Centrifugation based sample analysis cartridges and methods of using them

Abstract

Centrifugation based analysis cartridges for analyzing non-cellular constituents of a biological sample are provided. An analysis cartridge is centrifuged around a vertical axis in multiple steps including: a first centrifugation that moves a diluent from a diluent chamber into a diluent metering chamber, a second centrifugation that moves into a mixing chamber the diluent from the diluent metering chamber and a sample from a sample metering chamber; a third centrifugation that moves into one or more metering fingers the diluted samples from the mixing chamber; and a fourth centrifugation that moves into one or more detection chambers the diluted sample from the one or more metering fingers. The connections between the different chambers allow flow of liquids only under centrifugation forces above predetermined levels. Methods of analyzing biological samples using the analysis cartridges disclosed herein as well as analyzers that analyze biological samples by manipulating the analysis cartridges are provided.

Description

CENTRIFUGATION BASED SAMPLE ANALYSIS CARTRIDGES AND METHODS OF USING THEMCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit under 35 U.S.C. § 119(e) of provisional application Serial Nos. 63 / 704,316, filed October 7, 2024, and 63 / 800,328, filed on May 5, 2025, which applications are hereby incorporated by reference in their entirety.INTRODUCTION

[0002] Analyses of biological samples, such as blood samples, are limited by the availability of samples. Also, analyses of biological samples, such as blood samples, often involve testing non- cellular constituents. Therefore, analyses of biological samples for their non-cellular constituents using the smallest possible amounts are desirable.SUMMARY

[0003] In certain aspects, the disclosure provides analysis cartridges that allow analyzing small volumes of biological samples. In certain embodiments, the analysis cartridges allow analyses of non-cellular portions of biological samples.

[0004] In certain aspects, the disclosure provides an analysis cartridge for analyzing a biological sample, the analysis cartridge comprising:

[0005] a d iluent chamber fluidical ly connected to a diluent metering chamber, wherein a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0006] a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve and to a sample metering chamber via a sample valve such that, when the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0007] one or more detection chambers fluidically connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or moredetection chambers, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0008] In the analysis cartridges disclosed herein, a non-cellular portion of a biological sample, such as plasma, can be analyzed for comprehensive metabolic panel (CMP). Accordingly, certain aspects of the disclosure provide a method of analyzing a sample in an analysis cartridge, the method comprising:

[0009] conducting a first centrifugation of the analysis cartridge to move a diluent from a diluent chamber to a f luidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0010] conducting a second centrifugation of the analysis cartridge to move into a mixing chamber the diluent from the diluent metering chamber and the sample from the sample metering chamber; wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold, and

[0011] conducting a third centrifugation to move into a one or more detection chambers the diluted sample from the mixing chamber, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0012] Further provided herein are analyzers that use the analysis cartridges provided herein to analyze a sample, such as blood for non-cellular constituents. Thus, certain aspects of the disclosure provide an analyzer for analyzing a sample in an analysis cartridge, the analyzer comprising:

[0013] a rotor comprising a slot that holds the analysis cartridge such that, when the rotor rotates, the analysis cartridge is centrifuged around a vertical axis,

[0014] a processor, and

[0015] a non-transitory computer readable medium comprising instructions, which when executed by the processor, cause the analyzer to perform:

[0016] a first centrifugation of the analysis cartridge that moves a diluent from the diluent chamber into a f luidica I ly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0017] when a diluent valve and a sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from a sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on thediluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0018] a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more detection chambers, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0019] In a further aspect, the disclosure provides an analysis cartridge for analyzing a sample, the analysis cartridge configured to be centrifuged around a vertical axis, the analysis cartridge comprising:

[0020] a diluent chamber fluidical ly connected to a diluent metering chamber, wherein a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0021] a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve and to a sample metering chamber via a sample valve such that, when the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0022] one or more metering fingers fluidically connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0023] one or more detection chambers fluidically connected to the one or more metering fingers via capillary channels such that, a fourth centrifugation of the analysis cartridge moves the diluted sample from the one or more metering fingers into the one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold.

