Cell washing using acoustic waves

Disclosed is a device for separating a cellular component from a multicomponent fluid. The device can include a body, a first acoustic wave generator, and a second acoustic wave propagating component. The body can define a channel having a first surface, an opposing second surface, a first side, and an opposing second side. The channel can extend along a longitudinal axis from a first end to an opposing second end. The first acoustic wave generator can be coupled to the first surface. The second acoustic wave propagating component can be coupled to the second surface. The first acoustic wave generator and second acoustic wave propagating component can be configured to generate a bulk standing acoustic wave in the channel.

BACKGROUND

The present technology relates to separating components, such as red blood cells, from a mixture (such as a suspension), and particularly to separating a selected target component in a high concentration and purity using acoustic waves, such as bulk acoustic waves.

Blood transfusions are used to treat many disorders and injuries, such as in the treatment of accident victims and during surgical procedures. According to current American Red Cross statistics, about 5 million people receive blood transfusions each year, in the United States, alone. Thus, health care systems rely on the collection and distribution of blood. Typically, blood is obtained from a donor and then processed and stored; units of stored blood or blood products are then taken from storage as needed and transfused into a patient in need. In some cases, the blood may be an autologous donation, where an individual donates blood in expectation of receiving his or her own blood by transfusion during a medical procedure.

Donated blood is typically processed into components and then placed in storage until needed. When a subject is in need of a blood transfusion, a Unit of blood is commonly removed from storage, washed, and resuspended in an appropriate solution. The blood may also be treated with a red blood cell enhancement composition, to rejuvenate or improve aspects of red blood cell functionality, such as oxygen delivery capacity, that may be decreased during storage. In some instances, the red blood cells are lyophilized prior to storage, in which case they need to be resuspended, washed, and then resuspended again in an appropriate solution. The resuspended red blood cells are then transfused into the subject. In either scenario, washing the red blood cells is traditionally a tedious, time consuming and multistep process that requires a great deal of tubing, and the use of expensive centrifuges with rotating seals to separate the cells from the wash solution. Therefore, there remains a need to streamline and simplify the process for washing red blood cells prior to transfusion.

SUMMARY

To better illustrate the system disclosed herein, a non-limiting list of examples is provided here:

Example 1 can include a device for separating a cellular component from a multicomponent fluid. The device can comprise a body, a first acoustic wave generator, and a second acoustic wave propagating component. The body can define a channel having a first surface and a second surface opposite the first surface. The channel can extend along a longitudinal axis from a first end to a second end. The first acoustic wave generator can be coupled to the first surface. The first acoustic wave generator can be configured to generate an acoustic wave having a wavelength. The second acoustic wave propagating component can be coupled to the second surface. The second surface can be spaced an integer fractional multiple of the wavelength from the first surface and each integer factional multiple equals a number of pressure nodes within the channel.

In Example 2, the device of Example 1 can optionally include a central power generating region of the first acoustic wave generator being aligned with the second end of the channel and proximate a bifurcation region of the channel.

In Example 3, the device of any one of or any combination of Examples 1 and 2 can optionally include the integer fractional multiple being 0.5 and the number of pressure nodes is 1.

In Example 4, the device of any one of or any combination of Examples 1-3 can optionally include the first acoustic wave generator and the second wave propagating component being located proximate a midpoint of the channel.

In Example 5, the device of any one of or any combination of Examples 1-4 can optionally include the body comprises a phantom material forming at least a portion of one or both of the first surface and the second surface. The phantom material having acoustic properties similar to those of the multicomponent fluid and a thickness such that at least one of the pressure nodes is located proximate the phantom material.

In Example 6, the device of any one of or any combination of Examples 1-5, further comprising a first inlet and a second inlet proximate the first end, the first inlet having a higher elevation than the second inlet.

In Example 7, the device of Example 6 can optionally include a first outlet and a second outlet proximate the second end. The second outlet having a higher elevation than the first outlet.

In Example 8, the device of Example 7 can optionally include the first inlet being configured to receive a wash material and the second inlet is configured to receive a multicomponent mixture.

In Example 9, the device of Example 8 can optionally include the second outlet being arranged to receive the multicomponent mixture and the first outlet being arranged to receive the multicomponent mixture.

In Example 10, the device of any one of or any combination of Examples 1-9 can optionally include the channel having a cross-sectional width and height. An aspect ratio of width:height can be from about 1:11 to about 50:1. The first acoustic wave generator can produce waves having a frequency of from about 100 kHz to about 2000 kHz.

In Example 11, the device of any one of or any combination of Examples 1-10, can optionally include, during use, an antinode being formed at approximately the center of the channel and a first pressure node being formed at the first surface and a second pressure node being formed at the second surface.

Example 12 can include a device for separating a cellular component from a multicomponent fluid. The device can comprise a body, a first acoustic wave generator, and a second acoustic wave propagating component. The body can define a channel having a first surface and a second surface opposite the first surface. The channel can extend along a longitudinal axis from a first end to a second end. The channel can define a bifurcation region proximate the second end. The first acoustic wave generator can be coupled to the first surface. The first acoustic wave generator can be configured to generate an acoustic wave having a wavelength. The first acoustic wave generator can have a central power generating region aligned proximate the bifurcation region. The second acoustic wave propagating component can be coupled to the second surface. The second surface can be spaced a multiple of the half-wavelengths from the first surface such that, during use, an antinode is formed at approximately the center of the channel and a first pressure node is formed at the first surface and a second pressure node is formed at the second surface.

In Example 13, the device of Example 12 can optionally include the body comprising a phantom material forming at least a portion of one or both of the first surface and the second surface. The phantom material can have acoustic properties similar to those of the multicomponent fluid and a thickness such that at least one of the pressure nodes is located proximate the phantom material.

In Example 14, the device of any one of or any combination of Examples 12 and 13 can optionally include the first acoustic wave generator or the second wave propagating component being a resonator.

In Example 15, the device of any one of or any combination of Examples 12-14 can optionally include a first inlet and a second inlet proximate the first end. The first inlet can have a higher elevation than the second inlet.

In Example 16, the device of Example 15 can optionally include a first outlet and a second outlet proximate the second end. The second outlet can have a higher elevation than the first outlet.

In Example 17, the device of Example 16 can optionally include the first inlet being configured to receive a wash material and the second inlet being configured to receive a multicomponent mixture.

In Example 18, the device of Example 17 can optionally include the second outlet being arranged to receive the multicomponent mixture and the first outlet being arranged to receive the multicomponent mixture.

Example 19 can include a method of separating a cellular component from cellular component liquid stream. The method can comprise introducing the cellular component liquid stream and a wash material liquid stream into an acoustic wave separation device having a channel that defines a bifurcation region proximate a first outlet and a second outlet; contacting the cellular component liquid stream and the wash material liquid stream in the proximate a pressure node of a standing acoustic wave located proximate the bifurcation region thereby forcing the cellular component from the component liquid stream to the wash material liquid stream; and collecting the wash material liquid stream in the first outlet.

