no specific time aspects to deal with, as the case is when washing blood during surgical operation.
  • the drifting standing wave characteristics is adjusted in such a way that the platelets passes through the standing wave to an exit port (320) and that the remaining particulate matter is carried with the drifting standing wave to the window (310). At the window (310), the remaining particulate matter is removed to the container (324) for diluted blood.
  • this apparatus requires that all particulate matter that are not desired among the collected platelets are attached to the drifting standing wave and carried to the window (310). However, such a requirement is almost impossible to fulfill since different particle types have different acoustic properties and will thus, to a greater or less extent, attach to the drifting standing wave.
  • US 4,743,361 does not disclose an apparatus comprising washing to remove undesired particles in the collected liquid comprising the desired particles.
  • the US patent No. 5,484,537 shows another apparatus for manipulating particles in an ultrasonic field wherein a liquid is handled in an open system. Furthermore, the US patent No. 4,055,491 shows an apparatus and a method for separation of particles by means of ultrasound based on flocculation and sedimentation of particles to be separated.
  • the US patent No. 5,831,166 shows the manipulation of micro-particles by means of ultrasound, wherein a concave ultrasound transmitter is positioned in a solution containing micro-particles. Further a reflector is arranged in the focus of the concave ultrasound transmitter, and during operation standing waves are generated between the ultrasound transmitter and the reflector. Due to the fact that an ultrasound field is generated by a concave ultrasound transmitter, the amplitude of the standing wave will have its maximum in the vicinity of the reflector and thereby comparatively stable sonar field points are concentrated to the vicinity of the reflector. It is possible for the micro-particles to be collected in these more stable points and hence the particles are not affected to any particular extent by variations in the ultrasound frequency.
  • the general object of the present invention is to solve the problem of providing an increased separation of particles in blood, i.e. to separate with a higher degree of purification blood cells from blood plasma and possible pollutants.
  • the invention also aims to solve the following aspects of the problem: to provide separation of particles in a system not suffering of a blocked separation filter or decreased permeability with time due to particles clogging in the separation filter, to provide an increased process speed, to provide particle separation in a process, which to a high degree is automatic and requires a minimum of users and a minimum of a user's time for managing the equipment, to provide a system for separation of particles in blood such that the risk for contamination in the processed blood liquid is decreased, to provide a blood washing system enabling an automatic blood washing process, and to provide a system which is more gentle to the cells, as compared to existing techniques utilizing centrifugal force.
  • the stated problem is solved in accordance with the invention by generating in blood a ultrasonic standing wave pattern such that particles having a certain property is displaced by forces from the standing wave into a position in one of the nodes of the standing wave. In this manner there is collecting and holding in the nodes of such particles that for example has a certain size in relation to the wavelength of the generated ultrasonic wave pattern.
  • the blood or a rinsing fluid flows through the standing wave pattern one type of particles is held in the nodes while another type of particles is flowing through and past the standing wave pattern, which then achieves the function of a filter.
  • an acoustic filter is achieved, more specifically an ultrasonic filter that can be used to concentrate one of the particle types in the ultrasonic standing wave pattern and to collect the other particle types in another position.
  • the selection of particles to be caught by the acoustic filter can be based on controlling other parameters than sound wavelength, frequency, for example the distance between the ultrasound transmitter and reflector. Further, the selection of particles held in the nodes can be controlled dependent on their acoustical properties, e.g. their acoustic impedance, which depends on the density of the blood solution. Thus, dependent on the density, compressibility and size of the particles certain patterns of accumulations of particles will be formed in the blood solution exposed the ultrasonic field.
  • the acoustic filter is realized in what could be called a macro-dimension, with an ultrasound generator applied in or on a liquid container accommodating a magnitude of 10-100 ml liquid.
  • a larger liquid container can be used, in some embodiments of the invention the liquid container accommodates a magnitude of 10-500 ml.
  • Separated blood refers to the resulting solution or liquid after particle separation.
  • This liquid contains blood cells, preferably an increased concentration of blood cells, together with blood plasma or rinsing solution and possible contamination substances.
  • the rinsing solution is for example a sodium solution, such as a sodium chloride solution, a Ringer's solution or a glucose solution.
  • the contamination can include air bubbles, fat particles, coagulation products or other not desirable biological material.
  • the amount of blood plasma and possible contamination substances is related to the time during which the particle separation according to the present invention has been performed.
  • the liquid includes only blood cells and a possible carrier fluid, e.g. a rinsing fluid.
  • Collected blood is blood that has been collected from a living being, and comprising blood plasma and possible contamination substances such as fat emboli, complementary complexes, deranged coagulation factors, cytostatics and/or massive fibrinolysis.
  • Acoustic filter refers to the functional filter that occurs when standing waves are generated between an ultrasound transmitter and a reflector.
  • the filter When the filter is applied in a liquid, particles or substances comprised in or suspended in the liquid are subjected to forces exerting the particles towards the nodes or antinodes of the standing waves. The particles will thereby be arranged at different locations depending on their physical and acoustic properties. Particles having a certain size, density and/or compressibility are for example held or fixed in the nodes of the standing waves and particles having another size, density and/or compressibility can be carried with a rinsing fluid through the field of the standing waves.
  • the applied ultrasound has the function of a filter in which particles having a certain size are concentrated and fixed.
  • the size of the particles that are caught in the filter can be varied dependent on the distance between the ultrasound transmitter and the reflector or dependent on the ultrasound frequency. Furthermore, different particles having the same size can be separated dependent on their acoustic properties or density.
  • Nodes refer to pressure nodes of a generated ultrasonic standing wave, where particles of higher density than the solution or medium and/or lower compressibility will tend to accumulate, due to the inherent physical properties of the ultrasound standing wave.
  • Antinodes refer to pressure nodes of a generated ultrasonic standing wave, where particles of lower density than the solution or medium and/or higher compressibility will tend to accumulate, due to the inherent physical properties of the ultrasound standing wave.
  • Particles refers to cells, such as red and white blood cells, virus, proteins, molecules, or to accumulations of substances or matters
  • Micro-particles refer to particles having a diameter less than 15 ⁇ m.
  • Fig la and lb shows schematically flows of liquid to and from a separation container in accordance with the invention
  • Fig 2 shows a first exemplifying embodiment of the system in accordance with the present invention
  • Fig 3 shows a second exemplifying embodiment of the system in accordance with the present invention.
  • Fig 4 shows schematically an embodiment using gravity for the mechanism for generating a liquid flow.
  • the present invention is directed to the separation of particles or particulate matter suspended in a solution and particularly to the separation of particles, mostly micro-particles comprised in a solution. Further, the invention refers to separation and subsequent washing of particles.
  • the present invention is capable to separate particles of a first particle type from particles of a second particle type suspended in a solution, for example to separate cells in a suspension. More specifically the present invention can be used to separate for example bacteria, virus, protozoa, red and white blood cells, stem cells or another kind of cells from a living being. Further, the inventive concept can be used in other microbiological techniques for separating microorganism from a cell culture.
  • the present invention will be described by means of separation of micro-particles from a blood solution or blood collected from a living being.
  • the present invention can be used for separating other particles or particulate matters comprised or suspended in a solution.
  • Fig. la and lb show schematically flows of collected blood, rinsing fluid, blood plasma containing possible contaminating substances and separated blood to and from a separation container, which separation container is comprised in an apparatus in accordance with the present invention.
  • micro-particles such as blood cells can be separated from blood plasma, by means of a closed system in accordance with the invention where separated particles as well as remaining liquid or solution are collected in two separate containers. Since collected blood comprises contaminating substances these are separated together with the blood plasma from the blood cells, whereby the washing or purification of the blood is achieved whereupon the separated blood can be infused in a patient. In cases where collected blood from a living being is free from or only contains a small amount of contaminated substances, a separation of only blood plasma from blood cells can be performed and the separated blood plasma can be infused in a patient.
  • An embodiment of the present invention thus comprises a closed system for particle separation, by means of which particles in for example collected blood can be separated.
  • One embodiment of the particle separation comprises the steps of:
  • a sterile priming solution preferably a sodium chloride solution or a glucose solution
  • the apparatus thus comprises two fluid circuits. Firstly, a first circuit where collected blood is supplied to the separation container, whereby superfluous priming solution is forced to flow out of the separation container. Secondly, a second circuit where the rinsing fluid is supplied to the separation container and collected blood is flowing through the separation container and the acoustic filter, whereupon particles in the blood are caught in the filter and whereupon blood plasma and possible contaminating substances flow unaffected through the filter out in the separation container.
