Acoustophoretic device and method with conductive electrodes

Abstract

An acoustophoresis device having a sample vessel and a piezoelectric transducer is described. The sample vessel has an outer surface, a microchannel within the boundary of the outer surface, a first port extending through the outer surface into the microchannel, and a second port extending through the outer surface into the microchannel, and a blood sample can be inserted into the microchannel through the first port. The sample vessel has a conductive trace on the outer surface. The piezoelectric transducer is bonded to the outer surface of the sample vessel to form an integral structure. The piezoelectric transducer contacts at least one of the conductive traces, and the piezoelectric transducer is configured to generate ultrasonic waves in the sample within the microchannel. The piezoelectric transducer has an excitation signal input and a response signal output electrically connected to at least one of the conductive traces.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a national stage application under 35 U.S.C. 371 of PCT / US2022 / 077832, filed October 10, 2022, and claims the benefit under 35 U.S.C. 35 U.S.C. 119(e) of U.S. Provisional Application No. 63 / 366,552, filed June 17, 2022. The entire disclosures of the above-referenced patent applications are expressly incorporated herein by reference.

[0002] The present disclosure relates generally to devices, systems, and methods for testing blood samples. More particularly, the present disclosure relates to a lysing device configured to lyse red blood cells in a sample vessel by ultrasonic vibrations, shear forces, pressure, and / or fluid motion generated within the vessel by an acoustic transducer, such as a piezoelectric transducer, driven at one or more specific excitation frequencies or frequency ranges. In some non-limiting embodiments, the ultrasound waves are generated by a single piezoelectric transducer. The lysing device can be used with a blood sample test analyzer. [Background technology]

[0003] Point-of-care testing generally refers to medical testing that is performed at or near the point of patient care, such as in an emergency department or operating room. Often, the desired outcome of such testing is a rapid and accurate test result to determine the next course of action in the patient's care. Many such point-of-care testing involves the analysis of a blood sample from the patient. Many of these tests use whole blood, plasma, or serum.

[0004] Some tests involve disrupting (lysing) the cell walls of red blood cells in a blood sample to release hemoglobin. Lysis of red blood cells is sometimes called hemolysis. Typically, hemolysis is accomplished by chemical or mechanical means.

[0005] Some devices use ultrasound to lyse red blood cells. Some point-of-care rapid testing devices use spectrophotometric light absorption measurements to determine oximetry parameters on whole blood samples. Oximetry parameters are also known as CO-oximetry parameters. These devices are typically fluidic systems that test a patient's blood sample by placing it in a sample chamber. For example, one system described in U.S. Patent No. 6,213,999 ("Apparatus for Hemolyzing a Blood Sample and for Measuring at Least One Parameter Thereof," published August 4, 2015) uses two piezoelectric elements with two balanced resonating elements symmetrically surrounding the sample chamber and uses ultrasound to lyse red blood cells. However, these devices are difficult and expensive to manufacture, including requiring very precise symmetry with custom-made resonating elements and substructures to hold all elements aligned.

[0006] After lysing the red blood cells, the blood sample is then tested by a spectrophotometer to analyze the intensity of a given wavelength of light transmitted through the cartridge's sample vessel optical window. A spectrophotometer is a device for measuring the intensity of light in a portion of a spectrum of interest, specifically transmitted or emitted by a substance of particular interest. According to the Beer-Lambert law of absorption, a spectrophotometer determines how much light a chemical absorbs by measuring the intensity of light as a beam of light passes through a blood sample or other solution. Each compound in the sample or solution absorbs or transmits light in a specific wavelength range of interest.

[0007] Such tests can measure critical care blood parameters, including hematocrit, free and total hemoglobin, bilirubin, lipids, and oximetry (i.e., the form of hemoglobin known as the oximetry fraction). Physicians and clinicians rely on these measurements to make decisions during patient care. Often, these measurements are performed in central hematology laboratories on large analyzers that are complex to maintain. However, obtaining fast, accurate, and precise results in a point-of-care setting is preferable in many ways, saving time in critical diagnostic situations and avoiding specimen transport issues in the critical care room. While some blood gas analyzers offer point-of-care capabilities, are simpler and easier to manufacture than existing devices, and are easier for clinicians to use, no single solution offers the desired test time, accuracy, precision, and reliability. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] U.S. Patent No. 9,097,701 Summary of the Invention [Problem to be solved by the invention]

[0009] There is a need for a more easily manufactured, low cost dissolution device that provides improved accuracy and precision of measured parameters of a sample within a desired testing time at the point of care of the patient. [Means for solving the problem]

[0010] Acoustophoretic lysis devices, methods, and systems are disclosed. The problems of blood sample testing that are complex, slow, and inaccurate for clinical use are solved by a device uniquely configured to lyse red blood cells in a sample vessel by acoustophoretic forces in the sample from a single acoustic transducer, such as a piezoelectric transducer, driven to vibrate at one or more specific excitation frequencies or ranges of excitation frequencies.

[0011] Consistent with one aspect of the present disclosure, an exemplary acoustophoretic device can include a sample vessel having an exterior surface, a microchannel bounded by the exterior surface, a first port extending through the exterior surface into the microchannel, and a second port extending through the exterior surface into the microchannel, wherein a blood sample having red blood cells and plasma can be inserted into the microchannel through the first port, the sample vessel having conductive traces on the exterior surface; and an acoustic transducer bonded to the exterior surface of the sample vessel to form an integral structure, the piezoelectric transducer contacting at least one of the conductive traces and configured to generate ultrasound waves in the sample in the microchannel, the piezoelectric transducer having a power input electrically connected to at least one of the conductive traces. The microchannel has a length, a width, and a height, and the microchannel aspect ratio of width to height can be in the range of about 0.04 to about 0.175; the sample vessel has a width and a height, and the sample vessel aspect ratio of width to height can be in the range of about 0.5 to about 3.0. An acoustic transducer, such as a piezoelectric transducer, can be bonded to the outer surface of the sample vessel to form an integral structure. The piezoelectric transducer is configured to generate ultrasonic standing waves in the blood sample within the microchannel, vibrating the sample vessel and thereby inducing shear forces within the microchannel. The standing waves and shear forces cause cavitation within the blood sample, thereby rupturing the walls of red blood cells in the blood sample and releasing hemoglobin into plasma. The piezoelectric transducer has a length, width, and height, and the piezoelectric transducer's length can be within a range of 75% to 125% of the length of the channel. The piezoelectric transducer can have a height of 2 mm.

[0012] Consistent with one aspect of the present disclosure, an exemplary acoustophoresis system includes a sample vessel having an exterior surface, a microchannel within the boundary of the exterior surface, a first port extending through the exterior surface into the microchannel, and a second port extending through the exterior surface into the microchannel, wherein a sample is insertable into the microchannel through the first port, the sample vessel having conductive traces on the exterior surface; and an acoustic transducer, such as a piezoelectric transducer, bonded to the exterior surface of the sample vessel to form an integral structure, the piezoelectric transducer contacting at least one of the conductive traces and configured to generate ultrasound waves in the sample in the microchannel, causing the sample vessel to vibrate such that shear forces are induced in the microchannel; and an absorption spectrophotometer including a transmitter and a receiver located adjacent to the sample vessel. and a controller electrically connected to the piezoelectric transducer and configured to provide an electrical signal to the piezoelectric transducer, the electrical signal being received by the piezoelectric transducer causing the piezoelectric transducer to emit an ultrasonic wave, the piezoelectric transducer contracting and expanding and responsively producing magnitude and phase signals, the magnitude and phase signals being measured by a processor to calibrate and adjust the acoustophoresis system for optimal performance and to determine structural integrity. In some embodiments, the microchannel can have a length, a width, and a height, and the microchannel aspect ratio of width to height is in the range of about 0.04 to about 0.175, and the sample vessel has a width and a height, and the sample vessel aspect ratio of width to height is in the range of about 0.5 to about 3.0.The piezoelectric transducer is configured to generate ultrasonic standing waves in the blood sample within the microchannel and to vibrate the sample vessel such that shear forces are induced within the microchannel, the standing waves and shear forces causing cavitation within the blood sample, thereby rupturing cell walls and releasing hemoglobin into the blood sample.

[0013] The channel depth can be in the range of 50 to 150 microns. The channel width can be in the range of 1000 microns to 3500 microns. In one embodiment, the width can be in the range of 1000 microns to 2500 microns, and the length can be in the range of 8000 to 20000 microns. In some embodiments, the length can be in the range of 11000 to 20000 microns.

[0014] The accompanying drawings, which are incorporated into and constitute a part of this specification, illustrate one or more implementations described herein and together with the description, explain these implementations. The drawings are not intended to be drawn to scale and may be drawn to scale or diagrammatically, emphasizing particular configurations and particular views of the drawings for purposes of clarity and conciseness. Not all components are labeled in every drawing. The same reference numbers in the drawings may represent and refer to the same or similar elements or functions. [Brief explanation of the drawings]

[0015] [Figure 1] FIG. 1 is a perspective view of an acoustophoretic lysis device according to the present disclosure. [Figure 2] FIG. 1 is a top view of an acoustophoretic lysis device according to the present disclosure. [Figure 3] FIG. 1 is a bottom view of an acoustophoretic lysis device according to the present disclosure. [Figure 4] FIG. 1 is a first side view of an acoustophoretic lysis device according to the present disclosure. [Figure 5] FIG. 2 is a second side view of an acoustophoretic lysis device according to the present disclosure. [Figure 6] FIG. 1 is a first side view of an acoustophoretic lysis device according to the present disclosure. [Figure 7] 1 is a cross-sectional view of an exemplary acoustophoretic lysis device according to the present disclosure. [Figure 8] 1 is a cross-sectional view of an exemplary acoustophoretic lysis device according to the present disclosure. [Figure 9] FIG. 1 is a first side view of another exemplary acoustophoretic lysis device according to the present disclosure. [Figure 10] FIG. 10 is a first side view of yet another exemplary acoustophoretic lysis device according to the present disclosure. [Figure 11A] FIG. 1 is a perspective view of components of an exemplary sample vessel according to the present disclosure. [Figure 11B] FIG. 1 is a perspective view of components of another exemplary sample vessel according to the present disclosure. [Figure 12] 10 is a graph of total displacement of an exemplary dissolution device according to the present disclosure. [Figure 13] FIG. 1 is a plan view of pressure distribution in a microchannel of an exemplary sample vessel according to the present disclosure. [Figure 14] FIG. 1 is a plan view of fluid velocities in a microchannel of an exemplary sample vessel according to the present disclosure. [Figure 15] 1 is a perspective view of an exemplary analyzer according to the present disclosure. [Figure 16] 1 is a perspective view of components of an exemplary analyzer according to the present disclosure. FIG. [Figure 17] 1 is a perspective view of components of an exemplary analyzer according to the present disclosure. FIG. [Figure 18] FIG. 1 is a schematic diagram of components of an exemplary analyzer according to the present disclosure. [Figure 19] FIG. 1 is a schematic diagram of the determination of absorption spectra according to the present disclosure. [Figure 20] FIG. 10 is a diagram showing spectral profile coefficients of hemoglobin types. [Figure 21] FIG. 1 is a side perspective view of an assembly constructed in accordance with the present disclosure. [Figure 22] FIG. 22 is a bottom view of the assembly shown in FIG. 21. [Figure 23] FIG. 22 is a top view of the assembly shown in FIG. 21. [Figure 24] FIG. 22 is an exploded view of the assembly shown in FIG. 21. [Figure 25] FIG. 1 is a top perspective view of a sample vessel constructed in accordance with the present disclosure, including conductive traces extending across the top surface of the sample vessel. [Figure 26] FIG. 25 is a top perspective view of the sample vessel of FIG. 24 with electrical components bonded to the conductive traces to form circuits. [Figure 27] FIG. 1 is a top perspective view of a support substrate constructed in accordance with the present disclosure. [Figure 28] FIG. 28 is a bottom perspective view of the support substrate shown in FIG. 27. [Figure 29] FIG. 1 is a top perspective view of another support substrate constructed in accordance with the present disclosure. [Figure 30] FIG. 10 is a top perspective view of yet another support substrate constructed in accordance with the present disclosure. [Figure 31] FIG. 1 is a partial perspective view of an exemplary analyzer according to the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0016] The following detailed description refers to the accompanying drawings, in which the same reference numbers in different drawings may identify the same or similar elements.

[0017] The mechanisms proposed in this disclosure avoid the above-mentioned problems. This disclosure describes a lysis device, analyzer, and lysis method, including a lysis device configured to lyse red blood cells in a sample vessel by ultrasound, shear, pressure, and / or fluid motion generated in the sample vessel by an acoustic transducer, hereinafter referred to as a piezoelectric transducer, by way of example. The piezoelectric transducer is connected to the sample vessel and driven at one or more specific excitation frequencies or ranges of excitation frequencies. In one non-limiting embodiment, the piezoelectric transducer is a single piezoelectric transducer. This disclosure further describes an analyzer configured to receive and interact with the lysis device to test a sample in the sample vessel, as well as a method of use.

[0018] As used herein, the terms "comprises," "comprising," "includes," "including," "has," "having," or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a process, method, article, or apparatus that includes a list of elements is not necessarily limited to only those elements and may include other elements not expressly listed or inherent in such process, method, article, or apparatus. Furthermore, unless expressly stated to the contrary, "or" refers to an inclusive or, not an exclusive or. For example, condition A or B is satisfied by any one of the following: A is true (or present) and B is false (or absent); A is false (or absent) and B is true (or present); and both A and B are true (or present).