[0024] In an even further aspect, the disclosure provides a method of analyzing a sample in an analysis cartridge, the method comprising:

[0025] conducting a first centrifugation of the analysis cartridge to move a diluent from a diluent chamber to a f luidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0026] conducting a second centrifugation of the analysis cartridge to move into a mixing chamberthe diluent from the diluent metering chamberand the sample from a sample metering chamber; wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0027] conducting a third centrifugation of the analysis cartridge to move the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0028] conducting a fourth centrifugation to move into a one or more detection chambers the diluted sample from the one or more metering fingers, wherein the fourth centrifugation exerts on the diluted sample in the one or more metering fingers a fourth centrifugal force above a third threshold.

[0029] In certain aspects, the disclosure provides an analyzer for analyzing a sample in an analysis cartridge, the analyzer comprising:

[0030] a rotor comprising a slot that holds the analysis cartridge such that, when the rotor rotates, the analysis cartridge is centrifuged around a vertical axis,

[0031] a processor, and

[0032] non-transitory computer readable medium comprising instructions, which when executed by the processor, cause the analyzer to perform:

[0033] a first centrifugation of the analysis cartridge that moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold

[0034] when a diluent valve and a sample valve are open, a second centrifugation of the analysis cartridge that moves the diluent from the diluent metering chamber and a sample from a sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0035] when a diluted sample valve is open, a third centrifugation of the analysis cartridge that moves the diluted sample from the mixing chamber into the one or more metering fingers via adistribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0036] a fourth centrifugation of the analysis cartridge that moves the diluted sample from the one or more metering fingers into the corresponding one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold.BRIEF DESCRIPTION OF THE FIGURES

[0037] FIGS. 1A-1B depict views of an exemplary analysis cartridge.

[0038] FIGS. 2A-2G depict certain exemplary analysis cartridges and steps in the operation of such exemplary analysis cartridges.

[0039] FIG. 3 depicts an exemplary analysis cartridge that comprises one or more electrochemical sensors that electrochemically detect one or more analytes in a sample.

[0040] FIG. 4 depicts an exemplary analysis cartridge that comprises one or more electrochemical sensors that electrochemically detect one or more analytes in a sample.

[0041] FIG. 5 depicts an exemplary analysis cartridge that comprises one or more electrochemical sensors that electrochemically detect one or more analytes in a sample.

[0042] FIGS. 6A-6B depict an exemplary analysis cartridge that comprises one or more electrochemical sensors that electrochemically detect one or more analytes in a sample.

[0043] FIG. 7 depicts an exemplary analysis cartridge that comprises one or more electrochemical sensors that electrochemically detect one or more analytes in a sample.

[0044] FIG. 8 depicts an exemplary analysis cartridge that comprises detection chambers having two-compartments.

[0045] FIG. 9 depicts an exemplary analysis cartridge that comprises detection chambers having two-compartments that are joined together via a fluid-valve.

[0046] FIG. 10 depicts an exemplary analysis cartridge. This exemplary analysis cartridge is designed to analyze up to 20 analytes.

[0047] FIG. 11 provides a flow-chart of the operation of the exemplary analysis cartridge described in FIG. 10.

[0048] FIG. 12 shows exemplary fluidic volumes for each stage of sample processing in the analysis cartridge of FIG. 10.

[0049] FIG. 13 shows exemplary dimensions that could be used for the analysis cartridge of FIG.10.

[0050] FIGS. 14A-14H show different steps of sample processing in the analysis cartridge of FIG.10.

[0051] FIGS. 15A-15E show different stages during the separation of cellular components of a blood sample into the cellular components chamber and separation of the blood plasma in the sample metering chamber.

[0052] FIGS. 16A-16B d epict the connection between the sample metering chamber and the sample overflow chamber as well as the connection between the sample metering chamber and the cellular components chamber.

[0053] FIGS. 17A-17C depict separation of blood plasma from blood cells in the sample metering chamber and the cellular components chamber, respectively. B and C show collection of blood cells in the cellular components chamber and mechanisms that prevent the blood cells from traveling upstream into the sample metering chamber.

[0054] FIG. 18A-18D show different stages of the diluent movement from the diluent chamber into the diluent metering chamber.

[0055] FIGS. 19A-19C show an exemplary design of the diluent metering chamber. A shows a two-stage ramp that facilitates removal of metered fluid. B shows a diluent metering outlet, which is shallow with a sharp drop off designed to improve metering precision. C shows that a fluidic connection between the diluent metering chamber and a diluent siphon is not concentric to the vertical axis.

[0056] FIGS. 20A-20C show metering chamber and its outlet connection.