  • FIG. 2 shows a more detailed embodiment of an apparatus for performing the steps described above.
  • a sterile separation container 500 which has been treated with an anticoagulation agent, is filled with a sterile priming solution preferably in the form of a sodium chloride solution or a glucose solution, in order to avoid for example air or air bubbles in the system.
  • a sterile priming solution preferably in the form of a sodium chloride solution or a glucose solution, in order to avoid for example air or air bubbles in the system.
  • blood to be separated is collected, for example blood lost during a surgical operation.
  • the collected blood is put in a first sterile container 100, which preferably also have been treated with an anticoagulation agent.
  • Collected blood is thereafter arranged to flow by means of a flowing mechanism in the shape of a first activating device 120, for example a roller pump, into the separation container 500 through a first inlet 501 of the separation container 500 and a first sealed connection 110 to a predetermined level, preferably at level with the ultrasound transmitter 510 and the reflector 520.
  • a first activating device 120 for example a roller pump
  • only the first connection 110 and the first inlet 501, and the second sealed connection 210 and a second outlet 502 of the separation container 500 are open, when supplying collected blood to the separation container 500.
  • a part of the priming solution present in the separation container 500 is forced to flow out of the second outlet 502 of the separation container 500 through the second connection 210 to a second sterile container 200.
  • the supplying of collected blood to the separation container 500 is stopped manually, but it is also conceivable to provide one or several sensors arranged on the separation container 500 in order to control the supply.
  • the sensors can for example be arranged to register the amount of collected blood in the container 500 or the amount of collected blood supplied to the container 500 via the first connection 110.
  • the first connection 110 is closed by means of the first activating means 120.
  • An ultrasonic standing wave pattern 505 having a suitable frequency in the range of 0.5-10 MHz, preferably in the range of 2 MHz, is applied or generated by means of the ultrasound transmitter 510 over a predetermined part, area or space of the upper part of the separation container 500.
  • the generated ultrasonic standing wave pattern 505 is preferably a static or stationary standing wave pattern, i.e. the frequency of the ultrasonic field is static or constant.
  • the separation container 500 is preferably dimensioned such that an ultrasonic standing wave pattern 505 is occurring between the ultrasound transmitter 510 and the reflector 520, whereupon said part of the separation container 500 forms an acoustic filter.
  • a sterile rinsing fluid preferably a sodium chloride solution or a glucose solution, is arranged to flow from a third sterile container 300 by means of a third actuating mechanism 320, for example a roller pump, into a second inlet 503 of the separation container 500 through a third sealed connection 310.
  • the third connection 310 is in one embodiment of the invention arranged in the same part of the separation container 500 as the first connection 110.
  • the flow of rinsing fluid causes collected blood to flow upwards in the separation container 500 through the acoustic filter, whereby the generated ultrasonic standing wave pattern causes the blood cells to be concentrated and fixed in the acoustic filter, e.g. in the nodes of the generated standing ultrasonic wave pattern.
  • the second outlet 502 and the second connection 210 are open in addition to the open second inlet 503 and the open third connection 310, whereupon the rinsing fluid that flows through the separation container 500 causes collected blood plasma and possible contaminating substances to pass substantially unaffected by the ultrasonic standing wave pattern out of the separation container 500 through the second connection 210 to the second container 200.
  • aggregations or accumulations of blood cells are achieved in the nodes of the ultrasonic standing wave pattern, which accumulations of blood cells or particle clusters tend to move downwards due to the larger gravitational force.
  • the flow of rinsing fluid is stopped by means of the third activating mechanism 320, whereupon the separated blood by means of the fourth activating means 420, preferably a roller pump, is pumped out of a first outlet 504 of the separation container 500 through a fourth sealed connection 410 to a sterile container 400 for separated blood.
  • the fourth activating means 420 preferably a roller pump
  • the suitable level of flow rate depends on the fluid that is to be washed and its degree of contamination, but can in many cases be in the range of 100-1000 ml per hour and preferably in the range of 500 ml per hour.
  • the first and a second connection 110 and 210, respectively, are preferably arranged in a sealed connection with the lower and the upper part, respectively, of the separation container 500, or vice versa.
  • the first connection 110 and the first inlet 501 are preferably arranged in the lower part of the separation container 500, and the second connection 210 and the second outlet 502 in the upper part of the separation container 500.
  • the ultrasound transmitter 510 and the reflector 520 that are arranged on opposite sides of the separation container 500 are preferably designed with a plane and rectangular shape in order to generate an homogenous ultrasound field, which field generates transversal wave fronts in the separation container 500. Further, the generated homogenous ultrasonic standing wave pattern between the transmitter 510 and the reflector 520 will have a direction of propagation perpendicular to the flat surfaces of the transmitter 510 and the reflector 520.
  • the ultrasound generating means 510 and the reflecting means 520 have the same or approximately the same diametrical dimensions in the following ranges: width 5-100 mm, height 5-100 mm and depth/thickness 1-5 mm.
  • the thickness of the reflecting means depends on the material, which the reflecting means is constructed of, but the thickness should be one fourth of the wavelength or an even multiple of one fourth of the wavelength.
  • the reflector 520 can further be implemented as a second passive ultrasound transmitter or a reflecting material, such as a metal or a ceramic material.
  • the reflector may be integrated with the wall of the container 500 or even be a part of the container 500 wall.
  • the ultrasound transmitter 510 is preferably realized in a piezoelectric material vibrating in thickness mode, whereby mechanical oscillations are provided.
  • the frequency of the oscillations is provided in such a way that the wavelength is somewhat larger than the double diameter of the particles to be separated.
  • the frequency is in the interval from 0.5-10 MHz, preferably in the range of 2 MHz.
  • the frequency of 2 MHz has turned out to be advantageous in order to fix and concentrate micro-particles having a diameter in the range of 7 ⁇ m, such as red blood cells, in the acoustic filter. This frequency can however be varied dependent on the size of the particles to be fixed in the filter.
  • the applied frequency is preferably such that the diameter of the particles to be separated, that is the particles to be fixed, are less than half the wavelength of the waves.
  • the ultrasound transmitter 510 is connected to a control unit 550 applying a voltage over the ultrasound transmitter 510.
  • This voltage is in the range of 10 - 100 Vpp, and preferably in the range of 15- 50 Vpp.
  • the suitable order of magnitude of the applied voltage depends on the material of the ultrasound generating means, since a too large applied voltage can cause the ultrasound generating means 510 to get out of order.
  • a voltage of about 30 Vpp is applied when the frequency is about 2MHz and a voltage less than approximately 25 Vpp is applied when the frequency is about 4 MHz.
  • control unit 550 is arranged to control the ultrasound energy delivered by the ultrasound generating means 510 and arranged to deliver electrical energy of a certain waveform, frequency and power.
  • the waveform is controllable to be one of, but not limited to, sinus form, triangular form or square form.
  • the reflecting means or reflector 520 can be realized as passive ultrasonic generating means or as means consisting of an ultrasonic reflecting material, such as a metal or a ceramic material. Further, the reflectors 520 can be integrated with the wall of the container 500 or even be a part of the container 500 wall. In the latter case, the chamber 500 wall is preferably manufactured of a material having desired reflecting properties to reflect the generated ultrasonic wave substantially unmodified.
  • the ultrasound generating means 510 and the reflecting means 520 can be arranged in acoustic connection with the separation container 500.
  • the generating means 510 and the reflecting means 520 are separated a distance in the range of 5-100 mm, preferably a distance of about 30 mm.
  • the transmitters 510 and the reflectors 520 can also be arranged on the inside or outside of the separation container 500. If the transmitters 510 and the reflectors 520 are arranged on the inside of the container 500, they should be manufactured of a material that does not interact with or affect the solution in the separation container 500. If on the other hand the transmitters 510 and the reflectors 520 are arranged on the outside of the separation container 500, the walls of the separation container 500 should be manufactured of a material not affecting the generated ultrasonic standing wave pattern, i.e. of a material transparent or permeable for the generated ultrasonic field.
  • a second exemplifying embodiment is shown in fig 3, however being based on the same principle for particle separation as described above.
  • the flow from the container 100 containing collected blood is directed to the separation container 500 by means of a first 110a branch of the first connection 110 and the flow from the separation container 500 to the container 400 for separated blood is directed by means of a second 110b branch of the first connection 110.
  • the flow via the first 110a and second 110b branch is controlled by means of the activating means 120 and by the control devices 130a and 130b, which preferably are realized by means of electromagnetic valves.
  • a first online sensor 140 positioned close to the first connection 110.
  • This first on-line sensor 140 detects for example the concentration of red blood cells, that is haematocrit.
