High-throughput gravity-based multichannel separation assembly

Abstract

A high-throughput multichannel acoustic separator for separation and manipulation of microparticles, cells, or other small objects via gravity-based, siphon-mediated microfluidic flow and directed application of acoustic standing waves enabling efficient trapping of negative acoustic contrast particles (NACPs).

Description

[0001]HIGH-THROUGHPUT GRAVITY-BASED MULTICHANNEL SEPARATION ASSEMBLY CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of and priority to U.S. Provisional Application No. 63 / 664,140, filed June 25, 2024. The entire specification and figures of the above-referenced application are hereby incorporated, in their entirety by reference. STATEMENT OF GOVERNMENT INTEREST This invention was made with government support under grant number 1R21AI154266, awarded by the National Institutes of Health. The government has certain rights in the invention. TECHNICAL FIELD The present disclosure is directed to the field of acoustic separation of particles, and specifically a high-throughput device for the separation of analytes or cells using acoustic standing waves without the need for pumps or mechanical intervention. BACKGROUND Particles respond to acoustic radiation forces originating from an applied acoustic standing wave by being forced to specific locations along the wave, including the pressure node and pressure antinode. This relocation is dictated in part by the acoustic contrast factor, namely the positive or negative contrast factor, which originates from differences in density and elasticity between the particle and the surrounding media. For example, particles with positive contrast, such as incompressible positive acoustic contrast particles (PACPs) in aqueous media are generally transported to acoustic pressure nodes. On the other hand, compressible, negative acoustic contrast particles (NACPs) have a negative contrast property that is opposite to commonly used particles. Consequently, NACPs move to acoustic pressure anti-nodes when subjected to acoustic standing waves, which is a direction opposite from common, incompressible particles. These physical properties can make NACPs useful in techniques to rapidly separate and detect different types of cells and other analytes of interest from biological samples of interest in scientific research, the biotechnology industry, and healthcare. Notably, most biological particles, such as cells, in aqueous solutions appear to exhibit positive acoustic contrast. Moreover, functionalized NACPs have recently been developed to present biospecific ligands or other identification matrix on their surface. (Examples of fNACPs can be found in U.S. Patent Application No. 63 / 450,184, which is incorporated by reference herein). Exploiting these properties, acoustic radiation forces can be used to trap elastomeric NACPs, and any biological or chemical analytes captured or engaged therewith, or separate them from many biological components within aqueous or unmodified biological samples. As such, there exists a need for a simple, high-throughput, and inexpensive point-of-care system to separate particles, such as NACPs from biological or environmental samples. SUMMARY OF THE INVENTION The present disclosure describes systems, methods, and apparatus for the high-throughput acoustic separation of particles of interest in a fluid sample. In a preferred aspect, the acoustic separation is accomplished via siphon-mediated fluid flow and directed application of acoustic standing waves enabling efficient trapping of negative acoustic contrast particles (NACPs) within a capillary channel. In one preferred aspect, the disclosure describes a high-throughput acoustic separation assembly, the assembly including a siphon chamber array having a plurality of siphon chambers each adapted to receive a sample, and a trapping array securing a plurality of capillary channels, wherein each channel is in fluid communication with at least one of the siphon chambers through a transfer tube forming a plurality of fluid pathways, each adapted to generate a siphon effect. In this aspect, the siphon effect includes a system whereby a pressure differential created between each of the channel inlets and the channel outlets of the fluid pathways when the samples are added to the siphon chambers causing the samples to flow through the fluid pathways until an equilibrium point is reached, typically above the channel inlet. In another aspect the disclosure describes a high-throughput acoustic separation assembly, the assembly comprising one or more acoustic wave generator responsive each of the capillary channels, and further configured to generate a standing acoustic wave within each of the capillary channels. In this aspect, NACPs present in the same, that may preferably be functionalized to bind to one or more analytes of interest, are trapped at the antinode position and isolated from the sample fluid and non-acoustically reactive particles which flow and exit the capillary channel. The trapped NACPs can be further washed or contacted with additional solutions by the addition of wash buffer or other liquids to the siphon chambers. The trapped NACPs can be released from the capillary channel through the disengagement of the acoustic wave generator, and can be further collected through one or more washing steps performed using a buffer. Notably, the additional solutions can be added with the sample, or sequentially with the acoustic generator engaged or non-engaged depending on the desired application. In alternative embodiments, the solutions can be introduced to the NACPs that have been isolated and collected, for example in a wash buffer, by the systems and process described herein. In another aspect the disclosure describes a high-throughput magnetic separation assembly, the assembly comprising one or more engageable magnets capable of interfacing with each of the capillary channels, and further configured to generate a magnetic field within each of the capillary channels. In this aspect, magnetic particles present in the same, that may preferably be functionalized to bind to one or more analytes of interest, are magnetically trapped and isolated from the sample fluid and non-magnetic particles which flow and exit the capillary channel. The trapped NACPs can be released from the capillary channel through the disengagement of the magnet, and can be further collected through one or more washing steps performed using a buffer. Additional aspects of the disclosure will be evident from the specification, drawings and claims provided below. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1A-E. Multichannel acoustic separator and fNACP-based assay overview. (A) Schematic representation of fNACPs being trapped along the walls of a channel at the antinodes of an acoustic standing wave for purification from blood samples. (B) Schematic overview of the multichannel acoustic separator and the purification of fNACPs from whole blood in a 96-well plate format. (C) Bright-field microscopy image of fNACPs mixed with whole blood prior to purification using the multichannel acoustic separator. (D) Bright-field microscopy image of fNACPs after purification from whole blood using the multichannel acoustic separator. (E) Schematic representation of the workflow for fNACP-based biomarker detection. In step 1, the fluorescently barcoded fNACPs are mixed with whole-blood samples in individual wells of a 96- well plate for biomarker capture. In step 2, the fNACPs are purified in the multichannel acoustic separator and ejected into a clean well plate. In step 3, captured biomarkers on the fNACPs are fluorescently labeled within the 96-well plate. In step 4, the resultant fluorescence of the fNACPs is analyzed. Figure 2A-D. Overview of the multichannel acoustic separator design and function. (A) Image and details of the assembled multichannel acoustic separator. (B) Rendered images of the multichannel acoustic separator. (C) Schematic representation of the upper siphon chamber upon the addition (left) and after draining (right) of the sample. (D) Schematic representation of the fNACP purification procedure within a glass capillary of the trapping channel array. Figure 3A-H. Characterization of NACP washing and trapping performance in the multichannel acoustic separator. (A) Images of the multichannel acoustic separator at various stages of processing a blood sample. (B) Absorbance of drained samples upon consecutive washes with buffer after initially processing whole blood. N = 12; error bars indicate standard deviation (SD). (C) Estimated flow rate and volume per channel over time for water, buffer, and blood samples in the multichannel acoustic separator. Flow rate was estimated by linear fits of the first 5 s of each plot. Shaded regions indicate SD. (D) Fluorescence microscopy images of fluorescent NACPs and PS particles before and after engaging acoustics under an applied flow rate of 1 mL min−1. (E) Effective recovery of NACPs from water in each of the 12 channels of the separator two washes. n = 3; error bars indicate SD. (F) Effective recovery of NACPs from 3-fold- diluted whole porcine blood from each of the 12 channels of the separator after four washes. n = 3; error bars indicate SD. (G) Mean NACP fraction recovered during the initial addition of sample, first and second washes, collection of NACPs, and first and second rinses. N = 12; error bars indicate SD. (H) Bright-field microscopy images of NACPs before and after purification from 3- fold-diluted whole porcine blood via the multichannel acoustic separator. Figure 4A-F. Performance of the fNACP-based assay. (A) Schematic representation of the functionalization approach used to make biomarker-specific fNACPs. (B) Schematic representation of the BREs, biomarkers, and labels used to produce the three types of fNACPs used in this study. (C) Representative flow cytometry gating used to identify fluorescent SA- barcoded fNACPs. (D–F) fNACP fluorescence curves for the detection of anti-OVA, IgM, and IgA, respectively. n = 3; error bars indicate SD. Figure 5A-E. Multiplexed detection of anti-OVA, IgM, and IgA and detection of IgA from whole blood. (A) Representative fluorescence distributions of fNACPs incubated without or with anti-OVA, IgM, and IgA model biomarkers present. (B) Heatmap of particle fluorescence for all 8 multiplexing conditions evaluated. (C) Individual and combined fluorescence microscopy images of fNACPs after performing an assay on a sample containing anti-OVA, IgM, and IgA at the concentrations shown in the last row of (B). (D) Schematic representation of the workflow used to process a whole-blood-based fNACP assay using the multichannel acoustic separator. (E) fNACP fluorescence curve for the detection of IgA from whole blood. n = 3; error bars indicate SD. Figure 6. Absorbance scan of blood and wash buffer. Absorbance of ~40-times diluted whole blood and wash buffer over the range of 300-800 nm. Figure 7. Device washing characterization. Absorbance readings of samples collected after washing blue 1-dyed water with undyed deionized water. Figure 8A-B. NACP characterization. A) Representative microscopy image of NACPs after filtration, prior to functionalization. B) Histogram of NACP sizes obtained by analysis of captured microscopy images. N = 3357. Figure 9A-E: (a) shows a front perspective view of an acoustic separation assembly in one embodiment thereof; (b) shows a front view of an acoustic separation assembly in one embodiment thereof; (c) shows a top view of an acoustic separation assembly in one embodiment thereof; (d) shows a back view of an acoustic separation assembly in one embodiment thereof; (e) shows a front perspective view of an acoustic separation assembly in one embodiment thereof. Figure 10: shows a front perspective view of an acoustic separation assembly having a series of capillary channels individually coupled with a siphon tube that is in fluid communication with a siphon chamber in one embodiment thereof. Figure 11A-B. (A) shows a cross-sectional schematic view of the siphon-driven fluid pathway of an acoustic separation assembly in one embodiment thereof; (B) shows a schematic view of a capillary channel wherein NACPs (circles) are trapped along the antinodes at the walls of the channel and PACPs (squares) are focused at the nodes at the center of the channel. DETAILED DESCRIPTION OF THE INVENTION The following detailed description is provided to aid those skilled in the art practicing the various embodiments of the present disclosure, including all the methods, uses, compositions, etc., described herein. Even so, the following detailed description should not be construed to unduly limit the present disclosure, as modifications and variations in the embodiments herein discussed may be made by those of ordinary skill in the art without departing from the spirit or scope of the present discoveries. The present disclosure