Acoustic electrophoresis device and method having a conductive electrode

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 claims the benefit of U.S. Provisional Application No. 63 / 366,552, filed on June 17, 2022, under 35 U.S.C. § 119(e). The entire disclosure of the above - referenced patent application is hereby expressly incorporated herein by reference.

[0002] This disclosure generally relates to devices, systems, and methods for testing blood samples. More particularly, this disclosure relates to a lysis device configured to lyse red blood cells within 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 ultrasonic waves are generated by a single piezoelectric transducer. The lysis device can be used with a blood sample test analyzer.

Background Art

[0003] Point - of - care testing generally refers to medical tests performed at or near the site of patient treatment, such as in an emergency department or an operating room. Often, the desired result of such tests is a rapid and accurate test result for determining the next course of action in a patient's treatment. A number of such point - of - care tests involve the analysis of blood samples from patients. Many of these tests use whole blood, plasma, or serum.

[0004] In some tests, the cell walls of red blood cells in a blood sample are broken (lysed) to release hemoglobin. Lysis of red blood cells is sometimes referred to as hemolysis. Typically, hemolysis is performed by chemical or mechanical means.

[0005] Some devices use ultrasound to lyse red blood cells. Some point-of-care testing devices use spectrophotometric light absorption measurements to determine oximetry parameters for whole blood samples. The oximetry parameters are also known as CO oximetry parameters. These devices are typically fluid systems that place a patient's blood sample in a sample chamber and test the blood sample. For example, one system described in Patent Document 1 ("Apparatus for Hemolyzing a Blood Sample and for Measuring at Least One Parameter Thereof," issued on August 4, 2015) uses two piezoelectric elements having two equilibrium resonance elements that symmetrically surround the sample chamber and uses ultrasound to lyse red blood cells. However, these devices are difficult and expensive to manufacture, including requiring a very precise symmetry with specially made resonance elements and infrastructure to hold all elements in an aligned state.

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

[0007] In such tests, blood parameters for life-saving emergency treatment can be measured, 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 the treatment of patients. In many cases, these measurements are performed in a central blood laboratory using large, complex analyzers that are difficult to maintain. However, obtaining rapid, accurate, and precise results in a clinical setting is desirable in many respects to save time in critical diagnostic situations and avoid problems with specimen transportation in the emergency room. Some blood gas analyzers are capable of use in the clinical setting, can be manufactured more simply and easily than existing devices, and can be easily used by clinicians, but do not present one solution that provides the desired test time, accuracy, precision, and reliability.

Prior Art Documents

Patent Documents

[0008]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0009] There is a need for a lysis device that provides improved accuracy and precision of the measured parameters of a sample within the desired test time at the point of patient treatment, can be manufactured more easily, and is of low cost.

Means for Solving the Problems

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

[0011] Consistent with one aspect of the present disclosure, an exemplary acoustophoresis device is a sample vessel having an outer surface, a microchannel within the boundary of the outer surface, a first port extending through the outer surface to the microchannel, and a second port extending through the outer surface to the microchannel, wherein a blood sample having red blood cells and plasma can be inserted into the microchannel through the first port, and the sample vessel has a conductive trace on the outer surface; a piezoelectric transducer bonded to the outer surface of the sample vessel to form an integral structure, the piezoelectric transducer being configured to generate ultrasonic waves in a sample within the microchannel and 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 width-to-height microchannel aspect ratio can be in the range of about 0.04 to about 0.175; the sample vessel has a width and a height, and the width-to-height sample vessel aspect ratio 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, and the piezoelectric transducer is configured to generate ultrasonic standing waves in a blood sample within the microchannel, vibrate the sample vessel, thereby inducing shear forces within the microchannel, and the standing waves and shear forces cause cavitation within the blood sample, thereby destroying the red blood cell walls in the blood sample and releasing hemoglobin into the plasma. The piezoelectric transducer has a length, a width, and a height, and the piezoelectric transducer can have a length in the range of 75% to 125% of the length of the channel. The piezoelectric transducer can have a height of 2 mm.

[0012] In accordance with one aspect of the present disclosure, an exemplary acoustophoresis system includes a sample vessel having an outer surface, microchannels within the boundaries of the outer surface, a first port extending through the outer surface into the microchannels, and a second port extending through the outer surface into the microchannels, wherein a sample is insertable into the microchannels through the first port, and the sample vessel has a conductive trace on the outer surface; and an acoustic transducer, such as a piezoelectric transducer, bonded to the outer surface of the sample vessel to form an integral structure, the acoustic transducer being configured to contact at least one of the conductive traces and generate ultrasonic waves in the sample within the microchannels, the piezoelectric transducer being configured to vibrate the sample vessel to induce a shear force within the microchannels; an absorption spectrophotometer including a transmitter and a receiver positioned adjacent to the sample vessel, the transmitter being positioned to emit a light beam through the microchannels, and the receiver being positioned to receive that portion of the light beam after at least a portion of the light beam has passed through the microchannels; a fluid distribution system having an outlet connected to the first port and an inlet connected to the second port; and a controller electrically connected to the piezoelectric transducer and configured to provide an electrical signal to the piezoelectric transducer, such that when the electrical signal is received by the piezoelectric transducer, the piezoelectric transducer emits ultrasonic waves, the piezoelectric transducer contracts and expands, thereby generating magnitude and phase signals in response, and the magnitude and phase signals can be measured by a processor to calibrate and adjust the acoustophoresis system for optimal performance and determine structural integrity. In some embodiments, the microchannels can have a length, a width, and a height, the width-to-height microchannel aspect ratio is in the range of about 0.04 to about 0.175, the sample vessel has a width and a height, and the width-to-height sample vessel aspect ratio is in the range of about 0.5 to about 3.0.The piezoelectric transducer is configured to generate ultrasonic standing waves in a blood sample within a microchannel, and the sample vessel is configured to vibrate such that shear forces are induced within the microchannel. The standing waves and shear forces cause cavitation within the blood sample, thereby breaking 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 are incorporated herein and constitute a part of this specification, showing one or more implementations described herein together with the description, and explaining these implementations. These drawings are not intended to be drawn to scale, and for purposes of clarity and conciseness, may emphasize certain specific configurations and specific views of these figures, and may show to scale or schematically. Not all components are labeled in all the drawings. The same reference numbers in these figures represent the same or similar elements or functions, and can refer to them.

Brief Description of the Drawings

[0015]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11A

Figure 11B

Figure 12

Figure 13

Figure 14

Figure 15

Figure 16

Figure 17

Figure 18

Figure 19

Figure 20

Figure 21

Figure 22

Figure 23

Figure 24

Figure 25

Figure 26

Figure 27

Figure 28

Figure 29

Figure 30

Figure 31

DETAILED DESCRIPTION OF THE INVENTION

[0016] The following detailed description refers to the accompanying drawings. The same reference numerals in different drawings can identify the same or similar elements.

[0017] The mechanism proposed in the present disclosure avoids the above-described problems. The present disclosure describes, hereinafter by way of example, a lysis device, an analyzer, and a lysis method that include a lysis device configured to lyse red blood cells within a sample vessel by ultrasound, shear force, pressure, and / or fluid motion generated within the sample vessel by an acoustic transducer hereinafter referred to as a piezoelectric transducer. The piezoelectric transducer is connected to the sample vessel and is driven at one or more specific excitation frequencies or excitation frequency ranges. In one non-limiting embodiment, the piezoelectric transducer is a single piezoelectric transducer. The present disclosure further describes an analyzer configured to receive and interact with the lysis device for testing a sample within 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 comprises a list of elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Further, unless expressly stated to the contrary, “or” refers to an inclusive logical disjunction rather than an exclusive logical disjunction. For example, the 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] In addition, the use of "a" or "an" is employed herein to describe the elements and components of the embodiments. This is merely done for convenience to give a schematic meaning to the concept of the invention. This description should be read as including one or more, and the singular includes the plural unless it is clear otherwise.

[0020] Furthermore, the use of the term "plurality" means conveying "two or more" unless the contrary is clearly stated.

[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 the words modify, but also any slight deviation from the exact amount or value that can occur, for example, due to manufacturing tolerances, measurement errors, wear and tear, stresses applied to various components, and combinations thereof.

[0022] As used herein, the term "substantially" means that the parameter, event, or situation described below occurs completely, or that the parameter, event, or situation described below occurs to a large extent or degree. For example, the term "substantially" means that the parameter, event, or situation described below 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 the 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 dimension or measurement being referred to.

