Electromagnetic components for handling fluids

Abstract

The present disclosure relates to methods and apparatus for processing fluids through the use of a magnetic assembly, wherein the magnetic assembly includes at least one fluid chamber containing fluid and magnetic particles.

Description

[0001] Priority declaration

[0002] This patent application claims priority to U.S. Provisional Application Serial No. 63 / 004,913, filed on April 3, 2020, which is incorporated herein by reference in its entirety.

[0003] background

[0004] The preparation of samples is a key stage in chemical and biological analysis research. In order to achieve accurate and reliable analysis, the target compound must be processed from the complex original sample and delivered to the analytical equipment. For example, proteomics research usually focuses on a single protein or a group of proteins. Therefore, biological samples are processed to separate the target protein from other cell materials in the sample. Additional processing is usually required, such as protein separation (such as immunoprecipitation), matrix purification, digestion, desalination. Non-target substances are usually found in chemical and biological samples, such as salts, buffers, detergents, proteins, enzymes and other compounds. These non-target substances may interfere with analysis, for example, by causing a reduction in the target signal amount detected by the analytical equipment. Therefore, complex original samples are usually separated and / or extracted through one or more separation and / or extraction techniques to separate the compound of interest from the non-target substances.

[0005] Magnetic particles or beads are a technology that can be used for sample preparation for chemical and biological assays and diagnostics. A key element of magnetic particle separation and processing technology is efficient mixing to enhance the reaction rate between the target species and the particle surface, mass transfer from one substrate to another, or transfer of analytes from one medium to another.

[0006] A known technique for mixing fluids using magnetic particles involves moving a magnet relative to a stationary container or using a mechanical device to move the container relative to a stationary magnet to cause a relative displacement of the magnetic field gradient in the container. Another technique involves using two electromagnets facing each other, which surround a chamber with magnetic particles arranged therein. The two electromagnets are sequentially powered on and off (i.e., binary on / off control) at a sufficient frequency to suspend the magnetic particles in the fluid arranged in the chamber. Such a technique may require excessive power consumption and may cause the magnetic particles to separate slowly. Or such a technique may require an improved lens arrangement that may reduce the quality of mixing. However, these and other techniques known in the art have various disadvantages, including the aggregation of particles and the inefficiency of particle mixing. In addition, such a technique may require manual intervention between the stages of the process. The technique of improving mixed solutions using magnetic beads is to use an electromagnet around a sample container to generate a changing magnetic field.

[0007] However, the magnetic particles commonly used to capture and separate biomolecules are paramagnetic. Paramagnetic beads respond to an applied external magnetic field, but retain little or no residual magnetism when the field is removed. This low residual magnetism reduces or eliminates bead aggregation, allowing the beads to remain dispersed and suspended in solution and to be easily transferred through a pipette tip. However, paramagnetic beads are generally less responsive to external magnetic fields and are therefore more difficult to mix effectively using electromagnetic mixers, particularly in viscous solutions (such as those used to selectively precipitate and separate nucleic acids using magnetic beads). Therefore, there is a need to provide an arrangement of electromagnetic elements that more effectively induce efficient mixing of such magnetic particles. Summary of the invention

[0008] The devices, systems and methods described herein allow for the use of electromagnetic components to process sampling devices and fluids without being limited by known techniques. For example, the devices, systems and methods described herein allow for the use of electromagnetic components to process sampling devices and fluids on a sample volume without sample loss or magnetic particle loss. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] The following description is provided by way of example with reference to the following drawings. It will be understood that the drawings are provided as examples only and that all references to the drawings are made for illustrative purposes only and are not intended to limit the scope of the present disclosure in any way. For convenience, reference numerals may also be repeated (with or without offset) throughout the drawings to indicate similar parts or features.

[0010] Figures 1A to 1D is a schematic diagram of a fluid handling system according to various aspects described herein.

[0011] Figure 2A and Figure 2B is a schematic illustrative open well magnetic sample plate according to various aspects described herein.

[0012] Figure 3 is a schematic illustrative fluid handling system in accordance with various aspects described herein.

[0013] Figure 4 are schematic illustrative fluid handling structures and mixing modes thereof according to various aspects described herein.

[0014] Figure 5 are schematic illustrative fluid handling structures and mixing modes thereof according to various aspects described herein.

[0015] Figure 6 is a schematic illustrative fluid handling and analysis system in accordance with various aspects described herein.

[0016] FIG. 7A to FIG. 7Bis a schematic diagram of another example of a fluid handling system according to various aspects described herein.

[0017] Figure 8 is a representation of one example of z-direction mixing produced by the physical motion of the magnetic lenses described herein.

[0018] FIG. 9A to FIG. 9B is a representative fluid handling system according to various aspects described herein.

[0019] FIG. 10A to FIG. 10B It is a representation of a 4-point lens shape.

[0020] Fig.11 is a representation of an illustrative lens shape.

[0021] Fig.12 is a picture of an example magnetic lens assembly where the lens is fastened to an electromagnet core via a threaded nut.

[0022] FIG. 13A to FIG. 13C It is a representation of the permanent magnet track where such track components are moved in and out of the tube array for separation.

[0023] FIG. 14A to FIG. 14B is to move the sample tube 115 relative to the magnetic lens 730b created by the collection of lens components 730c ( Fig.14A ) and moving the entire magnetic assembly 900 relative to the sample tube 115 ( Fig. 14B ) is expressed as .

[0024] Fig.15 is a representation of one example of an assembly of a vertically oriented permanent magnet that can be reversibly positioned proximate a fluid sample. DETAILED DESCRIPTION

[0025] Those skilled in the art will appreciate that the methods, systems, and devices described herein are non-limiting examples, and that the scope of the applicant's disclosure is limited only by the claims. Although the applicant's teachings are described in conjunction with various aspects, it is not intended that the applicant's teachings be limited to such aspects. On the contrary, as will be appreciated by those skilled in the art, the applicant's teachings include various alternatives, modifications, and equivalents. Features shown or described in conjunction with an example may be combined with features of other aspects. Such modifications and variations are intended to be included within the scope of the applicant's disclosure.