[0019] Additionally, the use of "a" or "an" is employed herein to describe elements and components of embodiments. This is done merely for convenience to give a general sense of the inventive concept. This description should be read as including one or more, and the singular also includes the plural unless it is clear that something else is meant.

[0020] Furthermore, use of the term "plurality" is meant to convey "two or more" unless expressly stated to the contrary.

[0021] As used herein, modifiers such as "about," "approximately," and combinations and variations thereof are intended to include not only the exact amount or value that they modify, but also some slight deviations from the exact amount or value that may result from, for example, manufacturing tolerances, measurement errors, wear and tear, stresses exerted on various components, and combinations thereof.

[0022] As used herein, the term "substantially" means that the subsequently described parameter, event, or circumstance occurs entirely, or that the subsequently described parameter, event, or circumstance occurs to a large extent or degree. For example, the term "substantially" means that the subsequently described parameter, event, or circumstance occurs with a probability of at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or that a dimension or measurement is within at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% of the referenced dimension or measurement.

[0023] Use of the terms "at least one" or "one or more" will be understood to include one as well as any quantity greater than one. Additionally, use of the phrase "at least one of X, V, and Z" will be understood to include X alone, V alone, and Z alone, as well as any combination of X, V, and Z.

[0024] The use of ordinal numbers (i.e., "first," "second," "third," "fourth," etc.) is intended only to distinguish between two or more items and is not meant to imply any sequence or order or importance among the items or any additional order, unless expressly stated otherwise.

[0025] Finally, any reference herein to "one embodiment" or "an embodiment" means that a particular element, configuration, structure, or characteristic described in connection with that embodiment is included in at least one embodiment. Appearances of the phrase "in one embodiment" in various places herein do not necessarily all refer to the same embodiment.

[0026] As discussed above, typical conventional devices for testing blood samples used in clinical settings are complex, slow, and imprecise. The present disclosure addresses these deficiencies with devices, systems, and methods for lysing red blood cells in a sample vessel by ultrasound, shear, pressure, and / or fluid motion generated within the sample vessel by a single piezoelectric transducer connected to the sample vessel and driven at one or more specific excitation frequencies or ranges of excitation frequencies.

[0027] 1-8, there is shown an acoustophoretic lysis device 10. Generally, lysis device 10 includes a sample vessel 12 and a piezoelectric transducer 14 bonded to sample vessel 12. In one embodiment, lysis device 10 is a unitary structure, for example, formed by sample vessel 12 and piezoelectric transducer 14 being bonded together using a suitable bonding material, such as epoxy.

[0028] The sample vessel 12 has an exterior surface 20, a microchannel 22 within the boundaries of the exterior surface 20, a first port 24 extending through the exterior surface 20 into the microchannel 22 in fluid communication with the microchannel 22, and a second port 26 extending through the exterior surface 20 into the microchannel 22 in fluid communication with the microchannel 22. In one embodiment, the exterior surface 20 may have a mounting area for the piezoelectric transducer 14.

[0029] In one embodiment, the sample vessel 12 has a top surface 40, a bottom surface 42, a first end 44, a second end 46, a first side surface 48, and a second side surface 50, where the first side surface 48 and the second side surface 50 extend between the first end 44 and the second end 46 and between the top surface 40 and the bottom surface 42. In one embodiment, the top surface 40 and the bottom surface 42 are planar. In one embodiment, the first side surface 48 and the second side surface 50 are planar. In one embodiment, the first end 44 and the second end 46 are planar. In one embodiment, the top surface 40, the bottom surface 42, the first end 44, the second end 46, the first side surface 48, and the second side surface 50 cooperate to form a three-dimensional rectangular parallelepiped.

[0030] The sample vessel 12 can be partially, substantially, or completely transparent. In one embodiment, the sample vessel 12 is transparent at least above and below the microchannel 22, allowing a light beam to pass through the sample vessel 12, through the microchannel 22, interact with any material within the microchannel 22, and exit the sample vessel 12.

[0031] The sample vessel 12 can be constructed from glass. In one embodiment, the sample vessel 12 can be constructed from a material (glass or non-glass) having a Young's modulus in the range of about 50 GPa to about 90 GPa. The material property known as Young's modulus or elastic modulus is a measure of a material's ability to withstand a change in length when subjected to longitudinal tension or compression. Young's modulus is equal to the longitudinal stress divided by the strain. In one embodiment, the sample vessel 12 can be constructed from a plastic, such as a cyclic olefin copolymer, that has a stiffness and / or Young's modulus similar to that of glass. In one embodiment, the sample vessel 12 can be constructed from alkali borosilicate glass. An example of alkali borosilicate glass is made by Schott Advanced Optics, 400 York Avenue, Duryea, PA 18642, and is commercially available under the name "D263T ECO Thin Glass."

[0032] The sample vessel 12 has a length from a first end 44 to a second end 46, a width from a first side 48 to a second side 50, a thickness between the top surface 40 and the bottom surface 42, and an aspect ratio defining a proportional relationship between the length and the width. The sample vessel 12 has a longitudinal axis along the length and a latitudinal axis along the width.

[0033] In one embodiment, the aspect ratio of the sample vessel 12 is in the range of about 0.5 to about 3.0. In one embodiment, the aspect ratio of the sample vessel 12 is in the range of about 1.4 to about 1.9. In one embodiment, the length can be about 22 millimeters and the width can be about 12 millimeters. In one embodiment, the length can be about 17 millimeters and the width can be about 12 millimeters. In one embodiment, the length can be about 17 millimeters and the width can be about 6 millimeters. In one embodiment, the length can be about 12 millimeters and the width can be about 6 millimeters.

[0034] The microchannel 22 can be configured to receive a fluid sample 52 (including, but not limited to, a blood sample, a "blank" sample, and / or a wash solution sample) through the first port 24 and / or the second port 26. The microchannel 22 has a length, a width, and a height. Typically, the length of the microchannel 22 is oriented along the longitudinal axis of the sample vessel 12, and the width of the microchannel 22 is oriented along the latitudinal axis of the sample vessel 12. However, it will be understood that the microchannel 22 can be oriented at an angle or offset from the longitudinal and / or latitudinal axes of the sample vessel 12.

[0035] Microchannel 22 has an aspect ratio that defines a proportional relationship between the width and height of microchannel 22. In one embodiment, the width-to-height aspect ratio of microchannel 22 is within the range of about 0.04 to about 0.175. In one embodiment, the width-to-height aspect ratio of microchannel 22 is within the range of about 0.04 to about 0.125. In one embodiment, the width-to-height aspect ratio of microchannel 22 is about 0.05.

[0036] In one embodiment, the width of the microchannel 22 is approximately 2 millimeters. In one embodiment, the width of the microchannel 22 is greater than the illumination width of the light yield area of ​​the absorption spectrophotometer 102. The illumination width can be defined as the width of the cross section of the light yield along the optical path from the absorption spectrophotometer 102 that intersects with the microchannel 22. For example, when the illumination diameter is between 1 millimeter and 1.5 millimeters, the width of the microchannel 22 can be at least approximately 1.6 millimeters. The width of the microchannel 22 can be determined to allow sufficient mechanical alignment between the microchannel 22 and the optical path. For example, for an illumination width of between 1 millimeter and 1.5 millimeters, the width of the microchannel 22 can be approximately 2 millimeters.

[0037] In one embodiment, the length of microchannel 22 can be from about 10 millimeters to about 12 millimeters. In one embodiment, the length of microchannel 22 can be at least about 4 millimeters. In one embodiment, the length of microchannel 22 can be from about 4 millimeters to about 20 millimeters.

[0038] In one embodiment, the length of the microchannel 22 can be based at least in part on a predetermined desired number of nodes to be created within the microchannel 22. For example, if the microchannel 22 has a width of approximately 2 millimeters and the whole blood wave propagation velocity is approximately 1500 m / s, the calculated single node is 350 kHz. The nodes can be uniformly spaced and dispersed within the microchannel 22 along the length of the microchannel 22 (e.g., 2 x 2 mm = 4 mm), and the high pressure produces an even distribution of lysed blood. For example, if the predetermined desired number of nodes on each sidewall of the microchannel 22 is five nodes (see FIG. 13), the length of the microchannel 22 can be set to approximately 17 millimeters so that the first port 24 and the second port 26 are not located at one of these nodes.

[0039] The height of the microchannel 22 can vary, as discussed above. The height of the microchannel 22 can be based on the amount of absorption in the lysed blood of light yield from the absorption spectrophotometer 102 and the desired accuracy of the absorption. For example, the desired absorption can be about 1 optical density (OD).

[0040] In one embodiment, the height of microchannel 22 is about 100 micrometers. In one embodiment, the height of microchannel 22 is about 150 micrometers. In one embodiment, the height of microchannel 22 is about 250 micrometers. In one embodiment, the height of microchannel 22 is about 300 micrometers. In one embodiment, the height of microchannel 22 is about 80 micrometers to about 300 micrometers. In one embodiment, the height of microchannel 22 is about 80 micrometers to about 150 micrometers.

[0041] The first port 24 and the second port 26 are fluidly connected to the microchannel 22 and extend from the microchannel 22 through the exterior surface 20 of the sample vessel 12. In one embodiment, the first port 24 is fluidly connected to the microchannel 22 and may extend from the microchannel 22 to the top surface 40, the bottom surface 42, the first end 44, the second end 46, the first side 48, and / or the second side 50 of the sample vessel 12. In one embodiment, the second port 26 is fluidly connected to the microchannel 22 and may extend from the microchannel 22 to the top surface 40, the bottom surface 42, the first end 44, the second end 46, the first side 48, and / or the second side 50 of the sample vessel 12. The first port 24 and the second port 26 may extend to the same or different ones of the top surface 40, the bottom surface 42, the first end 44, the second end 46, the first side 48, and / or the second side 50.

[0042] In one embodiment, first port 24 and second port 26 each have a diameter of about 0.5 millimeters (500 micrometers) to about 1.5 millimeters (1500 micrometers). In one embodiment, first port 24 and second port 26 each have a diameter of about 0.8 millimeters (800 micrometers).

[0043] The sample vessel 12 may be monolithically fabricated in that the sample vessel 12 is formed from a single piece of material, or in that the sample vessel 12 is formed from multiple pieces of material that are interconnected to form an integrated, complete, or monolithic structure.

[0044] 4-8 , in one embodiment, the sample vessel 12 can include a single substrate 60 bounded by an exterior surface 20, the substrate 60 having a microchannel 22 therein and a first port 24 and a second port 26 fluidly connected to the microchannel 22 and extending to the exterior surface 20. For example, the sample vessel 12 can be a 3D printed substrate, such as a 3D printed glass substrate. The 3D printed substrate can be printed to include the microchannel 22, the first port 24, and the second port 26.

[0045] 9, in one embodiment, the sample vessel 12 can include a first substrate 70 and a second substrate 72. The second substrate 72 can be overlaid on the first substrate 70 to form a unitary structure. In one embodiment, the first substrate 70 and the second substrate 72 can be annealed to one another. In one embodiment, the first substrate 70 and the second substrate 72 can be thermal plasma bonded to one another. In one embodiment, the first substrate 70 and the second substrate 72 have the same length-to-width aspect ratio as the sample vessel 12.

[0046] The microchannel 22 can be located in the first substrate 70, the second substrate 72, and / or can be formed partially in the first substrate 70 and partially in the second substrate 72. In one embodiment, the microchannel 22, the first port 24, and the second port 26 are located in the first substrate 70. In one embodiment, the microchannel 22 is etched into the first substrate 70 and / or the second substrate 72. In one embodiment, the microchannel 22 is located in the first substrate 70, and one or both of the first port 24 and the second port 26 are located in the second substrate 72. One or both of the first port 24 and the second port 26 can be located in (and / or extend through) the first substrate 70 and / or the second substrate 72.

[0047] As shown in FIGS. 10 and 11A , in one embodiment, the sample vessel 12 can include a first substrate 70, a second substrate 72, and a third substrate 80 between the first substrate 70 and the second substrate 72. The first substrate 70, the second substrate 72, and the third substrate 80 can be stacked to form a unitary structure. In one embodiment, the first substrate 70, the second substrate 72, and the third substrate 80 can be thermal plasma bonded to one another. In one embodiment, the first substrate 70, the second substrate 72, and the third substrate 80 can be annealed to one another. One or both of the first port 24 and the second port 26 can be located in the first substrate 70. The first substrate 70 can have a thickness of 700 micrometers. The microchannel 22 can be located in the second substrate 72. The second substrate 72 can have a thickness of 700 micrometers and may not include the first port 24, the second port 26, and the microchannel 22. In one embodiment, the microchannel 22 is a slot located through the third substrate 80. In one embodiment, the third substrate 80 can have a thickness equal to the height of the microchannel 22. In one embodiment, the third substrate 80 can be 100 micrometers thick. In another embodiment, as shown in FIG. 11B (and FIG. 24), the sample vessel 212 has only two glass substrates 310, 312 bonded together, rather than the three substrates 70, 72, 80 shown in FIG. 11A. The use of two substrates 310, 312 bonded together, rather than three substrates 70, 72, 80, improves the manufacturability of the sample vessel 212 compared to the performance of the sample vessel 12, without changing the performance of the sample vessel 212 to lyse red blood cells and release hemoglobin from within the cells into plasma. 24, the first glass substrate 310 includes conductive traces 282 formed thereon, and the second glass substrate 312 includes the microchannel 222 and the first and second ports 224, 226. As shown in FIG. 11B, the first glass substrate 310 can be laminated to the second glass substrate 312 to form a unitary structure.In some embodiments, the conductive traces 282 (disposed on the first glass substrate 310) are formed from a conductive metallic material having a thickness sufficient to be optically opaque to light in the near-infrared, far-infrared, visible, and ultraviolet wavelength ranges.