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

[0024] The use of ordinal numbers (i.e., "first", "second", "third", "fourth", etc.) is for the sole purpose of distinguishing two or more items and does not imply any sequence or order or importance, or any additional ordering of the items, unless explicitly stated otherwise.

[0025] Finally, as used herein, any reference to "one embodiment" or "an embodiment" means that the particular element, configuration, structure, or characteristic described in connection with that embodiment is included in at least one embodiment. The recitation of the phrase "in one embodiment" in various places in this specification is not necessarily all referring to the same embodiment.

[0026] As discussed above, typical conventional devices for blood sample testing used in a clinical setting are complex, slow, inaccurate, and imprecise. The present disclosure addresses these deficiencies by a device, system, and method for lysing red blood cells within a sample vessel by ultrasonic waves, shear forces, 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 excitation frequency ranges.

[0027] Referring now to the drawings, and in particular to FIGS. 1 - 8, an acoustic phoresis lysis device 10 is shown. Generally, the lysis device 10 includes a sample vessel 12 and a piezoelectric transducer 14 joined to the sample vessel 12. In one embodiment, the lysis device 10 is a one-piece structure, formed, for example, by joining the sample vessel 12 and the piezoelectric transducer 14 together using a suitable bonding material such as epoxy.

[0028] The sample vessel 12 has an outer surface 20, a microchannel 22 within the bounds of the outer surface 20, a first port 24 extending through the outer surface 20 into the microchannel 22 and in fluid communication with the microchannel 22, and a second port 26 extending through the outer surface 20 into the microchannel 22 and in fluid communication with the microchannel 22. In one embodiment, the outer surface 20 can have an attachment 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 48, and a second side 50, with the first side 48 and the second side 50 extending 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 48 and the second side 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 48, and the second side 50 cooperate to form a three-dimensional rectangular prism.

[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, such that a light beam can pass through the sample vessel 12, through the microchannel 22, interact with some substance 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 the ability of a material to withstand a change in length when subjected to longitudinal tension or compression. Young's modulus is equal to the value obtained by dividing the longitudinal stress by the strain. In one embodiment, the sample vessel 12 can be constructed from a plastic such as a cyclic olefin copolymer having a rigidity 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 located at 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 a top surface 40 and a bottom surface 42, and an aspect ratio that defines a proportional relationship between the length and the width. The sample vessel 12 has a longitudinal axis along the length and a latitude 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 in a direction along the longitudinal axis of the sample vessel 12, and the width of the microchannel 22 is in a direction along the latitude axis of the sample vessel 12. However, it will be understood that the microchannel 22 can be angled or offset from the longitudinal axis and / or the latitude axis of the sample vessel 12.

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

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

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

[0038] In one embodiment, the length of the microchannel 22 can be at least partially based on a predetermined desired number of nodes to be created within the microchannel 22. For example, if the microchannel 22 has a width of about 2 millimeters and the whole blood wave propagation speed is about 1500 m / s, the calculated single node is 350 kHz. The nodes can be evenly spaced and dispersed along the length of the microchannel 22 within the microchannel 22 (e.g., 2×2 mm = 4 mm), and the high pressure produces an even distribution of the lysed blood. For example, if the predetermined desired number of nodes on each side wall of the microchannel 22 is 5 nodes (see FIG. 13), the length of the microchannel 22 can be set to about 17 millimeters so that the first port 24 and the second port 26 are not placed 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 absorption amount of the light yield from the absorptiometer 102 in the lysed blood 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 the microchannel 22 is about 100 micrometers. In one embodiment, the height of the microchannel 22 is about 150 micrometers. In one embodiment, the height of the microchannel 22 is about 250 micrometers. In one embodiment, the height of the microchannel 22 is about 300 micrometers. In one embodiment, the height of the microchannel 22 is from about 80 micrometers to about 300 micrometers. In one embodiment, the height of the microchannel 22 is from 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 outer surface 20 of the sample vessel 12. In one embodiment, the first port 24 is fluidly connected to the microchannel 22 and can extend from the microchannel 22 to the top surface 40, bottom surface 42, first end 44, second end 46, first side surface 48, and / or second side surface 50 of the sample vessel 12. In one embodiment, the second port 26 is fluidly connected to the microchannel 22 and can extend from the microchannel 22 to the top surface 40, bottom surface 42, first end 44, second end 46, first side surface 48, and / or second side surface 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, bottom surface 42, first end 44, second end 46, first side surface 48, and / or second side surface 50.

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

[0043] The sample vessel 12 can be integrally fabricated in that the sample vessel 12 is formed from a single piece of material or the sample vessel 12 is formed from a plurality of pieces of material that are interconnected and integrated to form a complete or integral structure.

[0044] As shown in FIGS. 4 to 8, in one embodiment, the sample vessel 12 can include a single substrate 60 bounded by an outer surface 20, and the substrate 60 has microchannels 22 within the single substrate 60 and a first port 24 and a second port 26 that are fluidly connected to the microchannels 22 and extend to the outer 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 microchannels 22, the first port 24, and the second port 26.

[0045] As shown in FIG. 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 stacked on the first substrate 70 to form an integral structure. In one embodiment, the first substrate 70 and the second substrate 72 can be annealed to each other. In one embodiment, the first substrate 70 and the second substrate 72 can be thermally plasma bonded to each other. 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 within the first substrate 70, within the second substrate 72, and / or can be partially formed within the first substrate 70 and partially formed within the second substrate 72. In one embodiment, the microchannel 22, the first port 24, and the second port 26 are located within 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 within the first substrate 70 and one or both of the first port 24 and the second port 26 are located within the second substrate 72. One or both of the first port 24 and the second port 26 can be located within (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 monolithic structure. In one embodiment, the first substrate 70, the second substrate 72, and the third substrate 80 can be thermally plasma bonded to each other. In one embodiment, the first substrate 70, the second substrate 72, and the third substrate 80 can be annealed to each other. One or both of the first port 24 and the second port 26 can be located within the first substrate 70. The first substrate 70 can have a thickness of 700 micrometers. The microchannel 22 can be located within 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 that passes through the third substrate 80. In one embodiment, the third substrate 80 can have the same thickness as the height of the microchannel 22. In one embodiment, the third substrate 80 can have a thickness of 100 micrometers. In other embodiments, as shown in FIG. 11B (and FIG. 24), the sample vessel 212 has only two glass substrates 310, 312 joined to each other, rather than the three substrates 70, 72, 80 shown in FIG. 11A. By using two substrates 310, 312 joined to each other instead of the three substrates 70, 72, 80, the manufacturability of the sample vessel 212 is improved without changing the performance of the sample vessel 212 that lyses red blood cells and releases hemoglobin from inside the cells into the plasma as compared to the performance of the sample vessel 12. As shown in FIG. 24, the first glass substrate 310 includes a conductive trace 282 formed on the first glass substrate 310, and the second glass substrate 312 includes a microchannel 222 and first and second ports 224, 226. As shown in FIG. 11B, the first glass substrate 310 can be stacked on the second glass substrate 312 to form a monolithic structure.In some embodiments, the conductive trace 282 (disposed on the first glass substrate 310) is formed from a conductive metal 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 (such as the attachment area of the outer surface 20) to form an integral structure of the dissolution device 10. The piezoelectric transducer 14 can have an attachment area that is attached to the attachment area of the outer 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 in relation to the microchannel 22 so as not to block the 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 as to allow light to enter the microchannel 22 from outside the sample vessel 12. In one embodiment, the piezoelectric transducer 14 has a length, and along that length, has a longitudinal axis 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 smaller than the length of the piezoelectric transducer 14.

[0049] The piezoelectric transducer 14 can be positioned on the sample vessel 12 on the opposite side of one or both of the first port 24 and the second port 26, or can be positioned 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 joined to the sample vessel 12. The joint can be made thin relative to the thicknesses of the piezoelectric transducer 14 and the sample vessel 12. The piezoelectric transducer 14 can be joined 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 then the piezoelectric transducer 14 can be attached to the sample vessel 12 through the liquid adhesive. For example, a liquid adhesive having a temperature stability of up to 350 °C, excellent adhesion to glass, and high hardness (rigidity) can be applied. In one example, the liquid adhesive can be an epoxy adhesive such as EPO-TEK 353ND (manufactured by Epoxy Technology, Inc., located at 14 Fortune Drive, Billerica, MA) that allows ultrasonic wave propagation and has a Shore D hardness of about 85. In one example, about 5 μl of the liquid adhesive can be applied. The piezoelectric transducer 14 can be clamped to the sample vessel 12 to cure the adhesive at about 150 °C. In one implementation, after curing, the thickness of the adhesive can be in the range of about 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 an ultraviolet light curable epoxy.