  • the second on-line sensor 240 e.g. a spectrophotometer, can be arranged in connection to the other connection 210 for detecting for example the concentration of contamina- ting substances being flushed out of the separation container 500. This second sensor 240 can thus control the time during which the separation takes place.
  • the other sensor 240 can also be arranged such that it detects haematocrit and possible the occurrence of air in the flushed fluid.
  • all components such as container 100, 200, 300, 400, 500, connections 110, 210, 310, 410 and branches 110a, 110b, comprised in the apparatus according to embodiments of the invention are preferably sterile and treated with an anti-coagulation agent, such as heparin, citrate or ACD (acid citrate dextrose).
  • an anti-coagulation agent such as heparin, citrate or ACD (acid citrate dextrose).
  • the present invention further comprises different embodiments of apparatuses, by means of which the above described particle separation can be carried out.
  • Embodiments of the apparatuses would comprise actuating mechanisms 120, 220, 320 and in one embodiment also actuating means 420 and control devices 130a, 130b, ultrasound transmitter 510 and reflector 520 and sensors 140, 240.
  • the separation container 500 can be connected to the apparatus, and the ultrasound transmitter 510 and the reflector 520 can be arranged in sealed connection against the separation container 500, the latter being separated at a mutual distance preferably in the range of 5-30 mm.
  • the containers 100, 200, 300, 400 are therefore arranged in sealed connection with the separation container 500 by means of said connections 110, 210, 310 and in a second embodiment by means of connection 410.
  • the sensors 140, 240 can be arranged at the apparatus such that they are positioned outside the connections 110, 210.
  • the apparatus may further comprise a not shown user interface, preferably a display and keyboard, by means of which the apparatus can present data to a user and/or the user can input data to the apparatus, for example the processing time of the separation or the degree of purity of the blood desired by the user.
  • Fig 4 shows schematically a very simple embodiment of the invention in which the flow of fluid is based on gravitation.
  • a container 401 which may be a rigid cup, bag or the like, is provided with an ultrasound generator 407 on the outside, the inside or integrated with the container wall.
  • the container 401 is provided with an outlet 403 and possible also an inlet 402, which possibly are provided with a closeable valve.
  • the container 401 is filled with a liquid in the shape of blood 405 and the ultrasound generator 407 generates a standing wave 409, in the nodes of which particles of a certain particle type having certain physical properties are collected. Due to gravitation fluid is flowing out of the container 401 while said particles are held in the standing wave. In this way the concentration of said particles can be increased in the fluid 405 and be flushed out or removed after disactivating the standing wave.
  • the container 401 can at the inlet 402 or the outlet 403 of course be connected to selectable containers for inputting blood or rinsing fluid, respectively, or for gathering rinsing fluid or particle concentration. It is considered that such simple embodiments of the invention can be used for alternative service conditions or as a low cost product.
  • the separation container 500 is in one embodiment realized in a suitable disposable material that is transparent to the applied or generated ultrasound field, that is a material which does not or to a low extent affect the applied ultrasound field.
  • the separation container 500 is preferably made out of plastic or a ceramic material.
  • the walls of the separation container 500 should be sufficiently thick in order to provide a required degree of stability and on the other hand be sufficiently thin in order to reduce the interfering effect on the material on the applied ultrasound field.
  • a suitable thickness of the walls should be in the range 0.2-3 mm, and in a preferred embodiment the thickness is 1 mm.
  • An embodiment of the separation container 500 has the diametrical dimensions in the following ranges: width 20-100 mm, height 20- 150 mm and depth 20-100 mm, and preferably 30 mm x 60 mm x 30 mm. Further, the separation container 500 is sterilized and treated with an anti-coagulation agent such as heparin, ACD (acid citrate dextrose) or citrate.
  • an anti-coagulation agent such as heparin, ACD (acid citrate dextrose) or citrate.
  • the ultrasound transmitter 510 and the reflector 520 are arranged to have corresponding shapes, and are preferably plane and rectangular in order to generate a homogenous ultrasound field.
  • the ultrasound transmitter 510 is preferably constituted by a piezoelectric material, and presents a suitable frequency in the range of 0.5-10 MHz, wherein the wavelength is somewhat larger than the double diameter of the particles to be separated. Preferably the applied frequency is about 2 MHz.
  • the ultrasound transmitter 510 is connected to a generator applying a voltage over the ultrasound transmitter 510. This voltage is in the range of 10 - 100 Vpp, preferably in the range of 15-50 Vpp.
  • the reflector 520 can be constituted by a second passive ultrasound transmitter or by a reflecting material, such as a metal or a ceramic material.
  • Embodiments of the ultrasound transmitter 510 and the reflector 520 are preferably designed with the same dimensions in the following ranges: width 10-100 mm, height 10-100 mm and depth 1-5 mm.
  • the ultrasound transmitter 510 and the reflector 520 are further arranged on opposite sides of the separation container 500.
  • Embodiments of the containers 100, 200, 300, 400 are preferably realized in a suitable sterile disposable material such as a plastic polymer or glass.
  • the embodiments of the containers 100, 200, 300, 400 are on their insides treated with an anti-coagulation agent and they have the shape of a bag or a bottle with a volume in a range of 50-5000 ml, preferably in the range of 500 ml.
  • connection 110, 210, 310, 410 are preferably realized as tubings made in a suitable disposable material such as polyurethane, PVC, silicone or other plastic.
  • the connections are further arranged to have a suitable diameter to provide a desired throughput.
  • the diameter of the tubings is preferably in the range of 1.5-2.5 mm.
  • the connections are preferably also sterile and treated with an anticoagulation agent.
  • An embodiment of the present invention realized as a kit of articles used for particle separation would comprise a separation container 500, a first and second container 100 and 200, respectively, and a third container 300 comprising a rinsing fluid and a fourth container 400 for separated blood.
  • An embodiment of the kit would further comprise the first connection 110 having a first and a second branch 110a and 110b or a first connection 110 and a fourth connection 410.
  • Embodiments of the kit would also comprise a second and a third connection 210 and 310, respectively. All the components of an embodiment of the kit would be sterile and treated with an anticoagulation agent. In one embodiment of the kit even the ultrasound transmitter 510 and the reflector 520 may be realized in suitable disposable materials.
  • the invention also comprises a separation product for example a blood product resulting from a process in accordance with the steps of the inventive method.

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Abstract

The present invention deals with separation of particles in a fluid comprising particulate matter, and especially the invention relates to a method and an apparatus for separating particles, and more specifically for separating particles in a blood solution by means of an acoustic filter using ultrasound. Embodiments of the method and system for separation of a blood solution uses a stationary ultrasonic standing wave pattern (505) for collecting particles of a first particle type in the nodes of the standing wave pattern (505); and generating a flow of liquid through the standing wave pattern (505) carrying away particles of a second particle type.

Description

PARTICLE SEPARATION
Field of the invention The present invention deals with separation of particles in a fluid comprising particulate matter. Especially the present invention relates to a method and an apparatus for separating particles, and more specifically for separating particles in a blood solution by means of an acoustic filter using ultrasound.
Background
It is well-known that particles or particulate matter comprised in a fluid subjected to an ultrasonic or acoustic standing wave field will be displaced to locations or positions at or in relation to the nodes or antinodes of the standing wave. Depending on the density, compressibility and size of the particles, certain patterns of accumulations of particles will be formed. This is an advantage when separating out particles or particulate matter of a certain weight and/or size from a medium containing a spectrum of particles or particulate matter of different density or acoustic impedance. Generally, particles having a density higher than the fluid without particles, accumulate in or in the proximity of the nodes, and particles having a density lower than the fluid without particles, accumulate in or in the proximity of the antinodes.
Several techniques for manipulating or separating particulate matter suspended in a suspension have been proposed within several fields of technology. Separating particles is for example important within several fields of medical technology and different separation methods are used for example in connection with blood donations, dialysis and laboratory analysis.
Further, particle separation is important when recycling blood in connection with major surgical operations, were the separation is carried out by means of so called blood washing. This method of recycling blood is particularly advantageous in surgical operations when the patient looses large amounts of blood and the need of expensive blood transfusions decreases. Furthermore, the risk for infection and immunization is decreased since the patient's own blood is recycled. At present blood washing is performed only in extreme cases, since the methods today suffers from low availability and a demand for comparatively expensive manual handling. Instead blood is often recycled unwashed, but more often expensive blood transfusions are used.