[0048] Referring again to FIG. 1 , the piezoelectric transducer 14 is attached to the sample vessel 12 (e.g., at a mounting area on the exterior surface 20) to form an integral part of the dissolution device 10. The piezoelectric transducer 14 can have a mounting area attached to the mounting area on the exterior surface 20. In one embodiment, the piezoelectric transducer 14 is at least partially attached to the top surface 40 of the sample vessel 12, although it will be understood that the piezoelectric transducer 14 can be attached to the top surface 40, the bottom surface 42, the first end 44, the second end 46, the first side surface 48, and / or the second side surface 50. The piezoelectric transducer 14 is positioned relative to the microchannel 22 so as not to obstruct light moving through the microchannel 22 from the top or bottom surface of the sample vessel 12. The piezoelectric transducer 14 can be offset from the microchannel 22 so that the piezoelectric transducer 14 allows light to enter the microchannel 22 from outside the sample vessel 12. In one embodiment, the piezoelectric transducer 14 has a length and a longitudinal axis along its length that is oriented substantially parallel to the longitudinal axis of the sample vessel 12. In one embodiment, the piezoelectric transducer 14 has a width that is less than its length.

[0049] The piezoelectric transducer 14 can be located on the sample vessel 12 opposite one or both of the first port 24 and the second port 26, or can be located on the same side as one or more of the first port 24 and the second port 26.

[0050] The piezoelectric transducer 14 can be bonded to the sample vessel 12. The bond can be thin relative to the thickness of the piezoelectric transducer 14 and the sample vessel 12. The piezoelectric transducer 14 can be bonded to the sample vessel 12 by an adhesive. The adhesive can be configured to allow wave propagation with low wave loss. In one embodiment, a liquid adhesive can be applied to the piezoelectric transducer 14, and the piezoelectric transducer 14 can then be attached to the sample vessel 12 via the liquid adhesive. For example, a liquid adhesive can be applied that has temperature stability up to 350°C, excellent adhesion to glass, and high hardness (rigidity). In one example, the liquid adhesive can be an epoxy adhesive such as EPO-TEK 353ND (manufactured by Epoxy Technology, Inc., 14 Fortune Drive, Billerica, MA), which allows ultrasonic propagation and has a Shore D hardness of approximately 85. In one example, approximately 5 μl of liquid adhesive can be applied. The piezoelectric transducer 14 can be clamped to the sample vessel 12 and the adhesive can be cured at approximately 150°C. In one implementation, after curing, the adhesive can have a thickness in the range of approximately 10 μm to 100 μm, more preferably in the range of 10 μm to 20 μm. In other embodiments, the adhesive can be a pre-fabricated adhesive layer film or a UV-curable epoxy.

[0051] The piezoelectric transducer 14 can be configured to convert voltage into another form of energy, such as sound waves having one or more frequencies and / or frequency ranges into coupled solids and liquids having different frequencies. The piezoelectric transducer 14 can be configured to oscillate when an alternating current is applied to the piezoelectric transducer 14, thereby producing sound waves that can be introduced into the sample vessel 12 and create one or more nodes in the blood sample 52 within the sample vessel 12. As shown in FIG. 1 , the piezoelectric transducer 14 can include a first electrode 90 and a second electrode 92 configured to connect to an alternating current source. In one embodiment, the piezoelectric transducer 14 can be a piezoelectric ultrasonic transducer.

[0052] The piezoelectric transducer 14 can be configured to generate ultrasonic activity and produce sound waves having frequencies by expanding and contracting when exposed to an alternating electric field, for example, when an electrical frequency and voltage is applied. Figure 12 shows a graph of an example of the total displacement of the piezoelectric transducer 14 during one exemplary operation of the piezoelectric transducer 14.

[0053] In one embodiment, the piezoelectric transducer 14 can be configured to generate ultrasonic waves having a resonant frequency that causes resonance in the blood sample 52 within the microchannel 22 of the sample vessel 12, thereby rupturing the walls of red blood cells in the blood sample 52. In one embodiment, the piezoelectric transducer 14 can be configured to generate ultrasonic sound waves (sometimes referred to herein as ultrasonic waves) having a resonant frequency that causes cavitation in the blood sample 52, which creates bubbles that collapse in the region of higher pressure, creating shock waves that then rupture the walls of the red blood cells, releasing hemoglobin into the plasma of the blood sample. In one embodiment, the piezoelectric transducer 14 has a first resonant frequency and the integral structure of the lysis device 10 has a second resonant frequency spectrally spaced from the first resonant frequency, the second resonant frequency being the frequency of the sound waves generated by the piezoelectric transducer 14 and introduced into the sample vessel 12, thereby causing cavitation in the blood sample 52, thereby rupturing the walls of the red blood cells.

[0054] 13 and 14, the second resonant frequency can induce one or more standing waves inside the microchannel 22 of the sample vessel 12, which can form in regions (called nodes) of the microchannel 22 with approximately zero force and approximately zero particle motion and the highest hydraulic pressure, thereby rupturing the walls of red blood cells in the blood sample 52. Standing waves, also known as stationary waves, are waves with a peak amplitude profile that oscillates in time but is stationary in space.

[0055] In one example, at the second resonance of the acoustophoretic dissolution device 10 (i.e., the sample vessel 12 bonded to the piezoelectric transducer 14), for example, when the sample vessel 12 is made of glass, the microchannel 22 has a width of approximately 2 millimeters and an aspect ratio of 0.05 to 0.125, and the sample vessel 12 has a width of approximately 12 millimeters and an aspect ratio of 1.4 to 1.9, the piezoelectric transducer 14 can be configured to generate ultrasound waves in the range of 330 kHz to 350 kHz, with a peak pressure of 5 MPa in the microchannel 22 (see FIG. 13) and a peak velocity of up to 8 m / s (see FIG. 14). An exemplary case of the pressure distribution ( FIG. 13 ) and fluid velocity ( FIG. 14 ) of the blood sample 52 in the microchannel 22 when the piezoelectric transducer 14 is activated is shown in FIGS. 13 and 14 .

[0056] However, the ultrasonic waves in the microchannel 22 and the ultrasonic piezoelectric transducer 14 may generate undesired heat, including undesired heat in the blood sample 52 in the microchannel 22. To avoid any overheating of the blood sample 52, the piezoelectric transducer 14 may be operated to generate a resonant frequency for a predetermined period of time. For example, the piezoelectric transducer 14 may be operated to generate sound waves having the second resonant frequency for about 1 second to about 2 seconds. In one embodiment, the piezoelectric transducer 14 may be operated to generate sound waves having the second resonant frequency for less than about 1.5 seconds. In one example, the lysing device 10 may be configured to operate the piezoelectric transducer 14 for 1.5 seconds or less, resulting in 99.99% lysis of red blood cells. In one example, the lysing device 10 may be configured to operate the piezoelectric transducer 14 for about 10 seconds or less.

[0057] In one embodiment, ultrasound within the microchannel 22 breaks down blood cells and cell walls into small particles that cause less light scattering during optical measurements of the blood sample 52 than larger particles.

[0058] In one embodiment, the piezoelectric transducer 14 may be configured to generate ultrasonic waves over a wide range of frequencies, and the second resonant frequency may be a wide range of frequencies.

[0059] In one embodiment, the piezoelectric transducer 14 may be configured to produce ultrasound waves in a frequency range greater than about 300 kHz.

[0060] The resonant frequency and / or frequency range can be determined based on one or more factors, including the size, shape, and material of the sample vessel 12; the size and shape of the microchannels in the sample vessel 12; the amount of fluid in the fluid sample 52; and / or the size, shape, and material of the piezoelectric transducer 14.

[0061] For example, when the sample vessel 12 is made of glass, the microchannel 22 has an aspect ratio of about 0.05 to about 0.125, and the sample vessel 12 has an aspect ratio of about 1.4 to about 1.9, the piezoelectric transducer 14 can be configured to generate ultrasound waves in the range of about 330 kHz to about 350 kHz.

[0062] The width of the microchannel 22 can be determined based at least on the wave propagation speed (e.g., about 1500 m / s) in the blood sample 52, using a predetermined desired number of nodes in the center of the microchannel 22 as one node to have a frequency of about 330 kHz to about 350 kHz. The following formula can be used to at least partially determine the first node (having an exemplary width of 2000 μm and a depth of 100 μm) in the microchannel 22, without considering any minor reflections or other reflections:

[0063] 2f=v / λ

[0064] where f is the frequency, v is the wave velocity in the fluid, and λ is the wavelength (where the wavelength is ½ the width of the microchannel 22).

[0065] Because the resonant frequency of the sample vessel 12 can be difficult to precisely calculate due to manufacturing and / or material dispersion, in one embodiment, the piezoelectric transducer 14 can be configured to sweep frequencies within a frequency range having multiple frequencies, starting with a first frequency and progressing through one or more second frequencies to a third frequency of the multiple frequencies. In one embodiment, the piezoelectric transducer 14 can be configured to sweep the frequency range in increments, such as 1 kHz increments. In one embodiment, sweeping the frequency range from the first frequency to the third frequency ensures that the resonant frequency for the lysis device 10 and blood sample 52 is reached, even when taking into account the geometry and material dispersion of the lysis device 10.

[0066] In one embodiment, the piezoelectric transducer 14 can be configured to sweep a frequency range from about 330 kHz to about 350 kHz, such as in increments of about 1 kHz. For example, the piezoelectric transducer 14 can be configured to sweep a frequency range starting from about 330 kHz to about 350 kHz, and / or the piezoelectric transducer 14 can be configured to sweep a frequency range starting from about 350 kHz to about 330 kHz.

[0067] In one embodiment, the piezoelectric transducer 14 can be configured to sweep a frequency range over a period of greater than 0 seconds and less than 5 seconds, less than 4 seconds, less than 3 seconds, less than 2 seconds, and / or less than 1 second. In one embodiment, the piezoelectric transducer 14 can be configured to sweep a frequency range within a period of about 1 to about 2 seconds.

[0068] In one embodiment, the lysis device 10 can additionally or alternatively lyse blood cells in a blood sample 52 by inducing shear and vibration modes within the microchannel 22 of the sample vessel 12. The displacement of the rigidly bonded ultrasonic piezoelectric transducer 14 can be primarily transverse, causing vibration and movement of the sample vessel 12 bonded to the piezoelectric transducer 14. When activated, the ultrasonic piezoelectric transducer 14 changes shape, contracting and expanding (transverse displacement), as shown in FIG. 12. The movement of the ultrasonic piezoelectric transducer 14 is translated to the sample vessel 12, changing the geometry and / or volume of the microchannel 22 and inducing shear and vibration within the microchannel 22 of the sample vessel 12. FIG. 12 shows a graph of an example of the total displacement of the piezoelectric transducer 14 during one exemplary operation of the piezoelectric transducer 14.

[0069] Displacement of the piezoelectric transducer 14 can result in bending of the sample vessel 12, vibrations and / or shear forces within the sample vessel 12, which in turn can cause and / or contribute to lysis of the blood sample 52 within the microchannel 22 of the sample vessel 12 through a combination of high pressure, shear forces, and / or fluid motion within the microchannel 22. Thus, in some implementations, lysis of the blood sample 52 within the microchannel 22 can be caused by a combination of standing waves, pressure, cavitation, shear forces, and / or fluid motion within the blood sample 52.

[0070] When the piezoelectric transducer 14 is activated, a shear stress is generated at the junction between the piezoelectric transducer 14 and the sample vessel 12. This shear stress can create a high pressure inside the microchannel 22. For example, in one embodiment, a preferred high pressure can be about 5 MPa. In one embodiment, the pressure can be in the range of about 3 MPa to about 7 MPa. The level of pressure can be controlled by the level of contraction / extension of the piezoelectric transducer 14, which can depend on the electric field strength of the piezoelectric transducer 14.

[0071] The combination of standing waves within the microchannel 22, along with shear and / or vibration of the sample vessel 12, causes significant cavitation in the whole blood sample 52 within the microchannel 22, thereby causing cell wall rupture.

[0072] 15-18, in some embodiments, the lysis device 10 can be a component of an analyzer 100. The analyzer 100 can include the lysis device 10, an absorption spectrophotometer 102, a fluid distribution system 104 (e.g., including a peristaltic pump), and / or a controller 106. In one embodiment, the lysis device 10 is removable and / or replaceable from other components of the analyzer 100. In one embodiment, the analyzer 100 can further include a mount 108 configured to receive and / or position the lysis device 10. In one embodiment, the lysis device 10 can be held (e.g., clamped) within the mount 108 to allow the lysis device 10 to vibrate and / or move within a range of vibration and / or movement.

[0073] In one embodiment, the analyzer 100 may further include one or more processors 140 and one or more non-transitory computer-readable media 142. In one embodiment, the one or more processors 140 and one or more non-transitory computer-readable media 142 may be part of the controller 106. However, it will be understood that one or more of the processors 140 and / or non-transitory computer-readable media 142 may be located external to the controller 106 and / or external to other components of the analyzer 100. In one implementation, the analyzer 100 may include and / or be connectable to one or more sensor cartridges 143 having blood gas sensors 144 and / or one or more reagent cartridges 145.