In Example 20, the method of Example 19 can optionally include the standing acoustic wave being a surface acoustic wave.

In Example 21, the devices or methods of any one of or any combination of Examples 1-20 are optionally configured such that all elements or options recited are available to use or select from.

Corresponding reference numerals indicate corresponding parts throughout the several views of the drawings. It should be noted that the figures set forth herein are intended to exemplify the general characteristics of materials, compositions, devices, and methods among those of the present technology, for the purpose of the description of certain embodiments and are not intended to limit the scope of the present disclosure. These figures may not precisely reflect the characteristics of any given embodiment, and are not necessarily intended to fully define or limit specific embodiments within the scope of this technology.

DETAILED DESCRIPTION

The following description of technology is merely exemplary in nature of the composition, manufacture and use of one or more inventions, and is not intended to limit the scope, application, or uses of any specific invention claimed in this application or in such other applications as may be filed claiming priority to this application, or patents issuing therefrom. A non-limiting discussion of terms and phrases intended to aid understanding of the present technology is provided at the end of this Detailed Description.

Although traditional methods for washing blood are largely effective, there remains a need to streamline the process for isolating blood cells from multicomponent fluids. It has been found that processing and washing blood may be performed using standing acoustic waves (SAWs), also referred to as stationary waves. Generally, standing waves are created by the interference between two intersecting sinusoidal waves having essentially identical frequencies, formed in a liquid or other medium. For example, opposing waves can be propagated laterally, parallel to the flow path in a channel through which fluid flows, creating an interfering standing acoustic wave pattern in the fluid. Such waves may be referred to as surface acoustic waves. Alternatively, opposing acoustic waves may be propagated on opposite sides of the channel (e.g., from the top and bottom, or from opposite ends) to form interfering standing wave patterns in the reservoir that may be referred to as bulk acoustic waves.

In both surface and bulk acoustic waves, pressure nodes and antinodes are formed in the fluid that can be used to manipulate a target particulate or other solid or semi-solid component, such as red blood cells, that is in the fluid. In particular, a pressure node of a SAW may be used to force a cell or other component in the fluid to a location within a fluid reservoir, based on the component's acoustical, physical, and mechanical properties. The present technology provides devices, systems and methods using SAWs to separate target components, such as cells, from multicomponent fluids. In some embodiments, the SAWs are surface acoustic waves. In various other embodiments, the SAWs are bulk acoustic waves.

In particular, the present technology provides devices (chips), systems, and methods for separating a component from a multicomponent fluid. As further described below, the devices comprise a channel or other reservoir in which the multicomponent fluid flows or is contained, wherein two or more wave propagating components are disposed on one or more surfaces of the reservoir, in acoustic communication with the reservoir. The wave propagating components generate standing acoustic waves that include pressure nodes and antinodes in the fluid.

The devices and systems of the present technology comprise at least one acoustic wave generator, and a second wave propagating component. Wave generators suitable for use in the present technology include acoustic wave generators among those known in the art. In various embodiments, acoustic wave generators comprise piezoelectric transducers, which convert electrical pulses to mechanical vibrations. Non-limiting examples of piezoelectric materials include quartz, quartz crystal, ceramic, ceramic composites, berlinite (AIPO4), lead titanate (PbTiO3), barium titanate (BaTiO3), lead zirconate titanate (Pb[ZrxTi1-x]O3, 0≤x≤1; “PZT”), potassium niobate (KNbO3), lithium niobate (LiNbO3), lithium tantalate (LiTaO3), sodium tungstate (Na2WO3), Ba2NaNb5O5, Pb2KNb5O15, zinc oxide (ZnO), sodium potassium niobate ((K,Na)NbO3), bismuth ferrite (BiFeO3), sodium niobate (NaNbO3), bismuth titanate (Bi4Ti3O12), sodium bismuth titanate (Na0.5Bi0.5TiO3), and polymers, such as polyvinylidene fluoride (PVDF). In various embodiments, the wave generators are operated at a frequency of from about 100 kHz to about 2000 kHz, from about 300 kHz to about 1000 kHz, from about 400 kHz to about 900 kHz, from about 500 kHz to about 800 kHz, or from about 600 kHz to about 700 kHz. In one embodiment, the frequency is from about 680 kHz to about 710 kHz.

In some embodiments, such as in devices for producing surface acoustic waves, the acoustic wave generator is an interdigital or interdigitated transducer (IDT), comprising interlocking comb-shaped arrays of electrodes disposed on the surface of a piezoelectric substrate. In some embodiments, such as for producing bulk acoustic waves, the acoustic wave generator comprises a monolithic ceramic piezoelectric material in a thin-film transducer, such as a thickness shear mode resonator (TSMR).

Second wave propagating components useful herein include acoustic wave generators (i.e., a second acoustic wave generator, as described above) and acoustic reflectors. Reflectors comprise acoustically reflective materials or surfaces, such as a slide, layer or membrane composed of glass, polymer, plastic, metal, or ceramic that is substantially reflective to acoustic waves. It will be appreciated that the reflectivity of the material may be a function of the density of the material relative to the fluid through which waves are propagated, as well as the frequency of the waves. As non-limiting examples, the reflective material can be biaxially-oriented polyethylene terephthalate (boPET) polyester film (such as Mylar® brand BoPET commercialized by DuPont; Wilmington, Del.), glass mica, polymers, or a combination thereof.

As briefly discussed above, devices of the present technology create standing acoustic waves by positioning an acoustic wave generator in proximity to a second wave propagating component, in a fluid reservoir substrate (e.g., a fluid channel), so as to create an interfering wave pattern in the fluid reservoir. For example, by positioning first and second wave generators, such as piezoelectric transducers, opposite each other on a substrate, a SAW can be generated when acoustic waves from each generator interfere with each other. Alternatively, a SAW can be generated by positioning a wave generator on one side of a substrate and positioning a reflective material (as the second wave propagating component) on a side of the substrate opposite the wave generator. By adjusting the distance between the wave generators (or wave generator and reflective surface) and/or by adjusting the frequencies of the acoustic waves, the position of a pressure node associated with a SAW can be manipulated, located and controlled, for example, within a channel positioned between the wave generators (or wave generator and reflective surface). As discussed further below, the position of the acoustic wave in the fluid is determined by the frequency of the wave and the dimensions of the reservoir (e.g., a channel), containing the fluid.