In the last few years there has been a plurality of scientific findings in favor of blood washing. For example, it has within heart surgery been found that blood that is lost in connection with a heart operation comprises blood plasma with a number of harmful substances, such as fat emboli. If this lost blood is recycled to the patient without preceding blood washing, these fat emboli can cause thrombus formation, i.e. clots of fat, in the brain. In the blood plasma, there has also been found activated complementary complex, deranged coagulation factors and massive fibrinolysis. Increased levels of a number of protein usually used as serum markers for different types of organ damages have also been registered, entailing that these markers cannot be used with the previously expected specificity when diagnosing.
Another important field for particle separation is the separation of blood plasma from blood cells, were the separated blood plasma can be used for example in dialysis treatment. Yet another important field for particle separation is the separation of blood plasma from blood cells in donated blood plasma. In the latter context there are generally no specific time aspects to deal with, as the case is when washing blood during surgical operation.
Prior Art
In prior art there are quite a few examples of particle separation by means of ultrasound. In the US patent No. 4,743,361 there is shown an apparatus for separation of biological particles in a liquid solution by means of ultrasound. An acoustic filter is arranged with standing sound waves generated by two separated ultrasound sources. The actual particle separation is achieved by means of controlling the flow of the liquid through the standing waves, attenuating the standing wave or controlling the intensity or frequency of the standing wave. An embodiment of the apparatus, as shown in figure 11, is used for the collection of e.g. a blood fraction, such as platelets. In this embodiment the drifting standing wave characteristics is adjusted in such a way that the platelets passes through the standing wave to an exit port (320) and that the remaining particulate matter is carried with the drifting standing wave to the window (310). At the window (310), the remaining particulate matter is removed to the container (324) for diluted blood. Further, this apparatus requires that all particulate matter that are not desired among the collected platelets are attached to the drifting standing wave and carried to the window (310). However, such a requirement is almost impossible to fulfill since different particle types have different acoustic properties and will thus, to a greater or less extent, attach to the drifting standing wave. Furthermore, US 4,743,361 does not disclose an apparatus comprising washing to remove undesired particles in the collected liquid comprising the desired particles.
The US patent No. 5,484,537 shows another apparatus for manipulating particles in an ultrasonic field wherein a liquid is handled in an open system. Furthermore, the US patent No. 4,055,491 shows an apparatus and a method for separation of particles by means of ultrasound based on flocculation and sedimentation of particles to be separated.
The US patent No. 5,831,166 shows the manipulation of micro-particles by means of ultrasound, wherein a concave ultrasound transmitter is positioned in a solution containing micro-particles. Further a reflector is arranged in the focus of the concave ultrasound transmitter, and during operation standing waves are generated between the ultrasound transmitter and the reflector. Due to the fact that an ultrasound field is generated by a concave ultrasound transmitter, the amplitude of the standing wave will have its maximum in the vicinity of the reflector and thereby comparatively stable sonar field points are concentrated to the vicinity of the reflector. It is possible for the micro-particles to be collected in these more stable points and hence the particles are not affected to any particular extent by variations in the ultrasound frequency. In prior art there is also blood washing using systems and methods for particle separation by centrifugal force. These systems, however, present several drawbacks compared to systems based on ultrasound. For example, the particles to be separated are damaged by the force exerted on the particles during centrifugation. The US patent No. 5,971,948 shows a system for collecting, washing and recycling to a patient blood lost during surgical operation. This system requires dilution of collected blood before centrifugation to allow lower centrifugal velocities in order to reduce the damages on the red blood cells. This dilution thus entail longer processing times for separating particles as compared to separation by means of ultrasound, which is of major importance during a surgical operation when the patient looses large amounts of blood that has to be replaced.
The US patent No. 5,964,724 shows another system for blood separation by means of centrifugation.
Object of the invention The general object of the present invention is to solve the problem of providing an increased separation of particles in blood, i.e. to separate with a higher degree of purification blood cells from blood plasma and possible pollutants. The invention also aims to solve the following aspects of the problem: to provide separation of particles in a system not suffering of a blocked separation filter or decreased permeability with time due to particles clogging in the separation filter, to provide an increased process speed, to provide particle separation in a process, which to a high degree is automatic and requires a minimum of users and a minimum of a user's time for managing the equipment, to provide a system for separation of particles in blood such that the risk for contamination in the processed blood liquid is decreased, to provide a blood washing system enabling an automatic blood washing process, and to provide a system which is more gentle to the cells, as compared to existing techniques utilizing centrifugal force.
Summary of the invention
The stated problem is solved in accordance with the invention by generating in blood a ultrasonic standing wave pattern such that particles having a certain property is displaced by forces from the standing wave into a position in one of the nodes of the standing wave. In this manner there is collecting and holding in the nodes of such particles that for example has a certain size in relation to the wavelength of the generated ultrasonic wave pattern. When the blood or a rinsing fluid flows through the standing wave pattern one type of particles is held in the nodes while another type of particles is flowing through and past the standing wave pattern, which then achieves the function of a filter. In this way an acoustic filter is achieved, more specifically an ultrasonic filter that can be used to concentrate one of the particle types in the ultrasonic standing wave pattern and to collect the other particle types in another position.
The selection of particles to be caught by the acoustic filter can be based on controlling other parameters than sound wavelength, frequency, for example the distance between the ultrasound transmitter and reflector. Further, the selection of particles held in the nodes can be controlled dependent on their acoustical properties, e.g. their acoustic impedance, which depends on the density of the blood solution. Thus, dependent on the density, compressibility and size of the particles certain patterns of accumulations of particles will be formed in the blood solution exposed the ultrasonic field.
In different applications of the invention and for different purposes a standing wave and a rinsing flow is configured in a suitable way, which are exemplified in the detailed description of different embodiments below.
According to different aspects of the invention the acoustic filter is realized in what could be called a macro-dimension, with an ultrasound generator applied in or on a liquid container accommodating a magnitude of 10-100 ml liquid. However, a larger liquid container can be used, in some embodiments of the invention the liquid container accommodates a magnitude of 10-500 ml.
Definitions In the present text the following terminology will be used:
Separated blood refers to the resulting solution or liquid after particle separation. This liquid contains blood cells, preferably an increased concentration of blood cells, together with blood plasma or rinsing solution and possible contamination substances. The rinsing solution is for example a sodium solution, such as a sodium chloride solution, a Ringer's solution or a glucose solution. The contamination can include air bubbles, fat particles, coagulation products or other not desirable biological material. The amount of blood plasma and possible contamination substances is related to the time during which the particle separation according to the present invention has been performed. In optimal particle separation the liquid includes only blood cells and a possible carrier fluid, e.g. a rinsing fluid.
Collected blood is blood that has been collected from a living being, and comprising blood plasma and possible contamination substances such as fat emboli, complementary complexes, deranged coagulation factors, cytostatics and/or massive fibrinolysis.
Acoustic filter refers to the functional filter that occurs when standing waves are generated between an ultrasound transmitter and a reflector. When the filter is applied in a liquid, particles or substances comprised in or suspended in the liquid are subjected to forces exerting the particles towards the nodes or antinodes of the standing waves. The particles will thereby be arranged at different locations depending on their physical and acoustic properties. Particles having a certain size, density and/or compressibility are for example held or fixed in the nodes of the standing waves and particles having another size, density and/or compressibility can be carried with a rinsing fluid through the field of the standing waves. Thereby the applied ultrasound has the function of a filter in which particles having a certain size are concentrated and fixed. The size of the particles that are caught in the filter can be varied dependent on the distance between the ultrasound transmitter and the reflector or dependent on the ultrasound frequency. Furthermore, different particles having the same size can be separated dependent on their acoustic properties or density.
Nodes refer to pressure nodes of a generated ultrasonic standing wave, where particles of higher density than the solution or medium and/or lower compressibility will tend to accumulate, due to the inherent physical properties of the ultrasound standing wave. Antinodes refer to pressure nodes of a generated ultrasonic standing wave, where particles of lower density than the solution or medium and/or higher compressibility will tend to accumulate, due to the inherent physical properties of the ultrasound standing wave. Particles refers to cells, such as red and white blood cells, virus, proteins, molecules, or to accumulations of substances or matters
Micro-particles refer to particles having a diameter less than 15 μm.
Brief description of the drawings The invention will be described below with reference to the accompanying drawings, in which:
Fig la and lb shows schematically flows of liquid to and from a separation container in accordance with the invention;
Fig 2 shows a first exemplifying embodiment of the system in accordance with the present invention;
Fig 3 shows a second exemplifying embodiment of the system in accordance with the present invention; and
Fig 4 shows schematically an embodiment using gravity for the mechanism for generating a liquid flow.