[0074] In one embodiment, the absorption spectrophotometer 102 can include a transmitter 112 and a receiver 114 located adjacent to the sample vessel 12, where the transmitter 112 is positioned to emit a light beam 116 through the top surface 40, the bottom surface 42, and the microchannel 22, and the receiver 114 is positioned to receive at least a portion of the light beam 116 after that portion has passed through the top surface 40, the bottom surface 42, and the microchannel 22. In one embodiment, the transmitter 112 can be a light source, and the light beam 116 can be light. The light source can be, for example, one or more light-emitting diodes. In one embodiment, the light can be white light having a wavelength in the range of approximately 450 to 700 nanometers.

[0075] The absorption spectrophotometer 102 can be configured to measure the intensity of light in a portion of the spectrum transmitted or emitted by particular substances in the fluid sample 52, particularly in the microchannel 22 of the sample vessel 12. The absorption spectrophotometer 102 can be configured to measure how much light a chemical absorbs by measuring the intensity of the light as a beam of light passes through the blood sample 52 or other fluid sample 52. Each compound in the sample or solution absorbs or transmits light within a particular range of wavelengths.

[0076] The fluid distribution system 104 (see FIG. 17 ) can have an inlet 120 (see FIG. 16 ) fluidly connectable to the first port 24 of the sample vessel 12 of the lysis device 10 and an outlet 122 (see FIG. 16 ) fluidly connectable to the second port 26. The fluid distribution system 104 can move one or more fluid samples 52, such as a blank sample, a blood sample, or a wash solution, through the inlet 120 and the first port 24 into the microchannel 22 of the sample vessel 12. In one embodiment, the fluid distribution system 104 can flow through the microchannel 22 and eject material within the microchannel 22 through the second port 26 of the sample vessel 12 and out the outlet 122. The fluid distribution system 104 can be operated automatically, manually, or a combination of automatically and manually.

[0077] The controller 106 may be electrically connected to the piezoelectric transducer 14 of the dissolution device 10. The controller 106 may be configured to provide an electrical signal to the piezoelectric transducer 14 that, when received by the piezoelectric transducer 14, causes the piezoelectric transducer 14 to emit ultrasonic waves at one or more frequencies and / or frequency ranges that include the resonant frequency of the combined structure of the dissolution device 10 and the fluid sample 52.

[0078] 16 , in one embodiment, the controller 106 may have a first electrical contact 130 and a second electrical contact 132. The first electrical contact 130 and the second electrical contact 132 may be electrically connectable to the first electrode 90 and the second electrode 92, respectively, of the piezoelectric transducer 14 of the dissolution device 10, and thus may provide an electrical potential to the piezoelectric transducer 14.

[0079] The mount 108 can hold the dissolving device 10 in a fixed position between the transmitter 112 and the receiver 114 and can position the dissolving device 10 for operative connection to the fluid distribution system 104 and the controller 106 (see FIG. 17 ). The mount 108 can be configured to stabilize the dissolving device 10 in position without applying forces that would significantly change the impedance of the integral structure of the dissolving device 10. For example, the mount 108 can include one or more fasteners that apply a clamping force of about 20 Newtons (N) or less.

[0080] In one embodiment, analyzer 100 can further include one or more digital temperature sensors and / or one or more thermal control elements (such as Peltier elements). In one embodiment, analyzer 100 includes a side holder with two digital temperature sensors (near the inlet and outlet) and two Peltier elements for thermal control.

[0081] In one embodiment, a method of analyzing blood includes obtaining or receiving a blood sample 52; introducing a lysis device 10 between a transmitter 112 and a receiver 114 of an absorption spectrophotometer 102; introducing the blood sample 52 into the microchannel 22 of the sample vessel 12 via an inlet 120 and a first port 24 by a fluid distribution system 104; activating a controller 106 to provide an electrical signal to the piezoelectric transducer 14, which, when received by the piezoelectric transducer 14, activates the lysis device 10. The method may include emitting ultrasonic waves at one or more frequencies and / or frequency ranges that include a resonant frequency of the integral structure of the lysed blood sample 52 and the blood sample 52, and / or causing the piezoelectric transducer 14 to expand and contract, thereby creating a shear force on the blood sample 52 within the microchannel 22; inducing cavitation in the blood sample 52, causing the walls of red blood cells in the blood sample 52 to break down; and activating the absorption spectrophotometer 102 to transmit a light beam 116 from the transmitter 112 through the lysed blood sample 52 to the receiver 114.

[0082] The method may further include reading the electrical signal generated by the receiver 114 to determine one or more oximetry parameters of the lysed blood sample 52 based at least in part on the signal indicative of the light received by the receiver 114 of the absorption spectrophotometer 102.

[0083] The absorption spectrum can be calculated based on known calculations for absorption of liquid media, as shown in Figure 19. Additionally, as shown in Figure 20, determining the one or more oximetry parameters can further include analyzing spectral profile coefficients of hemoglobin types, such as oxyhemoglobin (O2HB), deoxyhemoglobin (HHB), carboxyhemoglobin (COHB), methemoglobin (METHB), and plasma bilirubin (NBILI), as well as one or more of the interfering substances cyanmethemoglobin (CN_MET_B), sulfhemoglobin (SULF_HIGH), and methylene blue (METH_BLUE_A).

[0084] Determining one or more oximetry parameters can be based on spectrophotometric light absorption, ie, measuring the absorption of light by constituents in blood sample 52 .

[0085] Determining the one or more oximetry parameters can include measuring at least total hemoglobin (THB) and one or more of hemoglobin fractions, such as oxyhemoglobin (OHB), deoxyhemoglobin (HHB), carboxyhemoglobin (COHB), and methemoglobin (METHB).

[0086] In one embodiment, the method can include introducing and evacuating a wash solution into the microchannel 22 of the sample vessel 12 before and / or after introducing the blood sample 52 into the microchannel 22. The method can further include activating the piezoelectric transducer 14 to create wave and / or shear forces to agitate the wash solution in the microchannel 22. In one embodiment, the sample vessel 12 can be used, washed, and reused. In one embodiment, the lysing device 10 is not reusable and can be replaced for each new blood sample 52. In this embodiment, the lysing device 10 can be disposed of after a single use.

[0087] The method can further include calibrating the analyzer with a blank sample. In one embodiment, the fluid sample 52 can be a test sample, known as a "blank sample," that can be used to calibrate the analyzer 100 and has known optical characteristics / properties. The blank sample can include a die solution that can be used to measure the scattering of light transmission through a medium.

[0088] In one embodiment, the blood sample 52 may be approximately 12 microliters in volume. A blood sample typically contains plasma and red blood cells (which may make up 45% to 60% of the blood sample), and optionally lipids.

[0089] In one embodiment, blood sample 52 is maintained at a consistent temperature. In one embodiment, the temperature of blood sample 52 is 37 degrees Celsius plus or minus approximately 0.3 degrees Celsius. In one embodiment, the temperature of blood sample 52 is less than 45 degrees Celsius, preferably less than 40 degrees Celsius, to avoid damage to blood sample 52. In one embodiment, blood sample 52 is maintained at a substantially consistent temperature utilizing one or more digital temperature sensors and / or one or more thermal control elements. Thermal control can be used to maintain a patient's blood sample at a constant temperature for accurate measurement of blood sample oximetry analytes.

[0090] An example of the analyzer 100 and lysis device 10 in use will now be described. In one example, the sample vessel 12 can be made of glass and can have a length-to-width aspect ratio in the range of about 1.4 to about 1.9, and the microchannel 22 can have a height-to-width aspect ratio of about 0.05 (e.g., a height of about 100 micrometers and a width of about 2 millimeters). The sample vessel 12 can be inserted into a path along which a light beam travels between a transmitter 112 and a receiver 114 of an absorption spectrophotometer 102. It should be understood that the analyzer 100 can include various components, including mirrors and / or waveguides, for directing the light beam through the path. A fluid distribution system 104 can insert a blood sample 52 into the microchannel 22 of the sample vessel 12.

[0091] The controller 106 can be electrically connected to the piezoelectric transducer 14 of the sample vessel 12 and can provide electrical signals to the piezoelectric transducer 14 to cause the piezoelectric transducer 14 to emit ultrasonic waves in the frequency range of about 330 kHz to about 350 kHz in increments of about 1 kHz. This frequency range can be transmitted within a period of about 2 seconds.

[0092] In one embodiment, the non-transitory computer-readable medium 142 can store computer-executable instructions that, when executed by the one or more processors 140 of the controller 106, can cause the one or more processors 140 to pass signals to a piezoelectric transducer 14 connected to a sample vessel 12 having a microchannel 22 containing a whole blood sample 52 having blood cells and plasma, such that the signals cause the piezoelectric transducer 14 to emit ultrasonic waves into the sample vessel 12 at a frequency, intensity, and duration to induce cavitation in the blood sample, thus creating acoustic standing waves configured to disrupt, or lyse, the blood cells in the whole blood sample 52, thereby liberating hemoglobin from within the blood cells into the plasma. In some embodiments, the sample can be pre-separated plasma, in which case such a sample can be analyzed without lysing the red blood cells, i.e., without causing the piezoelectric transducer 14 to emit ultrasonic waves in the frequency range of about 330 kHz to about 350 kHz.

[0093] The frequency range includes a resonant frequency of the lysing device 10 relative to the integral structure of the blood sample 52, thereby inducing cavitation in the blood sample 52, thereby rupturing the cell walls of the blood cells in the blood sample 52. Additionally or alternatively, the controller 106 can cause the one or more processors 140 to pass signals to the piezoelectric transducers 14, which are configured to cause the piezoelectric transducers 14 to expand and contract, thereby inducing cavitation in the blood sample 52, i.e., creating shear forces, and thus acoustic standing waves, i.e., rupturing the cell walls of the blood cells in the blood sample 52 and releasing hemoglobin from within the blood cells into plasma.

[0094] It is capable of destroying a large portion (more than 50%) of the cell wall of blood cells.

[0095] The transmitter 112 of the absorption spectrophotometer 102 can be activated to transmit a light beam 116, such as light, through the sample vessel 12 and into the lysed blood sample 52. The receiver 114 can receive at least some portion of the light beam 116 that exits the lysed blood sample 52 and the sample vessel 12. The receiver 114 can include, for example, one or more photodiodes to generate an electrical signal upon receiving the light beam 116.

[0096] The analyzer 100, or one or more computer processors 140, can determine one or more analytes present in the lysed blood sample 52 based at least in part on the signal indicative of the light received by the receiver 114 of the absorption spectrophotometer 102. The analyzer 100, or one or more computer processors, can further analyze spectral profile coefficients of hemoglobin types, such as oxyhemoglobin (OH), deoxyhemoglobin (H), carboxyhemoglobin (CO), methemoglobin (MET), and plasma bilirubin (NBILI), as well as one or more of the interfering substances cyanmethemoglobin (CN_MET_B), sulfhemoglobin (SULF_HIGH), and methylene blue (METH_BLUE_A).

[0097] The analyzer 100, or one or more computer processors 140, can measure total hemoglobin (THB) and / or one or more of the hemoglobin fractions, such as oxyhemoglobin (OHB), deoxyhemoglobin (HHB), carboxyhemoglobin (COHB), and methemoglobin (METHB).

[0098] The analyzer 100, or one or more computer processors 140, can output the results of the analysis. The output can be shown on one or more displays. This output can be used to make treatment decisions for the patient.

[0099] 21-23, an assembly constructed in accordance with the present disclosure is shown and designated by the reference numeral 200. Assembly 200 comprises a support substrate 202 and an acoustophoretic device 204 that is preferably permanently bonded to support substrate 202 in a clamping-free manner. This clamping-free method is used to minimize vibration losses between support substrate 202 and acoustophoretic device 204, referred to as a near-zero-mass interface. Previous research has shown that adding additional mass to assembly 200 reduces its ability to hemolyze blood. When acoustophoretic device 204 is mechanically constrained by clamping (e.g., attached to support substrate 202), acoustic waves / energy are transmitted throughout the connected assembly, thus reducing the energy available for building a standing wave field in microchannel 22. A zero-mass interface refers to a connection that adds no additional mass to acoustophoretic device 204, which is a theoretically ideal situation; in practice, support substrate 202 adds additional mass to acoustophoretic device 204. As described in more detail below, acoustophoretic device 204 can be attached to support substrate 202 in a manner that allows acoustophoretic device 204 to vibrate more freely than if acoustophoretic device 204 were connected to support substrate 202 by a clamping method. Additionally, acoustophoretic device 204 can be attached to support substrate 202 in a manner that provides a near-massless interface that provides efficient excitation (e.g., voltages less than 150 Vp-p, in some embodiments 80 Vp-p to 100 Vp-p) and magnitude and phase response signals.

[0100] In some embodiments, acoustophoretic device 204 comprises a sample vessel 212 and a piezoelectric transducer 214 bonded to sample vessel 212. In some embodiments, piezoelectric transducer 214 extends through an opening 272 in support substrate 202 (see FIGS. 23, 24). In one embodiment, acoustophoretic device 204 is a unitary structure, formed by sample vessel 212 and piezoelectric transducer 214 bonded together using a suitable bonding material, such as, for example, epoxy.

[0101] The sample vessel 212 has an exterior surface 220, a microchannel 222 (shown in phantom in FIG. 21 ) within the boundaries of the exterior surface 220, a first port 224 extending through the exterior surface 220 into and in fluid communication with the microchannel 222, and a second port 226 extending through the exterior surface 220 into and in fluid communication with the microchannel 222. In one embodiment, the exterior surface 220 has an attachment area for a piezoelectric transducer 214.