For example,FIGS. 14A-14Cdepict a cross-section of a chip500having a body502that defines a channel504. The channel504has a channel ceiling506and an opposing channel floor508. A first wave generator510is positioned on an upper surface512of the chip500and a second wave generator514is positioned on an opposing lower surface516of the chip500. However, it is understood that a combination of a wave generator and an opposing reflective material or surface can also be utilized. InFIG. 14A, the wave generators510,514are tuned to generate a SAW518with a wavelength of 0.5λ. In this embodiment, the nodes, shown as filled-in circles, would push flowing cells toward the antinode, shown as an open circle. InFIG. 14B, the wave generators510,514are tuned to generate a SAW520with a wavelength of 1.5λ having two nodes and one antinode positioned in the channel504. Here, cells flowing through the channel504would be pushed away from the nodes towards the antinode in the center of the channel504and towards the antinodes within the chip body502. Therefore, if it is desired to direct cells toward the antinode in the center of the channel504, either a new chip can be manufactured with a channel having a different size or the current channel504can be modified.

As discussed above, the frequency of the wave generator(s) can vary, for example ranging from about 100 kHz to about 2000 kHz. The specific frequency may be determined in conjunction with the dimensions of the channel or other reservoir in which the standing wave is to be created, so as to produce pressure nodes in the desired locations. The position of a pressure node or antinode associated with a SAW in a chip is dependent on the thickness of the chip materials in between the wave generators (or between a wave generator and a reflective surface) and the speed of sound in the chip material. Thus, the fluid reservoir (channel) dimensions are preferably optimized in regard to the frequency of the wave generator. For example, whereas low frequencies can support large channel dimensions, high frequencies are typically used with small channel dimensions. Therefore, depending on the frequency of the wave generators, the chips of the present technology comprise channels having a cross-sectional aspect ratio (width:height) of from about 1:1 to about 50:1 or from about 1:1 to about 40:1, or from about 1:1 to about 30:1, or from about 1:1 to about 20:1, or from about 1:1 to about 10:1, or from about 1:1 to about 5:1. Moreover, the input voltage of the wave generators can be from about 1 V to about 120 V and is dependent on chip geometry, hematocrit, and flow rate.

As stated above, the wave propagating devices are disposed on the surface of the device reservoir, so as to be in acoustic communication with the multicomponent fluid in the reservoir. In embodiments comprising a channel, having a fluid inlet at a first end and a fluid outlet at the opposite second end, through which the fluid flows, the wave propagating devices may be disposed at any point laterally along a surface of the channel, parallel to the axis of fluid flow. In some embodiments, a first wave generator and a second fluid propagating component (a second wave generator or a reflector) may be essentially in the mid-point of the channel, between the inlet and outlet. In other embodiments, the wave generator and second fluid propagating component are disposed near the outlet of the channel. It has been found that, in some embodiments wherein the fluid propagating components are disposed near the outlet, cells in the multicomponent fluid may be disposed in the fluid more easily and using less power than in embodiments where the wave propagating components are disposed at or near the mid-point of the channel. It will be appreciated that the precise special orientation of a wave propagating component near the outlet of the channel will be affected by the length of the channel (i.e, in the dimension parallel to the fluid flow) and the size of the wave propagating component. In various embodiments, the mid-point of the wave propagating component is within 10%, within 20%, or within 30% of the outlet, as a percentage of the distance between the inlet and outlet.

In some embodiments, such as the chip602depicted inFIG. 15(which is further discussed below), a wave propagating component628comprises a central power generating region, defined by a first end point650and a second end point651on the longitudinal surface (e.g., top surface606) of the chip602. The length of the central power generating region, i.e., the distance652between the first end point651and the second end point652, consists of the middle 20%, 10% or 5% of the wave propagating component, as a percentage of the overall length653of the wave propagating component (i.e., the dimension that is parallel to the flow of fluid in the channel). In various embodiments, a point within the central power generating region of the wave propagating component is axially aligned with the outlet end (second end, as discussed above) of the separation channel. That is, in reference toFIG. 15, both a point that is within the central power generating region of the wave propagating component, and the outlet end622of the separation channel616, fall on a common axis632that is orthogonal to a surface (e.g., ceiling618) of the channel. In some embodiments, the second end point651of the central power generating region is axially aligned with the outlet end622of the separation channel616.

Devices

The present technology provides devices, such as fluidic chips, that comprise a channel or other reservoir in which standing acoustic waves may be used so as to apply forces to cell in a multicomponent fluid. As discussed above, such forces may be used to move the cells in the fluid, such as by forcing cells from the fluid into a second fluid within the device. In various embodiments, such movement of cells from a first multicomponent fluid effects washing of the cells, thereby creating a suspension of cells in a second fluid

The chips may be constructed of any of a variety of materials, including such materials known in the art. The materials are preferably compatible with physiological materials (e.g., blood cells) that are processed with the devices, and have appropriate acoustic characteristics. Examples include polyethylene terephthalate (PET) acrylics, such as poly(methyl methacrylate) (PMMA), and glasses.

FIG. 1Ashows a cross-sectional view of an exemplary device10acomprising a substrate or device body12, a first wave component14, and a second wave component16positioned on opposite sides15,17of the body12, wherein the body12defines a channel32with a square cross-sectional geometry. As discussed above, the first wave component14and the second wave propagating component16are individually either a wave generator or a reflective material or reflective surface or layer. However, when one of the wave components14,16is a reflective material or reflective surface or layer, the other wave component14,16is a wave generator. Alternatively, a side15,17of the device body12can be composed of a reflective material so long as the opposite side15,17comprises a wave generator. A SAW is generated between the first wave component14and the second wave component16along line18. A pressure node associated with the SAW, which is located within the channel32, forces a plurality of cells20into a plane perpendicular to the line18.

FIG. 1Bshows a cross-sectional view of another exemplary device10b, which is similar to device10a. However, the device10bfurther comprises a third wave propagating component22and a fourth wave propagating component24positioned on opposite sides23,25of the body12. The third wave component22and the fourth wave component24are individually either a wave generator or a reflective material or reflective surface or layer. However, when one of the wave components22,24is a reflective material or reflective surface or layer, the other wave component22,24is a wave generator. The third wave component22and the fourth wave component24are positioned orthogonal to the first wave component14and the second wave component16on sides23,25of the body12. A first SAW is generated between the first wave component14and the second wave component16along line18and a second SAW is generated between the third wave component22and the fourth wave component24along line26that is orthogonal to the first line18, such that the second SAW is orthogonal to the first SAW. Pressure nodes associated with the SAWs interest with each other and interact with the plurality of cells20in orthogonal directions to force the cells20into a linear configuration, as shown more clearly inFIG. 3.

FIG. 2provides a perspective view of a device30a, which is similar to the device10a. The device30acomprises a substrate or device body12, a first wave component14, and a second wave component16positioned on opposing sides15,17of the body12. As shown inFIG. 2, the first and second wave components14,16are wave generators. The device30acomprises a longitudinal channel32with a square cross-sectional geometry that extends along a longitudinal axis33. As shown inFIG. 2, the cells20are suspended in a plane that extends along the axis33and that is parallel to the wave components14,16by a pressure node associated with a SAW generated by the first wave component14and the second wave component16.