Detailed description of the invention
The present invention is directed to the separation of particles or particulate matter suspended in a solution and particularly to the separation of particles, mostly micro-particles comprised in a solution. Further, the invention refers to separation and subsequent washing of particles.
Generally, the present invention is capable to separate particles of a first particle type from particles of a second particle type suspended in a solution, for example to separate cells in a suspension. More specifically the present invention can be used to separate for example bacteria, virus, protozoa, red and white blood cells, stem cells or another kind of cells from a living being. Further, the inventive concept can be used in other microbiological techniques for separating microorganism from a cell culture.
By way of example only, the present invention will be described by means of separation of micro-particles from a blood solution or blood collected from a living being. However, as comprehended by a skilled person in the art, the present invention can be used for separating other particles or particulate matters comprised or suspended in a solution.
Fig. la and lb show schematically flows of collected blood, rinsing fluid, blood plasma containing possible contaminating substances and separated blood to and from a separation container, which separation container is comprised in an apparatus in accordance with the present invention.
With the inventive system, micro-particles such as blood cells can be separated from blood plasma, by means of a closed system in accordance with the invention where separated particles as well as remaining liquid or solution are collected in two separate containers. Since collected blood comprises contaminating substances these are separated together with the blood plasma from the blood cells, whereby the washing or purification of the blood is achieved whereupon the separated blood can be infused in a patient. In cases where collected blood from a living being is free from or only contains a small amount of contaminated substances, a separation of only blood plasma from blood cells can be performed and the separated blood plasma can be infused in a patient.
An embodiment of the present invention thus comprises a closed system for particle separation, by means of which particles in for example collected blood can be separated. One embodiment of the particle separation comprises the steps of:
1. filling a sterilized separation container with a sterile priming solution, preferably a sodium chloride solution or a glucose solution;
2. collecting blood from a patient to a first sterilized container;
3. filling a separation container with the collected blood to a predetermined level;
4. tapping of possible excess of priming solution to a second container;
5. applying an ultrasound, i.e. an ultrasonic field, in a part of the separation container, said part thereby forming an acoustic filter, entailing a concentration and fixation of blood cells in said part; 6. adding a sterile rinsing fluid, preferably a sodium chloride solution or a glucose solution from a first sterilized container to the separation container, whereupon blood plasma and possible contaminating substances unaffected by the ultrasound are flushed through the acoustic filter; 7. collecting blood plasma and possible contaminating substances in the second container;
8. after sufficient separation stopping the supply of rinsing fluid and the ultrasound wave can be discontinued to avoid further heating;
9. collecting the separated blood to a fourth sterilizing container for separated blood;
10. repeating steps 3-9 until the desired degree of concentration or separation has been obtained. 8 1 9 -04- 2002
The apparatus according to this embodiment thus comprises two fluid circuits. Firstly, a first circuit where collected blood is supplied to the separation container, whereby superfluous priming solution is forced to flow out of the separation container. Secondly, a second circuit where the rinsing fluid is supplied to the separation container and collected blood is flowing through the separation container and the acoustic filter, whereupon particles in the blood are caught in the filter and whereupon blood plasma and possible contaminating substances flow unaffected through the filter out in the separation container.
Fig. 2 shows a more detailed embodiment of an apparatus for performing the steps described above. A sterile separation container 500, which has been treated with an anticoagulation agent, is filled with a sterile priming solution preferably in the form of a sodium chloride solution or a glucose solution, in order to avoid for example air or air bubbles in the system. Thereafter blood to be separated is collected, for example blood lost during a surgical operation. The collected blood is put in a first sterile container 100, which preferably also have been treated with an anticoagulation agent.
Collected blood is thereafter arranged to flow by means of a flowing mechanism in the shape of a first activating device 120, for example a roller pump, into the separation container 500 through a first inlet 501 of the separation container 500 and a first sealed connection 110 to a predetermined level, preferably at level with the ultrasound transmitter 510 and the reflector 520. In one embodiment of the invention, only the first connection 110 and the first inlet 501, and the second sealed connection 210 and a second outlet 502 of the separation container 500 are open, when supplying collected blood to the separation container 500. Whereupon a part of the priming solution present in the separation container 500 is forced to flow out of the second outlet 502 of the separation container 500 through the second connection 210 to a second sterile container 200. Preferably the supplying of collected blood to the separation container 500 is stopped manually, but it is also conceivable to provide one or several sensors arranged on the separation container 500 in order to control the supply. The sensors can for example be arranged to register the amount of collected blood in the container 500 or the amount of collected blood supplied to the container 500 via the first connection 110.
When the separation container 500 has been filled with a desired amount of collected blood, the first connection 110 is closed by means of the first activating means 120. An ultrasonic standing wave pattern 505 having a suitable frequency in the range of 0.5-10 MHz, preferably in the range of 2 MHz, is applied or generated by means of the ultrasound transmitter 510 over a predetermined part, area or space of the upper part of the separation container 500. The generated ultrasonic standing wave pattern 505 is preferably a static or stationary standing wave pattern, i.e. the frequency of the ultrasonic field is static or constant. Further, the separation container 500 is preferably dimensioned such that an ultrasonic standing wave pattern 505 is occurring between the ultrasound transmitter 510 and the reflector 520, whereupon said part of the separation container 500 forms an acoustic filter. A sterile rinsing fluid, preferably a sodium chloride solution or a glucose solution, is arranged to flow from a third sterile container 300 by means of a third actuating mechanism 320, for example a roller pump, into a second inlet 503 of the separation container 500 through a third sealed connection 310. The third connection 310 is in one embodiment of the invention arranged in the same part of the separation container 500 as the first connection 110. The flow of rinsing fluid causes collected blood to flow upwards in the separation container 500 through the acoustic filter, whereby the generated ultrasonic standing wave pattern causes the blood cells to be concentrated and fixed in the acoustic filter, e.g. in the nodes of the generated standing ultrasonic wave pattern. During this flow of rinsing fluid only the second outlet 502 and the second connection 210 are open in addition to the open second inlet 503 and the open third connection 310, whereupon the rinsing fluid that flows through the separation container 500 causes collected blood plasma and possible contaminating substances to pass substantially unaffected by the ultrasonic standing wave pattern out of the separation container 500 through the second connection 210 to the second container 200. By having the rinsing fluid flowing continuously through the separation container 500 an efficient separation of blood plasma and possible contaminating substances from blood cells is achieved.
In one embodiment of the invention, aggregations or accumulations of blood cells are achieved in the nodes of the ultrasonic standing wave pattern, which accumulations of blood cells or particle clusters tend to move downwards due to the larger gravitational force.
When the desired separation or the desired degree of separation or washing has been achieved the flow of rinsing fluid is stopped by means of the third activating mechanism 320, whereupon the separated blood by means of the fourth activating means 420, preferably a roller pump, is pumped out of a first outlet 504 of the separation container 500 through a fourth sealed connection 410 to a sterile container 400 for separated blood.
Thereafter, new collected blood can be pumped or supplied into the separation container 500 in accordance with the description above and the procedure can be repeated. The suitable level of flow rate depends on the fluid that is to be washed and its degree of contamination, but can in many cases be in the range of 100-1000 ml per hour and preferably in the range of 500 ml per hour. The first and a second connection 110 and 210, respectively, are preferably arranged in a sealed connection with the lower and the upper part, respectively, of the separation container 500, or vice versa. The first connection 110 and the first inlet 501 are preferably arranged in the lower part of the separation container 500, and the second connection 210 and the second outlet 502 in the upper part of the separation container 500.
Ultrasound generating means and reflecting means
The ultrasound transmitter 510 and the reflector 520 that are arranged on opposite sides of the separation container 500 are preferably designed with a plane and rectangular shape in order to generate an homogenous ultrasound field, which field generates transversal wave fronts in the separation container 500. Further, the generated homogenous ultrasonic standing wave pattern between the transmitter 510 and the reflector 520 will have a direction of propagation perpendicular to the flat surfaces of the transmitter 510 and the reflector 520.
In one embodiment of the invention, the ultrasound generating means 510 and the reflecting means 520 have the same or approximately the same diametrical dimensions in the following ranges: width 5-100 mm, height 5-100 mm and depth/thickness 1-5 mm. However, the thickness of the reflecting means depends on the material, which the reflecting means is constructed of, but the thickness should be one fourth of the wavelength or an even multiple of one fourth of the wavelength.
The reflector 520 can further be implemented as a second passive ultrasound transmitter or a reflecting material, such as a metal or a ceramic material. The reflector may be integrated with the wall of the container 500 or even be a part of the container 500 wall.