[0051] The piezoelectric transducer 14 can be configured to convert a voltage into another form of energy, such as acoustic waves having one or more frequencies and / or frequency ranges to a coupled solid and liquid having different frequencies. The piezoelectric transducer 14 can be configured to oscillate when an alternating current is applied to the piezoelectric transducer 14, thereby generating an acoustic wave, and such an acoustic wave 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 be connected to an alternating current power source. In one embodiment, the piezoelectric transducer 14 can be a piezoelectric ultrasonic piezoelectric transducer.

[0052] The piezoelectric transducer 14 can be configured to generate ultrasonic activity and produce acoustic waves having a frequency by expansion and contraction when exposed to an alternating electric field, such as when an electrical frequency and voltage are applied. FIG. 12 shows a graph of an example of the total displacement of the piezoelectric transducer 14 in 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, thus rupturing the walls of the red blood cells in the blood sample 52. In one embodiment, the piezoelectric transducer 14 can be configured to generate ultrasonic sound waves (also referred to herein as ultrasonic waves) having a resonant frequency that causes cavitation to generate bubbles in the blood sample 52. The bubbles collapse within regions of higher pressure to produce 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 integrated structure of the lysis device 10 has a second resonant frequency that is spectrally separated from the first resonant frequency. The second resonant frequency is the frequency of the sound waves generated by the piezoelectric transducer 14, introduced into the sample vessel 12, thereby causing cavitation in the blood sample 52, thereby rupturing the walls of the red blood cells.

[0054] In one embodiment, as shown in FIGS. 13 and 14, the second resonant frequency can cause one or more standing waves inside the microchannel 22 of the sample vessel 12. Such standing waves can be formed within regions having approximately zero force and approximately zero particle motion as well as the highest water pressure (referred to as nodes) within the microchannel 22, thus rupturing the walls of the red blood cells in the blood sample 52. Standing waves are also known as stationary waves and are waves having a peak amplitude profile that oscillates in time but does not move spatially.

[0055] In one example, at the second resonance of the acoustophoresis lysis device 10 (i.e., the sample vessel 12 joined to the piezoelectric transducer 14), for example, when the sample vessel 12 is made of glass, the microchannel 22 has a width of about 2 millimeters, the aspect ratio is 0.05 to 0.125, the sample vessel 12 has a width of about 12 millimeters, and the aspect ratio is 1.4 to 1.9, the piezoelectric transducer 14 can be configured to generate ultrasonic waves within the range of 330 kHz to 350 kHz. The peak pressure in the microchannel 22 is 5 MPa (see FIG. 13), and the peak velocity is up to 8 m / s (see FIG. 14). One 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 unwanted heat, including unwanted heat in the blood sample 52 in the microchannel 22. To avoid any overheating of the blood sample 52, the piezoelectric transducer 14 can be operated to generate a resonance frequency over a predetermined period. For example, the piezoelectric transducer 14 can be operated to generate a sound wave having a second resonance frequency over about 1 second to about 2 seconds. In one embodiment, the piezoelectric transducer 14 can be operated to generate a sound wave having a second resonance frequency over less than about 1.5 seconds. In one example, the lysis device 10 can be configured to operate the piezoelectric transducer 14 for 1.5 seconds or less, as a result of which 99.99% of red blood cells are lysed. In one example, the lysis device 10 can be configured to operate the piezoelectric transducer 14 for about 10 seconds or less.

[0057] In one embodiment, the ultrasonic waves in the microchannel 22 break blood cells and cell walls into microparticles, reducing the light scattering that occurs during the optical measurement of the blood sample 52 compared to larger particles.

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

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

[0060] The resonance 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 of 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 ultrasonic waves in the range of about 330 kHz to about 350 kHz.

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

[0063] 2f = v / λ

[0064] In the above formula, f is the frequency, v is the wave speed in the fluid, and λ is the wavelength (where the wavelength is 1 / 2 of the width of the microchannel 22).

[0065] In one embodiment, since the resonant frequency of the sample vessel 12 may be difficult to precisely calculate due to manufacturing and / or material dispersions, the piezoelectric transducer 14 is configured to sweep frequencies within a frequency range having a plurality of frequencies, starting from a first frequency and proceeding from one or more second frequencies among the plurality of frequencies to a third frequency. In one embodiment, the piezoelectric transducer 14 can be configured to sweep the frequency range in units such as frequencies in 1 kHz increments. In one embodiment, by sweeping the frequency range from the first frequency to the third frequency, it is ensured that the resonant frequency for the lysis device 10 and the blood sample 52 is reached, even considering the geometry and material dispersions of the lysis device 10.

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

[0067] In one embodiment, the piezoelectric transducer 14 can be configured to sweep the frequency range over a period 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 the frequency range within a period of about 1 to about 2 seconds.

[0068] In one embodiment, additionally or alternatively, the lysis device 10 can lyse blood cells in the 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 sometimes be mainly a transverse displacement, 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 converted 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 in one exemplary operation of the piezoelectric transducer 14.

[0069] The displacement of the piezoelectric transducer 14 can result in bending of the sample vessel 12, vibration and / or shear forces within the sample vessel 12, which can then cause and / or contribute to the lysis of the blood sample 52 within the microchannel 22 of the sample vessel 12 by 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, shear stress is generated at the junction between the piezoelectric voltage transducer 14 and the sample vessel 12. This shear stress can create 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 / expansion of the piezoelectric transducer 14, which may depend on the electric field strength of the piezoelectric transducer 14.

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

[0072] Referring now to FIGS. 15-18, in some embodiments, the lysis device 10 can be a component of the analyzer 100. The analyzer 100 can include the lysis device 10, an absorption spectrophotometer 102, a fluid dispensing 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 (such as clamped) within the mount 108 such that the lysis device 10 is capable of vibrating and / or moving within a range of vibration and / or movement.

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

[0074] In one embodiment, the absorption spectrophotometer 102 can include a transmitter 112 and a receiver 114 positioned adjacent to the sample vessel 12. The transmitter 112 is positioned to emit a light beam 116 through the top surface 40, the bottom surface 42, and the microchannel 22. The receiver 114 is positioned to receive that portion of the light beam 116 after at least a portion of the light beam 116 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 about 450 to 700 nanometers.

[0075] The absorption spectrophotometer 102 can be configured to measure the intensity of a portion of the spectrum of light transmitted or emitted by a particular substance, particularly in the fluid sample 52 within the microchannel 22 of the sample vessel 12. The absorption spectrophotometer 102 can be configured to measure how much a chemical substance absorbs light by measuring the intensity of the light when 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 specific wavelength range.

[0076] The fluid distribution system 104 (see FIG. 17) can have an inlet 120 (see FIG. 16) that is fluidly connectable to the first port 24 of the sample vessel 12 of the lysis device 10 and an outlet 122 (see FIG. 16) that is 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 discharge the material within the microchannel 22 from the outlet 122 through the second port 26 of the sample vessel 12. The fluid distribution system 104 can be operated automatically, manually, or in a combination of automatic and manual.

[0077] The controller 106 can be electrically connected to the piezoelectric transducer 14 of the dissolution device 10. The controller 106 can be configured to provide an electrical signal to the piezoelectric transducer 14. When the electrical signal is received by the piezoelectric transducer 14, the piezoelectric transducer 14 emits ultrasonic waves of one or more frequencies and / or frequency ranges including the resonance frequency of the integrated structure of the dissolution device 10 and the fluid sample 52.

[0078] As shown in FIG. 16, in one embodiment, the controller 106 can have a first electrical contact 130 and a second electrical contact 132. The first electrical contact 130 and the second electrical contact 132 can 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 can provide a potential to the piezoelectric transducer 14.

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

[0080] In one embodiment, the 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, the analyzer 100 includes a side holder having 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 can include 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 a microchannel 22 of a sample vessel 12 through an inlet 120 and a first port 24 by a fluid distribution system 104; activating a controller 106 to provide an electrical signal to a piezoelectric transducer 14, and when the electrical signal is received by the piezoelectric transducer 14, the piezoelectric transducer 14 emits ultrasonic waves of one or more frequencies and / or frequency ranges including the resonance frequency of the integrated structure of the lysis device 10 and the blood sample 52, and / or the piezoelectric transducer 14 expands and contracts, thereby generating a shear force on the blood sample 52 within the microchannel 22; inducing cavitation in the blood sample 52 to break the walls of red blood cells in the blood sample 52; 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 can further include reading an electrical signal generated by the receiver 114 and determining one or more oximetry parameters of the lysed blood sample 52 based at least in part on a signal indicative of light received by the receiver 114 of the absorption spectrophotometer 102.