[0102] In one embodiment, the sample vessel 212 has a top surface 240, a bottom surface 242, a first end 244, a second end 246, a first side surface 248, and a second side surface 250, where the first side surface 248 and the second side surface 250 extend between the first end 244 and the second end 246 and between the top surface 240 and the bottom surface 242. In one embodiment, the top surface 240 and the bottom surface 242 are planar. In one embodiment, the first side surface 248 and the second side surface 250 are planar. In one embodiment, the first end 244 and the second end 246 are planar. In one embodiment, the top surface 240, the bottom surface 242, the first end 244, the second end 246, the first side surface 248, and the second side surface 250 cooperate to form a three-dimensional rectangular parallelepiped. In some embodiments, the piezoelectric transducer 214 matingly engages the exterior surface 220 of the sample vessel 212. For example, the piezoelectric transducer 214 and the exterior surface 220 can have flat surfaces configured to rest together.

[0103] Sample vessel 212 can be partially, substantially, or completely transparent. In one embodiment, sample vessel 212 is transparent at least above and below microchannel 222, such that a light beam can pass through sample vessel 212, travel through microchannel 222, interact with any material within microchannel 222, and exit sample vessel 212.

[0104] The sample vessel 212 can be constructed from glass. In one embodiment, the sample vessel 212 can be constructed from a material (glass or non-glass) having a Young's modulus in the range of about 50 GPa to about 90 GPa. The material property known as Young's modulus or elastic modulus is a measure of a material's ability to withstand a change in length when subjected to longitudinal tension or compression. Young's modulus is equal to the longitudinal stress divided by the strain. In one embodiment, the sample vessel 212 can be constructed from a plastic having a stiffness and / or Young's modulus similar to that of glass. In one embodiment, the sample vessel 212 can be constructed from alkali borosilicate glass. An example of alkali borosilicate glass is made by Schott Advanced Optics, 400 York Avenue, Duryea, PA 18642, and is commercially available under the name "D263T ECO Thin Glass."

[0105] The sample vessel 212 has a length L from the first end 244 to the second end 246, a width W from the first side 248 to the second side 250, a thickness between the top surface 240 and the bottom surface 242, and an aspect ratio defining a proportional relationship between the length and width. The sample vessel 212 has a longitudinal axis along the length and a latitudinal axis along the width.

[0106] In one embodiment, the aspect ratio of the sample vessel 212 is in the range of about 0.5 to about 3.0. In one embodiment, the aspect ratio of the sample vessel 212 is in the range of about 1.4 to about 1.9. In one embodiment, the length can be about 22 millimeters and the width can be about 12 millimeters. In one embodiment, the length can be about 17 millimeters and the width can be about 12 millimeters. In one embodiment, the length can be about 17 millimeters and the width can be about 6 millimeters. In one embodiment, the length can be about 12 millimeters and the width can be about 6 millimeters.

[0107] The microchannel 222 can be configured to receive a fluid sample (including, but not limited to, a blood sample, a "blank" sample, and / or a wash solution sample) through the first port 224 and / or the second port 226. The microchannel 222 has a length, a width, and a height. Typically, the length of the microchannel 222 is oriented along the longitudinal axis of the sample vessel 212, and the width of the microchannel 222 is oriented along the latitudinal axis of the sample vessel 212. However, it will be understood that the microchannel 222 can be oriented at an angle or offset from the longitudinal and / or latitudinal axes of the sample vessel 212.

[0108] Microchannel 222 has an aspect ratio that defines a proportional relationship between the width and height of microchannel 222. In one embodiment, the width-to-height aspect ratio of microchannel 222 is in the range of about 0.04 to about 0.175. In one embodiment, the width-to-height aspect ratio of microchannel 222 is in the range of about 0.04 to about 0.125. In one embodiment, the width-to-height aspect ratio of microchannel 222 is about 0.05.

[0109] In one embodiment, the width of the microchannel 222 is approximately 2 millimeters. In one embodiment, the width of the microchannel 222 is approximately 2.5 millimeters. In one embodiment, the width of the microchannel 222 is greater than the illumination width of the light yield area of ​​the absorbance spectrophotometer 102. The illumination width can be defined as the width of the cross section of the light yield along the optical path from the absorbance spectrophotometer 102 that intersects with the microchannel 222. For example, when the illumination diameter is between 1 millimeter and 1.5 millimeters, the width of the microchannel 222 can be at least approximately 1.6 millimeters. The width of the microchannel 222 can be determined to allow sufficient mechanical alignment between the microchannel 222 and the optical path. For example, for an illumination width of between 1 millimeter and 1.5 millimeters, the width of the microchannel 222 can be approximately 2 millimeters.

[0110] In one embodiment, the length of the microchannel 222 can be from about 10 millimeters to about 12 millimeters. In one embodiment, the length of the microchannel 222 can be at least about 4 millimeters. In one embodiment, the length of the microchannel 222 can be from about 4 millimeters to about 20 millimeters.

[0111] In one embodiment, the length of the microchannel 222 can be based at least in part on a predetermined desired number of nodes to be created within the microchannel 222. For example, if the microchannel 222 has a width of approximately 2 millimeters and the whole blood wave propagation velocity is approximately 1500 m / s, the calculated single node frequency is 350 kHz. The nodes can be uniformly spaced and dispersed within the microchannel 222 along the length of the microchannel 222 (e.g., 2 x 2 mm = 4 mm), and the high pressure produces an even distribution of lysed blood. For example, if the predetermined desired number of nodes on each sidewall of the microchannel 222 is five nodes (see FIG. 13), the length of the microchannel 222 can be set to approximately 17 millimeters.

[0112] The height of the microchannel 222 can vary, as discussed above. The height of the microchannel 222 can be based on the amount of absorption in the lysed blood of the light yield from the absorption spectrophotometer 102 and the desired accuracy of the absorption. For example, the desired absorption can be about 1 optical density (OD).

[0113] In one embodiment, the height of microchannel 222 is about 100 micrometers. In one embodiment, the height of microchannel 222 is about 150 micrometers. In one embodiment, the height of microchannel 222 is about 250 micrometers. In one embodiment, the height of microchannel 222 is about 300 micrometers. In one embodiment, the height of microchannel 222 is between about 80 micrometers and about 300 micrometers. In one embodiment, the height of microchannel 222 is between about 80 micrometers and about 150 micrometers.

[0114] The first port 224 and the second port 226 are fluidly connected to the microchannel 222 and extend from the microchannel 222 through the exterior surface 220 of the sample vessel 212. In one embodiment, the first port 224 is fluidly connected to the microchannel 222 and can extend from the microchannel 222 to the top surface 240, the bottom surface 242, the first end 244, the second end 246, the first side surface 248, and / or the second side surface 250 of the sample vessel 212. In the example shown in FIG. 21 , the first port 224 and the second port 226 extend from the bottom surface 242 of the sample vessel 212 to the microchannel 212. In one embodiment, the second port 226 is fluidly connected to the microchannel 222 and can extend from the microchannel 222 to the top surface 240, the bottom surface 242, the first end 244, the second end 246, the first side 248, and / or the second side 250 of the sample vessel 212. The first port 224 and the second port 226 can extend to the same or different ones of the top surface 240, the bottom surface 242, the first end 244, the second end 246, the first side 248, and / or the second side 250.

[0115] In one embodiment, the first port 224 and the second port 226 each have a diameter of about 0.5 millimeters (500 micrometers) to about 1.5 millimeters (1500 micrometers). In one embodiment, the first port 224 and the second port 226 each have a diameter of about 0.8 millimeters (800 micrometers). The microchannel 222 tapers toward the first port 224 and the second port 226, as shown in FIG. 11B. The taper aids in providing fluid to and / or from the first port 224. The cross-sectional widths (e.g., diameters) of the first port 224 and the second port 226 are smaller than the width of the microchannel 222. For example, the cross-sectional widths of the first port 224 and the second port 226 can be 50% to 100% of the width of the microchannel 222.

[0116] The sample vessel 212 can be monolithically fabricated, in that the sample vessel 212 is formed from a single piece of material, or in that the sample vessel 212 is formed from multiple pieces of material that are interconnected to form an integrated whole. As discussed in more detail with respect to FIG. 24, the sample vessel 212 can be formed from two substrates bonded together, as shown in FIG. 11B. Alternatively, the sample vessel 212 can be formed from three substrates bonded together, as shown in FIG. 11A. Referring again to FIG. 21, the piezoelectric transducer 214 is attached to the sample vessel 212 (e.g., to a mounting area on the exterior surface 220) after assembly to form the unitary structure of the acoustophoresis device 204. The piezoelectric transducer 214 can have a mounting area that is attached to the mounting area on the exterior surface 220. In one embodiment, the piezoelectric transducer 214 is at least partially attached to the top surface 240 of the sample vessel 212, although it will be understood that the piezoelectric transducer 214 can be attached to the top surface 240, the bottom surface 242, the first end 244, the second end 246, the first side 248, and / or the second side 250. The piezoelectric transducer 214 is positioned relative to the microchannel 222 such that the piezoelectric transducer 214 does not obstruct light moving through the microchannel 222 from the top or bottom surfaces of the sample vessel 212. The piezoelectric transducer 214 can be offset from the microchannel 222 such that the piezoelectric transducer 214 allows light to enter the microchannel 222 from outside the sample vessel 212. In one embodiment, the piezoelectric transducer 214 has a length and a longitudinal axis along its length that is oriented substantially parallel (e.g., within 5 degrees of parallel) to the longitudinal axis of the sample vessel 212. In one embodiment, the piezoelectric transducer 214 has a width that is less than the length of the piezoelectric transducer 214 .

[0117] The piezoelectric transducer 214 may be located on the opposite side of the sample vessel 212 from one or both of the first port 224 and the second port 226, or may be located on the same side as one or more of the first port 224 and the second port 226. The piezoelectric transducer 214 may be constructed and operated similarly to the piezoelectric transducer 14 described above.

[0118] The support substrate 202 includes a top surface 260, a bottom surface 262 opposite the top surface 260, and a peripheral edge 264 (see FIG. 22 ). The support substrate 202 also has an inner edge 270 defining an opening 272 that intersects the top surface 260 and the bottom surface 262. The sample vessel 212 is connected to at least one of the top surface 260 and the bottom surface 262. In the example shown, the piezoelectric transducer 214 passes through the opening 272. In some embodiments, the piezoelectric transducer 214 and the sample vessel 212 extend in opposite directions relative to the support substrate 202. For example, the piezoelectric transducer 214 can pass through the opening 272 and extend beyond the top surface 260 of the support substrate 202. As shown in FIG. 21, the sample vessel 212 is connected to and extends from the lower surface 262 but does not extend into or through the opening 272 in the support substrate 202 .

[0119] In some embodiments, the support substrate 202 includes a predetermined pattern of conductive traces 274 (see FIGS. 22 and 23 ) on at least one of the top surface 260 and the bottom surface 262. As described in more detail below, the conductive traces 274 allow for electrical connection to the heating elements and respective control systems, as well as providing a device configured for capacitive fluid detection. In some embodiments, the controller 106 is electrically connected to the piezoelectric transducer 214 and configured to simultaneously drive the piezoelectric transducer 214 and receive a response signal from the piezoelectric transducer 214. The controller 106 can be electrically connected to the piezoelectric transducer without the conductive traces 274 (alternatively, electrically connected via at least one wire), or the controller 106 can be electrically connected to the piezoelectric transducer 214 by at least one of the conductive traces 274. At least some of the conductive traces 274 include a connector portion 275 (see FIG. 23 ) adjacent the outer periphery 264 (see FIGS. 23 and 27 ). In the illustrated example, the conductive traces 274 include twelve connector portions 275, labeled 275a-l for clarity (see FIG. 27). At least some of the conductive traces 274 also include mounting pads 276 (see FIG. 28). The connector portions 275 adjacent the outer periphery edge 264 form a male edge connector 277. The mounting pads 276 can be connected to particular ones of the connector portions 275 by wires 278 (see FIG. 27). The wires 278 can extend to the top surface 260 or the bottom surface 262 of the support substrate 202. Additionally, the wires 278 can extend between the bottom surface 262 and the top surface 260, for example, by using through vias.

[0120] 28, support substrate 202 includes ten mounting pads 276, designated as mounting pads 276a-j, on bottom surface 262 and one mounting pad 276k on top surface 260 (see FIG. 27). Mounting pads 276a-k are electrically connected to corresponding ones of connector portions 275a-l to allow electrical signals to be transmitted to and / or received from mounting pads 276a-k. More or fewer mounting pads 276 may also be provided on support substrate 202. When sample vessel 212 is connected to support substrate 202, at least two of mounting pads 276 are bonded to sample vessel 212.

[0121] In some embodiments, the support substrate 202 can be a printed circuit board (PCB) constructed from plastic and fiber resin. Standard materials for the fabrication of printed circuit boards are FR4 G10 fiberglass epoxy, phenolic resin, polyimide, etc.

[0122] 25 and 26, in some embodiments, the sample vessel 212 has a plurality of conductive traces 282 on the exterior surface 220. In the example shown, the sample vessel 212 includes nine conductive traces 282a-i. Each of the conductive traces 282a-i includes at least one mounting pad 284. In the example shown, the conductive traces 282a-i include mounting pads 284a-t.