The ultrasound transmitter 510 is preferably realized in a piezoelectric material vibrating in thickness mode, whereby mechanical oscillations are provided. The frequency of the oscillations is provided in such a way that the wavelength is somewhat larger than the double diameter of the particles to be separated. In different embodiments of the invention, the frequency is in the interval from 0.5-10 MHz, preferably in the range of 2 MHz. The frequency of 2 MHz has turned out to be advantageous in order to fix and concentrate micro-particles having a diameter in the range of 7 μm, such as red blood cells, in the acoustic filter. This frequency can however be varied dependent on the size of the particles to be fixed in the filter. The applied frequency is preferably such that the diameter of the particles to be separated, that is the particles to be fixed, are less than half the wavelength of the waves.
In order to generate an ultrasonic field, the ultrasound transmitter 510 is connected to a control unit 550 applying a voltage over the ultrasound transmitter 510. This voltage is in the range of 10 - 100 Vpp, and preferably in the range of 15- 50 Vpp. However, the suitable order of magnitude of the applied voltage depends on the material of the ultrasound generating means, since a too large applied voltage can cause the ultrasound generating means 510 to get out of order. In an embodiment of the invention a voltage of about 30 Vpp is applied when the frequency is about 2MHz and a voltage less than approximately 25 Vpp is applied when the frequency is about 4 MHz.
Further, the control unit 550 is arranged to control the ultrasound energy delivered by the ultrasound generating means 510 and arranged to deliver electrical energy of a certain waveform, frequency and power. The waveform is controllable to be one of, but not limited to, sinus form, triangular form or square form.
As mentioned above, the reflecting means or reflector 520 can be realized as passive ultrasonic generating means or as means consisting of an ultrasonic reflecting material, such as a metal or a ceramic material. Further, the reflectors 520 can be integrated with the wall of the container 500 or even be a part of the container 500 wall. In the latter case, the chamber 500 wall is preferably manufactured of a material having desired reflecting properties to reflect the generated ultrasonic wave substantially unmodified.
Further, the ultrasound generating means 510 and the reflecting means 520 can be arranged in acoustic connection with the separation container 500. In one embodiment the generating means 510 and the reflecting means 520 are separated a distance in the range of 5-100 mm, preferably a distance of about 30 mm.
However, in other embodiments of the invention, the transmitters 510 and the reflectors 520 can also be arranged on the inside or outside of the separation container 500. If the transmitters 510 and the reflectors 520 are arranged on the inside of the container 500, they should be manufactured of a material that does not interact with or affect the solution in the separation container 500. If on the other hand the transmitters 510 and the reflectors 520 are arranged on the outside of the separation container 500, the walls of the separation container 500 should be manufactured of a material not affecting the generated ultrasonic standing wave pattern, i.e. of a material transparent or permeable for the generated ultrasonic field.
A second exemplifying embodiment is shown in fig 3, however being based on the same principle for particle separation as described above. In this embodiment however the flow from the container 100 containing collected blood is directed to the separation container 500 by means of a first 110a branch of the first connection 110 and the flow from the separation container 500 to the container 400 for separated blood is directed by means of a second 110b branch of the first connection 110. The flow via the first 110a and second 110b branch is controlled by means of the activating means 120 and by the control devices 130a and 130b, which preferably are realized by means of electromagnetic valves.
In order to check the quality of collected and separated blood with regard to for example the amount of possible contamination and the amount of certain type of particles, there is in some embodiments as shown in fig 2 and 3 arranged a first online sensor 140 positioned close to the first connection 110. This first on-line sensor 140 detects for example the concentration of red blood cells, that is haematocrit. The second on-line sensor 240, e.g. a spectrophotometer, can be arranged in connection to the other connection 210 for detecting for example the concentration of contamina- ting substances being flushed out of the separation container 500. This second sensor 240 can thus control the time during which the separation takes place. The other sensor 240 can also be arranged such that it detects haematocrit and possible the occurrence of air in the flushed fluid.
Furthermore, all components such as container 100, 200, 300, 400, 500, connections 110, 210, 310, 410 and branches 110a, 110b, comprised in the apparatus according to embodiments of the invention are preferably sterile and treated with an anti-coagulation agent, such as heparin, citrate or ACD (acid citrate dextrose).
The present invention further comprises different embodiments of apparatuses, by means of which the above described particle separation can be carried out. Embodiments of the apparatuses would comprise actuating mechanisms 120, 220, 320 and in one embodiment also actuating means 420 and control devices 130a, 130b, ultrasound transmitter 510 and reflector 520 and sensors 140, 240. Further, the separation container 500 can be connected to the apparatus, and the ultrasound transmitter 510 and the reflector 520 can be arranged in sealed connection against the separation container 500, the latter being separated at a mutual distance preferably in the range of 5-30 mm. The containers 100, 200, 300, 400 are therefore arranged in sealed connection with the separation container 500 by means of said connections 110, 210, 310 and in a second embodiment by means of connection 410. Furthermore, the sensors 140, 240 can be arranged at the apparatus such that they are positioned outside the connections 110, 210. The apparatus may further comprise a not shown user interface, preferably a display and keyboard, by means of which the apparatus can present data to a user and/or the user can input data to the apparatus, for example the processing time of the separation or the degree of purity of the blood desired by the user. Fig 4 shows schematically a very simple embodiment of the invention in which the flow of fluid is based on gravitation. A container 401, which may be a rigid cup, bag or the like, is provided with an ultrasound generator 407 on the outside, the inside or integrated with the container wall. The container 401 is provided with an outlet 403 and possible also an inlet 402, which possibly are provided with a closeable valve. In the figure the container 401 is filled with a liquid in the shape of blood 405 and the ultrasound generator 407 generates a standing wave 409, in the nodes of which particles of a certain particle type having certain physical properties are collected. Due to gravitation fluid is flowing out of the container 401 while said particles are held in the standing wave. In this way the concentration of said particles can be increased in the fluid 405 and be flushed out or removed after disactivating the standing wave. The container 401 can at the inlet 402 or the outlet 403 of course be connected to selectable containers for inputting blood or rinsing fluid, respectively, or for gathering rinsing fluid or particle concentration. It is considered that such simple embodiments of the invention can be used for alternative service conditions or as a low cost product.
Disposables
The separation container 500 is in one embodiment realized in a suitable disposable material that is transparent to the applied or generated ultrasound field, that is a material which does not or to a low extent affect the applied ultrasound field. The separation container 500 is preferably made out of plastic or a ceramic material. Further, the walls of the separation container 500 should be sufficiently thick in order to provide a required degree of stability and on the other hand be sufficiently thin in order to reduce the interfering effect on the material on the applied ultrasound field. A suitable thickness of the walls should be in the range 0.2-3 mm, and in a preferred embodiment the thickness is 1 mm. An embodiment of the separation container 500 has the diametrical dimensions in the following ranges: width 20-100 mm, height 20- 150 mm and depth 20-100 mm, and preferably 30 mm x 60 mm x 30 mm. Further, the separation container 500 is sterilized and treated with an anti-coagulation agent such as heparin, ACD (acid citrate dextrose) or citrate.
As mentioned above, the ultrasound transmitter 510 and the reflector 520 are arranged to have corresponding shapes, and are preferably plane and rectangular in order to generate a homogenous ultrasound field. The ultrasound transmitter 510 is preferably constituted by a piezoelectric material, and presents a suitable frequency in the range of 0.5-10 MHz, wherein the wavelength is somewhat larger than the double diameter of the particles to be separated. Preferably the applied frequency is about 2 MHz. In order to generate an ultrasonic field, the ultrasound transmitter 510 is connected to a generator applying a voltage over the ultrasound transmitter 510. This voltage is in the range of 10 - 100 Vpp, preferably in the range of 15-50 Vpp. As previously mentioned, the reflector 520 can be constituted by a second passive ultrasound transmitter or by a reflecting material, such as a metal or a ceramic material. Embodiments of the ultrasound transmitter 510 and the reflector 520 are preferably designed with the same dimensions in the following ranges: width 10-100 mm, height 10-100 mm and depth 1-5 mm. The ultrasound transmitter 510 and the reflector 520 are further arranged on opposite sides of the separation container 500.
Embodiments of the containers 100, 200, 300, 400 are preferably realized in a suitable sterile disposable material such as a plastic polymer or glass. The embodiments of the containers 100, 200, 300, 400 are on their insides treated with an anti-coagulation agent and they have the shape of a bag or a bottle with a volume in a range of 50-5000 ml, preferably in the range of 500 ml.