[0026] The present disclosure generally relates to a fluid processing method and system for mixing, separating, filtering or otherwise processing a fluid sample by utilizing magnetic particles dispersed therein. According to various aspects of the present disclosure, a fluid sample can be disposed in a fluid chamber. According to various aspects, the fluid can also be a viscous solution; however, the term fluid is generally used to describe any material in which a sample can be suspended. Multiple fluid chambers are accommodated and dispersed throughout the fluid container. The fluid chamber can be (e.g., open to the atmosphere) an open tube or similar device so that samples and / or reagents can be (e.g., via an automatic sampler or a pipette inserted through the open end of the fluid chamber) directly added to the open fluid chamber and can also be removed directly from the open fluid chamber after processing (e.g., via a capture device).

[0027] Magnetic particles arranged and dispersed in a fluid can be configured to be stirred under the action of a magnetic field (or gradient) generated by a magnetic assembly arranged adjacent to a fluid chamber (e.g., arranged around the periphery of a fluid container) to promote the movement of magnetic particles in the fluid. The magnetic assembly can include one or more magnetic structures arranged in a horizontal or substantially horizontal layer. Each of the magnetic structures can be formed by one or more magnets, such as electromagnets. The vertical position of one or more magnetic structures in the magnetic structure relative to the fluid can be movable or adjustable, for example, before, during or after promoting the movement of magnetic particles in the fluid. The adjustment of the vertical position of one or more magnetic structures in the magnetic structure before promoting the movement of magnetic particles can be used, for example, to process different sample volumes and / or to affect the characteristics of the magnetic field generated by the magnetic assembly. The vertical movement of the magnetic structure can add, for example, a vertical component of the movement of particles to provide more effective or efficient mixing of particles in the fluid while promoting the movement of magnetic particles. Additionally or alternatively, electrodes of various magnetic structures (e.g., different vertical spacing layers) can be selectively energized to process different sample volumes and / or affect the characteristics of the magnetic field generated by the magnetic assembly.

[0028] The magnetic assembly structure can be formed by a plurality of electromagnets arranged around a fluid chamber at one or more different vertical heights, wherein each electromagnet is individually controlled to generate a desired magnetic field in the fluid chamber that effectively affects the magnetic particles arranged therein. Based on the selective application of electrical signals to the plurality of electromagnets surrounding the fluid chamber, the magnetic particles can be affected by the combined effect of the magnetic field gradients generated by the various electromagnets and rotate, spin, horizontally move side-to-side and / or vertically move up and down or any combination of such movements in the fluid sample. By way of example, the signal applied to the electromagnets of each magnetic structure (e.g., in a single horizontal layer) can be configured to generate a magnetic field gradient substantially in the xy plane, while the signal applied to the electromagnets of different magnetic structures - if present (e.g., electromagnets in different horizontal layers) - may cause the magnetic field gradient to present a z-direction or vertical component. In this way, the combined effect of multiple electromagnets can generate a magnetic field with different characteristics (e.g., different strengths and / or directivities) in the sample container, so as to quickly and effectively mix the fluid and / or capture the target analyte in the fluid by way of non-limiting examples.

[0029] Reference Figure 8 , through the center of the coil 730 is placed an assembly 900 including a pin 901 made of a magnetically conductive metal, which extends above the coil 730. When the coil 730 is actuated, it generates a magnetic field that is transmitted to the pin 901 and in turn to the lens assembly 730a, which is also made of a magnetically conductive metal. The lens assembly 730a includes a plurality of magnetic lenses 730b created by a collection of lens members 730c (see Fig. 10A ), each lens member 730c focuses and shapes the magnetic field in a desired area, in this example within a sample tube 115 (not shown) including magnetic particles. The lens members 730c included in the assembly 730a can have any suitable shape. In various examples, the lens members 730c can have a circular shape. In various examples, the lens members 730c have a 4-point shape, such as Fig. 10B By way of other example, the lens member may be formed into any shape that is most efficient for the assembly, such as Fig.11 In various aspects, a magnetic lens is brought into contact with (or in close proximity to) a sample as described herein, such as sample tube 115 .

[0030] In various examples, the thickness of the lens is 0.25 mm to 20 mm. In another example, the thickness of the lens is 2 mm to 12 mm.

[0031] Although each lens member 730c is joined by the link member 730d, Fig. 10A and Fig. 10BThe lens assembly shown in is substantially formed as a single entity, but in various examples, one or more of the plurality of lens components 730c may be separate. See, for example Fig.12 , wherein each individual lens member 730c includes threads configured to receive and screw onto the threaded pin 901. In various aspects, the magnetic lens is formed by a single lens member 730c and a plurality of lens members 730c will constitute the lens assembly 730a.

[0032] In various examples, the coils for inducing the magnetic field are encapsulated in aluminum or copper. In various examples, the array of electromagnetic coils is completely encapsulated in an aluminum block or other high thermal conductivity material with low magnetic permeability. In addition, a small amount of thermal potting compound (not shown) can be placed between the block and the coils to create full contact between the coils and the block. In various aspects, the coil 730 and lens assembly 730a are encapsulated in a solid potting material (not shown).

[0033] In various examples, heat from the coil is isolated from the sample and removed from the device to maintain a suitable temperature for the sample.

[0034] In various examples, the samples can be heated or cooled so that they are maintained or thermally cycled at a different temperature than the surrounding environment. The heating or cooling can be accomplished using any suitable heating or cooling element. In one example, the samples can be heated using heat generated by a coil used to induce a magnetic field.

[0035] The lens assembly can be moved relative to the sample tube while actuating one or more coils in the coil to move the beads up and down in the sample fluid. The lens assembly can be physically moved while the sample tube remains stationary. The sample tube can be physically moved while the lens assembly remains stationary. Both the lens assembly and the sample tube can be physically moved. In various examples, the lens magnetic assembly and / or structure cause the particles (e.g., ferrimagnetic particles) to spin as defined by the presence of a magnetic field, or to travel back and forth in the x-direction, the y-direction, and the z-direction. By way of example, the signal of the electromagnets 110a to 110d applied to each magnetic structure 110 (e.g., in a single horizontal layer) can be configured to generate a changing magnetic field substantially in the xy plane, while the movement of the lens assembly relative to the sample tube generates a changing field in the z-direction or a mixed vertical component. In this way, the combined effect of multiple electromagnets can generate a magnetic field with different characteristics (e.g., different strengths and / or directivities) in the container 115, so as to quickly and effectively mix the sample and / or capture the target analyte in the sample by way of non-limiting examples. The vertical movement of the sample tube or lens assembly can be a single movement upward or downward, or can include any combination of continuous upward and downward movement. The vertical movement can start at any vertical position of the lens assembly relative to the sample tube. In some aspects, the upward vertical movement can start when the lens assembly is positioned near the bottom of the sample tube to cause the vertical re-suspension of magnetic particles that may have settled toward the bottom of the tube. In some examples, the vertical movement of the lens assembly or the sample tube can start when the lens assembly is positioned near the sedimentation layer or boundary layer between the components that can be separated in the sample fluid. In this way, while the coil is actuated, the vertical movement of the lens assembly or the sample tube can help destroy the sedimentation layer or boundary layer to provide more effective mixing of the entire sample fluid. The rate of vertical movement can be any suitable rate that maintains effective mixing in the xy plane while providing sufficient distribution of mixing along the z direction. The range of vertical movement can be any suitable range required to maintain sufficient mixing along the z direction.