[0123] Mounting pads 284a, 284b, 284c, 284d, and 284e are spaced apart and electrically isolated from one another. Mounting pads 284a, 284b, 284c, 284d, and 284e are located along first end 244 of sample vessel 212. Mounting pads 284a, 284b, 284c, 284d, and 284e are bonded, e.g., soldered, to mounting pads 276a-e to provide mechanical and electrical connections therebetween.

[0124] Mounting pads 284p, 284q, 284r, 284s, and 284t are spaced apart and electrically isolated from one another. Mounting pads 284p, 284q, 284r, 284s, and 284t are spaced apart a distance from mounting pads 284a, 284b, 284c, 284d, and 284e. For example, mounting pads 284p, 284q, 284r, 284s, and 284t can be located along second end 246 of sample vessel 212. Mounting pads 284p, 284q, 284r, 284s, and 284t are bonded, e.g., soldered, to mounting pads 276f-j on support substrate 202 to provide mechanical and electrical connections therebetween.

[0125] The conductive traces 282 on the sample vessel 212 can have different configurations and can be used for different purposes.

[0126] In particular, conductive trace 282a includes mounting pad 284a. Conductive trace 282a has a first portion 288 designated to receive piezoelectric transducer 214 within the mounting area of ​​top surface 240 and a second portion 290 outside the mounting area and electrically connected to mounting pad 284a.

[0127] The conductive trace 282b includes and extends from a mounting pad 284b in a first sensor portion 294. The first sensor portion 294 extends along the microchannel 222 and is used to form part of a capacitive sensor that can be used to determine the expected channel contents, e.g., the fluid type of a sample (e.g., blood), air, or aqueous solution, in the microchannel 222, by measuring the capacitance and correlating the capacitance with known dielectric properties of the expected channel contents in the microchannel 222. The dielectric properties of the expected channel contents can be determined empirically using techniques known in the art.

[0128] Conductive traces 282c are contained within mounting pads 284c and are used solely to connect the sample vessel 212 to the support substrate 202.

[0129] Conductive trace 282d includes two mounting pads 284d and 284f. Mounting pad 284d can be mechanically and electrically connected to mounting pad 276d. Mounting pad 284f can be connected to a lead of an electrical device. In the example shown, sample vessel 212 can include a thermistor 300 attached to mounting pads 284f and 284g (see FIG. 26). Thermistor 300 senses the temperature of sample vessel exterior surface 220 and provides a temperature signal to mounting pads 284d and 284q.

[0130] Conductive trace 282e includes six mounting pads 284e, 284j, 284k, 284l, 284m, and 284q. Mounting pads 284e and 284q can be mechanically and electrically connected to mounting pads 276e and 276g. Mounting pads 284j, 284k, 284l, and 284m can each be connected to a lead of an electrical device. In the example shown in FIG. 26, the sample vessel includes four electric heaters 302a-d. Electric heaters 302a-d can be used to generate and provide heat to sample vessel 212 by supplying electricity to electric heaters 302a-d. As discussed above, thermal control can be used to maintain a constant temperature of the patient's blood sample to accurately measure blood sample oximetry analytes. Electric heaters 302a-d can be connected in parallel by mounting pads 284j, 284h, 284k, 284i, 284l, 284o, 284m, and 284n. In this example, electric heater 302a is connected to mounting pads 284j and 284h; electric heater 302b is connected to mounting pads 284k and 284i; electric heater 302c is connected to mounting pads 284l and 284o; and electric heater 302d is connected to mounting pads 284m and 284n. Conductive trace 282e can also be used in combination with conductive trace 282b to form a capacitive sensor. In this regard, conductive trace 282e also includes a second sensor portion 304, which can extend within ±5 degrees of parallelism (preferably parallel) to first sensor portion 294. In the embodiment shown, both the first sensor portion 294 and the second sensor portion 304 are linearly configured and are adjacent to but do not cover the microchannel 222 so as not to obstruct the beam of light generated by the spectrophotometer, as discussed herein.In some embodiments, the first sensor portion 294 and the second sensor portion 304 may or may not be parallel, so long as the first sensor portion 294 and the second sensor portion 304 do not obstruct the beam 116, allowing capacitance readings from the first sensor portion 294 and the second sensor portion 304 to be correlated to expected channel contents in the microchannel 222 during calibration. In some embodiments, the first sensor portion 294 and / or the second sensor portion 304 may have a serpentine configuration, with one or more portions crossing over the microchannel 222 outside of the expected path of the beam 116 through the sample vessel 212.

[0131] Conductive trace 282f includes five mounting pads 284h, 284i, 284o, 284n, and 284p, the use of which is described above. Mounting pad 284p is configured to mechanically and electrically connect to mounting pad 276f.

[0132] Conductive traces 282g-I are contained within mounting pads 284r, 284s, and 284t and are used solely to mechanically connect sample vessel 212 to support substrate 202, for example, by reflow soldering in standard automated processing equipment used in electronics manufacturing.

[0133] In some embodiments, the conductive traces 282a-t are formed having conductive electrode patterns applied to the top surface 240 of the sample vessel 212 by any suitable technique, such as physical vapor deposition, sputter deposition, etc. The conductive electrode traces 282a-t can be made from any suitable conductive material, such as a metal, including but not limited to copper, aluminum, etc.

[0134] 24 and 11B, to create the assembly 200, first and second glass substrates 310, 312 for creating the sample vessel 212 can be fabricated on a wafer. The first glass substrate 310 includes a top surface 314 and a bottom surface 316. The top surface 314 of the first glass substrate 310 can be flat and forms the top surface 240 of the sample vessel 212 when the glass substrates 310, 312 are bonded together. The bottom surface 316 of the first glass substrate 310 can also be flat. In some embodiments, the first glass substrate 310 can be 700 micrometers thick. In some embodiments, the first glass substrate 310 does not include the microchannel 222 and the input and output ports 224 and 226. The second glass substrate 312 includes a top surface 320 on which the microchannel 222 is formed. The second glass substrate 312 is also formed with a first port 224 and a second port 226. The microchannel 222, the first port 224, and the second port 226 can be etched into a portion of the second glass substrate 312. The bottom surface 316 of the first glass substrate 310 is connected to the top surface 320 of the second glass substrate 312. In some embodiments, the bottom surface 316 of the first glass substrate 310 is connected to the top surface 320 of the second glass substrate 312 by annealing to completely encompass and surround the microchannel 222. In some embodiments, the second glass substrate 312 can be 700 micrometers thick.

[0135] A conductive electrode pattern is applied to a first glass substrate 310 (or 70 shown in FIG. 11A ), which is intended to be used as the top surface 240 when the glass substrates 310, 312 are part of a wafer. Electrical components such as thermistor 300 and electric heaters 302a-d are soldered to predetermined mounting pads 284 of the conductive traces 282. The substrates 310, 312 are then diced and singulated from the wafer. Matching pairs of substrates 310, 312 are bonded together such that the first substrate 310 covers the microchannel 222 in the second substrate 312 to form the sample vessel 212. 24 and 26 , the piezoelectric transducer 214 has a size smaller than that of the glass substrates 310, 312, and a peripheral portion of the piezoelectric transducer 214 is attached to a portion of the conductive electrode pattern including the first portion 288 of the conductive trace 282 a, such that some portions of the periphery of the piezoelectric transducer 214 are indirectly bonded to the top surface 314 of the first glass substrate 310, and most of the piezoelectric transducer 214 remains available for direct bonding to the top surface 314 within the bonding section 317 of the first glass substrate 310 shown in FIG. 25 . That is, the conductive electrode pattern covers a portion of the top surface 314 of the first glass substrate 310, and the bonding section 317, i.e., another portion of the top surface 314 of the first glass substrate 310, is left exposed to be directly bonded to the piezoelectric transducer 214. Figure 26 shows a sample vessel 212 in which the piezoelectric transducer 214 does not extend beyond the surface area of ​​the glass substrates 310, 312 when the integrated structure is formed. As shown in Figure 24, the piezoelectric transducer 214 is positioned within the mounting area so as to overlap the first portion 288 of the conductive trace 282a and is then bonded to the first glass substrate 310 using any suitable epoxy 318, such as EPO-Tek 353-ND. A sample inlet 320 is inserted into the first port 224 and bonded to the bottom surface 242 of the sample vessel 212 with an epoxy 330, such as an epoxy sold under the brand name LOCKTITE444.Sample outlet 322 is inserted into second port 226 and bonded to the bottom surface 242 of sample vessel 212 with epoxy 332, such as epoxy sold under the brand name LOCKTITE 444. After sample inlet 320 and sample outlet 322 are connected to sample vessel 212, mounting pads 284a-e and 284p-t are then soldered to mounting pads 276a-j to mechanically and electrically connect sample vessel 212 to support substrate 202. Flexible wire 340 is then soldered to mounting pad 276k and to a terminal on piezoelectric transducer 214.

[0136] In use, the assembly 200 can be a component of the analyzer 100. The analyzer 100 can include the assembly 200, the absorbance spectrophotometer 102, the fluid distribution system 104 (e.g., including a peristaltic pump), and / or the controller 106, as described above. In one embodiment, a male edge connector, e.g., connector portion 275 (shown in FIG. 27 ) adjacent the other periphery 264, is plugged into a female edge connector of the analyzer 100. In this position, the microchannel 222 (or 22 shown in FIG. 15 ) is located between the transmitter 112 and the receiver 114 of the absorbance spectrophotometer 102 of the analyzer 100. The assembly 200 can be removable and / or replaceable from the other components of the analyzer 100.

[0137] After the male edge connector is plugged into the female edge connector of the analyzer 100, the controller 106 forms a circuit with the piezoelectric transducer 214, the thermistor 300, the heaters 302a-d, the first sensor portion 294, and the second sensor portion 304. The controller 106 is configured to control the frequency and / or voltage of the AC signal supplied to the piezoelectric transducer 214. In addition, the controller 106 is configured to control the temperature of the sample vessel 212 by supplying current to the heaters 302a-d and receiving temperature feedback from the thermistor 300. The amount of current supplied to the heaters 302a-d can be varied to obtain a desired temperature of the sample vessel.

[0138] 29 shows another embodiment of a support substrate constructed in accordance with the present disclosure and is designated by the reference numeral 400. Support substrate 400 is identical in structure and use to support substrate 202, except that support substrate 400 includes at least one non-via opening 402 between an inner edge 404 and an outer edge 406, and this non-via opening 402 is not used to provide an electrical connection between layers or surfaces of support substrate 400, but rather refers to a means for restraining sample vessel 212 so as to provide minimally attenuated, sustained electrical contact.

[0139] 30 shows another embodiment of a support substrate constructed in accordance with the present disclosure and is designated by reference numeral 420. Support substrate 420 includes a first side 422, a second side 424, a first end 426, and a second end 428. Support substrate 420 is identical in structure and use to support substrate 202, except that support substrate 420 includes a first tab 430 connected to first side 422 and a second tab 432 connected to second side 424, with first tab 430 and second tab 432 extending toward each other. These tabs serve as a means of creating a minimal-mass interface contact with sample vessel 212 and help minimize acoustic attenuation.

[0140] Referring now to FIG. 31 , in some embodiments, assembly 200 can be a component of analyzer 100a. Analyzer 100a is similar in structure and function to analyzer 100 described above with reference to FIGS. 15-18 , except that analyzer 100a includes an edge connector 500, illustratively described herein as a female edge connector 500 configured to mechanically and electrically connect to a male edge connector 277 of support substrate 202. Elements common between analyzers 100 and 100a are labeled in FIG. 31 . Also, while FIG. 31 references support substrate 202, it should be understood that the support substrate can be support substrate 400 or 420. As discussed above with reference to FIGS. 23 and 27 , connector portion 275 is provided adjacent outer periphery 264. Connector portion 275 can be provided on top surface 260 and / or bottom surface 262 of support substrate 202. In the example shown in FIG. 31 , the connector portions 275 are provided on the top surface 260. In this embodiment, the connector portions 275 are coplanar. The connector portions 275 are spaced apart along at least a portion of the outer periphery 264. The female edge connector 500 can include a support member 501 connected to a plurality of spring contacts 502 constructed from a conductive material. In the example shown, the female edge connector 500 includes nine spring contacts 502, each positioned to contact a specific one of the connector portions 275. Each of the spring contacts 502 can be biased downward (in the example shown) and can be spaced apart from one or two adjacently disposed spring contacts 502. The spacing between the spring contacts 502 can be predetermined to match the spacing between the connector portions 275. The spring contacts 502 are conductive and electrically connected to other electrical components of the analyzer 100a, such as the controller 106. To electrically connect the assembly 200 to other electrical components of the analyzer 100a, the male edge connector 277 is biased against the spring contact 502. Continued movement of the male edge connector 277 toward the spring contact 502 allows the spring contact 502 to contact the connector portion 275.Moving the male edge connector 277 in the opposite direction, away from the spring contacts 502, allows the assembly 200 to be disconnected from the analyzer 100a.

[0141] To support assembly 200 within analyzer 100a, analyzer 100a is provided with a carriage 540 having an upper surface 542. Carriage 540 has a plurality of walls 546, at least partially defining a recess 548. In the illustrated embodiment, carriage 540 includes walls 546a, 546b, 546c, and 546d. Assembly 200 is positioned within recess 548 and supported by upper surface 542. More specifically, at least two of walls 546a-d are configured to matingly engage with outer periphery 264 of support substrate 202. In the illustrated example, walls 546b and 546d are in the form of tabs. Support substrate 202 is shaped such that outer periphery 264 forms first recess 550 and second recess 552. Wall 546b is positioned within first recess 550, and wall 546d is positioned within second recess 552. Wall 546 b is positioned within first recess 550 and wall 546 d is positioned within second recess 552 to precisely align assembly 200 within carriage 540 .