is explained in greater detail below. This disclosure is not intended to be a detailed catalog of all the different ways in which embodiments of this disclosure can be implemented, or all the features that can be added to the instant embodiments. For example, features illustrated with respect to one embodiment may be incorporated into other embodiments, and features illustrated with respect to a particular embodiment may be deleted from that embodiment. In addition, numerous variations and additions to the various embodiments suggested herein will be apparent to those skilled in the art in light of the instant disclosure, which variations and additions do not depart from the scope of the instant disclosure. Hence, the following specification is intended to illustrate some particular embodiments of the disclosure, and not to exhaustively specify all permutations, combinations, and variations thereof. Provided herein includes a multi-channel acoustic separation assembly (100), sometimes referred to as a siphon-driven multichannel acoustic separator, for separation and manipulation of microparticles, cells, or other small objects via gravity-based, siphon-mediated microfluidic flow and directed application of acoustic standing waves enabling efficient trapping of NACPs. The multi-channel acoustic separation assembly (100) can include a siphon chamber array (104) having a plurality of siphon chambers (106) each adapted to receive a sample (150). As shown in Figure 9, the siphon chamber array (104) can be secured to array frame (102) so as to be positioned approximately vertically. In additional embodiments, the siphon chamber array (104) can be secured to array frame (102) through a height adjustor (118) configured to allow the siphon chamber array (104) to be adjusted vertically or rotationally as detailed below. In the embodiment shown in Figure 10, the height adjustor (118) of the disclosure can include a slide aperture and a fastener that is configured to be inserted through the aperture and secure the siphon chamber array (104) at a desired height. As shown in Figure 11, each siphon chamber (106) of the siphon chamber array (104) can preferably include a vessel configured to receive a sample (150), such as a biological fluid, or environmental sample. In a preferred embodiment, each siphon chamber (106) of the array (104) can be formed as an integral vessel formed by a shaped cavity within the siphon chamber array (104). As noted above, the size, position, number and height of the siphon chambers (106) of the disclosure can be vary depending on the desired application and use for the multi-channel device. Moreover, during use, all of the siphon chambers (106) of the siphon chamber array (104) can be operated simultaneously, while in alternative embodiments one or a plurality of siphon chambers (106) of the siphon chamber array (104) can be operated. Notably, the siphon chambers (106) are not in fluid communication with one another, such that each can receive a different fluid sample (150) that may contain a distinct analyte of interest and / or NACPs. Notably, the siphon chamber array (104) and array frame (102) can be formed from any suitable material, such as plastic or thermoplastic and the like. In a preferred embodiment, the siphon chamber array (104) and / or array frame (102) can be injection molded, or even more preferably generated through additive 3-D printing processes. In certain embodiments, the siphon chamber array (104) and array frame (102) are separable components that can be secured to one another, while in alternative embodiments, they are a single integral component. The multi-channel acoustic separation assembly (100) further includes a trapping array (114) securing a plurality of capillary channels (112). The trapping array can be positioned directly adjacent, and preferably below the siphon chamber array (104). Again, the trapping array (114) can be formed from any suitable material, such as plastic or thermoplastic and the like, and is preferably generated via additive 3-D printing processes. As shown in Figure 10-11, each of the capillary channels (112) secured by the trapping array (114) is configured to be in fluid communication with at least one siphon chamber (106) through a transfer tube (108). Each capillary channel (112) and corresponding transfer tube (108) form a fluid pathway (A). As shown in Figure 11, an exemplary fluid pathway (A) includes a channel inlet (140) positioned at the terminal portion of a siphon chamber (106). In this embodiment, the terminal portion of a siphon chamber (106) forms a sample reservoir (144) having an angled surface generating a terminal point where the channel outlet (140) is positioned. The transfer tube (108) extends upward from the siphon chamber (106) and is coupled with a capillary channel (112) through a joint (110). The terminal portion of the capillary channel (112) includes a channel outlet (142). Notably, the siphon chamber (106) and channel inlet (140) are generally positioned higher than the channel outlet (144) of the capillary channel (112). In the above configuration, the components of the fluid pathway (A) generate a siphon effect where a pressure differential created between the channel inlet (140) and the channel outlet (142) of the fluid pathways (A) when the sample (150) is introduced to the siphon chamber (106). This siphon effect causes the sample (150) to flow through the fluid pathway (A) until an equilibrium point (138) is reached. In one embodiment, the siphon effect of the fluid pathway (A) can be generated by compression the transfer tube (108), which can help initiate the siphon effect, which when coupled with gravity allows the continuous flow of the sample (150) through the fluid pathway (A) until an equilibrium point is reached. The transfer tube (108) can facilitate transfer of the sample (150) from the siphon channel (106) via the siphon effect as described herein. In one embodiment of the disclosure the transfer tube (108) can be formed from a compressible material, such as silicone and the like. In this embodiment, the compression of the tube can be done manually by hand through pressing down on the transfer tube (108). In an alternative embodiment, a compression bar (not shown) can be positioned on top of the siphon chamber array (104) and above the transfer tubes (108). This compression bar can include a joint that allows it to transit downward and apply a compression force to the transfer tubes (108) thereby assisting the siphon effect. Finally, in still further embodiments, the transfer tubes (108) can be positioned within a slot (136) positioned along the top of the siphon chamber array (104) and adjacent to each siphon chamber (106). In this configuration, a transfer tube (108) exiting the siphon chamber (106) can be positioned with the slot (136) which is configured to compress the transfer tube (108) thereby assisting in the generation of the siphon effect of the fluid pathway (A). The multi-channel acoustic separation assembly (100) of the disclosure further includes one or more acoustic wave generators (116). In a preferred embodiment, a trapping array (114) secures one or more acoustic wave generator (116) adjacent to the capillary channels (112). In this configuration, when engaged, the acoustic wave generator(s) (116) generate a standing acoustic wave within each of the capillary channels (112). The acoustic wave generator (116) of the disclosure can include a plurality of piezoelectric transducers, responsive to a power source, for example through an electrical conduit, and configured to generate a standing acoustic wave within each of the capillary channels (112). In a preferred embodiment, engagement of the piezoelectric transducer at approximately 750 kHz leads to the establishment of a half-wavelength standing acoustic wave within each of the capillary channels (112), yielding a node along the centerline of the channels and antinodes along the walls of the channels. In this embodiment, the capillary channels (112) include glass capillaries, however any suitable material can be used within the context of this disclosure. As noted above, the standing acoustic wave generates a node (152) along the centerline of each of the capillary channels (112) and an antinode (154) along the walls of each of the capillary channels (112). As the sample flows through the capillary channel (112), NACPs (146) are relocated to the position of the antinode (154) positioned along the wall of the channel (112). These trapped NACPs (146) can be maintained at their position as long as the standing acoustic wave propagated by the acoustic wave generator (116) is maintained. Non-acoustically responsive particles, or positive acoustic contracting particles )PACPs) (148), such as cell or other biological materials align along the center (in the case of PACPs, at the nodes) and are allowed to flow through the capillary channel (112) an exit via the channel outlet (142) and can be collected in a container, such as a well plate (122) positioned within a plate receiver (120), to be discarded or further processed. In a preferred embodiment, the trapped NACPs (146) include functionalized NACPs, or fNACPs configured to separate one or more analytes of interest from the sample (150), for example through a ligand binding position presented on the surface of the fNACP that correspond with a target analyte. As noted elsewhere, an analyte of interest can include, but not be limited to: an ion, small molecule, antigen, protein, nucleic acid, carbohydrate, virus, cell or biomarker. Notably, the fNACPs can be coded for multiplex detection. example, of such coding include bar coding as well as color coded for detection. Various analytical techniques may be used to detect the NACPs (146) and / or the analyte of interest including, for example, but not limited to fluorometric analysis, colorimetric analysis, and image analysis. Once the sample (150) has passed through the capillary channel (112), the acoustic wave generator (116) can be disengaged thereby removing the standing acoustic wave within each channel (112). In this manner, the NACPs (146) and any bound analyte can now pass through the capillary channel (112) through the channel outlet (142) and collected for further processing or analysis. In a preferred embodiment, a buffer or other solution can be passed through the fluid pathway(s) (A) as described above without the presence of the standing acoustic wave thereby assisting in the removal of the NACPs (146) and any bound analytes from the capillary channel. In a preferred embodiment shown in Figure 10, a well plate (122) having a plurality of well positions can be positioned within a plate receiver (120) below the trapping array (114). In this configuration, each capillary channel (112) is positioned directly above a well allowing the fluid sample to be deposited into an individual well position. The well plate (122) can be slidably coupled with the plate receiver (12) allowing it to be horizontally adjusted or locked into place when collecting a processed sample. The collected sample can be discarded, or save for further processing., such as the introduction of one or more additional solutions having a reagent capable of binding to the analyte of interest or the NACPs, or a reagent capable of identifying to the analyte of interest, or the NACPs. In certain embodiments, the reagent of the disclosure can be selected from: antibodies, receptors, ligands, nucleic acids, aptamers, synthetic host molecules, enzymes, stains, dyes, or chromophores. As noted above, the number of fluid pathways is variable. As such, in certain embodiment the disclosure includes an acoustic separation device having a single fluid pathway having a siphon chamber (106) adapted to receive a sample (150), a transfer tube (108) in fluid communication with the siphon chamber (106) and a capillary channel (112) forming a fluid pathway (A) adapted to generate a siphon effect, and an acoustic wave generator in communication with the capillary channel. The operation of the above elements is generally consistent with the principals described above. In still further embodiments, the disclosure provides for a magnetic separation device. In this embodiment, the device includes one or more siphon chambers (106) each adapted to receive a sample (150) having one or more magnetic particles of interest. The device can further include a transfer tube (108) in fluid communication with each of the siphon chamber (106) and a capillary channel (112) forming a fluid pathway (A) adapted to generate a siphon effect, wherein the siphon effect comprises a pressure differential created between a channel inlet (140) and a channel outlet (142) of the fluid pathway (A) when the sample (150) is added to the siphon chamber (106) causing the sample (150) to flow through the fluid pathway (A) until an equilibrium point is reached. The device can include an engageable magnet (not