[0036] In various examples, the controller can be configured to actuate the electromagnets via one or more of a radio frequency (RF) signal, a direct current (DC) signal, an alternating current (AC) signal, an electrical frequency (EF), etc., and also include the application of any combination thereof. In various examples, the RF signals applied to a plurality of electromagnets can present phase delays different from each other to achieve the desired movement of the electromagnets in the sample fluid. In some aspects, the DC signal can effectively separate particles (e.g., magnetic particles are attracted to one side of the fluid chamber and / or the vertical level) so that the fluid can be withdrawn from the chamber by way of non-limiting examples without aspirating (aspiration) magnetic particles. In some examples, the constant voltage DC signal can be interspersed between alternating or varying actuation signals to provide more effective mixing of the sample fluid. The alternating or varying actuation signals around the constant voltage DC signal can be any suitable RF, AC, DC or EF signals, etc.

[0037] In various examples, the tube is held non-rotatable during the mixing process. For example, the tube can be mechanically fixed in place using an interference fit mechanism. The tube can also be screwed or similarly rotated into a locked position within the bracket. The tube can also be held in a non-rotatable manner by using a cover or similar feature associated with the bracket.

[0038] The fluid handling system described according to various examples can be configured to process fluids at a microscale or macroscale (including large volume formats). In general, macroscale relates to fluid volumes within a milliliter range, while microscale fluid handling relates to fluid volumes below the milliliter range such as microliters, picoliters or nanoliters. Large volume formats can relate to the processing of fluid volumes greater than 1mL. For example, the fluid handling system according to various aspects of this teaching can process fluid volumes of about 1 μL to about 15mL or even larger, including, for example, about 1.5mL, about 2mL, about 5mL, about 10mL or larger. In some aspects, the fluid chamber is configured to accommodate a volume within the range of about 20 μL to 200 μL.

[0039] In some examples, the fluid chamber is configured to extend from a lower closed end to an upper open end, the upper open end being configured to be open to the atmosphere to receive a fluid to be processed by the fluid chamber. In some examples, the fluid chamber includes a cover.

[0040] However, it will be appreciated in light of this disclosure that the fluid handling system can handle any fluid volume capable of operating as described herein.

[0041] The use of magnetic assemblies that affect magnetic particles according to various examples can provide multiple technical advantages, for example, compared with conventional magnetic particle processing systems. A non-limiting example of such advantages includes a significantly improved diffusion rate, for increasing the sample contact rate in various volumes of sample fluids, for example, to improve the analyte capture efficiency in magnetic immunoassays. Another non-limiting example of technical advantages includes increased sample mixing efficiency, because the magnetic structure of the magnetic assembly can affect the magnetic particles to provide faster and more effective sample mixing due to, for example, more robust magnetic particle motion and motion in multiple dimensions. For example, this can result in increased mass transfer between components.

[0042] The fluid handling structure processing sample using the configuration according to the applicant's teaching produces rapid reaction kinetics. For example, compared with the one or two day processing time of processing in the artificial tube, protein processing (including immunoaffinity pull-down, washing, elution / denaturation, reduction, alkylation and digestion steps) can be completed in about 10 minutes to 12 minutes. For example, due to overcoming the diffusion of the rate-limiting step (such as the rate-limiting step of LC) as fluid processing and the necessity of utilizing a small fixed volume in a known microfluidic platform, the increased processing speed can be achieved. In addition, since the fluid handling structure configured according to the applicant's teaching can be integrated into the array of a large sample reaction well, this fast and efficient sample processing can be realized in the array of a large sample reaction container at the same time, thereby enhancing sample processing and making it possible to realize automation via, for example, an automatic sampler. It will be understood according to the present disclosure that, in addition to the non-limiting examples mentioned above, the fluid handling system described herein also provides a plurality of other technical advantages.

[0043] Although the systems, devices, and methods described herein can be used in conjunction with many different fluid handling systems, Figure 1A An example of a suitable fluid treatment system 100 is schematically shown in FIG. It should be understood that the fluid treatment system 100 represents only one possible fluid treatment system for use in accordance with the systems, apparatuses, and methods described herein, and that fluid treatment systems and / or components thereof having other configurations and operating characteristics may also be used in accordance with the systems, apparatuses, and methods described herein.

[0044] In various aspects, in the solution that sample has been added to more viscous bead solution, two kinds of fluid can be partially separated, at least one boundary is formed between the partially separated fluid layers. The vertical motion of the magnetic assembly approaching or crossing such boundary to mix the sample of combination and bead solution while one or more electromagnets of actuating assembly can provide more effective or more thorough mixing of the sample of combination and bead solution. In some examples, the vertical position of the boundary can be estimated in advance based on the bead solution of known volume and the sample added. In other examples, the vertical motion of the magnetic assembly is programmed to contain most or all of the range of sample fluid or sample tube, so as to promote effective mixing regardless of the initial vertical position of the boundary.