[0057] FIGS. 21A-21F show filling of diluted sample into a plurality of metering fingers before the diluted sample is analyzed in the detection chambers.

[0058] FIG. 22 shows certain features of the distribution channel that improve timings and reliability of the distribution channel.

[0059] FIG. 23 shows a cone shaped (trumpet shaped) design for the siphon outlet.

[0060] FIGS. 24A-24E show certain metering finger geometry that facilitates sequential filling of the metering fingers with a diluted sample.

[0061] FIG. 25 shows capillary stop mechanism of the detection chamber.

[0062] FIGS. 26A-26B depict structure of a detection chamber for a two-step reaction. A shows the top view and B shows the side view.DETAILED DESCRIPTION

[0063] Certain aspects of the present disclosure provide analysis cartridges that allow analyzing non-cellular constituents of a sample, such as blood.

[0064] In certain embodiments, an analysis cartridge is configured to be centrifuged around a vertical axis and comprises:

[0065] a diluent chamber fluidical ly connected to a diluent metering chamber, wherein a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0066] a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve and to a sample metering chamber via a sample valve such that, when the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0067] one or more detection chambers fluidically connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more detection chambers, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0068] Also provided herein are methods of analyzing a biological sample, such as a blood sample in the analysis cartridges provided herein. In some cases, a method of analyzing a sample in an analysis cartridge comprises:

[0069] conducting a first centrifugation of the analysis cartridge to move a diluent from a diluent chamber to a fluidically connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0070] conducting a second centrifugation of the analysis cartridge to move into a mixing chamber the diluent from the diluent metering chamber and the sample from the sample metering chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold, and

[0071] conducting a third centrifugation to move into a one or more detection chambers the diluted sample from the mixing chamber, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0072] Further provided herein are analyzers that use the analysis cartridges provided herein to analyze a biological sample, such as blood for non-cellular constituents.

[0073] In certain embodiments, an analyzer for analyzing a sample in an analysis cartridge comprises:

[0074] a rotor comprising a slot that holds the analysis cartridge such that, when the rotor rotates, the analysis cartridge is centrifuged around a vertical axis,

[0075] a processor, and

[0076] a non-transitory computer readable medium comprising instructions, which when executed by the processor, cause the analyzer to perform:

[0077] a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into a fluidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0078] when a diluent valve and a sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from a sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0079] a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more detection chambers, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0080] In a further aspect, the disclosure provides an analysis cartridge for analyzing a sample comprising one or more detection chambers and, fluidically connected to the detection chambers, one or more metering fingers that measure a predetermined volume of a sample to be introduced into the one or more detection chambers. In certain such embodiments, the analysis cartridge is configured to be centrifuged around a vertical axis and comprises:

[0081] a diluent chamber fluidically connected to a diluent metering chamber, wherein a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0082] a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve and to a sample metering chamber via a sample valve such that, when the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0083] one or more metering fingers fluidically connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0084] one or more detection chambers fluidically connected to the one or more metering fingers via capillary channels such that, a fourth centrifugation of the analysis cartridge moves the diluted sample from the one or more metering fingers into the one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold.

[0085] In an even further aspects, the disclosure provides a method of analyzing a sample in an analysis cartridge comprising one or more metering fingers, the method comprising:

[0086] conducting a first centrifugation of the analysis cartridge to move a diluent from a diluent chamber to a fluidically connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0087] conducting a second centrifugation of the analysis cartridge to move into a mixing chamberthe diluent from the diluent metering chamberand the sample from a sample metering chamber; wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0088] conducting a third centrifugation of the analysis cartridge to move the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0089] conducting a fourth centrifugation to move into a one or more detection chambers the diluted sample from the one or more metering fingers, wherein the fourth centrifugation exertson the diluted sample in the one or more metering fingers a fourth centrifugal force above a third threshold.

[0090] In certain aspects, the disclosure provides an analyzer for analyzing a sample in an analysis cartridges comprising one or more metering fingers, the analyzer comprising:

[0091] a rotor comprising a slot that holds the analysis cartridge such that, when the rotor rotates, the analysis cartridge is centrifuged around a vertical axis,

[0092] a processor, and

[0093] non-transitory computer readable medium comprising instructions, which when executed by the processor, cause the analyzer to perform:

[0094] a first centrifugation of the analysis cartridge that moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold

[0095] when a diluent valve and a sample valve are open, a second centrifugation of the analysis cartridge that moves the diluent from the diluent metering chamber and a sample from a sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0096] when a diluted sample valve is open, a third centrifugation of the analysis cartridge that moves the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0097] a fourth centrifugation of the analysis cartridge that moves the diluted sample from the one or more metering fingers into the corresponding one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold.