shown) positioned adjacent to the capillary channel(s) (112), wherein the magnet exerts a magnetic force on the sample flowing through the capillary channel (112) thereby trapping magnetic particles in the sample within the capillary channel (112). The magnet of the disclosure can be engaged manually by being brought into proximity with the capillary channel (112), or via a power source coupled to an electromagnet capable of generating a directed magnetic field within the capillary channel (112). Similar to the prior disclosure, the trapped magnetic particles can be released through disengagement of the magnet, and further capture, preferably through the addition of one or more buffer washes as generally described herein. The above detailed description is provided to aid those skilled in the art in practicing the various embodiments of the present disclosure, including all the methods, uses, compositions, etc., described herein. Even so, the detailed description should not be construed to unduly limit the present disclosure, as modifications and variations in the embodiments herein discussed may be made by those of ordinary skill in the art without departing from the spirit or scope of the present discoveries. This disclosure is not intended to be a detailed catalog of all the different ways in which embodiments of this disclosure can be implemented, or all the features that can be added to the instant embodiments. For example, features illustrated with respect to one embodiment may be incorporated into other embodiments, and features illustrated with respect to a particular embodiment may be deleted from that embodiment. In addition, numerous variations and additions to the various embodiments suggested herein will be apparent to those skilled in the art in light of the instant disclosure, which variations and additions do not depart from the scope of the instant disclosure. Hence, the following specification is intended to illustrate some particular embodiments of the disclosure, and not to exhaustively specify all permutations, combinations, and variations thereof. Unless defined otherwise, all scientific and technical terms are to be understood as having the same meaning as commonly used in the art to which they pertain. As used herein, a particle that “display negative acoustic contrast” can also be referred to as a negative acoustic contrast particle (NACP). NACPs move to acoustic pressure antinodes when subjected to acoustic standing waves, which is a direction opposite from common, incompressible particles, such as cells in a complex biological sample. When acoustic radiation forces immobilize NACPs along the pressure antinodes, they are herein referred to as “acoustic trapped.” In a preferred embodiment, the functionalized non-spherical particles of the invention display negative acoustic contrast. In some embodiment, the acoustic separation assembly of the disclosure can be used to isolated functional NACPs (fNACPs). As used herein, fNACPs are NACPs having functionalized surface containing a biospecific ligand or other identification matrix. Examples of fNACPs can be found in U.S. Patent Application No. 63 / 450,184, which is incorporated by reference herein. As used herein, “siphon” or “siphon effect,” refers to conduit that uses the weight of a liquid to pull the liquid from the higher level to a lower level. More specifically, in one aspect a siphon of the disclosure refers to a tubular member having inlet and discharge ends that allows fluid to drain from a fluid reservoir through an intermediate point that is higher than the fluid reservoir with flow driven by a difference in hydrostatic pressure at the opposite ends of the tubular member. Depending on the siphon, a small pressure differential can cause flow through the siphon. A magnitude of pressure differential can make a difference in the flow rate through the siphon. The geometric cross-section shape of the tubular member is not limited, and can be any shape, such as circular, oval, square, and so forth. As used herein, “sample” includes a quantity of fluid containing a target analyte. A sample can preferably include a fluid sample having an analyte of interest. A Sample of the disclosure can include a biological as well as an environmental sample. As further used herein, the term “biological sample” includes a sample from any bodily fluid or tissue (e.g., serum, plasma, blood, cerebrospinal fluid, urine, saliva, cancer tissue, healthy tissue), preferably from an animal, and more preferably from a mammal, and even more preferably from a human subject. A biological sample thus can relate to any sub-portion of a body fluid removed from a subject prior to applying the body fluid sample to a test element. Samples can be obtained by well-known techniques including, for example, venous or arterial puncture, epidermal puncture, and the like. As used herein, “analyte” means a chemical compound present in a sample, and preferably a biological sample such as a body fluid. The analyte can be a small molecule (e.g., the analyte is not a biological macromolecule). In some instances, the analyte is an organic molecule or inorganic. Alternatively, an analyte can be a low molecular weight chemical compound such as, for example, a chemical compound with a molecular mass of less than 1000 u (1000 Da; 1.66×10−24kg). As used herein, a composition is referred to as “isolated” or “separated” when it has been separated from at least one component with which it is naturally associated. For example, a functionalized non-spherical particle can be considered isolated if it is separated from secondary components present in a sample, such as a biological sample, including cells, organelles, polysaccharides, lipids, polypeptides, polynucleotides, and other metabolites. Standard quantification methodologies known in the art can be employed to obtain and isolate the molecules of the invention. As used herein, a biological marker (e.g., a “biomarker”, a “marker”, or an “analyte”) is a characteristic that is objectively measured and evaluated as an indicator of normal biologic processes, pathogenic processes, or pharmacological responses to therapeutic interventions, consistent with NIH Biomarker Definitions Working Group (1998). Markers can also include patterns or ensembles of characteristics indicative of particular biological processes. Biomarker measurement can increase or decrease to indicate a particular biological event or process. In addition, if the biomarker measurement typically changes in the absence of a particular biological process, a constant measurement can indicate occurrence of that process. The present invention also encompasses recognition domains or elements, the two being generally interchangeable, which specifically bind the target molecules. As used herein, the term “binding” refers to the interaction between binding pairs (e.g., an antibody and an antigen or aptamer and its target). In other embodiments, the phrase “binds” refers to the specific binding of one protein to another (e.g., an antibody, fragment thereof, or binding partner to an antigen), wherein the level of binding, as measured by any standard assay (e.g., an immunoassay), is statistically significantly higher than the background control for the assay. For example, when performing an immunoassay, controls typically include a reaction well / tube that contains antibody or antigen binding fragments alone (e.g., in the absence of antigen), wherein an amount of reactivity (e.g., non-specific binding to the well) by the antibody or antigen binding fragments thereof in the absence of the antigen is considered to be background. Binding can be measured using a variety of methods standard in the art including enzyme immunoassays (e.g., ELISA, immunoblot assays, etc.). The molecules that may bind to one or more of the targets include antibodies, aptamers and antibody derivatives or fragments. As used herein, the term “antibody” refers to an immunoglobulin molecule capable of binding an epitope present on an antigen. The term is intended to encompass not only intact immunoglobulin molecules such as monoclonal and polyclonal antibodies, but also bi-specific antibodies, humanized antibodies, chimeric antibodies, anti-idiopathic (anti-ID) antibodies, single-chain antibodies, Fab fragments, F(ab') fragments, fusion proteins and any modifications of the foregoing that comprise an antigen recognition site of the required specificity. As used herein the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a compound” includes a plurality of such compounds, and reference to “the method” includes reference to one or more methods, method steps, and equivalents thereof known to those skilled in the art, and so forth. Similarly, the word “or” is intended to include “and” unless the context clearly indicates otherwise. Hence “comprising A or B” means including A, or B, or A and B. Furthermore, the use of the term “including”, as well as other related forms, such as “includes” and “included”, is not limiting. The term “about” as used herein is a flexible word with a meaning similar to “approximately” or “nearly”. The term “about” indicates that exactitude is not claimed, but rather a contemplated variation. Thus, as used herein, the term “about” means within 1 or 2 standard deviations from the specifically recited value, or ± a range of up to 20%, up to 15%, up to 10%, up to 5%, or up to 4%, 3%, 2%, or 1 % compared to the specifically recited value. The invention now being generally described will be more readily understood by reference to the following examples, which are included merely for the purposes of illustration of certain aspects of the embodiments of the present invention. The examples are not intended to limit the invention, as one of skill in the art would recognize from the above teachings and the following examples that other techniques and methods can satisfy the claims and can be employed without departing from the scope of the claimed invention. Indeed, while this invention has been particularly shown and described with references to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims. EXAMPLES Example 1: Operational Validation of Acoustic Separation System. The siphon-driven multichannel acoustic separator (100) preferably comprises a plurality of acoustic trapping channels, also referred to as capillary channels (112), for the parallel separation, washing, and collection of multiple samples in under 5 min. Fluid actuation through the channels is mediated by siphons, eliminating the need for automated pumps or plungers. Each of the channels supports a lateral half-wavelength bulk acoustic standing wave, yielding pressure nodes at the center of each channel and antinodes along the channel walls. The fNACPs are designed to specifically capture biomolecule targets from heterogeneous fluid samples, like whole blood. After capture, the fNACP samples are passed through the trapping channels of the device. Due to their negative acoustic contrast, the fNACPs strongly immobilize along the walls of the channel, whereas other objects, such as red and white blood cells, focus to the center of the channels and leave the channels as waste. After washing the fNACPs, the acoustics can be disengaged to collect the purified fNACPs in a well plate (Figures 1A and 1B). The biomolecules on the surfaces of fNACPs are then fluorescently labeled for downstream biomolecule quantification by fluorescence measurement. Applicants show that the acoustic separator can purify fNACPs from whole-blood samples (Figures 1C and 1D). Applicants additionally showed the simultaneous (e.g., multiplexed) isolation and detection of three biomolecules (e.g., IgA, IgM, and anti-ovalbumin IgG) by fluorescently barcoded fNACPs. Finally, Applicants showed rapid isolation and detection (e.g., in < 70 min) of IgA from whole blood using the multichannel acoustic separator (Figure 1E). By eliminating the need for pumps and enabling the rapid and simultaneous separation of multiple samples, the multichannel acoustic separator represents one of the fastest, simplest, and most accessible implementations of acoustofluidic separation technologies to date. Example 2: 3D-printed multichannel acoustic separator enables acoustic trapping. To develop a multichannel acoustic separator that is easily integrated into existing laboratory workflows, in one exemplary embodiment Applicants designed the device to interface with 96-well plates and 12-channel multichannel pipettes for the simultaneous processing of 12 samples. In this embodiment, separator body, also referred to herein as an array frame (102) consists of 5 printed parts: the upper siphon chamber array (104), a trapping channel array holder, also referred to herein as a trapping array (14) a left and right frame, and an electrical port cover (Figures 2A, 2B). In the embodiment shown in the figures, the trapping array consists of 12 square glass capillaries, also referred to as