[0142] Carriage 540 is movable toward and away from female edge connector 500, as indicated by arrow 560. More specifically, to install assembly 200 into analyzer 100a, carriage 540 is moved away from female edge connector 500, so that assembly 200 can be placed within recess 548 without male edge connector 277 engaging female edge connector 500. After assembly 200 is positioned within recess 548, carriage 540 is moved toward female edge connector 500, engaging male edge connector 277 with female edge connector 500.

[0143] In some non-limiting embodiments, the carriage 540 is supported by a guide assembly (not shown) and moved by a motorized control system. The guide assembly can include tracks, wheels, and / or bearings.

[0144] The following is a numerical list of non-limiting exemplary embodiments of the inventive concepts disclosed herein:

[0145] 1. An acoustophoretic device comprising: a sample vessel having an exterior surface, a microchannel within the confines of the exterior surface, a first port extending through the exterior surface into the microchannel, and a second port extending through the exterior surface into the microchannel, the microchannel configured to receive a blood sample through the first port, the sample vessel having conductive traces on the exterior surface; and a piezoelectric transducer formed on an exterior surface of the sample vessel to form an integral structure, the piezoelectric transducer contacting at least one of the conductive traces and configured to generate ultrasonic waves in the sample within the microchannel, the piezoelectric transducer having an excitation input and a response signal output electrically connected to at least one of the conductive traces.

[0146] 2. The acoustophoretic device of exemplary embodiment 1, wherein the sample vessel is constructed from glass.

[0147] 3. The acoustophoretic device of any one of exemplary embodiments 1-2, wherein the sample vessel includes a first substrate bonded to a piezoelectric transducer and a second substrate having a microchannel, a first port, and a second port.

[0148] 4. An acoustophoretic device as described in any one of exemplary embodiments 1 to 3, wherein the microchannel has a length, a width, and a height, the microchannel has a first side extending along the length of the microchannel and a second side extending along the length of the microchannel, a first conductive trace among the conductive traces includes a first mounting pad and a first sensor portion, the first sensor portion extending along the first side of the microchannel, and a second conductive trace among the conductive traces includes a second mounting pad and a second sensor portion, the second sensor portion extending along the second side of the microchannel.

[0149] 5. An acoustophoretic device as described in any one of exemplary embodiments 1 to 4, wherein the outer surface is a first outer surface having an attachment area, the attachment area having a first shape, at least one of the conductive traces having a first portion within the attachment area and a second portion outside the attachment area, the piezoelectric transducer having a second outer surface having a second shape corresponding to the first shape, and the second outer surface of the piezoelectric transducer is bonded to the attachment area.

[0150] 6. The acoustophoretic device of any one of exemplary embodiments 1-5, wherein at least one of the conductive traces is located between the piezoelectric transducer and the outer surface of the sample vessel.

[0151] 7. The acoustophoretic device of any one of exemplary embodiments 1-6, wherein the conductive traces are bonded in direct contact with the outer surface of the sample vessel.

[0152] 8. The acoustophoretic device of any one of exemplary embodiments 1-7, wherein the sample vessel has a periphery and the conductive trace includes a conductive mounting pad located adjacent the periphery.

[0153] 9. An acoustophoretic device as described in any one of exemplary embodiments 1-8, wherein the sample vessel has a first end and a second end, and the conductive trace includes a conductive mounting pad located adjacent to at least one of the first end and the second end of the sample vessel.

[0154] 10. An acoustophoretic device as described in any one of exemplary embodiments 1-9, further comprising an electrical component, wherein the sample vessel has an outer periphery, a first conductive trace among the conductive traces includes a first mounting pad and a second mounting pad electrically connected to the first mounting pad, a second conductive trace among the conductive traces includes a third mounting pad and a fourth mounting pad electrically connected to the third mounting pad, the first mounting pad and the third mounting pad are located adjacent to the outer periphery, and the electrical component has a first lead connected to the second mounting pad and a second lead connected to the fourth mounting pad.

[0155] 11. The acoustophoretic device of any one of exemplary embodiments 1-10, wherein the electrical component is a thermistor.

[0156] 12. An acoustophoretic device as described in any one of exemplary embodiments 1 to 11, wherein the second mounting pad has a first length and a first width, the first length being greater than the first width, the fourth mounting pad has a second length and a second width, the second length being greater than the second width, and the first length and second length extend within 5 degrees of parallelism.

[0157] 13. The acoustophoretic device of any one of exemplary embodiments 1-12, wherein the electrical components include a first electrical heater and a second electrical heater, the first electrical heater and the second electrical heater being connected in parallel to the second mounting pad and the fourth mounting pad.

[0158] 14. An assembly comprising: a support substrate; and an acoustophoretic device permanently bonded to a support substrate, the acoustophoretic device comprising: a sample vessel having an exterior surface, a microchannel within the confines of the exterior surface, a first port extending through the exterior surface into the microchannel, and a second port extending through the exterior surface into the microchannel, wherein a blood sample is insertable into the microchannel through the first port, the sample vessel having conductive traces on the exterior surface; and a piezoelectric transducer bonded to an outer surface of the sample vessel to form an integral structure, the piezoelectric transducer contacting at least one of the conductive traces and configured to generate ultrasonic standing waves in the blood sample in the microchannel, the piezoelectric transducer having a power input electrically connected to at least one of the conductive traces.

[0159] 15. An assembly according to any one of the preceding exemplary embodiments, wherein the support substrate includes an upper surface, a lower surface opposite the upper surface, and an outer peripheral edge, the support substrate having an inner edge defining an opening that intersects the upper and lower surfaces, and the sample vessel is connected to one of the upper and lower surfaces.

[0160] 16. An assembly according to any one of the preceding exemplary embodiments, wherein the support substrate includes a predetermined pattern of conductive traces on at least one of its upper and lower surfaces, at least some of the conductive traces including mounting pads, and at least two of the mounting pads are bonded to the sample vessel.

[0161] 17. The assembly of any one of the preceding exemplary embodiments, wherein the conductive trace is a first conductive trace, the mounting pad is a first mounting pad, the sample vessel has a plurality of second conductive traces on an exterior surface having second mounting pads, and the first mounting pad is bonded to the second mounting pad.

[0162] 18. The assembly of any one of the preceding exemplary embodiments, wherein the first mounting pad is soldered to the second mounting pad.

[0163] 19. The assembly of exemplary embodiment 17 or 18, wherein the first mounting pad is located adjacent the inner edge.

[0164] 20. The assembly of any one of the preceding exemplary embodiments, wherein the support substrate is a circuit board.

[0165] 21. The assembly of any one of the preceding exemplary embodiments, wherein the piezoelectric transducer extends through an opening in the support substrate.

[0166] 22. An analyzer comprising: 1. An acoustophoretic device comprising: a sample vessel having an exterior surface, a microchannel within the confines of the exterior surface, a first port extending through the exterior surface into the microchannel, and a second port extending through the exterior surface into the microchannel, wherein a sample is insertable into the microchannel through the first port, the sample vessel having conductive traces on the exterior surface; and an acoustophoretic device including a piezoelectric transducer bonded to an exterior surface of the sample vessel to form a unitary structure, the piezoelectric transducer contacting at least one of the conductive traces and configured to generate ultrasonic waves in the sample within the microchannel, causing the sample vessel to vibrate such that shear forces are induced within the microchannel; an absorption spectrophotometer including a transmitter and a receiver located adjacent to the sample vessel, the transmitter positioned to emit a light beam through the microchannel and the receiver positioned to receive at least a portion of the light beam after that portion has passed through the microchannel; a fluid distribution system having an outlet connected to a first port and an inlet connected to a second port; a controller electrically connected to the piezoelectric transducer and configured to provide an electrical signal to the piezoelectric transducer, wherein when the electrical signal is received by the piezoelectric transducer, the piezoelectric transducer emits ultrasonic waves and the piezoelectric transducer contracts and expands.

[0167] 23. The analyzer of exemplary embodiment 22, wherein the outer surface of the sample vessel has a first side and a second side opposite the first side, the transmitter is located on the first side of the sample vessel, the receiver is located on the second side of the sample vessel, and the sample vessel is constructed from a material that is transparent to the light beam.

[0168] 24. An analyzer according to any one of the preceding exemplary embodiments, wherein the outer surface of the sample vessel has a first side and a second side opposite the first side, and the first side and the second side are planar.

[0169] 25. The analyzer of any one of the preceding exemplary embodiments, wherein the sample vessel is constructed from glass.

[0170] 26. The analyzer of any one of the preceding exemplary embodiments, wherein the sample vessel is constructed from a non-glass material having a Young's modulus in the range of approximately 50 Gpa to 90 Gpa.

[0171] 27. An analyzer as described in any one of the preceding exemplary embodiments, wherein the outer surface of the sample vessel is a first outer surface having a mounting area, the mounting area having a first shape, the piezoelectric transducer has a second outer surface having a second shape corresponding to the first shape, and the second outer surface of the piezoelectric transducer is bonded to the mounting area.

[0172] 28. The analyzer of any one of exemplary embodiments 22-27, wherein the piezoelectric transducer matingly engages the outer surface of the sample vessel.

[0173] 29. An analyzer according to any one of exemplary embodiments 22-28, wherein the height of the microchannel is about 100 micrometers and the width of the microchannel is about 2 millimeters.

[0174] 30. An analyzer described in any one of exemplary embodiments 22 to 30, further comprising a support substrate including an upper surface, a lower surface opposite the upper surface, and an outer peripheral edge, the support substrate having an inner edge defining an opening that intersects the upper and lower surfaces, and the sample vessel being connected to one of the upper and lower surfaces.

[0175] 31. An analyzer according to any one of the preceding exemplary embodiments, wherein the support substrate includes a predetermined pattern of conductive traces on at least one of the upper and lower surfaces, at least some of the conductive traces include mounting pads, and at least two of the mounting pads are bonded to the sample vessel.

[0176] 32. An analyzer according to any one of the preceding exemplary embodiments, wherein the conductive trace is a first conductive trace, the mounting pad is a first mounting pad, the sample vessel has a plurality of second conductive traces on an outer surface having a second mounting pad, and the first mounting pad is bonded to the second mounting pad.

[0177] 33. The analyzer of any one of the preceding exemplary embodiments, wherein the first mounting pad is soldered to the second mounting pad.

[0178] 34. The analyzer of any one of the preceding exemplary embodiments, wherein the first mounting pad is located adjacent the inner edge.

[0179] 35. The analyzer of any one of the preceding exemplary embodiments, wherein the support substrate is a circuit board.

[0180] 36. The analyzer of any one of the preceding exemplary embodiments, wherein the piezoelectric transducer extends through an opening in the support substrate.

[0181] 37. An analyzer described in any one of the preceding exemplary embodiments, wherein the support substrate includes a predetermined pattern of conductive traces on at least one of the upper and lower surfaces, and at least some of the conductive traces include a plurality of connector portions provided adjacent the outer peripheral edge.

[0182] 38. The analyzer of any one of the preceding exemplary embodiments, further comprising an edge connector, the edge connector including a support member connected to a plurality of spring contacts, the spring contacts constructed from a conductive material in communication with the controller, and a specific one of the spring contacts engaging a specific one of the connector portions to provide an electrical connection between the conductive traces of the support substrate and the spring contacts.

[0183] 39. A method of fabricating an acoustophoretic device comprising: 1. A method comprising bonding a piezoelectric transducer to an exterior surface of a sample vessel to form an integral structure, wherein a first portion of a conductive trace extends between the piezoelectric transducer and the exterior surface of the sample vessel, the sample vessel having a microchannel within a boundary of the exterior surface, a first port extending through the exterior surface into the microchannel, and a second port extending through the exterior surface into the microchannel, the microchannel having a length, a width, and a height.

[0184] 40. bonding conductive traces to an exterior surface of the sample vessel; 40. The method of exemplary embodiment 39, further comprising connecting at least a portion of the conductive trace on the outer surface of the sample vessel to a mounting pad of the conductive trace on the piezoelectric transducer.

[0185] 41. The method of any one of the preceding exemplary embodiments, wherein the support substrate has an opening, and the step of connecting at least a portion of the conductive trace to the support substrate includes positioning the sample vessel to span the opening, and then connecting at least a portion of the conductive trace to the support substrate.

[0186] 42. The method of any one of the preceding exemplary embodiments, wherein mounting pads of conductive traces on the outer surface of the sample vessel are soldered to mounting pads of conductive traces on the support substrate.

[0187] conclusion As discussed above, assembly 200 comprises support substrate 202, 400, or 420 and acoustophoretic device 204 preferably permanently bonded in a non-clamping manner to support substrate 202, 400, or 420. Acoustophoretic device 204 can be attached to support substrate 202, 400, or 420 in a manner that allows acoustophoretic device 204 to vibrate more freely than if acoustophoretic device 204 were connected to support substrate 202, 400, or 420 in a clamping manner. Furthermore, acoustophoretic device 204 can be attached to support substrate 202 in a manner that provides a near-massless interface that provides efficient excitation (e.g., voltages less than 150 Vp-p, in some embodiments 80 Vp-p to 100 Vp-p) and a response signal with a high signal-to-noise ratio. The support substrate 202, 400, or 420 may be, for example, a printed circuit board and may include a predetermined pattern of conductive traces 274 on at least one of the top surface 260 and bottom surface 262 of the support substrate 202, 400, or 420 to enable electrical connection to components on the support substrate 202, 400, or 420 and to the acoustophoretic device 204 without wires.