[0045] Figure 1A An example of a fluid handling system 100 is schematically depicted. Figure 1A As shown in , the fluid handling system 100 includes a fluid handling structure or container 130 having a fluid chamber 115 and a magnetic structure 105, the magnetic structure 105 being configured to generate a magnetic field gradient or magnetic force within the fluid chamber as discussed in detail below. The fluid chamber 115 may generally include any type of vessel configured to contain a sample fluid, such as a sample well, a vial, a fluid reservoir, etc., defining a chamber containing a fluid therein. Figure 1B As best shown in FIG. 1 , the fluid chamber 115 extends from an open upper end 115a (open to the ambient atmosphere) to a bottom closed end 115b, such that the fluid within the fluid chamber 115 can be loaded and / or removed from the fluid chamber 115 by one or more fluid loading / collection devices 135 that can be inserted into the open upper end 115a. One skilled in the art will appreciate that the chamber 115 can include a removable cover that can be coupled to the open upper end 115a (e.g., an Eppendorf tube) during various processing steps, such as to prevent leakage, contamination, and / or evaporation of the fluid during mixing. Illustrative fluid loading / collection devices 135 can include, by way of non-limiting example, but are not limited to, a manual sample loading device (e.g., a pipette), a multichannel pipette device, an acoustic fluid handling device, and / or an autosampler.

[0046] Refer again Figure 1A , the sample fluid may have a plurality of magnetic particles 120 disposed therein and may be added to the sample fluid before transferring the sample fluid to the fluid chamber 115 , or may be added to the fluid chamber 115 before or after the sample fluid has been transferred to the fluid chamber 115 .

[0047] Suitable magnetic particles 120 for use in the systems and methods described herein include, but are not limited to, paramagnetic particles such as AMPure XP beads available from Beckman Coulter, Inc. of Brea, Calif. Suitable magnetic particles also include those described in U.S. Pat. Nos. 5,705,628; 5,898,071; and 6,534,262 and published PCT application No. WO 2020 / 018919, published on January 23, 2020, all of which are incorporated by reference as if fully set forth herein.

[0048] As used herein, "ferrimagnetic particles" refers to particles comprising ferrimagnetic materials. Ferrimagnetic particles can respond to an external magnetic field (e.g., a changing magnetic field), but can be demagnetized when the external magnetic field is removed. Thus, ferrimagnetic particles are effectively mixed through a sample by an external magnetic field, and effectively separated from a sample using a magnet or electromagnet, but can remain suspended without magnetically induced aggregation.

[0049] The magnetic particles 120 described herein have enough responses to the magnetic field so that they can effectively move through the sample. In general, the scope of field strength can be the same scope as any electromagnet, as long as it can move the particle. For example, the magnetic field has an intensity between about 10mT to about 250mT, between about 20mT to about 80mT and between about 30mT to about 50mT. In some examples, more powerful electromagnets can be used to mix the particles of lower responsiveness. In some examples, the magnetic field can be concentrated in the sample as much as possible. In addition, the electromagnet can be as close to the sample as possible because the intensity of the magnetic field decreases with the square of the distance.

[0050] The magnetic particles 120 can be various shapes that can be regular or irregular. In some examples, the shape maximizes the surface area of ​​the particles. For example, the magnetic particles 120 can be spherical, bar-shaped, elliptical, or any other suitable shape. The magnetic particles 120 can be various densities that can be determined by the composition of the core. In some examples, the density of the magnetic particles can be adjusted using a coating.

[0051] The magnetic structure 105 may include a plurality of electromagnets 110a to 110d. Figure 1A Four electromagnets 110a to 110d are depicted in the drawings, but the number and type of magnets are not limited thereto, and any number of electromagnets capable of operating in accordance with various aspects of the applicant's teachings may be used. The four electromagnets 110a to 110d may operate the same or substantially similar to a quadrupole magnet structure. For example, the magnetic structure 105 may include two electromagnets, three electromagnets, or a plurality of electromagnets. Figure 1A; however, there may be more electromagnets if desired. The electromagnets 110a to 110d may include any electromagnet known to those skilled in the art, including, for example, ferromagnetic core electromagnets. The electromagnets 110a to 110d may have a variety of shapes, including square, rectangular, circular, oval, or any other shape capable of operating in accordance with various aspects of the applicant's teachings.

[0052] like Figure 1A As shown in, the fluid handling system 100 additionally includes a controller 125, which is operably coupled to the magnetic structure 105 and is configured to control the magnetic field generated by the electromagnets 110a to 110d. In various aspects, the controller 125 can be configured to control one or more power supplies (not shown), which are configured to supply electrical signals to multiple electromagnets 110a to 110d. The electrical signal can be in the form of a radio frequency (RF) waveform, a DC current, an AC current, etc. Although the RF waveform is generally used herein as an example of a waveform that can be applied to the electromagnets 110a to 110d to promote the mixing of the fluid sample, the type of electrical signal is not limited thereto, and any type of current that can be operated according to various aspects of the applicant's teachings is contemplated herein. By way of example, a DC signal can be additionally or alternatively applied to one or more of the electromagnets so as to attract magnetic particles to one side of the fluid chamber. Another example may include a DC signal that may be supplied between the RF and / or AC signals to promote sample mixing, or supplied after the RF and / or AC signals to help transfer fluid from the chamber after the mixing step and / or prevent the suction of magnetic particles. In various examples, the controller 125 may be any type of device and / or electrical component capable of actuating an electromagnet. The controller 125 may be operable to adjust the magnetic field generated by each electromagnet in the electromagnets 110a to 110d by controlling the current of the solenoid or coil passing through each electromagnet in the electromagnet. The controller 125 may include or be coupled to a logic device (not shown) and / or a memory, such as a computing device configured to execute an application that is configured to provide instructions for controlling the electromagnets 110a to 110d. The application may provide instructions based on operator input and / or feedback from the fluid handling system 100. The application may include and / or the memory may be configured to store one or more sample processing protocols for execution by the controller 125.

[0053] In various aspects, each electromagnet 110a to 110d can be individually addressed and actuated by the controller 125. For example, the controller 125 can supply RF electrical signals of different phases to each of one or more of the electromagnets 110a to 110d, so that one or more of the electromagnets produce different magnetic fields. In this way, the magnetic field gradient generated by the magnetic structure 105 in the fluid chamber 115 can be quickly and effectively controlled to manipulate the movement of the magnetic particles 120 in the sample fluid. The RF waveform and its characteristics (e.g., phase shift) can be applied to the electromagnets 110a to 110d according to the sample processing protocol. It will be understood from the present disclosure that the magnetic structure 105 can be used to manipulate the magnetic particles 120 in the sample fluid in various processes, including but not limited to protein determination, sample derivatization (e.g., steroid derivatization, sample derivatization of gas chromatography, etc.), and / or sample purification and desalination. After this treatment, the treated fluid can be delivered to various analytical equipment 140, such as a mass spectrometer (MS) for analysis. A single layer of electromagnets 110a-110d (e.g., arranged around the periphery of the fluid container at a height above the bottom 115b of the fluid chamber) can be actuated to generate a magnetic field within the fluid chamber 115 that captures and / or suspends magnetic particles 120 in a specific plane within the fluid chamber. For example, the magnetic particles 120 can be suspended in a specific plane to move the magnetic particles away from the bottom of the fluid chamber during a fluid collection process and / or for processing fluids (e.g., reagents) in a plane above a material (e.g., cells adhered to the bottom surface of the fluid chamber), where contact with the material on the bottom surface of the fluid chamber is to be avoided.