[0083] As shown in FIG. 19, based on known calculations regarding the absorption of a liquid medium, an absorption spectrum can be calculated. Further, as shown in FIG. 20, determining one or more oximetry parameters can further include analyzing hemoglobin-type spectral profile coefficients, such as one or more of oxyhemoglobin (O2HB), deoxyhemoglobin (HHB), carboxyhemoglobin (COHB), methemoglobin (METHB), and plasma bilirubin (NBILI), as well as 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 measurement of light absorption, i.e., the absorption of light by components in the blood sample 52.

[0085] Determining one or more oximetry parameters can include measuring at least total hemoglobin (THB) and one or more of the hemoglobin fractions such as oxyhemoglobin (O2HB), deoxyhemoglobin (HHB), carboxyhemoglobin (COHB), methemoglobin (METHB).

[0086] In one embodiment, the method can include introducing and evacuating a cleaning 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 generate waves and / or shear forces to agitate the cleaning solution within the microchannel 22. In one embodiment, the sample vessel 12 can be used, cleaned, and reused. In one embodiment, the lysis device 10 is not reusable and can be replaced for each new blood sample 52. In this embodiment, the lysis device 10 can be discarded each time it is used.

[0087] This 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 features / characteristics. The blank sample can include a die solution that can be used to measure the scattering of light transmission through the medium.

[0088] In one embodiment, the blood sample 52 can be a volume of about 12 microliters. The blood sample typically includes plasma and red blood cells (which can account for 45% - 60% of the blood sample), and optionally lipids.

[0089] In one embodiment, the blood sample 52 is maintained at a consistent temperature. In one embodiment, the temperature of the blood sample 52 is 37 degrees Celsius ± about 0.3 degrees. In one embodiment, the temperature of the blood sample 52 is less than 45 degrees Celsius, preferably less than 40 degrees Celsius, to avoid damaging the blood sample 52. In one embodiment, the blood sample 52 is maintained at a substantially consistent temperature using one or more temperature sensors and / or one or more thermal control elements. Thermal control can be used to keep the patient's blood sample at a constant temperature to accurately measure the blood sample oximetry analyte.

[0090] An example of the analyzer 100 and the lysing device 10 in use will now be described. In one example, the sample vessel 12 can be made of glass, 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 (for example, having a height of about 100 micrometers and a width of about 2 millimeters). The sample vessel 12 can be inserted into the path through which the light beam travels between the transmitter 112 and the receiver 114 of the absorption spectrophotometer 102. It should be understood that the analyzer 100 can include various devices including mirrors and / or waveguides for guiding the light beam through the path. The fluid distribution system 104 can insert the 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 provide an electrical signal 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 1 kHz increments. 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 one or more processors 140 of the controller 106, cause the one or more processors 140 to pass a signal to the piezoelectric transducer 14 connected to the sample vessel 12 having the microchannel 22 containing the whole blood sample 52 including blood cells and plasma. These signals cause the piezoelectric transducer 14 to emit ultrasonic waves into the sample vessel 12 at a frequency, intensity, and duration for inducing cavitation in the blood sample, such that an acoustic standing wave is configured to break, i.e., lyse, the blood cells in the whole blood sample 52, thereby releasing hemoglobin from 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 red blood cells, i.e., without causing the piezoelectric transducer 14 to emit ultrasonic waves in a frequency range of about 330 kHz to about 350 kHz.

[0093] The frequency range includes the resonant frequency with respect to the integral structure of the lysis device 10 with the blood sample 52, thereby causing cavitation in the blood sample 52 and thereby breaking 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 a signal to the piezoelectric transducer 14, which signals cause the piezoelectric transducer 14 to expand and contract, thereby inducing cavitation in the blood sample 52, i.e., generating shear forces, such that an acoustic standing wave is configured to break, i.e., the cell walls of the blood cells in the blood sample 52 and release hemoglobin from the blood cells into the plasma.

[0094] Greater than 50% of the cell walls of the blood cells can be broken.

[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 into the lysed blood sample 52. The receiver 114 can receive at least some portions of the light beam 116 that have exited the lysed blood sample 52 and the sample vessel 12. The receiver 114 can include one or more photodiodes, for example, to generate an electrical signal upon reception of 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 a 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 hemoglobin type spectral profile coefficients, such as one or more of oxyhemoglobin (O2HB), deoxyhemoglobin (HHB), carboxyhemoglobin (COHB), methemoglobin (METHB), and plasma bilirubin (NBILI), as well as interfering substances cyanmethemoglobin (CN_MET_B), sulfhemoglobin (SULF_HIGH), 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 (O2HB), deoxyhemoglobin (HHB), carboxyhemoglobin (COHB), methemoglobin (METHB).

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

[0099] Referring to FIGS. 21 - 23, an assembly constructed in accordance with the present disclosure is shown and designated by reference numeral 200. Assembly 200 includes a support substrate 202 and an acoustophoretic device 204 preferably permanently joined to the support substrate 202 in a non-clamping manner. This non-clamping method is performed to minimize vibration loss between the support substrate 202 and the acoustophoretic device 204, which is referred to as an interface of substantially zero mass. Conventional studies have shown that when additional mass is added to the assembly 200, the ability to hemolyze blood decreases. If the acoustophoretic device 204 is mechanically constrained by clamping (e.g., attached to the support substrate 202), acoustic waves / energy are transmitted throughout the connected assembly, thus reducing the energy available for the accumulation of a standing wave field in the microchannel 22. A zero mass interface means a connection that does not add additional mass to the acoustophoretic device 204, which is theoretically an ideal state, but in practice the support substrate 202 adds additional mass to the acoustophoretic device 204. As will be described in more detail below, the acoustophoretic device 204 can be attached to the support substrate 202 such that the acoustophoretic device 204 can vibrate more freely compared to when the acoustophoretic device 204 is connected to the support substrate 202 by a clamping method. Further, the acoustophoretic device 204 can be attached to the support substrate 202 to achieve a substantially zero mass interface that provides efficient excitation (e.g., a voltage of less than 150 Vp-p, 80 Vp-p - 100 Vp-p in some embodiments) as well as a response signal of magnitude and phase.

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

[0101] The sample vessel 212 has an outer surface 220, a microchannel 222 (shown by an imaginary line in FIG. 21) within the boundary of the outer surface 220, a first port 224 that extends through the outer surface 220 to the microchannel 222 and is in fluid communication with the microchannel 222, and a second port 226 that extends through the outer surface 220 to the microchannel 222 and is in fluid communication with the microchannel 222. In one embodiment, the outer surface 220 has an attachment area for the 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, and 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 fits and engages with the outer surface 220 of the sample vessel 212. For example, the piezoelectric transducer 214 and the outer surface 220 can have flat surfaces configured to be positioned together.

[0103] The sample vessel 212 can be partially, substantially, or completely transparent. In one embodiment, the sample vessel 212 is transparent at least above and below the microchannel 222, such that a light beam can pass through the sample vessel 212, through the microchannel 222, interact with some substance within the microchannel 222, and exit the 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 the ability of a material to withstand a change in length when subjected to longitudinal tension or compression. Young's modulus is equal to the value obtained by dividing the longitudinal stress by the strain. In one embodiment, the sample vessel 212 can be constructed from a plastic having a rigidity 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 located at 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 a first end 244 to a second end 246, a width W from a first side 248 to a second side 250, a thickness between a top surface 240 and a bottom surface 242, and an aspect ratio that defines a proportional relationship between the length and the 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 in a direction along the longitudinal axis of the sample vessel 212, and the width of the microchannel 222 is in a direction along the latitude axis of the sample vessel 212. However, it will be understood that the microchannel 222 can be angled or offset from the longitudinal axis and / or the latitude axis of the sample vessel 212.