FIG. 3provides a perspective view of another exemplary device30b, which is similar to the device10b. The device30bcomprises a substrate or device body12, a first wave component14and a second wave component16positioned on opposing sides15,17of the body12, and a third wave component22and a fourth wave component24positioned on opposing sides23,25of the body12that are orthogonal to the sides15,17that include the first and second wave components14,17. Again, the device30bcomprises a longitudinal channel32with a square cross-sectional geometry that extends along the axis33. As shown inFIG. 3, the cells20are suspended in a cylindrical line along the axis33of the channel32by a first pressure node associated with a first SAW generated by the first wave component14and the second wave component16and by a second pressure node associated with a second SAW generated by the third wave component22and the fourth wave component24, wherein the second SAW is orthogonal to the first SAW.

Referring now toFIG. 1C, a device10cis shown, which is similar to device10b. However, device10cfurther comprises a channel28awith a circular cross-sectional shape. Because the channel28ais centered in the substrate12, and because the wave components14,16,22,24are centered on their respective sides of the substrate12, the cells20are suspended in a line central to the channel28a. As shown inFIG. 1D, a device10dcomprises a channel28b, which is offset relative to the center of the substrate12. The cells20are positioned in a line extending along a midpoint of a cross-section of the substrate12because the pressure nodes force the cells20to that position. In other words, the cells20are positioned based upon the pressure node or nodes and not upon the positioning of the channel28a,28b,32.

In various embodiments, channels or other reservoirs may comprise a phantom material so as to alter the flow of fluid within the reservoir relative to the SAW and, in some embodiments, inlet and outlet regions of the device. As used herein, a “phantom material” is a material that mimics the acoustic properties of the fluid through which acoustic waves are propagated. In various embodiments, the phantom material mimics the acoustic properties of water with a low attention coefficient. Therefore, an acoustic wave travels through phantom materials substantially as it would, such as with the same speed, through water. For example, sound travels through water at a rate of from about 1450 m/s to about 1570 m/s. Similarly, sound travels through the phantom materials at a rate of from about 1200 m/s to about 1600 m/s, or at a rate of from about 1400 m/s to about 1500 m/s. Non-limiting examples of suitable phantom materials include Solid Water® phantom material from CNMC Co. Inc. (Nashville, Tenn.), Virtual Water™ phantom material from CNMC Co. Inc., and Plastic Water® phantom material from Computerized Imaging Reference Systems, Inc. (Norfolk, Va.). Various plastics, acrylics, and glasses detrimentally affect how acoustic waves travel. Because phantom materials do not affect how an acoustic wave travels, separation devices with complex geometries, such as single chips or devices having multiple channels, can be generated. Therefore, phantom materials may be included in channels to alter a flow path without affecting the position of an acoustic node or antinode. Additionally, in some embodiments, two or more devices of the current technology may be multiplexed to reduce surface area and to increase efficiency.

For example, the devices10a,10b,10c,10dshown inFIGS. 1A-1Dcomprise, at least partially, a phantom material. Similarly,FIG. 14C, depicts a device having channels comprising a phantom material. In particular reference toFIG. 14C, the channel504may comprise a first sheet of phantom material522along the ceiling506of the channel504and a second sheet of phantom material524along the floor508of the channel504. The SAW520travels through phantom materials substantially as it would, such as with the same speed, through water. Therefore, placement of the sheets of phantom material522,524does not affect the location of the nodes and antinodes. By using the sheets of phantom materials522,524, nodes can be located at interfaces between the sheets of phantom materials522,524and the channel504so that the nodes force cells only towards the antinode in the center of the channel504. In other words, phantom materials can be used to manipulate the dimensions of the channel504without affecting the location of the nodes generated by the SAW520. Further exemplary chip and separation embodiments are provided below.

FIG. 4shows a cross-section view of another exemplary device40for washing a multicomponent mixture comprising cells, such as, for example, red blood cells. The device40comprises a body42defining a channel44, a first wave component46positioned on or near a first side47of the body42and a second wave component48positioned opposite to the first wave component46on or near a second opposing side49of the body42. The first wave component46and the second wave component48are individually either a wave generator or a reflective material or reflective surface or layer. However, when one of the wave components46,48is a reflective material or reflective surface or layer, the other wave component46,48is a wave generator. A SAW is generated between the first wave component46and the second wave component48such that a pressure node is located within the channel44. The channel44comprises a first horizontal section50, a second connecting section52, and a third horizontal section54, such that the first horizontal section50is offset from the second horizontal section54. The third section52is bifurcated into a first collection channel56and a second collection channel58by a planar shelf60defined by the body42. The wave components46,48are positioned on the first side47and on the second opposing side49of the body42, respectively, which are parallel to the channel44at the connecting section52and the second horizontal section54, such that the channel44is positioned between the first and second wave components46,48. In this embodiment, the wave propagating components46,48(e.g., wave generators) are positioned close to the collection channel58to promote efficient separation. When a multicomponent mixture comprising red blood cells62and a wash material64are introduced into the device40, they mix at the first lower horizontal section50. However, upon reaching the pressure node, the red blood cells62are forced into a plane at the connecting section52corresponding to a pressure antinode. Simultaneously, the wash material, flow thereof represented by arrows64, passes through the red blood cells62, thereby washing the red blood cells62. The red blood cells62are then collected from the second collection channel58and the wash material64and other waste is collected from the first collection channel56. In other embodiments, the multicomponent mixture comprising red blood cells62and the wash material62are mixed prior to being introduced into the device40.

FIG. 5, depicts another exemplary device a device70for washing a multicomponent mixture. The device70comprises a body72having a first surface74, a second opposing surface76, a first end region78, and a second end region80. The body72defines a channel82extending along a longitudinal axis84from the first end region78to the second end region80. The device70further comprises a first inlet86, a second inlet88, a first outlet90, and a second outlet92, all in fluid communication with the channel82.FIG. 6Ais an exploded, cross-sectional perspective of the device70taken along line6A ofFIG. 5when the device70is generated by stacking a plurality of layers together as shown inFIG. 6B. As shown inFIGS. 6A and 6B, the channel82is bifurcated at the first end region78by a first planar shelf94defined by the body72, which keeps components that are introduced into the device70through the inlets86,88separate. However, in some embodiments (not shown) there is only one inlet and no shelf to separate components. Also, the channel82is bifurcated at the second end region80by a second planar shelf96defined by the body72, which keeps the components separated for collection through the outlets90,92by way of a first collection channel97and a second collection channel99, respectively.