capillary channels (112) that preferably have 1 × 1 mm inner dimensions and are approximately 25 mm long, all affixed to a large, rectangular piezoelectric transducer, also referred to as acoustic wave generator (116), preferably possessing a resonance frequency of ∼750 kHz. Two three-dimensionally (3D)-printed transducer clips hold the transducer in position during use. Each capillary is connected to a corresponding upper siphon chamber by a compressible tube (108), which preferably comprises short segments of compressible silicone tubing. Electrical leads for transducer operation are routed through the device to the electrical ports, which accommodate banana plugs for operation. When liquid samples are present in the channels (112) of the separator (100), actuation of the piezoelectric transducer by application of a ∼750 kHz alternating current (AC) sine wave results in establishment of half-wavelength standing pressure waves within each channel (112). Because of this, acoustic radiation forces are exerted on objects smaller than the acoustic wavelength within each channel. The axial component of the primary acoustic radiation force (e.g., acting perpendicularly to the length of the channel) pushes objects toward either the node, located along the centerline of the channel, or the antinodes of the standing wave, located at the walls of the channel (Figure 1A). The direction of this force is dependent on the acoustic contrast factor of the objects, which can be positive or negative and describes the density and compressibility of the objects relative to that of the liquid in which they are suspended. Positive acoustic contrast particles (PACPs) are forced to the nodes of the standing wave, whereas negative acoustic contrast particles (NACPs) are forced to the antinodes. Most cells and commercial polymeric particles exhibit positive acoustic contrast when suspended in water or blood. Here, to enable rapid separation from whole-blood samples, Applicants designed the fNACPs from an elastomeric polymer to have strong negative acoustic contrast. Importantly, the standing waves also exert primary acoustic radiation forces in the lateral direction (i.e., parallel to the length of the channel), as well as secondary acoustic radiation forces, which are attractive between particles at small distances. Because the fNACPs are forced to the channel walls, where flow rate is low relative to the center of the channel, and because of the secondary and lateral primary acoustic radiation forces, fNACPs can be strongly trapped against the channel walls at high flow rates, while blood components are washed from the channels (Figures 1C and 1D). Such trapping of NACPs results in the ability to conduct acoustofluidic separation and purification at flow rates much higher than those of traditional microfluidic systems, enabling the implementation of microfluidic siphons. Example 3: Fluid actuation driven by siphon. Large-scale siphons with a non-submerged outlet at a height below the inlet will drain fluid from the upper chamber until it reaches the height of the inlet, resulting in the introduction of air to the siphon and an inability to restart the siphon without repriming. However, the miniaturization of this siphon allows for the siphon stopping point to occur above the inlet before the complete draining of the chamber (Figure 2C). Because the liquid in each channel exits the channels as discrete droplets, the pressure driving fluid flow must overcome the Laplace pressure associated with the liquid-air interface of the droplet at the bottom of each capillary, which is proportional to the surface tension of the liquid. By tuning the height of the channel outlet relative to the channel inlet (ΔH in Figure 2C), the Laplace pressure can be balanced with the hydrostatic pressure in the upper siphon chamber to arrest fluid flow before the height of the fluid in the upper siphon chamber reaches that of the inlet. To minimize the volume of fluid retained at the siphon stopping point, , Applicants designed the upper siphon chambers to taper toward the tubing inlet at the bottom of the chamber. Upon the addition of fluid to the upper chamber, the siphon effect is recovered, and the fluid again drains until reaching the equilibrium height of the liquid. To confirm that the Laplace pressure at the outlet is the primary mechanism responsible for arresting the siphon effect prior to complete draining, Applicants tuned the ΔH of the separator such that the siphon stopping point was above the inlet when liquid was added to the upper siphon chamber, as described previously. Applicants then submerged the end of the capillary in water to remove the liquid-air interface, and, as a result, the upper chamber completely drained and introduced air to the channel. Because the microfluidic siphon effect in this device is semi-continuous, meaning it only requires one priming step, washing samples or transferring them to new fluids is simplified (Figure 2D). For example, after priming a channel, an initial sample of fNACPs in whole blood can be added to the upper siphon chamber. As the sample is drawn through the channel via siphon, fNACPs are trapped within the capillary array. Upon cessation of flow at the siphon stopping point, wash buffer or other fluids can be added to the upper siphon chamber to expel blood components into a waste container positioned below the device. This process can be repeated until the fNACP samples are sufficiently clean. The acoustics can then be disengaged, and additional fluid can be added to the upper siphon chamber to release the fNACPs, which are collected in a well plate positioned below the device. While the trapping and fluid transfer process was used to wash fNACPs in this example, it could also be used to accomplish other tasks, such as transferring particles into labeling solutions or functionalizing particles (e.g., through layer-by-layer coatings). Example 4: Siphon-based flow for washing blood from channels. To assess the ability of the device to purify NACPs from whole blood, Applicants adjusted ΔH such that the equilibrium liquid level was minimized so as to be below the sloped section of the chamber but above the channel inlet when blood was present in the device and the channels were primed. Applicants added 300 μL of 3-fold-diluted porcine blood to the upper siphon chambers and allowed the channels to reach equilibrium (Figure 3A). Applicants then washed the channels by the repeated addition of wash buffer to the siphon chambers and collected the fluid leaving the channel outlets. Applicants diluted the collected samples by 13.3-fold to measure the absorbance of the samples at 542 nm, an absorbance that is characteristic of whole blood (Figure 6). Because samples collected from the first wash were highly concentrated, absorbance values were saturated; thus, only the absorbance values of subsequent washes are reported (Figure 3B). Applicants repeated this process with blue dye samples to quantify the performance of the initial wash (Figure 7). Overall, Applicants found that the fluid remaining in the channels was sufficiently devoid of blood components after 4 washes. The absorbance of the collected was only 3.2% ± 0.3% of that collected after the second wash, whereas the absorbance of the buffer control was 2.1% ± 0.3% of that collected after the second wash. Therefore, the channel was washed 4 times before collecting fNACPs for all subsequent studies. Example 5: Siphon-based flow for processing disparate fluid types. Blood and buffer differ in both surface tension and viscosity, resulting in different behaviors when actuated by the siphon. To characterize these differences and ensure the separator can process disparate types of fluid, ΔH was manually adjusted until the fluid equilibrium height in all siphon chambers was minimized for water, buffer, and blood. Because the surface tension of water is greater than that of blood (e.g., ∼71 versus ∼51 mN m−1), the Laplace pressure at the outlet is more substantial for water samples, resulting in a minimization of the fluid equilibrium height at larger ΔH values (∼14 mm for water and buffer versus ∼11 mm for blood). Applicants observed all three fluids draining by siphon after adding 300 μL of each fluid to the upper siphon chambers with the acoustics engaged. The chambers were considered fully drained when the dripping of fluid in all channels had ceased for at least 10 s. Upon analysis, Applicants plotted the average volume drained from each channel over time (Figure 3C). It should be noted that the flow is not continuous but pulsatile, with the periods between droplets increasing as the fluid level in the upper chamber, and therefore the hydrostatic pressure, drops. Applicants found that water was drained by the siphons more quickly (∼17 s) than buffer (∼26 s) and blood (∼79 s). One potential limitation of acoustofluidic systems is that high flow rates can diminish the ability to trap NACPs, resulting in a loss of captured biomolecules. Previously, Applicants showed that large NACPs (24.3 μm) are trapped at the pressure antinodes and retained in the presence of flows up to 5.0 mL min−1at efficiencies >95%.24The maximum flow rates in the multichannel acoustic separator were found to be 1.6 mL min−1for water, 1.3 mL min−1for buffer, and 0.9 mL min−1for blood, indicating that the flow rates driven by the siphon in this device are within the expected regime to enable effective trapping of NACPs. Flow rates were estimated by finding the slope of linear fits of the recorded volume per time plots to the first 5 s (Figure3C). Example 6: NACP trapping in the multichannel acoustic separator. To produce NACPs of low size polydispersity, Applicants followed a previously described protocol known in the art. In short, NACPs were prepared by homogenizing a polydimethylsiloxane (PDMS) precursor with 0.1% v / w triethoxyvinylsilane (TEOVS) submerged in water with 1.0% Pluronic F-108 surfactant. After curing the homogenized droplets, Applicants filtered the mixture to isolate a low polydispersity fraction of NACPs. Resultant NACPs had a mean diameter of 23.0 μm and a coefficient of variance of 11.8%. To evaluate the isolation of NACPs from PACPs within the separator device, Applicants first mixed NACPs coated with fluorescent protein in wash buffer at ∼1.7 × 105particles mL−1. To this mixture, Applicants added ∼5 μL of a 5 wt % solution of 5.1 μm fluorescent polystyrene (PS) particles as a PACP model. Applicants passed the particle solution through a trapping channel situated on a fluorescent microscope at 1 mL min−1via syringe pump. Upon engaging the acoustics, the NACPs were rapidly trapped against the walls of the channel, while the PS PACPs continued through the channel without being trapped (Figure 3D). To quantitatively evaluate the trapping of NACPs, Applicants studied the trapping efficiency of NACPs from both water and blood (Figures 3E and 3F). In biomolecule detection experiments, fNACPs are collected in a single step upon the addition of 300 μL of wash buffer after the acoustics are disengaged. To quantify the fraction of total NACPs collected during this step and determine the percentage lost during the wash steps, Applicants added 5 × 104NACPs to 300 μL samples of water and blood. Applicants used the multichannel acoustic separator to trap, wash, and collect the NACPs. The samples in water were washed twice, whereas the samples in blood were washed four times, whereby a wash constitutes flowing 300 μL of wash buffer through each channel while the NACPs are trapped. After NACP collection, Applicants washed the channels two additional times to flush any residual NACPs from the channels. Applicants collected the fluid leaving the channels during each step (i.e., the initial addition of sample, the individual washes, the collection, and the final rinses). Applicants enumerated the NACPs via flow cytometry and found that the effective NACP recovery was >90% from both water and blood samples. Applicants determined that the initial addition of the sample was the step at which the majority of particle loss occurred (Figure 3G). Applicants speculate that this is likely due to the absence of already trapped NACPs in the channels at that point. Since these initial particles would not be attracted to particles already trapped by secondary acoustic radiation forces, they are more prone to passing through the device. Consequently, this loss could be mitigated using trapping channels pre-seeded with NACPs along the channel walls during the initial step, though implementation of this trapping channel feature is left to future work. Upon analysis of samples prior to and after purification via microscopy, Applicants found that the purification method was successful in removing the vast majority of contaminants (e.g., red and white blood cells) from the samples (Figure 3H). Example 7: NACP functionalization for tunable biomarker specificity. The detection and enumeration