[0098] Before the present analysis cartridges, methods, and analyzers are described in greater detail, it is to be understood that the present disclosure is not limited to particular embodiments described, as such may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0099] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the analysis cartridges and methods. The upper and lower limitsof these smaller ranges may independently be included in the smaller ranges and are also encompassed within the analysis cartridges and methods, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the analysis cartridges and methods.

[0100] Certain ranges are presented herein with numerical values being preceded by the term "about." The term "about" is used herein to provide literal support for the exact number that it precedes, as well as a number that is near to or approximately the number that the term precedes. In determining whether a number is near to or approximately a specifically recited number, the near or approximating un-recited number may be a number which, in the context in which it is presented, provides the substantial equivalent of the specifically recited number.

[0101] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in practice or testing of the present disclosure. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.

[0102] The present disclosure may be understood more readily by reference to the following detailed description of desired embodiments and the examples included therein. In the following specification and the claims which follow, reference will be made to a number of terms which shall be defined to have the following meanings.

[0103] The terms "comprise(s)," "include(s)," "having," "has," "can," "contain(s)," and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms "a," "an" and "the" include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments "comprising," "consisting of" and "consisting essentially of," the embodiments or elements presented herein, whether explicitly set forth.

[0104] The term "comprising" is used herein as requiringthe presence of the named component and allowing the presence of other components. The term "comprising" should be construed to include the term "consisting essentially of" and "consisting of." The "consisting essentially of" allows the presence of the named component(s), along with other components which do notchange the function / structure of the named component(s). The "consisting of" allows the presence of the named component(s), along with any adhesives or other bonding means for attaching the listed component(s).

[0105] Numerical values should be understood to include numerical values which are the same when reduced to the same number of significant figures and numerical values which differ from the stated value by less than the experimental error of conventional measurement technique of the type described in the present application to determine the value.

[0106] For the recitation of numeric ranges herein, each intervening number therebetween with the same degree of precision is explicitly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated.

[0107] All ranges disclosed herein are inclusive of the recited endpoint and independently combinable (for example, the range of "from 2 to 10" is inclusive of the endpoints, 2 and 10, and all the intermediate values). The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value; they are sufficiently imprecise to include values approximating these ranges and / or values.

[0108] The modifier "about" used in connection with a quantity is inclusive of the stated value and has the meaning dictated by the context. When used in the context of a range, the modifier "about" should also be considered as disclosing the range defined by the absolute values of the two endpoints. For example, the range of from about "2 to about 10" also discloses the range "from 2 to 10." The term "about" may refer to plus or minus 10% of the indicated number. For example, "about 10%" may indicate a range of 9% to 11%, and "about 1" may mean from 0.9- 1.1.

[0109] It should be noted that many of the terms used herein are relative terms. For example, the terms "upper" and "lower" are relative to each other in location, i.e., an upper component is located at a higher elevation than a lower component in a given orientation, but these terms can change if the component is flipped. The terms "inlet" and "outlet" are relative to a fluid flowing through them with respect to a given structure, e.g., a fluid flows through the inlet into the structure and flows through the outlet out of the structure.

[0110] The terms "horizontal" and "vertical" are used to indicate direction relative to an absolute reference, i.e., ground level. Also, when used in the context of describing the features of the analysis cartridges described herein, the terms "horizontal" and "vertical" refer to the analysis cartridge when in operation, i.e., when the analysis cartridge is centrifuged around avertical axis, i.e., an axis vertical to the ground level. However, these terms should not be construed to require structures to be absolutely parallel or absolutely perpendicular to each other. For example, the first vertical structure and the second vertical structure are not necessarily parallel to each other. The terms "top" and "bottom" are used to refer to surfaces where the top is always higher than the bottom relative to an absolute reference, i.e., the surface of the earth. The terms "upwards" and "downwards" are also relative to an absolute reference; upwards is always against the gravity of the earth while downwards is always towards the gravity of the earth.

[0111] The term "parallel" should be construed in its lay sense of two surfaces that maintain a generally constant distance between them, and not in the strict mathematical sense that such surfaces will never intersect when extended to infinity.