Additionally, the device70comprises a first wave component110positioned adjacent and parallel to the channel82and a second wave component112positioned adjacent and parallel to the channel82, such that the channel82is positioned between the first and second wave components110,112. In various embodiments, the separating region100of the channel82is positioned between the first and second wave components110,112. The first wave component110and the second wave component112are individually either a wave generator or a reflective material or reflective surface or layer. However, when one of the wave components110,112is a reflective material or reflective surface or layer, the other wave component110,112is a wave generator. In embodiments where the second wave component112is a reflective surface, the reflective surface can be the second surface76of the device70, or it can be a reflective film, sheet, slide, or membrane coupled to the second surface76. As discussed further below regarding systems of the present technology, in some embodiments the first wave component110is an electrical contact that couples to a wave generator on a base unit. Therefore, when the first wave component110is a wave generator or an electrical contact, the second wave component112is either a second wave generator or a reflective surface or layer or material.

When the device70is activated, a SAW is generated between the first wave component110and the second wave component112, whereby a pressure node114(seeFIG. 7B) associated with the SAW is located within the separation region100of the channel82. In various embodiments, the SAW is generated from the wave components86,88operating at a low frequency range of from about 300 kHz to about 1000 kHz, or from about 400 kHz to about 600 kHz, or from about 450 kHz to about 500 kHz, in order to isolate components from a multicomponent mixture in the channel62with such a large volume. Even though this low frequency range results in a low pressure gradient, surprisingly, component isolation is achieved. In other embodiments, not shown inFIG. 4, the device70further comprises third and fourth wave components as or on opposing sides of the device70such that the third and fourth wave components generate a second SAW orthogonal to the SAW generated by the first and second wave components110,112, wherein the second SAW provides a second pressure node located in the separation region100of the channel82.

The device70can be manufactured by any means known in the art, including, for example, injection molding, compression molding, or 3-dimensional printing (3-D printing). In some embodiments, as shown inFIG. 6B, the device70is manufactured by stacking together a plurality of layers116a-116h, wherein each layer is bonded to an adjacent layer with an adhesive. With the optional exception described below in regard to the layer116g, the layers116a-116hare composed of any material known in the art. Non-limiting examples of materials for the layers116a-116hinclude plastics, such as polyethylene terephthalate (PET) acrylics, such as poly(methyl methacrylate) (PMMA), and glasses. Combining the layers116a-116hresults in the device70with the cross-sectional geometry shown inFIG. 6A. The layer116ghas two longitudinal protrusions118that form the two side walls106of the channel82. In various embodiments, the layer116gis composed of a phantom material (as described above) that mimics how acoustic waves travel through water to provide the device70with the channel82having phantom side walls106and a phantom floor104. In some embodiments, not shown inFIG. 6A or 6B, the first wave component110is coupled to a bottom surface120of the layer116gand the layer below it, layer116h, is optional. In other embodiments, the first wave component110is coupled to a bottom surface122of the layer116h. A first layer116acan either be composed of a reflective material or the second wave component112can be coupled to a surface76of the layer116a. Moreover, the first layer116acan be composed of a phantom material in various embodiments.

FIG. 7is a cross-sectional illustration of the device70when the device70is manufactured by a means other than by stacking together a plurality of layers, such as by injection molding, compression molding, or 3-D printing. The components ofFIG. 7are the same as those shown inFIGS. 6A and 6B, but the dimensions may be slightly different.

With reference toFIGS. 5-7B, the device70is configured to wash a multicomponent mixture. As described above, in various embodiments the multicomponent mixture comprises red blood cells124or red blood cells124and a rejuvenation solution. The multicomponent mixture is introduced to the device70through a first conduit coupled to an inlet86,88. As shown inFIG. 7B, a first conduit128is coupled to the second inlet88. Likewise, a wash material126is introduced to the device70through a second conduit coupled to the inlet86,88that is not coupled to the first conduit128. As shown inFIG. 7, a second conduit130is coupled to the first inlet86. Flow of the multicomponent mixture comprising red blood cells124and the wash material126can be established, by pumps, such as peristaltic pumps, optionally coupled to pulse dampeners or pulse suppressors. Examples of suitable pumps, pulse dampeners, and pulse suppressors that can be used for any embodiments provided herein are described in U.S. Patent Publication No. 2015/0111195, Hamman et al., published on Apr. 23, 2015, which is incorporated herein by reference. Upon entry into the device70, the multicomponent mixture comprising red blood cells124and the wash material126are mixed together at the receiving or mixing region98of the channel82. In other embodiments, the multicomponent mixture and washing material are combined prior to be introduced into the device70. In such embodiments, the device70may have a single input, as described above or the multicomponent mixture and wash material can be delivered into the device by either inlet86,88of the device70.

Referring again toFIGS. 5-7B, as the multicomponent mixture comprising red blood cells124and the wash material126flow through the channel82, they interact with a pressure node114, generated by the wave components110,112, in the separation region100of the channel82. In various embodiments, the pressure node114is located at or near the channel ceiling108and/or the channel floor104, such that an antinode is positioned at a location to which the red blood cells124are directed. Although the wave components110,112are shown positioned in the middle of the first and second surfaces74,76inFIGS. 5, 6A, 7A, and7B, in some embodiments (as discussed above), the wave components110,112are positioned near the outlets90,92, such that a strong pressure wave pushes the cells124towards the collection channel99easier and with less power; rather than aligning the cells124the length of the channel82. The pressure node114pushes, forces, isolates, or moves a component of the multicomponent mixture, such as the red blood cells124, adjacent to the shelf96and into the second collection channel99while the remainder of the multicomponent mixture and wash material126flow to the first collection channel97. The shelf96is thin and rigid so as to minimize turbulence within the channel82. The component pushed, forced, isolated, or moved to the second collection channel99is collected through a third conduit132coupled to the second outlet92and the remaining materials are collected through a fourth conduit134coupled to the first outlet90.

In various embodiments, chips are designed so have a particular spacial orientation, such as in systems (as described below) in which the devices are placed in a base unit. Thus, the gravity may have an effect on the flow of materials, such as cellular materials, through the chip. In some embodiments, the outlet of the chip is oriented lower than the inlet (i.e., at a location at a position lower than the inlet relative to the vertical axis of the chip, it being understood that the inlet and outlet are substantially at opposing ends of the chip relative to the orthogonal horizontal axis of the chip). In other embodiments, the outlet of the device is oriented higher than inlet. For example,FIGS. 6B and 7Bshow red blood cells124flowing downward to the second collection channel99. In other embodiments the pressure antinode is located such that the component is forced upward to the first collection channel97. In such embodiments, the red blood cells124are preferably introduced through the first inlet86and the wash material126is introduced through the second inlet88. As the red blood cells124flow against the channel floor104, the wash material126contacts an upper surface of the flow of red blood cells124. When the red blood cells124interact with the nodes of a SAW generated between the wave components110,112, the red blood cells124are forced upward, against gravity, through the wash material126and isolated at the first collection channel97as washed red blood cells124. The isolated and washed red blood cells are then collected through the first outlet90and the remaining materials are collected through the second outlet92. Another example of such an embodiment, where cells are forced upward against gravity by a node of a SAW is shown inFIG. 12, which is described in more detail below.