[0054] According to various examples of the present disclosure, the magnetic structure 105 can be incorporated into various fluid handling systems and fluid handling devices. Figure 1B , examples of the magnetic structure 105 are depicted as stand-alone mixing devices. For example, the magnetic structure 105 can be used as a mixing element of a magnetic mixer or a mixing element of a vortex-type mixer (i.e., in place of a motor-driven mixing element). In accordance with the applicant's teachings, a fluid chamber 115 (e.g., a sample well of a single vial and / or a sample plate) can be pressed against an actuator 150 to activate a controller 125 to actuate the electromagnets 110a to 110d. In other examples, the magnetic structure 105 can be used to mix magnetic particles 120 within a sample well of a sample plate (e.g., a conventional 4, 8, 12, or 96-well sample plate). The magnetic structure 105 can be configured to mix magnetic particles 120 within a sample well of an open well sample plate (i.e., open to the atmosphere, sealed with a removable cover or hood, and / or partially enclosed). As Figure 1C As shown in , the fluid chamber 115 (ie, the sample well) of the sample plate 160 can be mounted downwardly within the cavity formed between the electromagnets 110a to 110d. Figure 1D As shown in , the sample plate 160 can be placed on a portion of the fluid handling system 100 , such as on a planar surface 170 thereof, such that the sample well 115 can be arranged adjacent to the electromagnets 110 a - 110 d .

[0055] Figure 2A An example of an open trap magnetic sample plate is depicted. Figure 2A As shown in , the 96-well sample plate 205 may include a plurality of sample wells 215. Figure 2A 20 . A diamond-shaped sample well 215 is depicted in FIG. 20 , but it will be understood that the fluid chamber according to the present disclosure is not limited thereto. For example, the sample well 215 may have a variety of shapes, including square, rectangular, circular, oval, or any other shape capable of operating according to various examples of the applicant's teachings. Each sample well 215 may be surrounded around its periphery by a magnetic structure 210 including a plurality of electromagnets 220 a to 220 d. The magnetic structure 210 and the method of mixing magnetic particles using an RF-driven oscillating magnetic field according to various aspects of the applicant's teachings may be incorporated into existing sample plate devices, including sample plate devices configured as large, open arrays of sample wells 215. For example, the magnetic structure 210 may be configured to receive a standard sample plate device, such as an industry-standard 96-sample well array 205. This may be achieved, for example, by using electromagnets 220 a to 220 d and a magnetic structure 210 configuration having a geometry corresponding to a standard sample well plate. In this way, fluid channels and pumps are not required, thereby reducing or even eliminating fluid handling problems associated with these elements, including but not limited to non-specific binding and carryover (i.e., using disposable sample plates). In addition, the use of open well sample systems provides a more efficient method for sample loading and collection, such as integration with autosamplers and other automated fluid handling systems. In this way, various exemplary fluid handling systems according to the applicant's teachings can allow large sample arrays to be processed simultaneously, which is simple and efficient from the perspective of fluid manipulation and mechanical complexity.

[0056] Figure 2BAn example of a partial view of a container including a layout of a plurality of sample wells 215a to 215d and an associated magnetic structure including electromagnets 220a to 220f is depicted, which demonstrates the sharing of electromagnets 220a to 220f between the plurality of sample wells 215a to 215d. In this example, sample well 215d is surrounded by a magnetic structure including electromagnets 220a, 220b, 220c, and 220d. Electromagnets 220a and 220c also surround sample well 215c, which itself is also surrounded by electromagnets 220e and 220f. Electromagnets 220a and 220c can generate a magnetic field that penetrates both sample wells 215c and 215d. Similarly, sample wells 215b and 215d share electromagnets 220a and 220b, and sample wells 215a and 215c share electromagnets 220e and 220f. Electromagnet 220a is shared by sample wells 215a to 215d and can generate a magnetic field in all four sample wells. It should be understood that this structure can be similarly repeated throughout sample well plate 205 for all sample wells.

[0057] Figure 3 Illustrative fluid handling systems according to various aspects are schematically depicted. Figure 3 As shown in , the fluid treatment system 300 includes a plurality of magnetic structures 305a to 305f configured to generate a magnetic field gradient within an associated fluid chamber 315a to 315f. Each magnetic structure 305a to 305f may include a plurality of electromagnets 310a to 310l, wherein some of the electromagnets 310a to 310l are shared between the magnetic structures 305a to 305f. The electromagnets 310a to 310l may be controlled via application of an RF signal having any suitable phase delay thereto.

[0058] like Figure 3 320, the electromagnets 310a-310l are labeled AD. The phase delay of the electromagnets 310a-310l of the magnetic structure 305a-305f can produce a 90° phase shift for adjacent electromagnets. However, the present disclosure is not limited in this regard, as other phase shift values, such as 180° phase delay, 270° phase delay, etc., can be used in accordance with various aspects of the applicant's teachings. In various aspects, actuation of the electromagnets 310a-310l according to the phase delay equation 320 causes magnetic particles (not shown) in sample wells 315a, 315e, and 315c to mix in a clockwise motion and magnetic particles in sample wells 315b, 315d, and 315f to mix in a counterclockwise motion.

[0059] Mixing the fluid using agitated magnetic particles according to various examples of applicants' teachings causes the magnetic particles to be evenly dispersed within each fluid chamber, thereby providing optimal exposure and enhanced mixing with the fluid.