[0112] The term "proximate location" or "proximate end" or a grammatical variation thereof in the context of the location on an analysis cartridge refers to a location that is closer to the vertical axis around which the analysis cartridge is centrifuged or configured to be centrifuged. Similarly, the term "distal location" or "distal end" or a grammatical variation thereof in the context of the location on an analysis cartridge refers to a location that is closer to the periphery of the analysis cartridge. When two locations on an analysis cartridge are compared, a location closer to the vertical axis around which the analysis cartridge is centrifuged or configured to be centrifuged is "proximate" to the location that is closer to the periphery and, conversely, a location closer to the periphery is "distal" to the location that is closer to the vertical axis around which the analysis cartridge is centrifuged or configured to be centrifuged. When two locations are equidistant from the vertical axis around which the analysis cartridge is centrifuged or configured to be centrifuged, such locations are referenced as "concentric," i.e., indicating that they fall on the circumference of an imaginary circle with the center from which passes the vertical axis around which the analysis cartridge is centrifuged or configured to be centrifuged.

[0113] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates, which may need to be independently confirmed.

[0114] The claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely," "only," and the like in connection with the recitation of claim elements or use of a "negative" limitation.

[0115] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present analysis cartridges and methods. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.ANALYSIS CARTRIDGES

[0116] As summarized above, certain aspects of the present disclosure provide analysis cartridges for analyzing samples, particularly, analyzing non-cellular constituents of a biological sample, such as blood. Also, certain aspects of the disclosure provide analysis cartridges that allow analysis of small volumes of samples.

[0117] In some cases, certain analysis cartridges described herein comprise one or more detection chambers that receive a sample directly from a mixing chamber that houses a diluted sample. In certain such cases, the volume of the detection chambers is predetermined such that the concentration of an analyte analyzed in a detection chamber is calculated based on the predetermined volume and the signal generated in the reaction mixture.

[0118] Alternatively, in some cases, certain analysis cartridges described herein comprise one or more detection chambers that receive samples from a mixing chamber that houses a diluted sample via intermediate metering fingers. The metering fingers have predetermined volume such that a predetermined volume of a diluted sample is introduced into the detection chambers. Thus, the concentration of an analyte analyzed in a detection chamber is calculated based on the predetermined volume of a metering finger and the signal generated in the reaction mixture.ANALYSIS CARTRIDGES WITHOUT METERING FINGERS

[0119] In certain aspects, the disclosure provides an analysis cartridge for analyzing a sample. An analysis cartridge is configured to be centrifuged around a vertical axis, and comprises:

[0120] a d iluent chamber fluidical ly connected to a diluent metering chamber, wherein a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluentmetering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0121] a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve and to a sample metering chamber via a sample valve such that, when the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0122] one or more detection chambers fluidically connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more detection chambers, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0123] The term "configured to be centrifuged around a vertical axis" indicates that the analysis cartridge has a suitable shape that can be fitted onto a slot of a centrifuge rotor. Also, different components of the analysis cartridge are located such that centrifugation of the cartridge moves fluids from certain compartments into other compartments as described in this disclosure. For example, a diluent metering chamber is fluidically connected to a diluent chamber such that, compared to the diluent chamber, the diluent metering chamber is located away from the vertical axis, i.e., a diluent metering chamber is distal to the diluent chamber. Thus, centrifuging the analysis cartridge around the vertical axis would exert centrifugation force on the diluent in the diluent chamber and move it to the diluent metering chamber.

[0124] The vertical axis can be external to the cartridge, i.e., the cartridge rotates around an axis outside the body of the cartridge. In such cases, multiple analysis cartridges are arranged with a common vertical axis of rotation that is external to the bodies of the multiple analysis cartridges. The rotor can have multiple slots to accommodate the multiple analysis cartridges. Typically, two or more such analysis cartridges are fitted into two or more slots of a centrifuge rotor. For example, a centrifuge rotor can comprise between 2 to 10 slots that are radially arranged around a vertical axis.

[0125] In some cases, the slots are pie-shaped thereby allowing pie shaped analysis cartridges to be placed therein. An example of such pie-shaped analysis cartridge is shown in FIGS. 1A-1B. However, the slots can be of any suitable shape to hold analysis cartridges of the correspondingshape. For example, a rectangular slot can hold a rectangular analysis cartridge or a triangular slot can hold a triangular analysis cartridge.