Embodiments of the connections 110, 210, 310, 410 are preferably realized as tubings made in a suitable disposable material such as polyurethane, PVC, silicone or other plastic. The connections are further arranged to have a suitable diameter to provide a desired throughput. The diameter of the tubings is preferably in the range of 1.5-2.5 mm. The connections are preferably also sterile and treated with an anticoagulation agent.
An embodiment of the present invention realized as a kit of articles used for particle separation would comprise a separation container 500, a first and second container 100 and 200, respectively, and a third container 300 comprising a rinsing fluid and a fourth container 400 for separated blood. An embodiment of the kit would further comprise the first connection 110 having a first and a second branch 110a and 110b or a first connection 110 and a fourth connection 410. Embodiments of the kit would also comprise a second and a third connection 210 and 310, respectively. All the components of an embodiment of the kit would be sterile and treated with an anticoagulation agent. In one embodiment of the kit even the ultrasound transmitter 510 and the reflector 520 may be realized in suitable disposable materials.
The invention also comprises a separation product for example a blood product resulting from a process in accordance with the steps of the inventive method.
The present invention has been explained by means of exemplifying embodiments, but other implementations of the invention within the scope of the accompanying claims are also conceivable.

Claims

1. Method for separating particles in a solution of particulate matter comprising particles of a first and a second particle type, said method comprising the steps of: - receiving said solution of particulate matter in a separation container (500);
- generating a stationary ultrasonic standing wave pattern (505) in a part of said separation container (500) such that said particles of said first particle type, having a first property dependent on the characteristics of said generated ultrasonic standing wave pattern (505), is collected and held at the nodes of said ultrasonic standing wave pattern;
- establishing a flow of a rinsing fluid through said generated ultrasonic standing wave pattern, said rinsing fluid carrying said particles of said second particle type, having a second property dependent on the characteristics of said generated ultrasonic standing wave pattern, such that said particles of said second particle type passes through said generated ultrasonic standing wave pattern (505);
- removing said particles of said second particle type from said separation container (500);
- removing said ultrasonic standing wave pattern (505); and
- emptying said separation container (500), whereby said particles of said first particle type are removed from said separation container (500).
2. The method as recited in claim 1, further comprising the step of increasing the concentration of said particles of said first particle type at said nodes of said generated ultrasonic standing wave pattern (505) by conducting a flow of said solution of particulate matter to said ultrasonic standing wave pattern (505).
3. Method for separating particles in a blood solution comprising particles of a first and a second particle type, said method comprising the steps of:
- receiving said blood solution in a separation container (500); - generating a stationary ultrasonic standing wave pattern (505) in a part of said separation container (500) such that said particles of said first particle type, having a first property dependent on the characteristics of said generated ultrasonic standing wave pattern (505), is collected and held at the nodes of said ultrasonic standing wave pattern (505); - establishing a flow of a rinsing fluid through said generated ultrasonic standing wave pattern (505), said rinsing fluid carrying said particles of said second particle type, having a second property dependent on the characteristics of said generated ultrasonic standing wave pattern (505), such that said particles of said second particle type passes through said generated ultrasonic standing wave pattern (505);
- removing said particles of said second particle type from said separation container (500);
- removing said ultrasonic standing wave pattern (505); and - emptying said separation container (500), whereby said particles of said first particle type are removed from said separation container (500).
4. The method as recited in claim 3, further comprising the step of increasing the concentration of said particles of said first particle type at said nodes of said generated ultrasonic standing wave pattern (505) by conducting a flow of said blood solution to said ultrasonic standing wave pattern (505).
5. The method as recited in claim 1 or 3, further comprising the step of generating said stationary ultrasonic standing wave pattern (505) in such a way that said particles of said first particle type tend to accumulate, whereby particle clusters of said particles of said particle types are created, which particle clusters tend to move downwards due to the gravitational force.
6. The method as recited in claim 1 or 3, further comprising the step of controlling the size of said particles of said first particle type dependent on the distance between an ultrasonic generating means (510) and an ultrasonic reflecting means (520) between which means (510,520) said ultrasonic standing wave pattern (505) is generated.
7. The method as recited in claim 1 or 3, further comprising the step of controlling the size of said particles of said first particle type dependent on the ultrasound frequency at which said ultrasonic standing wave pattern (505) is generated.
8. The method as recited in claim 1 or 3, further comprising the step of controlling the separation of said particles of said first particle type from said particles of said second particle type dependent on the acoustic properties of each particle type, respectively.
9. The method as recited in claim 1 or 3, further comprising the step of controlling the separation of said particles of said first particle type from said particles of said second particle type dependent on the density of each of the particles types, respectively.
10. The method as recited in claim 1 or 3, wherein said particles of said first particle type comprises red blood cells and said particles of said second particle type comprises blood plasma.
11. An apparatus for separating particles in a solution of particulate matter comprising particles of a first and a second particle type, said apparatus comprising:
- a separation container (500) for receiving said solution of particulate matter;
- an ultrasound source (510,520) arranged for generating a stationary ultrasonic standing wave pattern (505) in a part of said separation container (500), whereby said particles of said first particle type, having a first property dependent on the characteristics of said generated ultrasonic standing wave pattern (505), is collected and held at the nodes of said generated ultrasonic standing wave pattern (505);
- a liquid flow mechanism (320) arranged to flow a rinsing fluid through said generated ultrasonic standing wave pattern (505), said rinsing fluid carrying said particles of said second particle type, having a second property dependent on the characteristics of said generated ultrasonic standing wave pattern (505), such that said particles of said second particle type passes through said generated ultrasonic standing wave pattern (505); - a container (200) for receiving said particles of said second particle type;
- means for removing said ultrasonic standing wave pattern (505); and
- an activating means (420) arranged to empty said separation container (500), whereby said particles of said first particle type are removed from said separation container (500).
12. An apparatus for separating particles in a blood solution comprising particles of a first and a second particle type, said apparatus comprising:
- a separation container (500) for receiving said blood solution;
- an ultrasound source (510,520) arranged for generating a stationary ultrasonic standing wave pattern (505) in a part of said separation container (500), whereby said particles of said first particle type, having a first property dependent on the characteristics of said generated ultrasonic standing wave pattern (505), is collected and held at the nodes of said generated ultrasonic standing wave pattern (505); - a liquid flow mechanism (320) arranged to flow a rinsing fluid through said generated ultrasonic standing wave pattern (505), said rinsing fluid carrying said particles of said second particle type, having a second property dependent on the characteristics of said generated ultrasonic standing wave pattern (505), such that said particles of said second particle type passes through said generated ultrasonic standing wave pattern (505);
- a container (200) for receiving said particles of said second particle type;
- means for removing said ultrasonic standing wave pattern (505); and
- an activating means (420) arranged to empty said separation container (500), whereby said particles of said first particle type are removed from said separation container (500).
13. The apparatus as recited in claim 11 or 12, wherein said ultrasound source (510,520) is arranged to generated said stationary ultrasonic standing wave pattern (505) in such a way that said particles of said first particle type tend to accumulate, whereby particle clusters of said particles of said particle types are created, which particle clusters tend to move downwards due to the gravitational force.
14. The apparatus as recited in claim 11 or 12, wherein said ultrasound source (510,520) comprises an ultrasound generating means (510) and an ultrasound reflecting means (520) arranged at opposite sides of said separation container (500).
15. The apparatus as recited in claim 14, wherein said ultrasound generating means (510) and said ultrasound reflecting means (520) are designed with a plane and rectangular shape in order to generate an homogenous ultrasound field, which field generates transversal wave fronts in said separation container (500).
16. The apparatus according to claim 14, wherein said ultrasound generating means (510) and said ultrasound reflecting means (520) are arranged to generate an ultrasonic standing wave pattern (505) having a direction of propagation perpendicular to the flat surfaces of said ultrasound generating means (510) and said ultrasound reflecting means (520).
17. The apparatus as recited in claim 14, wherein said ultrasound generating means (510) and said ultrasound reflecting means (520) are arranged to have approximately the same diametrical dimensions in the following ranges: width 5- 100 mm, height 5-100 mm and depth/thickness 1-5 mm.
18. The apparatus as recited in claim 14, wherein said ultrasound reflecting means (520) is realized as a second passive ultrasound generating means.
19. The apparatus as recited in claim 14, wherein said ultrasound reflecting means (520) is constructed of a ultrasound reflecting material, such as a metal or a ceramic material.
20. The apparatus as recited in claim 14, wherein said ultrasound generating means (510) is realized in a piezoelectric material vibrating in thickness mode, whereby mechanical oscillations are provided.