of multiple antibodies and antibody isotypes in a clinical setting can be informative for assessing the state of, and immunological response to, infectious diseases in patients. Multiplexed assays, which simultaneously detect multiple biomarkers in a single test, can significantly enhance diagnostic confidence in such situations. However, conventional detection assays, such as ELISA, are generally not suitable for rapid, multiplexed detection; such assays are typically designed for the detection of a single analyte, necessitating the completion of multiple assays to detect multiple analytes. Detection of multiple biomarkers using mainstay techniques thus requires considerable user engagement, long processing times, and complex workflows. Here, Applicants created fNACPs with distinct biorecognition elements to develop a multiplexed fNACP assay that can be quantified using common laboratory-based fluorescence detection methods (e.g., flow cytometry). To produce fNACPs, Applicants incubated NACPs with biotin-polyethylene glycol (PEG)- silane (BPS), followed by streptavidin (SA), and then by various biotinylated biorecognition elements (BREs) with wash steps between each incubation. The PEG chains of the BPS provide antifouling properties and allow greater flexibility of the BREs to capture biomolecules relative to that of BREs immobilized on solid surfaces. Since SA contains 4 biotin-binding sites, its use in the functionalization scheme allows for modular tuning of fNACP specificity by attaching different biotinylated BREs (Figure 4A). In this example, Applicants functionalized fNACPs to capture three distinct biomolecules: anti-OVA IgG, IgM, and IgA. To detect anti-OVA, IgM, and IgA, Applicants functionalized fNACPs with OVA, anti-IgM, and anti-IgA BREs, respectively. The target biomolecules were selected to serve as diverse model biomarkers by varying in species type (e.g., mouse and rabbit), structure (e.g., monomeric anti-OVA, pentameric IgM and dimeric IgA), and size (e.g., ∼150 kDa for anti-OVA, ∼900 kDa for IgM, and ∼500 kDa for IgA). To measure captured biomarkers by flow cytometry, Applicants used red fluorescent anti-IgG, anti- IgM, and anti-IgA secondary labels (Figure 4B). Applicants additionally created control NACPs that can be included in each fNACP assay to reveal assay errors. Instead of being modified with biotinylated BREs, control NACPs were modified with biotinylated bovine serum albumin (BSA) as a blocking protein. As the specific binding of target biomolecules or secondary labels to BSA is not anticipated, the use of control NACPs alongside fNACP assays allows users to identify assay inaccuracies during fluorescence analysis. For example, if control NACPs exhibit red fluorescence on the order of that recorded for fNACPs, the assay would be considered invalid and would require retesting. To distinguish the control NACPs and the three types of fNACPs, Applicants functionalized NACPs with varied ratios of blue-fluorescent SA to nonfluorescent SA during the SA incubation step. This yielded particle groups that are fluorescently barcoded for facile distinction by flow cytometry. By gating particles based on fluorescence and a size parameter (e.g., forward scatter), each of the four types of fNACPs can be individually assessed for secondary- label fluorescence quantification (Figure 4C). Example 8: fNACP assays for detection over a large dynamic range. To evaluate the sensitivity of the three fNACP assays, Applicants mixed 104anti-OVA-, IgM-, or IgA-specific fNACPs and 104control NACPs with 100 μL of PBS spiked with anti-OVA, IgM, or IgA, respectively, over multiple orders of magnitude of concentration (e.g., yielding sub- picomolar to micromolar concentrations). After a 30-min incubation period in a 96-well plate, Applicants washed the particles and resuspended them in their respective labeling solutions at 10 μg mL−1. After an additional 30-min incubation period, Applicants again washed the particles and then analyzed particle fluorescence via flow cytometry. For all three assays, the generated standard curves exhibited an expected logarithmic increase in fluorescence as the biomolecule concentration increased (Figures 4D–4F). While the control NACPs displayed some elevation in fluorescence as the biomolecule concentration increased, their fluorescence intensities were multiple orders of magnitude below those of the fNACPs, indicating acceptable assay function. This increase in fluorescence could be mitigated by future optimization of the particle antifouling layer or the use of other antifouling layers (e.g., polyphosphoesters or oligoethylene glycol and sulfobetaine methacrylate copolymers). To determine assay sensitivity, Applicants fit the data with 5-parameter logistic (5PL) curves and determined the limit of detection (LOD), defined as the biomolecule concentration that yields fNACP fluorescence intensities 3 standard deviations above that of the background (e.g., nonfluorescent NACPs), based on the fit curves. The LODs of the anti-OVA, IgM, and IgA assays were 0.02 nM, 3.0 pM, and ∼0.25 pM, respectively. These sensitivities are competitive with (on the same order of magnitude as) those of commercial ELISA assays, a finding that highlights the potential for implementation of these assays in laboratory workflows. In the case of IgA detection, the sensitivity of the fNACP assay was one order of magnitude lower than that of commercial ELISA. Through these results, Applicants demonstrate the ability to capture a range of biomarker types through simple modifications to the fNACP functionalization. Example 9: Barcoded fNACPs for multiplexed biomarker detection. To evaluate the multiplexing performance of the fNACP assay, Applicants mixed 104of each fNACP type (e.g., to capture anti-OVA, IgM, or IgA) with 104control NACPs and incubated all four particle types with PBS spiked with all possible combinations of 0 or 10 nM anti-OVA, 0 or 0.2 nM IgM, and 0 or 3.2 nM IgA. Applicants selected these concentrations as they fell in the middle of the detectible range, as discovered during individual standard curve experiments (Figures 4D–4F). Control NACPs were functionalized with fluorescent green SA to enable simple visualization during later analyses. Applicants followed the same general steps of the previous experiments, including incubating the particles for 30 min for biomolecule capture, washing the particles, and incubating once more in labeling solution. Here, the labeling solution consisted of 10 μg mL−1of each of the secondary labels (e.g., fluorescent anti-IgG, anti-IgM, and anti-IgA). Upon fluorescence analysis, Applicants found that all particle types exhibited an expected increased fluorescence only when the corresponding biomolecules were present (Figure 5A). This indicates that the capture and detection of the biomolecules was specific and that assay components do not cross-react at levels that would impact multiplexing assays (Figure 5B). Applicants analyzed the samples by fluorescence microscopy and confirmed that, for the samples with all biomarkers present, the control particles did not exhibit red fluorescence from the secondary labels but that each of the three fNACP types did (Figure 5C). These experiments confirm the ability to create multiplexed fNACP-based assays, which could substantially decrease assay timescales in laboratory-based biomolecule detection protocols. Example 10: Multichannel acoustic separator for simplified detection from complex biofluids. To evaluate integration of the fNACP assay with the multichannel acoustic separator for detection of biomolecules from whole blood, Applicants conducted an assay to detect IgA at concentrations ranging 4 orders of magnitude. Applicants incubated 104IgA-specific fNACPs and 104control NACPs with 100 μL of porcine blood spiked with IgA over a range of 0–32 nM in a 96-well plate. After a 30-min incubation, Applicants used the multichannel acoustic separator to trap and purify the fNACPs and control NACPs from whole blood after diluting with buffer 3- fold. Applicants then incubated the particles in a labeling solution for 30 min, washed the particles thrice, and analyzed particle fluorescence by flow cytometry. The entire assay took 70 min, and most of this time was consumed by the two 30-min incubation steps (Figure 5D). Applicants anticipate that these incubations could be shortened through optimization of the protocol or use of higher-affinity BREs and labels. Applicants found that the IgA detection assay in whole blood performed similarly to that conducted in buffer, where the LOD of the assay increased from ∼0.25 to ∼1.6 pM (Figure 5E). Applicants believe this increase in LOD is reasonable for detection in whole blood, as it is still competitive with commercially available ELISA kits (e.g., with an LOD of ∼3 pM). These results demonstrate the ability of both the fNACP assay and the multichannel acoustic separator to detect low-concentration biomolecules from complex samples containing large amounts of off-target molecules and particulates. Example 10: Additional Validation Data. Functionalization of control NACPs and anti-OVA-, IgM-, and IgA-specific fNACPs NACPs were functionalized by conjugating to the particles a biotin-PEG-silane, followed by SA, followed by a biotinylated BRE, whereby washing was performed between each conjugation. Typically, 5x106 NACPs were washed thrice with 0.05% v / v Tween 20 (Sigma-Aldrich®) in DIW and resuspended in 50 μL of a 30 mg mL-1 solution of 2 kDa silane-PEG-biotin (Laysan Bio®) in DIW. Particles were incubated for 2 hr at room temperature while mixing on a VorTemp 56 shaking incubator (Labnet) at ~1200 rpm. NACPs were washed thrice with 0.05% v / v Tween 20 in PBS (Sigma-Aldrich®). NACPs were resuspended in 100 μL of 1.7 mg mL1 SA (ThermoFisher®) and incubated for 90 min at room temperature while mixing. After incubation, NACPs were washed thrice with 0.05% v / v Tween 20 in PBS and resuspended in solutions containing biotinylatedBREs. For anti-OVA-specific fNACPs, NACPs were resuspended in 200 μL of 2.5 mg mL-1 biotin-conjugated OVA (Nanocs) in PBS. For IgM-specific fNACPs, NACPs were resuspended in 1000 μL of 0.5 mg mL-1 biotin-conjugated goat anti-Mouse IgM (ThermoFisher) in PBS. For IgA-specific fNACPs, NACPs were resuspended in 1000 μL of 0.5 mg mL-1 biotin-conjugated goat anti-mouse IgA (Southern Biotech®) in PBS. In all cases, particles were incubated for 90 min at room temperature while mixing. Finally, fully functionalized fNACPs were washed thrice in storage buffer comprising 1% w / v BSA (Sigma-Aldrich®) in PBS. fNACPs were then resuspended in storage buffer and stored at 4˚C until use. Control NACPs were functionalized identically to fNACPs through the conjugation of SA, after which point, control NACPs were incubated with 450 μL of a 2 mg mL-1 solution of biotin-conjugated BSA (ThermoFisher®) in PBS. Fluorescently barcoding fNACPs and control NACPs To fluorescently barcode fNACPs and control NACPs, particles were incubated with varied ratios of nonfluorescent SA to fluorescent Alexa Fluor 405-conjugated SA (ThermoFisher) during the SA functionalization step. Ratios utilized include 1:0, 99:1, 9:1, and 0:1. During some experiments, control particles were instead functionalized with fluorescent Alexa Fluor 488 SA (ThermoFisher®) alone. Evaluation of NACP trapping To evaluate NACP trapping, samples of 5x104 NACPs were added to 300 μL of either wash buffer or 3- times diluted whole blood. The multichannel acoustic separator was primed with wash buffer and the acoustics were engaged. NACP samples were added to the upper siphon chambers and the draining fluid was collected for analysis. The channels were then washed two (e.g., for separation from buffer) or four (e.g., for separation from blood) times and the drained fluid was collected in a 96-well plate. The acoustics were then disengaged and the NACPs were collected. To determine the extent of residual NACPs in the channels after NACP collection, channels were then washed two more times, and the rinse fluid was collected. The volume of each collected sample was adjusted to the same approximate volume by pipette. Finally, all samples were analyzed by flow cytometry (BD FACSCelesta with a high-throughput sampler), and the number of NACPs in each sample were recorded. Notably, the samples from the initial addition of particles in blood, as well as the corresponding first two washes, had to be diluted 200, 100, or 10 times, respectively, to evaluate NACP numbers with flow cytometry. These dilution factors were considered when completing data analysis. Applicants then calculated trapping efficiency by the following: Individual fNACP assays For anti-OVA, IgM, and IgA assays, 104 fNACPs specific to either anti-OVA, IgM, or IgA were mixed with 104 control NACPs. The particle mixtures were then resuspended in 100 μL PBS spiked with either antiOVA spanning 0.001–1000 nM, IgM spanning 0.0002–200 nM, or IgA spanning 0.00032–320 