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

[0109] In one embodiment, the width of the microchannel 222 is about 2 millimeters. In one embodiment, the width of the microchannel 222 is about 2.5 millimeters. In one embodiment, the width of the microchannel 222 is greater than the illumination width of the light collection region of the absorption spectrophotometer 102. The illumination width can be defined as the width of the cross-section of the light collection along the optical path from the absorption spectrophotometer 102 that intersects the microchannel 222. For example, when the illumination diameter is from 1 millimeter to 1.5 millimeters, the width of the microchannel 222 can be at least about 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 1 millimeter to 1.5 millimeters, the width of the microchannel 222 can be about 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 at least partially based on a predetermined desired number of nodes to be created within the microchannel 222. For example, if the microchannel 222 has a width of about 2 millimeters and the whole blood wave propagation speed is about 1500 m / s, the calculated single node is 350 kHz. The nodes can be evenly spaced and dispersed along the length of the microchannel 222 within the microchannel 222 (e.g., 2×2 mm = 4 mm), and the high pressure produces an even distribution of the lysed blood. For example, if the predetermined desired number of nodes on each sidewall of the microchannel 222 is 5 nodes (see FIG. 13), the length of the microchannel 22 can be set to about 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 of the light yield from the absorptiometer 102 in the lysed blood, 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 the microchannel 222 is about 100 micrometers. In one embodiment, the height of the microchannel 222 is about 150 micrometers. In one embodiment, the height of the microchannel 222 is about 250 micrometers. In one embodiment, the height of the microchannel 222 is about 300 micrometers. In one embodiment, the height of the microchannel 222 is from about 80 micrometers to about 300 micrometers. In one embodiment, the height of the microchannel 222 is from about 80 micrometers to 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 outer 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 surface 248, and / or the second side surface 250 of the sample vessel 212. The first port 224 and the second port 226 may 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 surface 248, and / or the second side surface 250.

[0115] In one embodiment, the first port 224 and the second port 226 each have a diameter of from about 0.5 millimeter (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 millimeter (800 micrometers). The microchannel 222 tapers towards the first port 224 and the second port 226, as shown in FIG. 11B. The taper assists in providing fluid from and / or to the first port 224. The cross-sectional width (e.g., diameter) of the first port 224 and the second port 226 is smaller than the width of the microchannel 222. For example, the cross-sectional width 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 integrally 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 a plurality of pieces of material that are interconnected and integrated to form an integrated whole. As will be discussed in more detail with respect to FIG. 24, the sample vessel 212 can be formed from two substrates joined together as shown in FIG. 11B. Alternatively, the sample vessel 212 can be formed from three substrates joined together as shown in FIG. 11A. Referring again to FIG. 21, the piezoelectric transducer 214 is attached to the sample vessel 212 (such as the attachment area of the outer surface 220) after assembly to form an integral structure of the acoustic levitation device 204. The piezoelectric transducer 214 can have an attachment area that is attached to the attachment area of the outer surface 220. In one embodiment, the piezoelectric transducer 214 is at least partially attached to the top surface 240 of the sample vessel 212, but 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 surface 248, and / or the second side surface 250. The piezoelectric transducer 214 is positioned in relation to the microchannel 222 such that the piezoelectric transducer 214 does not block light moving through the microchannel 222 from the top or bottom surface of the sample vessel 212. The piezoelectric transducer 214 can be offset from the microchannel 222 such that light can enter the microchannel 222 from outside the sample vessel 212. In one embodiment, the piezoelectric transducer 214 has a length, and along that length, has a longitudinal axis that is oriented substantially parallel (e.g., within a range of 5 degrees from parallel) to the longitudinal axis of the sample vessel 212. In one embodiment, the piezoelectric transducer 214 has a width that is smaller than the length of the piezoelectric transducer 214.

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

[0118] The support substrate 202 includes an upper surface 260, a lower surface 262 opposite to the upper surface 260, and an outer peripheral edge 264 (see FIG. 22). The support substrate 202 also has an inner edge 270 that defines an opening 272 that intersects the upper surface 260 and the lower surface 262. The sample vessel 212 is connected to at least one of the upper surface 260 and the lower 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 with respect to the support substrate 202. For example, the piezoelectric transducer 214 can pass through the opening 272 and extend beyond the upper surface 260 of the support substrate 202. As shown in FIG. 21, the sample vessel 212 is connected to the lower surface 262 and extends from the lower surface 262, but does not enter or pass through the opening 272 within the support substrate 202.

[0119] In some embodiments, the support substrate 202 includes a conductive trace 274 (see FIGS. 22 and 23) in a predetermined pattern on at least one of the upper surface 260 and the lower surface 262. As will be described in more detail below, the conductive trace 274 enables electrical connection to the heating element and each control system, and provides an apparatus configured for capacitive fluid detection. In some embodiments, the controller 106 is electrically connected to the piezoelectric transducer 214 and is 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 trace 274 (alternatively, it is 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 to the outer peripheral edge 264 (see FIGS. 23 and 27). In the example shown, the conductive trace 274 includes twelve connector portions 275 indicated by reference numerals 275a-l for clarity (see FIG. 27). At least some of the conductive traces 274 also include attachment pads 276 (see FIG. 28). The connector portion 275 adjacent to the outer peripheral edge 264 forms a male edge connector 277. The attachment pad 276 can be connected to a specific one of the connector portions 275 by a wire 278 (see FIG. 27). The wire 278 can extend to the upper surface 260 or the lower surface 262 of the support substrate 202. Further, the wire 278 can extend between the lower surface 262 and the upper surface 260, for example, by using a through via.

[0120] In the example shown in FIG. 28, the support substrate 202 includes ten attachment pads 276 indicated by reference numerals 276a to 276j on the lower surface 262, and one attachment pad 276k on the upper surface 260 (see FIG. 27). The attachment pads 276a to 276k are electrically connected to corresponding ones of the connector portions 275a to 275l so as to enable transmission of electrical signals to and / or reception from the attachment pads 276a to 276k. The number of attachment pads 276 provided on the support substrate 202 can be more or less. When the sample vessel 212 is connected to the support substrate 202, at least two of the attachment pads 276 are joined to the 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 production of printed circuit boards include FR4 G10 fiberglass epoxy, phenolic resin, polyimide, and the like.

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

[0123] The attachment pads 284a, 284b, 284c, 284d, and 284e are spaced from each other and electrically isolated. The attachment pads 284a, 284b, 284c, 284d, and 284e are located along the first end 244 of the sample vessel 212. The attachment pads 284a, 284b, 284c, 284d, and 284e are joined to the attachment pads 276a to 276e, for example, soldered, to provide a mechanical and electrical connection therebetween.

[0124] The attachment pads 284p, 284q, 284r, 284s, and 284t are spaced from each other and electrically isolated. The attachment pads 284p, 284q, 284r, 284s, and 284t are spaced apart from the attachment pads 284a, 284b, 284c, 284d, and 284e. For example, the attachment pads 284p, 284q, 284r, 284s, and 284t can be located along the second end portion 246 of the sample vessel 212. The attachment pads 284p, 284q, 284r, 284s, and 284t are joined, for example soldered, to the attachment pads 276f - j on the support substrate 202 to provide a mechanical and electrical connection therebetween.

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

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

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

[0128] The conductive trace 282c is included within the attachment pad 284c and is used only to connect the sample vessel 212 to the support substrate 202.

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

[0130] The conductive trace 282e includes six attachment pads 284e, 284j, 284k, 284l, 284m, and 284q. The attachment pads 284e and 284q can be mechanically and electrically connected to the attachment pads 276e and 276g. Each of the attachment pads 284j, 284k, 284l, and 284m can be connected to a lead of an electrical device. In the example shown in FIG. 26, the sample vessel includes four electrical heaters 302a - d. The electrical heaters 302a - d can be used to generate heat and provide it to the sample vessel 212 by supplying electricity to the electrical heaters 302a - d. As discussed above, in order to accurately measure a blood sample oximetry specimen, thermal control can be used to keep the patient's blood sample at a constant temperature. The electrical heaters 302a - d can be connected in parallel by the attachment pads 284j, 284h, 284k, 284i, 284l, 284o, 284m, and 284n. In this example, electrical heater 302a is connected to attachment pads 284j and 284h; electrical heater 302b is connected to attachment pads 284k and 284i; electrical heater 302c is connected to attachment pads 284l and 284o; electrical heater 302d is connected to attachment pads 284m and 284n. The conductive trace 282e can also be used in combination with the conductive trace 282b to form a capacitance sensor. In this regard, the conductive trace 282e also includes a second sensor portion 304, and the second sensor portion 304 can extend within a range of ±5 degrees from parallel (preferably, extend parallel) to the first sensor portion 294. In the illustrated embodiment, both the first sensor portion 294 and the second sensor portion 304 are in a linear configuration and are adjacent to but do not cover the microchannel 222 so as not to block 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 as long as the first sensor portion 294 and the second sensor portion 304 do not block the beam 116 and it is possible to correlate the capacitance readings from the first sensor portion 294 and the second sensor portion 304 with the expected channel contents within the microchannel 222 during calibration. In some embodiments, the first sensor portion 294 and / or the second sensor portion 304 can also have a meandering configuration, and one or more portions cross over the microchannel 222 outside the expected path of the beam 116 through the sample vessel 212.