With reference toFIGS. 8-10, the present technology provides another device150for washing a multicomponent mixture. The device150comprises a body152having a first surface154, a second opposing surface156, a first end region158, and a second end region160. The body152defines a channel162extending along a longitudinal axis163from the first end region158to the second end region160. The device150further comprises a first pair of inlets164, a second pair of inlets166, a first pair of outlets168, and a second pair of outlets170, all in fluid communication with the channel162.FIG. 10is an exploded cross-sectional perspective of the device150taken along line9A ofFIG. 8when the device150is generated by stacking a plurality of layers together as shown inFIG. 9B. As shown inFIGS. 9A and 9B, the channel162is trifurcated at the first end region158by a first shelf181defined by the body152and a second shelf182defined by the body152, which keeps components that are introduced into the device150through the pairs of inlets164,166separate. Also, the channel162is trifurcated at the second end region160into a first collection channel187, a second collection channel189, and a third collection channel191by a third shelf183defined by the body152and fourth shelf184defined by the body152, wherein the first collection channel187is located between the second surface156and the third shelf183, the second collection189channel is located between the third and fourth shelves183,184, and the third collection channel191is located between the fourth shelf184and the first surface154. The collection channels187,189,191, keep the components separated for collection through the pairs of outlets168,170, such that the second collection channel189is in fluid communication with the first pair of outlets168and the first and third collection channels187,191are in fluid communication with the second pair of outlets170.

Additionally, the device150comprises a first wave component186positioned adjacent to the channel162on or near the first side154of the device150and a second wave component188positioned adjacent to the channel162on or near the second side156of the device150such that the channel162is positioned between the first and second wave components186,188. In various embodiments, the separation region174of the channel162is positioned between the first and second wave components186,188. Unless described otherwise, the first wave component186and the second wave component188are individually either a wave generator or a reflective material or reflective surface. However, when one of the wave components186,188is a reflective material or reflective surface, the other wave component186,188is a wave generator. Therefore, at least one of the wave components186,188is a wave generator. In embodiments where the second wave component188is a reflective surface, the reflective surface can be the second surface156of the device150, or it can be a reflective film, sheet, slide, or membrane. As discussed further below, in some embodiments the first wave component186is an electrical contact that couples to a wave generator on a base unit. Therefore, when the first wave component186is a wave generator or an electrical contact, the second wave component188is either a second wave generator or a reflective surface or material. When the device150is activated, a SAW is generated between the first wave component186and the second wave component188, whereby a pressure node196(seeFIG. 10) associated with the SAW is positioned within the separation region174of the channel162. In various embodiments, the SAW is generated from the wave components186,188operating at a low frequency range of from about 300 kHz to about 1000 kHz, or from about 400 kHz to about 600 kHz, or from about 450 kHz to about 500 kHz, in order to isolate components from a multicomponent mixture in the channel162with such a large volume. Even though this low frequency range results in a low pressure gradient, surprisingly, component isolation is achieved. In other embodiments, not shown inFIG. 8, the device150further comprises third and fourth wave components as or on opposing sides of the device150such that the third and fourth wave components generate a second SAW orthogonal to the SAW generated by the first and second wave components186,188, wherein the second SAW provides a second pressure node located in the separation region174of the channel162.

The device150can be manufactured by any means known in the art, including, for example, injection molding, compression molding, or 3-dimensional printing (3-D printing). In some embodiments, as shown inFIG. 11, the device150is manufactured by stacking together a plurality of layers190a-1901, wherein each layer is bonded to an adjacent layer with an adhesive. With the optional exception described below in regard to a phantom layer, the layers190a-1901are composed of any material known in the art. Non-limiting examples of materials for the layers190a-1901include plastics, such as polyethylene terephthalate (PET) acrylics, such as poly(methyl methacrylate) (PMMA), and glasses. Combining the layers190a-1901results in the device150with the cross-sectional geometry shown inFIG. 10. Optionally, an optional layer equivalent to layer90gofFIG. 7, but configured to provide communication between layer190kand the second input166and second output170, is positioned between layer190kand1901and has two longitudinal protrusions that form the two side walls180of the channel162. In various embodiments, the optional layer is composed of a phantom material that mimics how acoustic waves travel through water to results in the device150with the channel162having phantom side walls180and a phantom floor178. In some embodiments, not shown inFIG. 11, the first wave component186is coupled to a bottom surface of the optional layer. In other embodiments, the first wave component186is coupled to a bottom surface198of the layer190l. In yet other embodiments, layer190lis composed of a phantom material and comprises two longitudinal protrusions that form the two side walls180of the channel162. In such embodiments, the first wave component186is coupled to the bottom surface198of the layer190l. A first layer190acan either be composed of a reflective material or the second wave component188can be coupled to the second surface156of the layer190a. Moreover, the first layer190ais composed of a phantom material in various embodiments.

FIG. 10is a cross-sectional illustration of the device150when the device150is manufactured by a means other than by stacking together a plurality of layers, such as by injection molding, compression molding, or 3-D printing. The components ofFIG. 10are the same as those shown inFIGS. 9A and 9B, but the dimensions may be slightly different.

With reference toFIGS. 8-10, the device150is configured to wash a multicomponent mixture. As described above, in various embodiments the multicomponent mixture comprises red blood cells210or red blood cells210and a rejuvenation solution. The multicomponent mixture comprising red blood cells210is introduced to the device150through a pair of first conduits coupled to the pair of second inlets166. Likewise, a wash material212is introduced to the device150through a pair of second conduits coupled to the pair of first inlets164. Flow of the multicomponent mixture210and the wash material212can be established, by pumps, such as peristaltic pumps, optionally coupled to pulse dampeners or pulse suppressors, as described above. Upon entry into the device150, the multicomponent mixture210and the wash material212are mixed together at the receiving or mixing region172of the channel162. In other embodiments, the multicomponent mixture comprising red blood cells210and the washing material212are combined prior to be introduced into the device150to generate a pre-mixed composition. In such embodiments, the device150may have a single input, as described above, or the pre-mixed composition can be delivered into the device150by any inlet or combination of inlets164,166. Referring again toFIGS. 8-10, as the multicomponent mixture comprising red blood cells210and the wash material212flows through the channel162, they interact with a pressure node196, generated by the wave components186,188, in the separation region174of the channel162. In various embodiments, the pressure node196is located at or near the channel ceiling179and/or the channel floor178, such that an antinode is positioned at a location to which the red blood cells210are directed. As discussed above, although the wave components186,188are shown positioned in the middle of the first and second surfaces154,156inFIGS. 8 and 10, in some embodiments, the centers of the wave components186,188are positioned near to the outlets168,170, such that a strong pressure wave pushes the cells210towards the collection channel189easier and with less power; rather than aligning the cells210the length of the channel162. The pressure node196pushes, forces, isolates, or moves a component of the multicomponent mixture, such as red blood cells, between the third and fourth shelves183,184and into the second collection channel189while the remainder of the multicomponent mixture and wash material flow into the first and third collection channels187,191. The third and fourth shelves183,184are thin and rigid so as to minimize turbulence within the channel162. The component pushed, forced, isolated, or moved into the second collection channel180is collected through a third pair of conduits coupled to the first pair of outlets168and the remaining materials are collected through a fourth pair of conduits coupled to the second pair of outlets170.