21. The apparatus as recited in claim 20, wherein the frequency of said oscillations is arranged in such a way that the wavelength of the ultrasonic wave pattern (505) is somewhat larger than the double diameter of the particles to be separated.
22. The apparatus as recited in claim 21, wherein said frequency is in the range of 0,5- 10 MHz.
23. The apparatus as recited in claim 21, wherein said frequency is in the range of 2 MHz.
24. The apparatus as recited in claim 11 or 12, further comprising a control unit (550) arranged to control the ultrasound energy delivered by the ultrasound generating means (510).
25. The apparatus according to claim 24, wherein said control unit (550) is arranged to deliver electrical energy of a certain waveform, frequency and power.
26. The apparatus according to claim 25, wherein said waveform is controllable to be one of sinus form, triangular form or square form.
27. The apparatus as recited in claim 24, wherein said control unit (550) is arranged to apply a voltage in the range of 10-100 Vpp over said ultrasound generating means (510).
28. The apparatus as recited in claim 27, wherein said control unit (550) is arranged to apply a voltage in the range of 15-50 Vpp.
29. The apparatus as recited in claim 11, further comprising; - a first inlet (501) for inputting said solution of particulate matter to said separation container (500);
- a second inlet (503) for inputting said rinsing fluid to said separation container (500);
- a first outlet (504) for outputting a first separation product, comprising said particles of said first particle type, from said separation container (500);
- a second outlet (502) for outputting a second separation product, comprising said particles of said second particle type, from said separation container (500).
30. The apparatus as recited in claim 12, further comprising;
- a first inlet (501) for inputting said blood solution to said separation container (500);
- a second inlet (503) for inputting said rinsing fluid to said separation container (500); - a first outlet (504) for outputting a first separation product, comprising said particles of said first particle type, from said separation container (500);
- a second outlet (502) for outputting a second separation product, comprising said particles of said second particle type, from said separation container (500).
31. The apparatus as recited in claim 29 or 30, wherein said first inlet (501) and said second outlet (502) are arranged in a first and second part of said separation container (500), respectively, and in that said ultrasonic standing wave pattern (505) is generated in a third part of said separation container (500), which third part is arranged in between said first and second part.
32. The apparatus as recited in claim 11 or 12, wherein the liquid flow mechanism (320) is arranged such that gravitation causes the rinsing fluid to flow through the ultrasonic standing wave pattern (505).
33. The apparatus as recited in claim 11 or 12, wherein said particles of said first particle type comprises red blood cells and said particles of said second particle type comprises blood plasma.
34. A separation product produced through a method of separating particles from a solution of particulate matter comprising particles of a first and a second particle type, the method comprising the steps of:
- receiving said solution of particulate matter in a separation container (500);
- generating a stationary ultrasonic standing wave pattern (505) in a part of said separation container (500) such that said particles of said first particle type having a first property depending of the characteristics of said ultrasonic standing wave pattern (505) are collected and held at the nodes of said ultrasonic standing wave pattern (505);
- establishing a flow of a rinsing fluid through said ultrasonic standing wave pattern (505), said rinsing fluid carrying said particles of said second particle type, having a second property dependent on the characteristics of said generated ultrasonic standing wave pattern (505), such that said particles of said second particle type passes through said generated ultrasonic standing wave pattern (505);
- removing said particles of said second particle type from said separation container (500);
- removing said ultrasonic standing wave pattern (505); and
- emptying said separation container (500), whereby said particles of said first particle type are removed from said separation container (500).
35. The separation product as recited in claim 34, further comprising the step of increasing the concentration of said particles of said first particle type at said nodes of said ultrasonic standing wave pattern (505) by conducting a flow of said solution of particulate matter to said ultrasonic standing wave pattern (505).
36. A separation product produced through a method of separating particles from a blood solution comprising particles of a first and a second particle type, the method comprising the steps of:
- receiving said blood solution in a separation container (500);
- generating a stationary ultrasonic standing wave pattern (505) in a part of said separation container (500) such that said particles of said first particle type having a first property depending of the characteristics of said ultrasonic standing wave pattern (505) are collected and held at the nodes of said ultrasonic standing wave pattern (505);
- establishing a flow of a rinsing fluid through said ultrasonic standing wave pattern (505), said rinsing fluid carrying said particles of said second particle type, having a second property dependent on the characteristics of said generated ultrasonic standing wave pattern (505), such that said particles of said second particle type passes through said generated ultrasonic standing wave pattern (505);
- removing said particles of said second particle type from said separation container (500);
- removing said ultrasonic standing wave pattern (505); and
- emptying said separation container (500), whereby said particles of said first particle type are removed from said separation container (500).
37. The separation product as recited in claim 36, further comprising the step of increasing the concentration of said particles of said first particle type at said nodes of said ultrasonic standing wave pattern (505) by conducting a flow of said blood solution to said ultrasonic standing wave pattern (505).
38. The separation product as recited in claim 34 or 36, further comprising the step of generating said stationary ultrasonic standing wave pattern (505) in such a way that said particles of said first particle type tend to accumulate, whereby particle clusters of said particles of said particle types are created, which particle clusters tend to move downwards due to the gravitational force.
39. The separation product as recited in claim 34 or 36, further comprising the step of controlling the size of said particles of said first particle type dependent on the distance between an ultrasound generating means (510) and an ultrasound reflecting means (520) between which means (510,520) said ultrasonic standing wave pattern (505) is generated.
40. The separation product as recited in claim 34 or 36, further comprising the step of controlling the size of said particles of said first particle type dependent on the ultrasound frequency at which said ultrasonic standing wave pattern (505) is generated.
41. The separation product as recited in claim 34 or 36, further comprising the step of controlling the separation of said particles of said first particle type from said particles of said second particle type dependent on the acoustic properties of each particle type, respectively.
42. The separation product as recited in claim 34 or 36, further comprising the step of controlling the separation of said particles of said first particle type from said particles of said second particle type dependent on the density of each particle type, respectively.
43. The separation product as recited in claim 34 or 36, wherein said particles of said first particle type comprises red blood cells and said particles of said second particle type comprises blood plasma.
44. A set of disposables for use in an apparatus for separating particles in a solution of particulate matter comprising particles of a first and a second particle type, said set of disposables comprising: - a separation container (500);
- a container (100) for said solution of particulate matter and connectable to a first inlet (501) of said separation container (500);
- a container (300) for a rinsing fluid and connectable to a second inlet (503) of said separation container (500); - a container (400) for a first separation product comprising said particles of said first particle type and connectable to a first outlet (504) of said separation container (500); - a container (200) for a second separation product comprising said particles of said second particle type and connectable to a second outlet (502) of said separation container (500); and
- a set of tubings (110,210,310,410) arranged to connect said containers (100,200,300,400) to said separation container (500).
45. A set of disposables for use in an apparatus for separating particles in a blood solution comprising particles of a first and a second particle type, said set of disposables comprising:
- a separation container (500); - a container (100) for said blood solution and connectable to a first inlet
(501) of said separation container (500);
- a container (300) for a rinsing fluid and connectable to a second inlet (503) of said separation container (500);
- a container (400) for a first blood product comprising said particles of said first particle type and connectable to a first outlet (504) of said separation container
(500);
- a container (200) for a second blood product comprising said particles of said second particle type and connectable to a second outlet (502) of said separation container (500); and - a set of tubings ( 110,210,310,410) arranged to connect said containers
(100,200,300,400) to said separation container (500).
46. The set of disposables as recited in claim 44 or 45, further comprising an ultrasound generating means (510) and an ultrasound reflecting means (520).
47. The set of disposables as recited in claim 44 or 45, wherein said containers (100,200, 300,400) are realized in a disposable material such as a plastic polymer or glass.
48. The set of disposables as recited in claim 44 or 45, wherein said containers (100,200, 300,400) are arranged to comprise a volume in the range of 50-5000 milliliters, preferably in the range of 500 milliliters.
49. The set of disposables as recited in claim 44 or 45, wherein said tubings
(110,210, 310,410) are realized in a disposable material, such as polyurethane, PVC, silicone or another plastic-like material.
50. The set of disposables as recited in claim 44 or 45, wherein said tubings (110,210, 310,410) are arranged to have a diameter in the range of 1.5-2.5 millimetres.
51. The set of disposables as recited in claim 44 or 45, wherein said tubings (110) is arranged to have a first branch (110a) and a second branch (110b).
52. The set of disposables as recited in any of the claims 44-51, wherein said disposables are sterile and treated with an anti-coagulation agent, such as heparin, ACD (acid citrate dextrose) or citrate.
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