nM in a 96- well plate. Samples were incubated for 30 min while mixing at 1200 rpm at room temperature. After incubation, samples were washed thrice in the well plate and resuspended in labeling solutions. The labelling solutions for anti-OVA, IgM, and IgA detection were 10 μg mL-1 Alexa Fluor Plus 594-conjugated donkey anti-rabbit IgG IgG (ThermoFisher®) in PBS, 10 μg mL-1Alexa Fluor 594-conjugated goat anti- mouse IgM IgG (Southern Biotech®) in PBS, or 10 μg mL-1 Alexa Fluor Plus 555-conjugated goat anti-mouse IgA IgG (Southern Biotech®) in PBS, respectively. After incubating for 30 min while mixing at 1200 rpm at room temperature, particles were washed thrice in the well plate. Finally, the particles were resuspended in 200 μL wash buffer and analyzed by flow cytometry. Multiplexed fNACP assays To evaluate multiplexing of the fNACP assay, 104 of each of the three fNACP types and 104 control fNACPs were mixed and resuspended in 100 μL PBS spiked with all 8 possible combinations of 0 or 10 nM anti-OVA, 0 or 0.2 nM IgM, and 0 or 3.2 nM IgA. Samples were incubated for 30 min while mixing at 1200 rpm at room temperature. After incubation, samples were washed thrice in the well plate and were resuspended in a labelling solution containing 10 μg mL-1 of each of the 3 fluorescent labels used for the individual fNACP assays. After incubating for 30 min while mixing at 1200 rpm at room temperature, the particles were washed thrice in the well plate. Finally, the particles were resuspended in 200 μL wash buffer and analyzed by flow cytometry. Example 11: Materials and Methods. Multichannel acoustic separator body design and fabrication The multichannel acoustic separator was designed in Autodesk Fusion and 3D printed on a masked stereolithography (STL) 3D printer (SL1S, Prusa Research®) using Fast Mecha White resin (Siraya Tech). Printed parts include the upper siphon chamber, left side, right side, trapping array, electrical port cover, and two transducer clips. Printed parts were washed in ethanol (VWR), dried, and cured for 1 min using a curing chamber (CW1S, Prusa Research®) before device assembly. Imperfections in the printed parts, such as lipped edges originating from the placement of parts on the print platform, were removed with sandpaper (Norton Abrasives). Trapping array fabrication To assemble the trapping array, 65-mm sections of 1.02 inner diameter (ID) × 2.16 outer diameter (OD) silicone tubing (Freudenberg Medical) were attached to one end of 12 square glass capillaries that had 1 × 1 mm inner dimensions and were ∼25 mm long (VitroCom®). The junctions were sealed with cyanoacrylate glue (Gorilla Glue Company®) and Parafilm (VWR®). Capillaries were then glued to a custom thickness-mode piezoelectric transducer with a resonance frequency of 750 kHz (Chengdu Chengyao Technology®, PST4, 2.7 × 10 × 105 mm) using cyanoacrylate glue. During attachment of the capillaries to the transducer, care was taken to ensure that the capillaries were evenly distributed along the length of the transducer, attached to the transducer at the middle of the capillaries, and that the open ends of each capillary were aligned. 24 American wire gauge (AWG®) wires (Digikey®), used as transducer leads, were soldered to either side of the transducer. The terminal ends of the wires were soldered to 4-mm female banana jack ports (Bei Qian®). Device fabrication and setup The assembled trapping array was placed on the trapping array holder. The lead from the underside of the trapping array was passed through the lead hole. The two transducer clips were placed over the trapping array to keep the array assembly in one piece during subsequent assembly steps. The trapping array was fastened to the left and right side socket screws (Grainger, M3-0.5, l = 10 mm) and thin hex nuts (M3-0.50, w = 5.5 mm, h = 1.8 mm). Both banana ports were then passed through the lead through-holes of the left side and were seated in the banana port holder. Both wires were seated in the wire guides of the left side to avoid interference with well plates. The electrical port cover was attached to the left side to contain both banana ports using a screw and nut. The upper siphon chamber was attached to the device with two screws and two nuts. The ends of the silicone tubes extending from each capillary were positioned at the bottom of the siphon chamber wells, and the lengths of the tubes in the chambers were pushed into the tubing guides to be immobilized. The external tips of the outlets of each glass capillary were coated with a hydrophobic silicone solution (Rain-X glass water repellent) to ensure clean droplet separation during siphoning. The ΔH of the device was adjusted by loosening the two screws connected to the upper siphon chamber array and manually sliding the upper siphon chamber array along the continuous screw slots. Once the upper siphon chamber array was positioned, the screws were tightened to lock the upper siphon chamber array into place. Multichannel acoustic separator operation The device was connected to a function generator and amplifier via banana port. An oscilloscope (InfiniiVision®, Keysight®) was attached to each lead via banana port to alligator clip wiring. To engage acoustics, a 750 kHz AC sine wave was applied at 25 Vpp. To prime the siphon, 600 μL of water or buffer was added to each of the 12 siphon chambers via a multichannel pipette. To complete priming, the compressible tubing of each channel was depressed and manipulated with a rolling motion to draw liquid from the siphon chambers through the entirety of each channel. Flow through each channel was then controlled by the gravity-based siphon effect. Liquids in the siphon chambers were siphoned out of the chamber until an equilibrium in pressure between the inlet and outlet of each channel was reached. The height, and therefore the volume, of the fluid remaining in the siphon chambers after reaching equilibrium was tuned by adjusting the relative inlet and outlet height (ΔH). When fluid remained in the chambers after an equilibrium was reached, flow through each of the channels was reinitiated by adding other liquids to the siphon chambers, typically 300 μL of water, buffer, or 3-fold-diluted whole blood (porcine blood with sodium heparin, Lampire). While the siphon was active, liquid drained through the channels and dripped from the trapping channel outlets into a waste container or 96-well plate for sample collection. Prior to use, water and wash buffer were degassed for at least 1 h to avoid the generation of air bubbles in the trapping channels. Evaluation of channel washing To analyze the effectiveness of device washing when samples suspended in water were present in the device, all 12 channels of the multichannel acoustic separator were primed with a solution of blue 1 gel food coloring (General Mills) in water, and the acoustics were engaged. The channels were washed 5 times. For every wash, the liquid leaving each channel was collected in a 96-well plate. After this, 200 μL of each sample was transferred to clear, flat-bottom 96-well plates (Grenier). Finally, each sample was analyzed by recording absorbance at 630 nm using a plate reader (Infinite 200Pro, Tecan) (Figure 7). To analyze the effectiveness of washing when samples suspended in diluted whole blood were present in the device, the same process was repeated after priming the channels with 3-times-diluted whole blood. Because the optical density of diluted whole blood was high, 15 μL of each collected sample was diluted to 200 μL with PBS prior to the measurement of absorbance at 542 nm. Evaluation of flow rate To evaluate the drained volume over time and, subsequently, the flow rate when different fluids pass through the device, the device was primed with either water, buffer, or 3-fold-diluted whole blood. The acoustics were then engaged, and 300 μL of the respective liquid was added to the upper siphon chamber while being recorded by video. The experiment continued until all channels had stopped dripping for at least 10 s. These videos were analyzed to determine the number of drops per channel as well as the time points for each droplet. Then, the drained volume from each channel was determined by pipette. This volume was divided by the total number of drops per channel to yield an average drop volume per channel. The fluid volume drained over time was then plotted assuming a constant volume per droplet for each individual channel. NACP production and characterization NACPs were produced as previously described. Briefly, 0.1% v / w TEOVS (Sigma- Aldrich) was thoroughly mixed with a standard 10:1 weight ratio mixture of Sylgard 184 base and crosslinker (Dow Chemical). This mixture was submerged in a solution of 1% w / w Pluronic F- 108 (Sigma-Aldrich) in deionized water (DIW), and all components were homogenized for 1 min, producing polydisperse, uncured NACPs. The mixture was cured for 2 h at 80°C. The cured NACPs were then filtered through a series of cell strainers (PluriStrainer) while vortexing at speeds sufficient to agitate the liquid in the cell strainer to isolate a low size polydispersity population of NACPs between 20 and 30 μm. NACP washing during production and functionalization was performed by centrifugation at 3,000 × g for 3 min, followed by aspiration and resuspension. NACP washing in 96-well plates was performed by centrifugation of the well plate at 1,500 × g for 1 min, also followed by aspiration and resuspension. For sizing, NACPs were pipetted onto a glass slide, and images were captured by a Zeiss AxioVert A1 TL / RL inverted fluorescence microscope equipped with an Axiocam 305 mono camera (Zeiss, Germany). NACP sizes were determined by analyzing images with Fiji / ImageJ software (Figures 8A and 8B). fNACP trapping and purification Before fNACP purification, the multichannel acoustic separator was primed with wash buffer and acoustics were engaged. Then, 300 μL samples containing 4–5 × 104particles were added to the upper siphon chambers via multichannel pipette. After draining, the channels were washed 2–4 times by adding 300 μL of wash buffer to the upper siphon chambers and allowing the chambers to drain into a waste container until reaching equilibrium. Then, the acoustics were disengaged. After waiting 1–2 min, 300 μL of wash buffer was added to the upper siphon chambers, and the fNACP-rich purified samples were collected on a 96-well plate. To visually assess NACP trapping, ∼5 μL of a 5 wt % solution of 5.1 μm red fluorescent carboxylated PS particles (Magsphere) was added to a 1.7 × 105particle mL−1solution of NACPs coated with green-fluorescent SA in wash buffer. The particle mixture was passed through a trapping channel situated on the fluorescent microscope at 1 mL min−1via syringe pump. Images of the trapping channels were captured before and after engaging the acoustics. fNACP assay in whole blood using the multichannel acoustic separator For each sample, 104IgA-specific fNACPs and 104control NACPs were suspended in 100 μL of whole porcine blood with spiked IgA spanning 0.0032–32 nM in a 96-well plate. Samples were incubated for 30 min while mixing at 1,200 rpm at room temperature. Samples were then diluted to 300 μL with wash buffer and were purified using the multichannel acoustic separator. The purified samples were collected in a round-bottom 96-well plate and were pelleted, aspirated, and resuspended in 100 μL of a labeling solution containing 10 μg mL−1Alexa Fluor Plus 555-conjugated goat anti-mouse IgA IgG (Southern Biotech) in PBS. After incubating for 30 min while mixing at 1,200 rpm at room temperature, particles were washed thrice in the well plate. Finally, particles were resuspended in 200 μL wash buffer and analyzed by flow cytometry. Assay curve fitting Assay standard curves, shown in Figures 4D–4F and 5E, were fit with a 5PL curve to determine the assay LOD. To do so, experimental data were fitted to the standard 5PL equation, given by where ^^ is the fluorescence intensity, intensities at the minimum and maximum biomarker concentrations, respectively, ^^ is the biomarker concentration, ^^ is the mid- range concentration, ^^ is the slope factor, and ^^ is the asymmetry factor. ^^ was constrained to be the mean background fluorescence measured during experiments (e.g., of nonfluorescent particles). MATLAB was used for all fittings. LOD was determined by identifying the concentration at which the response was 3 standard deviations above the background response, as predicted by the curve fit. Fit parameters may be found in Table 1. Statistical analysis Differences between groups were compared by one-tailed Student’s t tests, and p < 0.05 was considered significant in the analyses. All quantitative data are expressed as the mean ± standard deviation. TABLE 1: 5PL parameters. Parameter Anti-OVA IgM IgA IgA in blood a 152.7 148 96.4 69.9 b 177990.3 384062.0 1079091.3 635792.0 g 0.17 0.0092 0.0037 0.0051 c 69.50 0.082 0.097 1.94 d 0.86 1.39 1.28 1.00