With further reference toFIG. 15, the present technology provides another device600for washing a multicomponent mixture. The device600comprises a separation chip602having a body604. The body604has a top surface606and an opposing bottom surface608. Additionally, the body604defines an upper inlet channel610and a lower inlet channel612that merge into a first end614of a separation channel616due to an incline path of the lower inlet channel612. The separation channel616has a channel ceiling618and a channel floor620. The separation channel616bifurcates at a second end622into an upper outlet channel624and a lower outlet channel626, wherein the lower outlet channel626has a declined path relative to the separation channel616. A first wave component628is positioned on the top surface606of the chip602and a second wave component630is positioned on the bottom surface608of the chip602. The first wave component628and the second wave component630are individually either a wave generator or a reflective material or reflective surface or layer. However, when one of the wave components628,630is a reflective material or reflective surface or layer, the other wave component628,630is a wave generator. The first and second wave components628,630have a center or central region represented by a dotted line axis632. As discussed above, the wave components628,630are positioned on the respective surfaces606,608of the chip602such that their center or central power generating region, defined by a first end point650and a second end point651, is aligned with the second end622of the separation channel616prior to the separation channel's616bifurcation into the upper and lower outlet channels624,626. In some embodiments, it has been found that this alignment can result in high separation efficiency because a resulting SAW is strongest in a region between the center regions of the wave components628,630and because cells do not have to be suspended throughout the whole length of the separation channel616as discussed further below. However, in other embodiments, the wave components628,630are positioned relative to other sections of the separation channel616, with the proviso that the separation channel616is positioned between the first and second wave components628,630.

The device600is configured to wash a multicomponent mixture comprising cells634. As described above, in various embodiments the multicomponent mixture comprises red blood cells or red blood cells and a rejuvenation solution. The multicomponent mixture comprising red blood cells634is introduced to the device600through a lower inlet port636that is in fluid communication with the lower inlet channel612. Likewise, a wash material638is introduced to the device600through an upper inlet port640that is in fluid communication with the upper inlet channel610. Flow of the multicomponent mixture634and the wash material638can be established, by pumps, such as peristaltic pumps, optionally coupled to pulse dampeners or pulse suppressors, as described above. As the multicomponent mixture634and the wash material638merge at the separation channel, the multicomponent mixture634flows adjacent to the channel floor620and the wash material638flows adjacent to the channel ceiling618. As such, the wash material638flows over the multicomponent mixture634to create an interface between the wash material638and the multicomponent mixture634. There is little or no mixing between the wash material638and the multicomponent mixture640near the first end614of the separation channel. The first and second wave components628,630generate a SAW with an antinode positioned near the upper outlet channel624. As the multicomponent mixture comprising cells634and the wash material638flow relative to the SAW at the second end622of the separation channel616, pressure nodes pushes, forces, isolates, or moves the cells634toward the antinode positioned near the upper outlet channel624. Accordingly, the cells634are forced upward through the wash material638and toward the upper outlet channel624, whereby the cells634are washed and cleaned. This movement of the cells634displaces the wash material638and remaining components of the initial multicomponent mixture comprising cells634into the lower outlet channel644. Red blood cells634that are washed and clean can be collected at an upper outlet port642that is in fluid communication with the upper outlet channel624and remaining wash material638along with other components, such as, for example, rejuvenation solution, can be collected at a lower outlet port644that is in fluid communication with the lower outlet channel626.

Systems

The present technology provides systems for separating of cells from a multicomponent fluid, comprising a device of the present technology (as described above) and a base unit that facilitates the function of the device. In some embodiments, the device is a disposable chip, operable for a limited number of uses (e.g., a single use). Preferably in such embodiments the base unit comprises components that are operable for multiple uses.

The base unit350comprises at least one of a plurality of coupling members352and a third wave component354. The coupling members can be any coupling members known in the art. Non-limiting examples of connecting members include snaps, clips, clasps, screws, adhesives, fasteners, etc. The third wave component354is either a wave generator or an electrical contact. In embodiments where the first wave component310of the disposable separation device302is a wave generator, the third wave component354is an electrical contact. In one embodiment the disposable separation device302comprises a first wave component310, which is a wave generator. In such embodiments, the third wave component354of the base unit350is an electrical contact. The coupling members352are then configured to couple and hold the disposable separation device302to the base unit350such that the wave generator of the disposable separation device302contacts and communicates with the electrical contact. In another embodiment, the disposable device302does not comprise a first wave component310. In this embodiment, the third wave component354of the base unit350is a wave generator. The snaps352are then configured to snap the disposable separation device302to the base unit350such that the separation channel308is positioned between the wave generator on the base unit350and the second wave component314of the disposable separation device302. Nonetheless, in all embodiments a SAW is generated in the disposable separation device302with power provided by the base unit350.

The disposable separation device302can be prepackaged and sterilized. When ready for use, the disposable separation device302is removed from the packaging and snapped onto the base unit350. A wash material is then pumped through the device and the base unit is activated to generate an SAW. A multicomponent mixture, such as a red blood cell composition, is then pumped through the separation device302, wherein the blood is washed and separated from undesired components.

Methods

The present technology provides devices, systems, and methods for separating a target component from a multicomponent fluid. For example, the target component may be red blood cells or other cells. In various embodiments, the multicomponent fluid comprises a physiologically-acceptable carrier for the target component, such as saline or plasma. Methods include those comprising separating the red blood cells from one more second components of the multicomponent fluid. In some embodiments, the second component comprises at least a portion of the carrier; in some embodiments, the second component comprises essentially all of the carrier. The second component may be used in other processes, or may be discarded. In some embodiments wherein the target material is red blood cells, the second component comprises cells and cell debris, such as white blood cells, platelets, dead cells, or cell debris.

In various embodiments, the present technology provides methods for washing red blood cells that have been suspended in a storage solution or other carrier that is not suitable for administration to a human or other animal in a transfusion. In such methods, the red blood cells are substantially removed from storage solution, and resuspended in a wash solution in a device of the present technology.