[0131] The conductive trace 282f includes five attachment pads 284h, 284i, 284o, 284n, and 284p. The use of the attachment pads 284h, 284i, 284o, 284n has been described above. The attachment pad 284p is configured to mechanically and electrically connect to the attachment pad 276f.

[0132] The conductive traces 282g - I are included within the attachment pads 284r, 284s, and 284t and are used only to mechanically connect the sample vessel 212 to the support substrate 202, for example, by reflow soldering in standard automated processing equipment used in the manufacture of electronic devices.

[0133] In some embodiments, the conductive traces 282a - t are formed to have a conductive electrode pattern 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] Referring next to FIGS. 24 and 11B, to fabricate the assembly 200, the first and second glass substrates 310, 312 for fabricating 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 a flat surface, which forms the top surface 240 of the sample vessel 212 when the glass substrates 310, 312 are joined together. The bottom surface 316 of the first glass substrate 310 can also be a flat surface. In some embodiments, the first glass substrate 310 can have a thickness of 700 micrometers. 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 to have the first port 224 and the 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 so as to completely enclose and surround the microchannel 222. In some embodiments, the second glass substrate 312 can have a thickness of 700 micrometers.

[0135] When the glass substrates 310 and 312 are part of a wafer, the conductive electrode pattern is applied to a first glass substrate 310 (or 70 shown in FIG. 11A) intended to be used as the top surface 240. Electrical components such as the thermistor 300 and the electrical heaters 302a - d are soldered to predetermined attachment pads 284 of the conductive trace 282. The substrates 310 and 312 are then diced from the wafer and singulated. Corresponding pairs of the substrates 310 and 312 are joined together such that the first substrate 310 covers the microchannel 222 in the second substrate 312 to form the sample vessel 212. As shown in FIGS. 24 and 26, the piezoelectric transducer 214 has a size smaller than the size of the glass substrates 310 and 312, and a peripheral portion of the piezoelectric transducer 214 is attached to a portion of the conductive electrode pattern that includes a first portion 288 of the conductive trace 282a. Thus, some portions of the periphery of the piezoelectric transducer 214 are indirectly joined to the top surface 314 of the first glass substrate 310, and most of the piezoelectric transducer 214 remains available for direct joining 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, remains exposed to be directly joined to the piezoelectric transducer 214. FIG. 26 shows the sample vessel 212 in which the piezoelectric transducer 214 does not extend beyond the surface area of the glass substrates 310 and 312 when the integrated structure is formed. As shown in FIG. 24, the piezoelectric transducer 214 is positioned within the attachment area so as to overlap the first portion 288 of the conductive trace 282a and is then joined 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 is joined to the bottom surface 242 of the sample vessel 212 by an epoxy 330 such as an epoxy sold under the brand name LOCKTITE444.The sample outlet 322 is inserted into the second port 226 and joined to the bottom surface 242 of the sample vessel 212 by an epoxy 332, such as an epoxy sold under the brand name LOCKTITE 444. After the sample inlet 320 and the sample outlet 322 are connected to the sample vessel 212, the attachment pads 284a - e and 284p - t are then soldered to the attachment pads 276a - j to mechanically and electrically connect the sample vessel 212 to the support substrate 202. Next, the flexible wire 340 is soldered to the attachment pad 276k and the terminals on the piezoelectric transducer 214.

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

[0137] After the male edge connector is inserted 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 alternating current 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 the desired temperature of the sample vessel.

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

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

[0140] Referring now to FIG. 31, in some embodiments, the assembly 200 can be a component of the analyzer 100a. The analyzer 100a is similar in structure and function to the analyzer 100 described above with reference to FIGS. 15 - 18, except that the analyzer 100a includes an edge connector 500, which is described herein as an example as a female edge connector 500 configured to be mechanically and electrically connected to the male edge connector 277 of the support substrate 202. Common elements between the analyzers 100 and 100a are indicated by reference numerals in FIG. 31. Also, although the support substrate 202 is referenced in FIG. 31, it should be understood that the support substrate can be that of the support substrate 400 or 420. As discussed above with reference to FIGS. 23 and 27, the connector portion 275 is provided adjacent to the outer peripheral edge 264. The connector portion 275 can be provided on the upper surface 260 and / or the lower surface 262 of the support substrate 202. In the example shown in FIG. 31, the connector portion 275 is provided on the upper surface 260. In this embodiment, the connector portions 275 are in the same plane. The connector portions 275 are spaced along at least a portion of the outer peripheral edge 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, and each of the spring contacts 502 is positioned to contact a particular one of the connector portions 275. Each of the spring contacts 502 can be biased downward (in the example shown) and spaced from one or two adjacent spring contacts 502. The spacing of the spring contacts 502 can be predetermined to match the spacing of the connector portions 275. The spring contacts 502 are conductive and are 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 contacts 502. By continuously moving the male edge connector 277 towards the spring contacts 502, it becomes possible for the spring contacts 502 to contact the connector portions 275.Moving the male edge connector 277 away from the spring contact 502 in the opposite direction enables the disconnection of the assembly 200 from the analyzer 100a.

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

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

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

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

[0145] 1. An acoustophoresis device comprising: A sample vessel having an outer surface, microchannels within the boundaries of the outer surface, a first port extending through the outer surface to the microchannels, and a second port extending through the outer surface to the microchannels, wherein the microchannels are configured to receive a blood sample through the first port, and the sample vessel has a conductive trace on the outer surface; A piezoelectric transducer formed on the outer surface of the sample vessel to form an integral structure, contacting at least one of the conductive traces, configured to generate ultrasonic waves in a sample within the microchannels, and having an excitation input and a response signal output electrically connected to at least one of the conductive traces.

[0146] 2. The acoustophoresis device according to exemplary embodiment 1, wherein the sample vessel is constructed from glass.

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

[0148] 4. 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. Among the conductive traces, the first conductive trace includes a first attachment pad and a first sensor portion, and the first sensor portion extends along the first side of the microchannel. Among the conductive traces, the second conductive trace includes a second attachment pad and a second sensor portion, and the second sensor portion extends along the second side of the microchannel. The acoustic levitation device according to any one of exemplary embodiments 1 to 3.

[0149] 5. The outer surface is a first outer surface having an attachment area. The attachment area has a first shape. At least one of the conductive traces has a first portion within the attachment area and a second portion outside the attachment 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 joined to the attachment area. The acoustic levitation device according to any one of exemplary embodiments 1 to 4.

[0150] 6. At least one of the conductive traces is located between the piezoelectric transducer and the outer surface of the sample vessel. The acoustic levitation device according to any one of exemplary embodiments 1 to 5.

[0151] 7. The conductive trace is joined in direct contact with the outer surface of the sample vessel. The acoustic levitation device according to any one of exemplary embodiments 1 to 6.

[0152] 8. The sample vessel has an outer peripheral portion, and the conductive trace includes a conductive attachment pad located adjacent to the outer peripheral portion. The acoustic levitation device according to any one of exemplary embodiments 1 to 7.

[0153] 9. The sample vessel has a first end and a second end, and the conductive trace includes a conductive attachment pad positioned adjacent to at least one of the first end and the second end of the sample vessel, the acoustophoretic device according to any one of exemplary embodiments 1-8.

[0154] 10. Further comprising an electrical component, the sample vessel has an outer peripheral portion, and the first conductive trace of the conductive traces includes a first attachment pad and a second attachment pad electrically connected to the first attachment pad, and the second conductive trace of the conductive traces includes a third attachment pad and a fourth attachment pad electrically connected to the third attachment pad, the first attachment pad and the third attachment pad are positioned adjacent to the outer peripheral portion, and the electrical component has a first lead connected to the second attachment pad and a second lead connected to the fourth attachment pad, the acoustophoretic device according to any one of exemplary embodiments 1-9.

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

[0156] 12. The second attachment pad has a first length and a first width, the first length is greater than the first width, the fourth attachment pad has a second length and a second width, the second length is greater than the second width, and the first length and the second length extend within a range of 5 degrees from parallel, the acoustophoretic device according to any one of exemplary embodiments 1-11.

[0157] 13. The electrical component includes a first electric heater and a second electric heater, and the first electric heater and the second electric heater are connected in parallel to the second attachment pad and the fourth attachment pad, the acoustophoretic device according to any one of exemplary embodiments 1-12.