Claims

CLAIMS What is claimed is:

1. A multi-channel acoustic separation assembly comprising: − a siphon chamber array having a plurality of siphon chambers each adapted to receive a sample; − a trapping array securing a plurality of capillary channels, wherein each of said channels is in fluid communication with at least one of the siphon chambers through a transfer tube forming a plurality of fluid pathways, each adapted to generate a siphon effect; and − wherein each of the capillary channels is responsive to an acoustic wave generator configured to generate a standing acoustic wave within each of the capillary channels.

2. The assembly of claim 1, wherein the siphon effect comprises a pressure differential created between each of the channel inlets and the channel outlets of the fluid pathways when the samples are added to the siphon chambers causing the samples to flow through the fluid pathways until an equilibrium point is reached.

3. The assembly of claim 1, wherein the acoustic wave generators comprise piezoelectric transducers configured to generate a standing acoustic wave within each of the capillary channels.

4. The assembly of any of claims 1- 3, wherein the standing acoustic wave generates a node along the centerline of each of the capillary channels and an antinode along the walls of each of the capillary channels.

5. The assembly of claim 1, further comprising a power source responsible for actuating the acoustic wave generators.

6. The assembly of any of claims 1-5, wherein the sample(s) contains negative acoustic contrast particles (NACPs).

7. The assembly of claim 6, wherein said NACPs comprise functionalized NACPs (fNACPs) configured to separate one or more analytes of interest.

8. The assembly of claim 7, wherein said fNACPs are coded for multiplex detection of biomolecules or other molecules.

9. The assembly of claim 8, wherein said fNACPs are bar coded.

10. The assembly of claim 8, wherein said fNACPs are color coded.

11. The assembly of claim 1, further comprising an array frame securing the siphon chamber array and the trapping array, and wherein the trapping array further secures the acoustic wave generator(s).

12. The assembly of claim 11, wherein said array frame comprises a plate receiver configured to accept a well plate positioned below the capillary channels.

13. The assembly of claim 11, wherein said array frame comprises height adjustors configured to allow vertical adjustment of the upper siphon chamber array, trapping array, or capillary channels.

14. The assembly of claim 11, wherein said array frame comprises a lock configured to slidably secure a well plate positioned below the capillary channels.

15. The assembly of claim 1, wherein each of the siphon chambers comprise a sample reservoir, each in fluid communication with the transfer tubes.

16. The assembly of claim 1, wherein said sample comprises a biological sample.

17. The assembly of claim 16, wherein said biological sample comprises a human biological sample.

18. The assembly of claim 1, further comprising a slot positioned on the top surface of each of the siphon chambers and configured to secure the transfer tube.

19. The assembly of claim 1, wherein the capillary channels comprise glass capillary tubes.

20. The assembly of claim 1, wherein the capillary channels each comprise a channel outlet.

21. The assembly of claim 1, wherein each of the channel outlets are positioned above a well plate.

22. The assembly of claim 21, wherein said well plate comprises a slidably adjustable well plate having a plurality of wells.

23. The assembly of claim 1, further comprising one or more additional solutions comprising a reagent capable of binding to the analyte of interest or the NACPs, or a reagent capable of identifying to the analyte of interest, or the NACPs.

24. The assembly of claim 23, wherein the reagent is selected from: antibodies, receptors, ligands, nucleic acids, aptamers, synthetic host molecules, enzymes, stains, dyes, or chromophores.

25. The assembly of claim 23, wherein the analyte is selected from: an ion, small molecule, antigen, protein, nucleic acid, carbohydrate, virus, cell or biomarker.