[0060] Figure 4 Illustrative fluid handling structures and mixing patterns thereof in accordance with various examples of the applicant's teachings are depicted. Graph 405 depicts magnetic fields 410a, 410b generated by applying current to electromagnets 420a to 420d of the fluid handling structure 400 at time intervals T1 to T5 in accordance with various aspects of the applicant's teachings. In various examples, the waveforms of the magnetic fields 410a, 410b represent sine waves that produce exemplary schematic motion 425 of magnetic particles within a container to promote continuous magnetic particle mixing and improved mixing efficiency. The magnetic fields 410a, 410b have a 90° phase shift relative to each other, wherein the magnetic field 410a corresponds to the electromagnets 420a and 420d, and the magnetic field 410b corresponds to the electromagnets 420b and 420c. In Figure 4 In the illustrative description of FIG. 4 , it will be appreciated that the electromagnets 420a to 420d are arranged at different positions relative to the fluid sample so that the orientation of the magnetic field generated by each electromagnet when the same electrical signal is applied thereto is generally different. Likewise, because the electromagnetic pairs (i.e., 420a and 420d, and 420b and 420c) are arranged on opposite sides of the fluid sample, the magnetic fields generated by the electrodes in each pair when electrical signals of the same amplitude and opposite phase are applied to the electromagnets in each pair are in the same direction 430. Thus, when the exemplary sinusoidal electrical signals of equations (1) to (4) are applied to the electromagnets 420a to 420d, respectively, the resulting magnetic field in the sample fluid will be as follows: Figure 4 , where a pair of electromagnets 420a and 420d together generate a magnetic field 410a, and a pair of electromagnets 420b and 420c together generate a magnetic field 410b (magnetic field 410b is delayed by 90° relative to magnetic field 410a), causing the fluid to undergo mixing due to the generally counterclockwise movement 425 and alignment 435 of the particles at various points in time as schematically depicted.

[0061] Therefore, it will be appreciated from this disclosure that different hybrid modes can be achieved by controlling the RF waveform applied to the electromagnets of the magnetic structure. Figure 5 , depicted according to various aspects of the applicant's teachings Figure 4 Another illustrative mixing mode of the fluid handling structure of FIG. As shown, the fluid mixing mode is Figure 4 The fluid mixing pattern shown in FIG. 4 differs in that, for example, the controller is configured to apply differently phase-delayed RF signals to the electromagnets 420a to 420d.

[0062] like Figure 5As shown in , when sinusoidal electrical signals are applied to electromagnets 420a to 420d, respectively, the resulting magnetic field in the sample fluid will vary over time as schematically depicted, where a pair of electromagnets 420a and 420d together generate magnetic field 410a, and a pair of electromagnets 420b and 420c together generate magnetic field 410b. In this case, in contrast, magnetic field 410a is delayed by 90° relative to magnetic field 410b, so that the fluid is mixed in a generally clockwise manner due to the movement 425 of the particles at various points in time schematically depicted.

[0063] Although applied to the surrounding Figures 3 to 5 The sinusoidal RF waveform of each of the four electromagnets of the container presents a ±90° offset relative to the adjacent electromagnet, but the present disclosure is not limited thereto. In fact, it will be understood that any type of waveform can be supplied to an electromagnet capable of operating according to the applicant's teachings. By way of non-limiting example, the number of electromagnets surrounding each fluid chamber, the phase shift between adjacent electromagnets (e.g., 30°, 60°, 90°, 120°, 150°, 180°, 210°, 240°, 270°, 300°, and 330° phase shifts) and the waveform shape can vary according to the changing aspects of the present disclosure. Non-limiting examples of current waveforms may include square, rectangular, triangular, asymmetric, sawtooth, or any combination thereof. The type of current supplied to the electromagnet can be modified during the operation of a fluid treatment system configured according to some embodiments. For example, at least a portion of the electromagnet can receive an RF waveform with a 90° phase shift, while another portion can receive an RF waveform with a 180° phase shift. In such embodiments, the phase shift of each portion can be modified during operation of the fluid treatment system (e.g., the phase shift can be switched, synchronized, etc.). At least a portion of the electromagnets can be operated in parallel, sequentially, pulsed, etc. In various aspects, the current supplied to the electromagnets can be controlled according to the treatment protocol. The treatment protocol can be dynamically changed during operation of the fluid treatment system based on various factors (e.g., feedback, operator input, detection of mixing efficiency, analytical results, etc.).

[0064] In various examples, the waveform may include different segments with different amplitudes. For example, the waveform may include an initial segment of relatively short duration with a higher amplitude (boost), followed by a sustained segment of lower amplitude. In various aspects, the amplitude of the sustained segment is lower than the amplitude that will overheat the sample. In various embodiments, the boost amplitude is higher but can be tolerated at the beginning of actuation. In various aspects, the sustained segment may be followed by a constant segment. The constant portion may include a DC signal of a constant voltage, which includes zero voltage. A combination of boost, sustained and constant segments or any sub-combination thereof may be repeated sequentially. In various examples, the boost amplitude may be 1% to 50% higher than the sustained amplitude. In various aspects, the boost amplitude may be 10% to 30% higher than the sustained amplitude. In various aspects, the boost amplitude may be 20% higher than the sustained amplitude.

[0065] In another example, if Fig.15 As shown in , the vertically oriented neodymium magnet 330 is an example of a separate permanent magnet for attracting beads or pulling down beads in a chamber. Magnet 330 can be used in a tray 340 or other holding mechanism. For example, when using a neodymium magnet, such magnets are arranged in a single row on opposite sides of at least one chamber or a row of chambers. In such an example, a row of magnets 330 will be arranged so that the north pole is oriented upward, and in the opposite row, the south pole will be oriented upward. A plate 350, such as made of steel, can be placed below the magnet 330 to connect the magnet 330 to the magnetic circuit. In addition, a motor 360 can be coupled to one or both of the tray 340 and the plate 350 so that when one or both of the tray 340 and the plate 350 are inserted into the guide bracket 370, the motor 360 can cause the movement of the tray 340. This movement is to the position of the adjacent chamber to pull down the beads. During mixing, the tray 340 moves the magnet 330 away from the chamber to allow the beads to remain suspended.

[0066] FIG. 13 is another example of a single permanent magnet for pulling down beads. The single permanent magnet is shown here in the pull-down position. The permanent magnet is the bar closest to the bottom of the tube (the conical portion of the cone shown inverted in the current figure). In the retracted position, the tray pulls the magnetic bar away from the sample tube. Fig. 13B This is the view from the top. Fig.13A is the view from the right, and Fig. 13C This is the view from the front.