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

[0159] 15. 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 intersecting the upper and lower surfaces, the sample vessel being connected to one of the upper and lower surfaces, the assembly according to any one of the exemplary embodiments described above.

[0160] 16. The support substrate includes conductive traces of a predetermined pattern on at least one of the upper and lower surfaces, at least some of the conductive traces including attachment pads, at least two of the attachment pads being bonded to the sample vessel, the assembly according to any one of the exemplary embodiments described above.

[0161] 17. The conductive trace is a first conductive trace, the attachment pad is a first attachment pad, the sample vessel has a plurality of second conductive traces on an outer surface having a second attachment pad, the first attachment pad being bonded to the second attachment pad, the assembly according to any one of the exemplary embodiments described above.

[0162] 18. The first attachment pad is soldered to the second attachment pad, the assembly according to any one of the exemplary embodiments described above.

[0163] 19. The first attachment pad is the assembly according to exemplary embodiment 17 or 18, which is located adjacent to the inner edge portion.

[0164] 20. The support substrate is a circuit board, and the assembly according to any one of the above exemplary embodiments.

[0165] 21. The piezoelectric transducer is the assembly according to any one of the above exemplary embodiments, which extends through an opening in the support substrate.

[0166] 22. An analyzer comprising: An acoustophoresis device comprising: A sample vessel having an outer surface, a microchannel within the boundary of the outer surface, a first port extending through the outer surface to the microchannel, and a second port extending through the outer surface to the microchannel, wherein the sample can be inserted into the microchannel through the first port, and the sample vessel has a conductive trace on the outer surface; and A piezoelectric transducer bonded to the outer surface of the sample vessel to form an integral structure, contacting at least one of the conductive traces, configured to generate ultrasonic waves in the sample within the microchannel, and configured to vibrate the sample vessel so as to induce a shear force within the microchannel; and an acoustophoresis device including the piezoelectric transducer; An absorptiometric spectrophotometer including a transmitter and a receiver located adjacent to the sample vessel, wherein the transmitter is positioned to emit a light beam through the microchannel, and the receiver is positioned to receive that portion of the light beam after at least a portion of the light beam has passed through the microchannel; and 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 a 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, and the analyzer includes the controller.

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

[0168] 24. The outer surface of the sample vessel has a first side surface and a second side surface opposite to the first side surface, and the first side surface and the second side surface are flat. The analyzer according to any one of the above exemplary embodiments.

[0169] 25. The sample vessel is constructed of glass. The analyzer according to any one of the above exemplary embodiments.

[0170] 26. The sample vessel is constructed of a non - glass material having a Young's modulus in the range of about 50 GPa to 90 GPa. The analyzer according to any one of the above exemplary embodiments.

[0171] 27. The outer surface of the sample vessel is a first outer surface having a mounting area. The mounting area has 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 is joined to the mounting area. The analyzer according to any one of the above exemplary embodiments.

[0172] 28. The piezoelectric transducer fits and engages with the outer surface of the sample vessel. The analyzer according to any one of exemplary embodiments 22 - 27.

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

[0174] 30. Further comprising a support substrate including an upper surface, a lower surface opposite to the upper surface, and an outer peripheral edge portion, the support substrate having an inner edge portion defining an opening intersecting the upper surface and the lower surface, and the sample vessel being connected to one of the upper surface and the lower surface. The analyzer according to any one of exemplary embodiments 22 to 30.

[0175] 31. The support substrate includes conductive traces of a predetermined pattern on at least one of the upper surface and the lower surface, at least some of the conductive traces including attachment pads, and at least two of the attachment pads being joined to the sample vessel. The analyzer according to any one of the exemplary embodiments described above.

[0176] 32. The conductive trace is a first conductive trace, the attachment pad is a first attachment pad, the sample vessel has a plurality of second conductive traces on an outer surface having a second attachment pad, and the first attachment pad is joined to the second attachment pad. The analyzer according to any one of the exemplary embodiments described above.

[0177] 33. The first attachment pad is soldered to the second attachment pad. The analyzer according to any one of the exemplary embodiments described above.

[0178] 34. The first attachment pad is located adjacent to the inner edge portion. The analyzer according to any one of the exemplary embodiments described above.

[0179] 35. The support substrate is a circuit board. The analyzer according to any one of the exemplary embodiments described above.

[0180] 36. The piezoelectric transducer extends through an opening in the support substrate. The analyzer according to any one of the exemplary embodiments described above.

[0181] 37. The support substrate includes conductive traces of a predetermined pattern 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 to the outer peripheral edge, the analyzer according to any one of the exemplary embodiments.

[0182] 38. Further including an edge connector, the edge connector includes a support member connected to a plurality of spring contacts, the spring contacts are constructed from a conductive material that communicates with the controller, and a specific one of the spring contacts engages a specific one of the connector portions to provide an electrical connection between the conductive trace of the support substrate and the spring contact, the analyzer according to any one of the exemplary embodiments.

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

[0184] 40. The step of bonding the conductive trace to the outer surface of the sample vessel; The method according to exemplary embodiment 39, further including 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 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 so as to reach the opening and then connecting at least a portion of the conductive trace to the support substrate, the method according to any one of the exemplary embodiments.

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

[0187] Conclusion As discussed above, the assembly 200 includes a support substrate 202, 400, or 420 and an acoustic levitation device 204 preferably permanently joined to the support substrate 202, 400, or 420 in a manner that does not clamp the support substrate 202, 400, or 420. The acoustic levitation device 204 can be attached to the support substrate 202, 400, or 420 such that the acoustic levitation device 204 can vibrate more freely compared to when the acoustic levitation device 204 is connected to the support substrate 202, 400, or 420 by a clamping method. Further, the acoustic levitation device 204 can be attached to the support substrate 202 to achieve a substantially zero-mass interface that provides an efficient excitation (e.g., a voltage of less than 150 Vp-p, 80 Vp-p to 100 Vp-p in some embodiments) and a response signal with a high signal-to-noise ratio. The support substrate 202, 400, or 420 can be, for example, a printed circuit board and includes a predetermined pattern of conductive traces 274 on at least one of the upper surface 260 and the lower surface 262 of the support substrate 202, 400, or 420, enabling electrical connection to components on the support substrate 202, 400, or 420 and the acoustic levitation device 204 without wires.

[0188] Assembly 200 can be a consumable that can be replaced by the user if desired, or a disposable cartridge for single use that is disposed of once used. Additionally, Assembly 200 enables the entire microchannel 222 to be self - contained with respect to heating, hemolysis, and fluid detection. The inclusion of microchannel 222 within Assembly 200 enhances the uniformity of microchannel 222, thereby enhancing the accuracy of reading blood samples performed within microchannel 222 by analyzers 100, 100a. Finally, Assembly 200 electrically connects conductive trace 282 without wires that are not joined to sample vessel 212 along the length of the wire while enabling a substantially zero - mass interface connection of acoustic - electrophoresis device 204 to support substrates 202, 400, or 420. This provides the advantages of easier and less expensive fabrication, as well as a more reliable electrical interface and enhanced signal integrity.

[0189] The above description is provided by way of example and illustration and is not intended to be exhaustive or to limit the invention to the precise form disclosed. Modifications and variations are possible in light of the above teachings, or they may be acquired from practice of the methods described in this disclosure.

[0190] Certain combinations of configurations and steps are described and / or disclosed in the claims and / or this specification, but these combinations are not intended to limit the disclosure. In fact, many of these configurations and steps can be combined in forms not specifically described and / or disclosed in the claims or this specification. Each of the dependent claims listed below may depend directly only on one other claim, but the disclosure includes each dependent claim in combination with all other claims in the claim set.

[0191] Elements, acts, or instructions used in this application should not be construed as essential or integral to the present invention unless explicitly stated to be outside the scope of the preferred embodiments. Further, the phrase "based on" is intended to mean "based, at least in part, on" unless otherwise explicitly stated.