For example, before transfusions, red blood cells are often rejuvenated with a rejuvenation or enhancement solution, such as Rejuvesol® red blood cell processing solution commercialized by Citra Labs, LLC (Braintree, Mass.). Such enhancement solutions and methods of use are described in U.S. Pat. No. 9,066,909, Alan Gray, issued Jun. 30, 2015; U.S. Patent Publication No. 2014/0212400, Alan Gray published Jul. 31, 2014, and U.S. Patent Publication No. 2014/0212397, Alan Gray et al., published Jul. 31, 2014, incorporated by reference herein. After rejuvenation, the red blood cells are washed with a wash solution, such as water, saline, dextrose, saline with 5% dextrose, phosphate buffered saline, and other wash liquids to remove excess rejuvenation solution from the red blood cells. Therefore, the rejuvenation solution and/or the wash solution need to be removed from the red blood cells prior to transfusion.

In some embodiments, methods for washing a multicomponent fluid comprising cells comprises delivering, such as by pumping or flowing, a composition comprising cells and a wash material into a separation device comprising a separation channel having a receiving or mixing region, a separation region and a collection region. In various embodiments, the composition comprising cells is a composition comprising red blood cells. The composition may also comprise materials to be washed away from the cells, including other cell types, dead cells, cell debris, rejuvenation solution, or combinations thereof. The wash material is selected from the group consisting of water, saline, dextrose, saline with 5% dextrose, and phosphate buffered saline. The separation device can be any separation device described above.

The method also comprises mixing the composition comprising cells with the wash material. Mixing occurs when the composition comprising cells contacts the wash material in the receiving or mixing region of the channel. Alternatively, the composition comprising cells can be mixed with the wash material outside of the device to generate a pre-mixed composition. In such embodiments, the pre-mixed composition is delivered into the separation device. Then, the method comprises isolating or separating a component from the composition comprising cells. The component can be a desired type of cell, such as, for example, red blood cells. Isolating or separating a component comprises passing, such as by pumping or flowing, the composition comprising cells and the wash material relative to a pressure node generated by a SAW, wherein a pressure node associated with the SAW is located within the separation region of the channel. The SAW is generated by wave components operating at a frequency range of from about 300 kHz to about 1000 kHz, or from about 680 kHz to about 710 kHz.

After the component is isolated or separated, the method comprises collecting the component at an outlet of the device that is in fluid communication with the collection region of the channel. In embodiments where the composition comprising cells is a composition comprising red blood cells, the red blood cells can be washed and isolated by this method, and then transfused into a human or non-human subject in need thereof.

An exemplary embodiment of the present technology is depicted inFIG. 12. As shown, a wash material liquid stream401is introduced in the mixing region or chamber405of a device400that is operable to separate a component from a multi-component solution using standing acoustic waves, such as described above. Such devices and methods are also described in U.S. patent application Ser. No. 14/519,284, Leach et al., filed Oct. 21, 2014, and U.S. Provisional Patent Application Ser. No. 62/095,480, Abeskaron, filed Dec. 22, 2014, the disclosures of which are incorporated by reference herein.

In further reference to exemplaryFIG. 12, a cellular component liquid stream402, such as comprising red blood cells (RBC), is introduced into the region405, in contact with the wash material liquid stream. Application of acoustic waves causes the red blood cells to be moved to the wash material stream, forming a washed component liquid stream403. While, as depicted inFIG. 4, the cellular component liquid stream402is introduced to the mixing region405below the wash material liquid stream401, the relative orientation of the streams may be varied, e.g., such that the cellular component liquid stream402may be introduced above the wash material liquid stream401.

In various embodiments, the interfacial tension between the cellular component liquid stream and the wash material liquid stream is near zero. The interfacial tension may be controlled by selection of the components of the respective streams. For example, one or both of the density and viscosity of the streams may be adjusted by inclusion of an interfacial adjustment material is preferably biocompatible materials suitable for intravenous administration to a human or animal subject. For example, the wash material liquid may comprise dextrose, sucrose or hydrophilic polysaccharide polymers (e.g., dextran and Ficoll) so as to effect a desired density or viscosity. Preferably, the wash material liquid comprises salt, and is isotonic with the cellular component, so as to avoid damage to the cells (e.g., through osmotic shock). In some embodiments, the wash material liquid comprises sucrose. For example, the wash material may be an isotonic mixture of saline and sucrose, having a sucrose concentration of about 9.25%.

As shown inFIG. 13A, generating a standing wave with a node positioned in a channel of a device forces particles to align into a standard band. In contrast,FIG. 13Eshows free floating particles in a channel. However, the thickness of the band shown inFIG. 13Acan be manipulated or tightened by adjusting various parameters. Input offset voltage (Vos) is a parameter that define a differential DC voltage required between inputs of an amplifier, such as an operational amplifier (op-amp), to make the output zero (for voltage amplifiers, 0 V with respect to ground or between differential outputs, depending on the output type). When an input offset voltage is applied, the band of particles flowing through a channel is tighter, i.e., thinner, relative to the standard band shown inFIG. 13A. Another parameter that may be adjusted is phase shift or phase offset. This parameter creates a change in the initiation point of a waveform. As shown inFIG. 13C, when a phase shift or phase offset is applied, the band of particles flowing through a channel is tighter, i.e., thinner, relative to the standard band shown inFIG. 13A. Also, a user may employ a dithered or swept signal. This process generates a signal for several given frequencies over a given time interval. As shown inFIG. 13D, when a dithered or swept signal is applied, the band of particles flowing through a channel is tighter, i.e., thinner, relative to the standard band shown inFIG. 13A. Moreover, channel volume, hematocrit and flow rate can also be adjusted to increase efficiency.

NON-LIMITING DISCUSSION OF TERMINOLOGY

The disclosure of all patents and patent applications cited in this disclosure are incorporated by reference herein.

The description and specific examples, while indicating embodiments of the technology, are intended for purposes of illustration only and are not intended to limit the scope of the technology. Equivalent changes, modifications and variations of specific embodiments, materials, compositions and methods may be made within the scope of the present technology, with substantially similar results. Moreover, recitation of multiple embodiments having stated features is not intended to exclude other embodiments having additional features, or other embodiments incorporating different combinations of the stated features. Specific examples are provided for illustrative purposes of how to make and use the compositions and methods of this technology and, unless explicitly stated otherwise, are not intended to be a representation that given embodiments of this technology have, or have not, been made or tested.

As used herein, the words “prefer” or “preferable” refer to embodiments of the technology that afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, and is not intended to exclude other embodiments from the scope of the technology.

A” and “an” as used herein indicate “at least one” of the item is present; a plurality of such items may be present, when possible. “About” when applied to values indicates that the calculation or the measurement allows some slight imprecision in the value (with some approach to exactness in the value; approximately or reasonably close to the value; nearly). If, for some reason, the imprecision provided by “about” is not otherwise understood in the art with this ordinary meaning, then “about” as used herein indicates at least variations that may arise from ordinary methods of measuring or using such parameters.