26. A method of acoustically separating a particle, the method comprising: − introducing one or more samples into a siphon chamber array having a plurality of siphon chambers, wherein each sample contains an analyte of interest and negative acoustic contrast particles (NACPs) capable of biospecific recognition of the analyte; − establishing trapping array having a plurality of capillary channels, wherein each of the channels is in fluid communication with at least one of the siphon chambers through a transfer tube forming a plurality of fluid pathways having a pressure differential between a channel inlet and a channel outlet of the fluid pathway causing each of the samples to flow through the fluid pathway until the equilibrium point is reached; and − generating a standing acoustic wave within each of the capillary channels forming a node along the centerline of each of the capillary channels and an antinode along the walls ofeach of the capillary channels, and wherein the NACPs are trapped at the antinode position in response to the acoustic wave and the remining sample and non-acoustically responsive particles and / or PACPs pass through the capillary channel; − removing the standing acoustic waves within each of the capillary channels thereby releasing the trapped NACPs; − collecting the NACPs from each of the capillary channels.

27. The method of claim 26, further comprising the step of compressing each of the transfer tubes.

28. The method of claim 26, wherein the step of generating a standing acoustic wave comprises engaging one or more acoustic wave generators to generate acoustic forces in each of the capillary channels.

29. The method of claim 28, wherein said acoustic wave generator comprises a piezoelectric transducer.

30. The method of claim 26, wherein said NACPs comprise functionalized NACPs (fNACPs) configured to separate one or more analytes of interest.

31. The method of claim 30, wherein said fNACPs are coded for multiplex detection of biomolecules or other molecules.

32. The method of claim 31, wherein said fNACPs are bar coded.

33. The method of claim 31, wherein said fNACPs are color coded.

34. The method of claim 26, wherein said sample comprises a biological sample.

35. The method of claim 34, wherein said biological sample comprises a human biological sample.

36. The method of claim 26, further comprising securing the siphon chamber array and the trapping array to an array frame, and wherein the trapping array further secures the acoustic wave generator(s).

37. The method of claim 36, wherein said array frame comprises a plate receiver configured to accept a well plate positioned below the capillary channels.

38. The method of claim 36, further comprising adjusting the height of the capillary channels.

39. The method of claim 36, further comprising positioning a well plate positioned below the capillary channels.

40. The method of claim 26, wherein said step of collecting comprises introducing one or more buffers to the siphon chambers of the siphon chamber array in the absence of the standing acoustic wave.

41. The method of claim 26, wherein said step of collecting comprises collecting the NACPs and the analyte of interest in a well plate positioned below the channel outlet of the capillary channel.

42. The method of claim 26, further comprising contacting the NACPs and the analyte of interest with one or more additional solutions comprising a reagent capable of binding to the analyte of interest, or a reagent capable of identifying to the analyte of interest, or the NACPs.

43. The method of claim 42, wherein the reagent is selected from: antibodies, receptors, ligands, nucleic acids, aptamers, synthetic host molecules, enzymes, stains, dyes, or chromophores.

44. The method of claim 42, wherein the analyte is selected from: an ion, small molecule, antigen, protein, nucleic acid, carbohydrate, virus, cell or biomarker.

45. The method of claim 26, further comprising the step of introducing the NACPs and the analyte of interest into the siphon chamber and repeating the steps for claim 26.

46. An acoustic separation device comprising: − a siphon chamber adapted to receive a sample; − a transfer tube in fluid communication with the siphon chamber and a capillary channel forming a fluid pathway adapted to generate a siphon effect; and − an acoustic wave generator in communication with the capillary channel.

47. The device of claim 46, wherein the siphon effect comprises a pressure differential created between a channel inlet and a channel outlet of the fluid pathway when the sample is added to the siphon chamber causing the sample to flow through the fluid pathway until an equilibrium point is reached.

48. The device of claim 46, wherein said acoustic wave generator generates a standing acoustic wave within the capillary channel.

49. The device of claim 46, wherein said acoustic wave generator comprises a piezoelectric transducer.

50. The device of any of claims 46-49, wherein said standing acoustic wave generates a node along the centerline of the capillary channel and an antinode along the walls of the capillary channel.

51. The device of claim 46, further comprising a power source responsible for actuating the acoustic wave generator.

52. The device of any of claims 46-51, further comprising negative acoustic contrast particles (NACPs).

53. The device of claim 52, wherein said NACPs comprise functionalized NACPs (fNACPs) configured to separate one or more analytes of interest.

54. The device of claim 53, wherein said fNACPs are coded for multiplex detection of biomolecules or other molecules.

55. The device of claim 53, wherein said fNACPs are bar coded.

56. The device of claim 53, wherein said fNACPs are color coded.

57. The device of claim 46, wherein said siphon chamber comprises a sample reservoir in fluid communication with the transfer tube.

58. The device of claim 46, wherein said sample comprises a biological sample.

59. The device of claim 58, wherein said biological sample comprises a human biological sample.

60. The device of claim 46, further comprising a slot positioned on the top surface of the siphon chamber and configured to secure the transfer tube.

61. The device of claim 46, wherein said capillary channel comprises a glass capillary tube.

62. The device of claim 46, wherein said capillary channel comprises a channel outlet.

63. The device of claim 62, wherein said channel outlet is positioned above a well plate.

64. The device of claim 63, wherein said well plate comprises a slidably adjustable well plate having a plurality of wells.

65. The device of claim 46, further comprising one or more additional solutions comprising a reagent capable of binding to the analyte of interest, or a reagent capable of identifying to the analyte of interest, or the NACPs.

66. The device of claim 65, wherein the reagent comprises antibodies, receptors, ligands, nucleic acids, aptamers, synthetic host molecules, enzymes, stains, dyes, or chromophores.

67. The device of claim 65, wherein the analyte comprises an ion, small molecule, antigen, protein, nucleic acid, carbohydrate, virus, cell or biomarker.

68. A method of acoustically separating a particle, the method comprising: − introducing a sample into a siphon chamber, wherein the sample contains an analyte of interest and negative acoustic contrast particles (NACPs) capable of biospecific recognition of the analyte; − positioning a transfer tube in fluid communication with the siphon chamber and a capillary channel forming a fluid pathway having a pressure differential between a channel inlet and a channel outlet of the fluid pathway causing the sample to flow through the fluid pathway until an equilibrium point is reached; and − generating a standing acoustic wave within the capillary channel forming a node along the centerline of the capillary channel and an antinode along the walls of the capillary channel, and wherein the NACPs are trapped at the antinode position in response to the acoustic wave; − removing the standing acoustic wave thereby releasing the trapped NACPs; − collecting the NACPs from the capillary channel.

69. The method of claim 68, further comprising the step of compressing the transfer tube.

70. The method of claim 68, wherein the step of generating a standing acoustic wave comprises engaging an acoustic wave generator.

71. The method of claim 70, wherein said acoustic wave generator comprises a piezoelectric transducer.

72. The method of claim 68, wherein said NACPs comprise functionalized NACPs (fNACPs) configured to separate one or more analytes of interest.

73. The method of claim 72, wherein said fNACPs are coded for multiplex detection of biomolecules or other molecules.

74. The method of claim 73, wherein said fNACPs are bar coded.

75. The method of claim 73, wherein said fNACPs are color coded.

76. The method of claim 73, wherein said sample comprises a biological sample.

77. The method of claim 68, wherein said biological sample comprises a human biological sample.

78. The method of claim 68, wherein said step of collecting comprises introducing one or more buffers to the siphon chamber in the absence of the standing acoustic wave.

79. The method of claim 68, wherein said step of collecting comprises collecting the NACPs and the analyte of interest in a well plate positioned below the channel outlet of the capillary channel.

80. The method of claim 79, further comprising contacting the NACPs and the analyte of interest with one or more additional solutions comprising a reagent capable of binding to the analyte of interest, or a reagent capable of identifying to the analyte of interest, or the NACPs.

81. The method of claim 80, wherein the reagent comprises antibodies, receptors, ligands, nucleic acids, aptamers, synthetic host molecules, enzymes, stains, dyes, or chromophores.

82. The method of claim 80, wherein the analyte comprises an ion, small molecule, antigen, protein, nucleic acid, carbohydrate, virus, cell or biomarker.

83. The method of claim 68, further comprising the step of introducing the NACPs and the analyte of interest into the siphon chamber and repeating the steps for claim 68.

84. A magnetic separation device comprising: − a siphon chamber adapted to receive a sample; − a transfer tube in fluid communication with the siphon chamber and a capillary channel forming a fluid pathway adapted to generate a siphon effect, wherein the siphon effect comprises a pressure differential created between a channel inlet and a channel outlet of the fluid pathway when the sample is added to the siphon chamber causing the sample to flow through the fluid pathway until an equilibrium point is reached.; and − an engageable magnet positioned adjacent to the capillary channel, wherein the magnet exerts a magnetic force on the sample flowing through the capillary channel thereby trapping magnetic particles in the sample within the capillary channel.

85. The device of claim 84, wherein the magnet comprises an electromagnet responsive to a power source or a permanent magnet 86. The device of claim 85, wherein the permanent magnet is a neodymium magnet 87. A multi-channel magnetic separation assembly comprising: − a siphon chamber array having a plurality of siphon chambers each adapted to receive a sample; − a trapping array securing a plurality of capillary channels, wherein each channel is in fluid communication with at least one of the siphon chambers through a transfer tube forming a plurality of fluid pathways, each adapted to generate a siphon effect wherein the siphon effect comprises a pressure differential created between each of the channel inlets and the channel outlets of the fluid pathways when the samples are added to the siphon chambers causing the samples to flow through the fluid pathways until an equilibrium point is reached; and − wherein each of the capillary channels is responsive to a magnetic field within each of the capillary channels, wherein the magnetic field exerts a magnetic force on the sample flowing through the capillary channels thereby trapping magnetic particles within their respective capillary channels.

88. The assembly of claim 87, wherein the magnetic field is generated by a magnet.

89. The assembly of claim 88, wherein the magnet comprises an electromagnet responsive to a power source.

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