[0067] In addition, as noted herein, by way of non-limiting example, electromagnets 420a to 420d may alternatively have a DC signal applied to generate a static magnetic field so that magnetic particles are attracted to one side of the fluid chamber (and away from bulk fluid) to help transfer from the fluid of the chamber after the mixing step and / or prevent the suction of magnetic particles. In various aspects, a separate magnet is used to attract particles to one side of the chamber. In some examples, the separate magnet is a permanent magnet. In another example, the separate magnet is movable to be positioned adjacent to the container at a desired height relative to the bottom of the container to attract particles. In some examples, the separate magnet may be configured to slide horizontally to a position adjacent to the container. In some examples, the separate magnet may align its magnetic axis perpendicular to the vertical axis of the container. In another example, the separate magnet may align its magnetic axis parallel to the vertical axis of the container.

[0068] Now refer to FIG. 7A to FIG. 7B , which provide examples of fluid handling systems 700 according to various examples of the present disclosure. Fig. 7A , the fluid handling system 700 depicted in an exploded view includes a substrate 710, a printed circuit board (PCB) 720, a plurality of electromagnetic structures 730, and an upper plate 740 defining a plurality of sample wells 740 extending from a substantially planar upper surface 740a thereof. Those skilled in the art will appreciate that although the upper plate 740 is Fig. 7A 740 is depicted as a 96-well format with sample wells having a substantially circular cross-sectional shape, but the upper plate 740 may include any number of sample wells 742 presenting various cross-sectional shapes and maximum volumes as discussed above. For example, in accordance with the present disclosure, each of the open sample wells 742 may be filled or partially filled with various volumes of fluid sample, thereby allowing for a reduction or expansion of the sample volume to be processed depending on, for example, the availability or cost of the sample and / or the requirements of a particular assay. It should also be understood that the upper plate 740 may be manufactured from any material known in the art or developed below in accordance with the present disclosure, such as a polymeric material (e.g., polystyrene or polypropylene), all by way of non-limiting example. In addition, as is known in the art, the surface may be coated with various surface coatings to provide increased hydrophilicity, hydrophobicity, passivation, or increased binding to cells or other analytes. In some examples, the bottom surface 740b of the upper plate 740 may be configured to engage (permanently or removably) with a lower portion of a fluid handling system as discussed below. For example, in some aspects, bottom surface 740 b may include a depression formed therein for engaging upper end 730 a of electromagnetic structure 730 or an aperture through which a portion of the electromagnetic structure may extend to be disposed around and about each of sample wells 742 .

[0069] Referring now to the lower portion of the fluid handling system 700, Fig. 7A PCB 720, substrate 710, and multiple electromagnetic structures 730 are depicted. As shown, PCB 720 includes multiple electrical contacts 722 to which electrical signals can be applied by a power source (not shown) and to which electromagnetic structures 730 can be electrically coupled. As discussed elsewhere herein, PCB 720 can be wired so that each electromagnetic structure can be individually addressed and actuated by a controller by selectively applying electrical signals thereto. Additionally, PCB 720 includes multiple holes 724 through which portions of the electromagnetic structures can extend to make electrical contact with substrate 710. For example, as Fig. 7A As shown in , the electromagnetic structure 730 may include a mounting post 732 that extends through the hole 724 when the electromagnetic structure 730 is placed on the electrical contact 722, and allows the conductive lead associated with the mounting post 732 to be electrically coupled to the substrate 710. As shown, the substrate 710 may include a hole corresponding to the mounting post 732 to ensure that the mounting post 732 is securely engaged therewith. The substrate 710 may also be coupled to a power source (or ground) to complete the circuit, so that one or more electrical signals can be applied to the plurality of electrical contacts 722 of the PCB 720 to allow current to flow through the electromagnetic structure 730 according to the present disclosure. As shown in Fig. 7A As shown in , the electromagnetic structure 730 may include an upper post around which a wire 734 electrically coupled to the contact 722 is coiled and the upper post terminates at an upper end 730a. It should be understood that when current flows between the electrical contact 722, the lead coil 734, the upper end 730a and the metal substrate 710 (the direction of the current depends on the voltage of the signal applied to the specific contact 722 of the PCB 720), the lead coil 734 acts as a solenoid, thereby generating a magnetic field through and around the lead coil 734, and the directionality of the magnetic field depends on the direction of the current. The upper end 730a of the electromagnetic structure 730 can have a variety of shapes (e.g., substantially the same cross-sectional shape as the post around which the lead is coiled), however it has been found that the upper end 730a can be preferably formed of a conductive material and shaped to correspond to the outer peripheral surface of the sample well so as to act as a lens to concentrate the magnetic field and / or increase the uniformity of the magnetic field within the sample well. It should be understood that from FIG. 1 to FIG. Figure 5 The examples embodied in FIG. 7 relate to apparatus and methods in which a magnetic structure is arranged around a fluid container in only a single horizontal layer. In this configuration, the generation of a magnetic field results in mixing of particles substantially in the xy plane, which describes only one aspect of the present disclosure. As will be further described in detail in the present disclosure, such systems and methods may also be modified in a manner that an additional magnetic field is generated to result in mixing of particles in the z direction.

[0070] Therefore, it will be appreciated in light of this disclosure that different hybrid modes may be achieved by controlling the RF waveform applied to the electromagnets of the magnetic structure.

[0071] Although cylindrical members have been described above in describing the tube 115, it should be understood that other shapes having different cross-sectional shapes may be utilized, including triangular, square, rectangular, or any other polygonal shape.

[0072] The magnetic assembly and / or magnetic structure including the electromagnet may be placed on the exterior of the metal tube, or may be part of the metal tube itself and integrated directly with the metal at or near the tip.

[0073] It will be appreciated that the teachings described herein may be modified and adapted to meet specific needs as may be determined by one of ordinary skill.