Claims

**Claim 1** An acoustophoresis device comprising: A sample vessel having an outer surface, microchannels within the boundaries of the outer surface, a first port extending through the outer surface to the microchannels, and a second port extending through the outer surface to the microchannels, wherein the microchannels are configured to receive a blood sample through the first port; the sample vessel having an electrically conductive trace on the outer surface; A piezoelectric transducer formed on the outer surface of the sample vessel to form an integral structure, the piezoelectric transducer being configured to generate ultrasonic waves in a blood sample within the microchannels by contacting at least one of the electrically conductive traces and having an excitation input and a response signal output electrically connected to at least one of the electrically conductive traces, the acoustophoresis device comprising the piezoelectric transducer. **Claim 2** The acoustophoresis device according to claim 1, wherein the sample vessel is constructed from glass. **Claim 3** The acoustophoresis device according to claim 1, wherein the sample vessel includes a first substrate bonded to the piezoelectric transducer and a second substrate having the microchannels, the first port, and the second port. **Claim 4** The microchannels have a length, a width, and a height, and the microchannels have a first side extending along the length of the microchannels and a second side extending along the length of the microchannels. The first electrically conductive trace of the electrically conductive traces includes a first attachment pad and a first sensor portion, the first sensor portion extending along the first side of the microchannels. The second electrically conductive trace of the electrically conductive traces includes a second attachment pad and a second sensor portion, the second sensor portion extending along the second side of the microchannels. The acoustophoresis device according to claim 1. **Claim 5** The outer surface is a first outer surface having an attachment area, the attachment area having a first shape. At least one of the electrically conductive traces has a first portion within the attachment area and a second portion outside the attachment 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 attachment area. The acoustophoresis device according to claim 1. **Claim 6** The acoustic phoretic device according to claim 1, wherein at least one of the conductive traces is located between the piezoelectric transducer and the outer surface of the sample vessel.

7. The acoustic phoretic device according to claim 1, wherein the conductive trace is joined in direct contact with the outer surface of the sample vessel.

8. The acoustic phoretic device according to claim 1, wherein the sample vessel has an outer peripheral portion, and the conductive trace includes a conductive attachment pad located adjacent to the outer peripheral portion.

9. The acoustic phoretic device according to claim 1, wherein the sample vessel has a first end and a second end, and the conductive trace includes a conductive attachment pad located adjacent to at least one of the first end and the second end of the sample vessel.

10. The acoustic phoretic device according to claim 1, further comprising an electrical component, wherein the sample vessel has an outer peripheral portion, the first conductive trace of the conductive traces includes a first attachment pad and a second attachment pad electrically connected to the first attachment pad, the second conductive trace of the conductive traces includes a third attachment pad and a fourth attachment pad electrically connected to the third attachment pad, the first attachment pad and the third attachment pad are located adjacent to the outer peripheral portion, and the electrical component has a first lead connected to the second attachment pad and a second lead connected to the fourth attachment pad.

11. The acoustic phoretic device according to claim 10, wherein the electrical component is a thermistor.

12. The second attachment pad has a first length and a first width, the first length is greater than the first width, the fourth attachment pad has a second length and a second width, the second length is greater than the second width, and the first length and the second length extend within a range of 5 degrees from parallel. The acoustic phoretic device according to claim 10.

13. The acoustic phoretic device according to claim 12, wherein the electrical component includes a first electrical heater and a second electrical heater, and the first electrical heater and the second electrical heater are connected in parallel to the second attachment pad and the fourth attachment pad.

14. An assembly comprising: a support substrate; and an acoustic phoretic device permanently joined to the support substrate, the acoustic phoretic device comprising: A sample vessel having an outer surface, microchannels within the range of the outer boundary, a first port extending through the outer surface to the microchannels, and a second port extending through the outer surface to the microchannels, wherein a blood sample can be inserted into the microchannels through the first port, and the sample vessel has a conductive trace on the outer surface, the sample vessel; A piezoelectric transducer joined to the outer surface of the sample vessel to form an integral structure, configured to generate an ultrasonic standing wave in a blood sample within the microchannels by contacting at least one of the conductive traces, and having an excitation signal input and a response signal output electrically connected to at least one of the conductive traces, the piezoelectric transducer, included in the assembly.

15. The support substrate includes an upper surface, a lower surface opposite the upper surface, and an outer peripheral edge, the support substrate has an inner edge defining an opening intersecting the upper surface and the lower surface, and the sample vessel is connected to one of the upper surface and the lower surface, the assembly according to claim 14.

16. The support substrate includes conductive traces of a predetermined pattern on at least one of the upper surface and the lower surface, at least some of the conductive traces include attachment pads, and at least two of the attachment pads are joined to the sample vessel, the assembly according to claim 15.

17. The conductive trace is a first conductive trace, the attachment pad is a first attachment pad, the sample vessel has a plurality of second conductive traces on an outer surface having a second attachment pad, and the first attachment pad is joined to the second attachment pad, the assembly according to claim 16.

18. The first attachment pad is soldered to the second attachment pad, the assembly according to claim 17.

19. The first attachment pad is located adjacent to the inner edge, the assembly according to claim 17.

20. The support substrate is a circuit board, the assembly according to claim 14.

21. The piezoelectric transducer extends through an opening in the support substrate, the assembly according to claim 15.

22. An analyzer: An acoustophoresis device: A sample vessel having an outer surface, microchannels within the range of the outer boundary, a first port extending through the outer surface to the microchannels, and a second port extending through the outer surface to the microchannels, wherein the sample is insertable into the microchannels through the first port, and the sample vessel has a conductive trace on the outer surface; and A piezoelectric transducer bonded to the outer surface of the sample vessel to form an integral structure, contacting at least one of the conductive traces, configured to generate ultrasonic waves in the sample within the microchannels, and configured to vibrate the sample vessel to induce a shear force within the microchannels, the acoustic phoresis device including the piezoelectric transducer; An absorption spectrophotometer including a transmitter and a receiver positioned adjacent to the sample vessel, the transmitter being positioned to emit a light beam through the microchannels, and the receiver being positioned to receive that portion of the light beam after at least a portion of the light beam has passed through the microchannels; 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, the analyzer including the controller.

23. 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 of a material transparent to the light beam, the analyzer according to claim 22.

24. 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, the analyzer according to claim 22.

25. The sample vessel is constructed of glass, the analyzer according to claim 22.

26. The sample vessel is constructed of a non-glass material having a Young's modulus in the range of about 50 GPa to 90 GPa, the analyzer according to claim 22.

27. The outer surface of the sample vessel is a first outer surface having an attachment area, the attachment area has a first shape, and 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 joined to the attachment area. The analyzer according to claim 22.

28. The piezoelectric transducer fits and engages with the outer surface of the sample vessel. The analyzer according to claim 22.

29. The height of the microchannel is about 100 micrometers, and the width of the microchannel is about 2 millimeters. The analyzer according to claim 22.

30. The analyzer further includes a support substrate including an upper surface, a lower surface opposite the upper surface, and an outer peripheral edge portion, the support substrate has an inner edge portion defining an opening intersecting the upper surface and the lower surface, and the sample vessel is connected to one of the upper surface and the lower surface. The analyzer according to claim 22.

31. The support substrate includes conductive traces in a predetermined pattern on at least one of the upper surface and the lower surface, at least some of the conductive traces include attachment pads, and at least two of the attachment pads are joined to the sample vessel. The analyzer according to claim 30.

32. The conductive traces are first conductive traces, the attachment pads are first attachment pads, the sample vessel has a plurality of second conductive traces on an outer surface having second attachment pads, and the first attachment pads are joined to the second attachment pads. The analyzer according to claim 31.

33. The first attachment pads are soldered to the second attachment pads. The analyzer according to claim 32.

34. The first attachment pads are located adjacent to the inner edge portion. The analyzer according to claim 32.

35. The support substrate is a circuit board. The analyzer according to claim 29.

36. The piezoelectric transducer extends through an opening in the support substrate. The analyzer according to claim 30.

37. The support substrate includes conductive traces in a predetermined pattern on at least one of the upper surface and the lower surface, and at least some of the conductive traces include a plurality of connector portions provided adjacent to the outer peripheral edge portion. The analyzer according to claim 30.

38. Further comprising an edge connector, the edge connector includes a support member connected to a plurality of spring contacts, the spring contacts are constructed from a conductive material that communicates with the controller, and a particular one of the spring contacts engages a particular one of the connector portions to provide an electrical connection between the conductive trace of the support substrate and the spring contact. The analyzer according to claim 37.

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

40. Bonding a conductive trace to the outer surface of the sample vessel; Connecting at least a portion of the conductive trace on the outer surface of the sample vessel to the attachment pad of the conductive trace on the piezoelectric transducer The method according to claim 39, further comprising.

41. 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 so as to reach the opening and then connecting at least a portion of the conductive trace to the support substrate. The method according to claim 40.

42. The attachment pad of the conductive trace on the outer surface of the sample vessel is soldered to the attachment pad of the conductive trace on the support substrate. The method according to claim 40.

Images