[0188] The assembly 200 can be a user-replaceable consumable item, if desired, or a single-use, disposable cartridge that is disposed of after a single use. Additionally, the assembly 200 allows the entire microchannel 222 to be self-sustaining with respect to heating, hemolysis, and fluid detection. Containment of the microchannel 222 within the assembly 200 enhances the uniformity of the microchannel 222, thereby enhancing the accuracy of blood sample readings made within the microchannel 222 by the analyzer 100, 100a. Finally, the assembly 200 allows for near-zero-mass interfacing of the acoustophoresis device 204 to the support substrate 202, 400, or 420, while still electrically connecting the conductive traces 282 without wires that are bonded to the sample vessel 212 along their length. This offers the advantages of easier and less expensive fabrication, a more reliable electrical interface, and enhanced signal integrity.

[0189] The foregoing description provides illustration and description, and is not intended to be exhaustive or to limit the inventive concepts to the precise form disclosed. Modifications and variations are possible in light of the above teachings or may be acquired from practice of the methods described in the present disclosure.

[0190] Although particular combinations of features and steps are recited in the claims and / or disclosed herein, these combinations are not intended to limit the disclosure. Indeed, many of these features and steps can be combined in ways not specifically recited in the claims and / or disclosed herein. Although each dependent claim listed below may depend directly on only one other claim, the present disclosure includes each dependent claim in combination with all other claims in the claim set.

[0191] No element, act, or instruction used in this application should be construed as essential or essential to the invention unless expressly described as being outside the scope of the preferred embodiments. Further, the phrase "based on" is intended to mean "based, at least in part, on," unless expressly described otherwise.

Claims

1. 1. An acoustophoretic device comprising: a sample vessel having an exterior surface, a microchannel within the boundary of the exterior surface, a first port extending through the exterior surface into the microchannel, and a second port extending through the exterior surface into the microchannel, the microchannel configured to receive a blood sample through the first port; the sample vessel having conductive traces on the exterior surface; a piezoelectric transducer formed on an exterior surface of the sample vessel to form a unitary structure, the piezoelectric transducer contacting at least one of the conductive traces and configured to generate ultrasound waves in the blood sample in the microchannel, the piezoelectric transducer having an excitation input and a response signal output electrically connected to at least one of the conductive traces; the microchannel has a length, a width, and a height, the microchannel has a first side extending along the length of the microchannel and a second side extending along the length of the microchannel, a first one of the conductive traces different from the conductive trace contacting the piezoelectric transducer includes a first mounting pad and a first sensor portion, the first sensor portion extending along the first side of the microchannel, a second one of the conductive traces different from the conductive trace contacting the piezoelectric transducer includes a second mounting pad and a second sensor portion, the second sensor portion extending along the second side of the microchannel; the first sensor portion of the first conductive trace and the second sensor portion of the second conductive trace form part of a capacitive sensor; The acoustophoretic device.

2. The acoustophoretic device of claim 1 , wherein the sample vessel is constructed from glass.

3. 10. The acoustophoretic device of claim 1, wherein the sample vessel comprises a first substrate bonded to a piezoelectric transducer and a second substrate having a microchannel, a first port, and a second port.

4. The exterior surface is a first exterior surface having an attachment area, the attachment area having a first shape.

2. The acoustophoretic device of claim 1, wherein at least one of the conductive traces has a first portion within the mounting area and a second portion outside the mounting area, the piezoelectric transducer has a second outer surface having a second shape corresponding to the first shape, and the second outer surface of the piezoelectric transducer is bonded to the mounting area.

5. The acoustophoretic device of claim 1 , wherein at least one of the conductive traces is located between the piezoelectric transducer and an exterior surface of the sample vessel.

6. 10. The acoustophoretic device of claim 1, wherein the conductive traces are bonded in direct contact with the exterior surface of the sample vessel.

7. The acoustophoretic device of claim 1 , wherein the sample vessel has a perimeter and the conductive trace includes a conductive mounting pad located adjacent the perimeter.

8. 10. The acoustophoretic device of claim 1, wherein the sample vessel has a first end and a second end, and the conductive trace includes a conductive mounting pad located adjacent at least one of the first end and the second end of the sample vessel.

9. 10. The acoustophoretic device of claim 1, further comprising an electrical component, wherein the sample vessel has a periphery, a first one of the conductive traces includes a first mounting pad and a second mounting pad electrically connected to the first mounting pad, a second one of the conductive traces includes a third mounting pad and a fourth mounting pad electrically connected to the third mounting pad, the first and third mounting pads being located adjacent the periphery, and the electrical component has a first lead connected to the second mounting pad and a second lead connected to the fourth mounting pad.

10. The acoustophoretic device of claim 9 , wherein the electrical component is a thermistor.

11. 10. The acoustophoretic device of claim 9, wherein the second mounting pad has a first length and a first width, the first length being greater than the first width, the fourth mounting pad has a second length and a second width, the second length being greater than the second width, and the first length and second length extending within 5 degrees of parallelism.

12. 12. The acoustophoretic device of claim 11, wherein the electrical components include a first electrical heater and a second electrical heater, the first electrical heater and the second electrical heater connected in parallel to the second mounting pad and the fourth mounting pad.

13. An assembly comprising: a support substrate; and an acoustophoretic device permanently bonded to the support substrate, the acoustophoretic device comprising: a sample vessel having an exterior surface, a microchannel within the confines of the exterior surface, a first port extending through the exterior surface into the microchannel, and a second port extending through the exterior surface into the microchannel, wherein a blood sample is insertable into the microchannel through the first port, the sample vessel having conductive traces on the exterior surface; a piezoelectric transducer bonded to an exterior surface of the sample vessel to form a unitary structure, the piezoelectric transducer contacting at least one of the conductive traces and configured to generate an ultrasonic standing wave in the blood sample in the microchannel, the piezoelectric transducer having an excitation signal input and a response signal output electrically connected to at least one of the conductive traces; the microchannel has a length, a width, and a height, the microchannel has a first side extending along the length of the microchannel and a second side extending along the length of the microchannel, a first one of the conductive traces different from the conductive trace contacting the piezoelectric transducer includes a first mounting pad and a first sensor portion, the first sensor portion extending along the first side of the microchannel, a second one of the conductive traces different from the conductive trace contacting the piezoelectric transducer includes a second mounting pad and a second sensor portion, the second sensor portion extending along the second side of the microchannel; the first sensor portion of the first conductive trace and the second sensor portion of the second conductive trace form part of a capacitive sensor; the support substrate includes an upper surface, a lower surface opposite the upper surface, and an outer periphery, the support substrate having an inner edge defining an opening that intersects the upper and lower surfaces, the sample vessel being connected to one of the upper and lower surfaces; The piezoelectric transducer extends through an opening in the support substrate. The assembly.

14. 14. The assembly of claim 13, wherein the support substrate includes a predetermined pattern of conductive traces on at least one of its top and bottom surfaces, at least some of the conductive traces on the support substrate include mounting pads, and at least two of the mounting pads are bonded to the sample vessel.

15. The assembly described in claim 14, wherein the conductive trace on the support substrate is a fifth conductive trace, the mounting pad is a fifth mounting pad, the sample vessel has a plurality of sixth conductive traces on an outer surface having a sixth mounting pad, and the fifth mounting pad is bonded to the sixth mounting pad.

16. 16. The assembly of claim 15, wherein the fifth mounting pad is soldered to the sixth mounting pad.

17. The assembly of claim 15 , wherein the fifth mounting pad is located adjacent the inner edge.

18. The assembly of claim 13 , wherein the support substrate is a circuit board.

19. 1. An analyzer comprising:

1. An acoustophoretic device comprising: a sample vessel having an exterior surface, a microchannel within the confines of the exterior surface, a first port extending through the exterior surface into the microchannel, and a second port extending through the exterior surface into the microchannel, wherein a sample is insertable into the microchannel through the first port, the sample vessel having conductive traces on the exterior surface; and a piezoelectric transducer bonded to an exterior surface of the sample vessel to form an integral structure, the piezoelectric transducer contacting at least one of the conductive traces and configured to generate ultrasonic waves in the sample within the microchannel, causing the sample vessel to vibrate such that shear forces are induced within the microchannel; the microchannel has a length, a width, and a height, the microchannel has a first side extending along the length of the microchannel and a second side extending along the length of the microchannel, a first one of the conductive traces different from the conductive trace contacting the piezoelectric transducer includes a first mounting pad and a first sensor portion, the first sensor portion extending along the first side of the microchannel, a second one of the conductive traces different from the conductive trace contacting the piezoelectric transducer includes a second mounting pad and a second sensor portion, the second sensor portion extending along the second side of the microchannel; an acoustophoretic device, wherein a first sensor portion of the first conductive trace and a second sensor portion of the second conductive trace form a portion of a capacitive sensor; an absorption spectrophotometer including a transmitter and a receiver located adjacent to the sample vessel, the transmitter positioned to emit a light beam through the microchannel and the receiver positioned to receive at least a portion of the light beam after that portion has passed through the microchannel; a fluid distribution system having an outlet connected to the first port and an inlet connected to the second port; a controller electrically connected to the piezoelectric transducer and configured to provide an electrical signal to the piezoelectric transducer, wherein when the electrical signal is received by the piezoelectric transducer, the piezoelectric transducer emits ultrasonic waves and the piezoelectric transducer contracts and expands; a support substrate including an upper surface, a lower surface opposite the upper surface, and a peripheral edge, the support substrate having an inner edge defining an opening intersecting the upper and lower surfaces, the sample vessel being connected to one of the upper and lower surfaces; The piezoelectric transducer extends through an opening in the support substrate. The analyzer.

20. 20. The analyzer of claim 19, wherein the exterior surface of the sample vessel has a first side and a second side opposite the first side, the transmitter is located on the first side of the sample vessel and the receiver is located on the second side of the sample vessel, and the sample vessel is constructed from a material that is transparent to the light beam.

21. 20. The analyzer of claim 19, wherein the outer surface of the sample vessel has a first side and a second side opposite the first side, the first side and the second side being planar.

22. 20. The analyzer of claim 19, wherein the sample vessel is constructed from glass.

23. 20. The analyzer of claim 19, wherein the sample vessel is constructed from a non-glass material having a Young's modulus in the range of about 50 Gpa to 90 Gpa.

24. 20. The analyzer of claim 19, wherein the outer surface of the sample vessel is a first outer surface having a mounting area, the mounting area having a first shape, and the piezoelectric transducer has a second outer surface having a second shape corresponding to the first shape, the second outer surface of the piezoelectric transducer being bonded to the mounting area.

25. 20. The analyzer of claim 19, wherein the piezoelectric transducer matingly engages an exterior surface of the sample vessel.

26. 20. The analyzer of claim 19, wherein the height of the microchannel is about 100 micrometers and the width of the microchannel is about 2 millimeters.

27. 20. The analyzer of claim 19, wherein the support substrate includes a predetermined pattern of conductive traces on at least one of its top and bottom surfaces, at least some of the conductive traces on the support substrate include mounting pads, and at least two of the mounting pads are bonded to the sample vessel.

28. The conductive trace on the support substrate is a fifth conductive trace, and the mounting pad is a fifth conductive trace. 5 mounting pad, and the sample vessel has a plurality of sixth conductive traces on an exterior surface having the sixth mounting pad, the fifth mounting pad bonded to the sixth mounting pad.

29. 30. The analyzer of claim 28, wherein the fifth mounting pad is soldered to the sixth mounting pad.

30. 30. The analyzer of claim 28, wherein the fifth mounting pad is located adjacent the inner edge.

31. 20. The analyzer of claim 19, wherein the support substrate is a circuit board.

32. 20. The analyzer of claim 19, wherein the support substrate includes a pattern of conductive traces on at least one of its upper and lower surfaces, at least some of the conductive traces including a plurality of connector portions disposed adjacent an outer periphery.

33. 33. The analyzer of claim 32, further comprising an edge connector, the edge connector including a support member connected to a plurality of spring contacts, the spring contacts constructed from a conductive material in communication with the controller, a specific one of the spring contacts engaging a specific one of the connector portions to provide an electrical connection between the conductive traces of the support substrate and the spring contacts.

34. 10. A method of fabricating the acoustophoretic device of claim 1, comprising: bonding a piezoelectric transducer to an exterior surface of a sample vessel to form a unitary structure, a first portion of the conductive trace extending between the piezoelectric transducer and the exterior surface of the sample vessel, the sample vessel having a microchannel within a boundary of the exterior surface, a first port extending through the exterior surface into the microchannel, and a second port extending through the exterior surface into the microchannel, the microchannel having a length, a width, and a height; The method, wherein the support substrate has an opening, and the step of connecting at least a portion of the conductive trace to the support substrate includes positioning the sample vessel to span the opening and then connecting at least a portion of the conductive trace to the support substrate.

35. bonding conductive traces to an exterior surface of the sample vessel; connecting at least a portion of the conductive trace on the exterior surface of the sample vessel to a mounting pad of the conductive trace on the piezoelectric transducer; 35. The method of claim 34, further comprising:

36. 35. The method of claim 34, wherein mounting pads of the conductive traces on the exterior surface of the sample vessel are soldered to mounting pads of the conductive traces on the support substrate.

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