[0074] The magnetic structures and fluid handling systems described in accordance with the applicant's disclosure can be used in combination with various analytical equipment such as LC, CE or MS devices known in the art and developed and modified thereafter in accordance with the present disclosure. Figure 6 , schematically depicts an illustrative fluid handling and analysis system in accordance with various aspects of the applicant's teachings. Figure 6 As shown in , according to some embodiments, the fluid handling system 610 can be configured to process fluid samples using a magnetic structure and an open well sample plate. Manual sample loading devices (e.g., pipettes, multichannel pipettes) or various automated systems (e.g., fluid handling robots, autosamplers) or acoustic fluid handling devices (e.g., manufactured by LabCyte, Inc. of Sunnyvale, California) can be used, all by way of non-limiting example. 525 fluid processors) collect the processed fluid from the fluid handling system 610. Various fluid transfer devices, such as vortex-driven sample transfer devices, can be used to transfer the processed fluid. As noted above, the sample removed from one sample well can be added to a different sample well on the plate for further processing steps, or can be delivered to a downstream analyzer. For example, in some aspects, the processed sample can be delivered to the LC column 615 for online LC separation, and the eluent is delivered to the ion source 620 for ionization of the processed analyte, which can then be analyzed by the DMS 625 and / or the mass spectrometer 630, the DMS 625 analyzes the ions based on the mobility of the ions through the carrier gas, and the mass spectrometer 630 analyzes the ions based on the m / z ratio of the ions. In some aspects, the processed sample can be directly transferred to the ion source 615, where the separation is provided by a differential mobility spectrometer (DMS) component (e.g., consistent with the MS described in U.S. Pat. No. 8,217,344). The combination of the fluid handling system described according to the applicant's disclosure and the DMS assembly for chemical separation can eliminate the need for LC (or HPLC) columns for treating samples for MS analysis. In various fields, surface acoustic wave atomization (SAWN) equipment, electrospray ionization (ESI) devices and matrix assisted inlet ionization (MAII) sources can be used to introduce processed samples into analytical equipment such as MS.

[0075] It should be understood that the above disclosed aspects and other features and functions or their alternatives are expected to be combined into many other different systems or applications. It should also be understood that those skilled in the art may subsequently make various currently unforeseen or unanticipated alternatives, modifications, changes or improvements, which are also intended to be covered by the appended claims.

Claims

1. A method for treating a fluid, comprising: providing at least one fluid container having a fluid chamber containing a fluid and a plurality of magnetic particles; The at least one fluid container is disposed on a magnetic assembly, the magnetic assembly comprising a plurality of electromagnets positioned around a periphery of the fluid chamber, each of the plurality of electromagnets having: (i) a conductive coil positioned about a centerline extending in the z-direction, and (ii) a magnetic lens configured and arranged to direct the magnetic field generated by the conductive coil toward the fluid chamber, wherein at least one of the fluid chamber or the magnetic lens is movable along the z direction; providing an electrical signal to each of the plurality of electromagnets to generate a magnetic field within the fluid chamber, wherein the magnetic field is configured to affect the plurality of magnetic particles; and The electrical signal is adjusted to modify the magnetic field within the fluid chamber, while at least one of the fluid chamber or the magnetic lens is moved along the z-direction to further modify the magnetic field within the fluid chamber.

2. The method according to claim 1, wherein: The at least one fluid container comprises a plurality of fluidically separated fluid chambers, wherein at least one electromagnet of the plurality of electromagnets is configured to generate the magnetic field within two or more fluid chambers of the plurality of fluidly separated fluid chambers.

3. The method according to claim 1, wherein: The at least one fluid container includes a plurality of sample wells disposed within a sample plate.

4. The method according to claim 3, wherein: The magnetic assembly is configured to simultaneously affect the magnetic particles disposed within the plurality of sample wells.

5. The method according to claim 3, wherein: The sample plate includes a bottom surface configured to removably engage at least a portion of the magnetic assembly.

6. The method according to claim 1, wherein: Adjusting the electrical signal to modify the magnetic field within the fluid chamber includes applying at least one electrical frequency waveform to each of the plurality of electromagnets.

7. The method according to claim 6, wherein: The at least one electrical frequency waveform applied to each of the plurality of electromagnets has a phase delay.

8. The method according to claim 1, wherein: The fluid chamber is configured to hold a maximum volume in the range of 1 μL to 15 mL.

9. The method according to claim 1, wherein: The plurality of electromagnets are arranged to surround the fluid chamber at a plurality of vertical locations.

10. The method according to claim 1, wherein: The magnetic lens is moved while adjusting the electrical signal to the coil.

11. The method according to any one of claims 1 to 10, further comprising: Fluid is added to, mixed in, removed from, or heated in the fluid chamber.

12. The method of any one of claims 1 to 10, further comprising removing the magnetic particles from the fluid.

13. A fluid processing system comprising: at least one fluid container, the at least one fluid container defining a fluid chamber in the at least one fluid container for containing a fluid and a plurality of magnetic particles; A magnetic assembly comprising a plurality of electromagnets disposed around a periphery of the fluid chamber, each of the plurality of electromagnets having: (i) a conductive coil positioned about a centerline extending in the z-direction, and (ii) a magnetic lens configured and arranged to direct the magnetic field generated by the conductive coil toward the fluid chamber, wherein at least one of the fluid chamber or the magnetic lens is movable along the z direction; as well as A control component coupled to the magnetic assembly, the control component being configured to: (i) controlling the magnetic field generated by each of the plurality of electromagnets to generate a plurality of magnetic field gradients within the fluid chamber sufficient to magnetically affect the plurality of magnetic particles within the fluid chamber, and (ii) controlling the movement of at least one of the fluid chamber or the magnetic lens along the z-direction while generating the plurality of magnetic field gradients within the fluid chamber.

14. The system according to claim 13, wherein: The control component is configured to move the magnetic lens relative to the conductive coil.

15. The system according to claim 13 or 14, wherein: The magnetic particles are paramagnetic or ferrimagnetic.

16. The system of claim 13 or 14, further comprising a permanent magnet that generates a magnetic field within the fluid chamber that attracts the magnetic particles to an inner surface of the fluid chamber.

17. The system of claim 16, further comprising at least one mechanical device for moving the magnetic assembly into a position adjacent the fluid chamber.

18. The system of claim 16, wherein: The magnetic assembly has at least one electromagnet of the plurality of electromagnets, the at least one electromagnet having a magnetic axis oriented along the z-direction.

19. The system according to claim 13 or 14, wherein: The control component is configured to control the magnetic field generated by each of the plurality of electromagnets via applying at least one electrical frequency waveform to each of the plurality of electromagnets.

20. The system of claim 19, wherein: The electrical frequency waveform includes an alternating waveform and a constant waveform.

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