[0126] The vertical axis can be internal to the cartridge, i.e., the cartridge rotates around an axis that goes through the body of the cartridge. In such cases, the rotor can have only one slot to accommodate a single analysis cartridge that rotates around itself.

[0127] The term "fluidically connected" indicates that the two components of an analysis cartridge are directly or indirectly connected so that a fluid present in one component can be transferred to the other component. For example, a diluent chamber fluidically connected to a diluent metering chamber indicates that the diluent chamber has a direct fluidic connection to the diluent metering chamber or an indirect connection through intervening connections or chambers.

[0128] In some cases, the analysis cartridge comprises a diluent chamber fluidically connected to a diluent metering chamber. The connection can be microfluidic channel with or without a valve. A diluent chamber is fluidically connected to a diluent metering chamber such that a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber. The first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold. For example, a centrifugation force above a first threshold can overcome the surface tension of the diluent in a microfluidic channel or force open a valve to allow the diluent from the diluent chamber to flow to the diluent metering chamber.

[0129] A diluent metering chamber has a pre-determined volume and is fluidically connected to a diluent overflow chamber. Thus, upon centrifugation of the analysis cartridge, a diluent from a diluent chamber fills a predetermined volume of the diluent in the diluent metering chamber, with the excess diluent flowing into the diluent overflow chamber. Thus, a first centrifugation of an analysis cartridge loads a predetermined volume of the diluent in the diluent metering chamber. This volume can be used in calculating concentrations of the one or more analytes in the sample.

[0130] An analysis cartridge disclosed herein also comprises a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve. The mixing chamber is also fluidically connected to a sample metering chamber via a sample valve. When the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber. The second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above asecond threshold. For example, a centrifugation force above the second threshold can overcome the surface tension of the diluent in a microfluidic channel or force open a valve to allow the diluent from the diluent metering chamber to flow into the mixing chamber. Similarly, a centrifugation force above the second threshold can overcome the surface tension of the sample in a microfluidic channel or force open a valve to allow the sample from the sample metering chamber to flow to the mixing chamber.

[0131] While the second centrifugation can move both the diluent and the sample into the mixing chamber, in some cases, the device can be configured so that the diluent is moved into the mixing chamber in a centrifugation step and the sample is moved into the mixing chamber in a separate centrifugation step.

[0132] An analysis cartridge disclosed herein can also further comprise one or more detection chambers. The one or more detection chambers are fluidically connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more detection chambers. The third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold. For example, a centrifugation force above the third threshold can overcome surface tension of the diluted sample in a microfluidic channel or force open a valve to allow the diluted sample from the mixing chamber to the one or more detection chambers.

[0133] In some cases, the sample metering chamber is fluidically connected to a sample chamber. A sample can be applied to the sample chamber through a fluidic connection to a sample reservoir. A sample can also be applied to the sample chamber by a user, for example, using a pipette.

[0134] A sample loaded into the sample chamber can be moved to the sample metering chamber by subjecting the analysis cartridge to a fourth centrifugation. The fourth centrifugation exerts on the sample in the sample chamber a fourth centrifugation force above a fourth threshold. For example, a centrifugation force above the fourth threshold can overcome surface tension of the sample in a microfluidic channel or force open a valve to allow the sample from the sample chamber to the sample metering chamber.

[0135] In some cases, the third threshold is higher than the second threshold and the second threshold is higher than the first threshold. Also, in some cases, the fourth threshold is lower than the first and / or the second threshold.

[0136] Thus, in some cases, the fourth centrifugation only allows a sample from the sample chamber to flow to the sample metering chamber. However, the fourth centrifugation does not exert enough centrifugation force on the sample in the sample metering chamber to move it from the sample metering chamber to the mixing chamber. Similarly, in some cases, the first centrifugation only allows a diluent from the diluent chamber to flow to the diluent metering chamber. However, the first centrifugation does not exert enough centrifugation force on the diluent in the diluent metering chamber to move it from the diluent metering chamber to the mixing chamber.

[0137] The second centrifugation then exerts enough centrifugation force on the diluent in the diluent metering chamber to move it from the diluent metering chamber to the mixing chamber. The second centrifugation also exerts enough centrifugation force on a sample in the sample metering chamber to move it from the sample metering chamber to the mixing chamber. However, the second centrifugation does not exert enough centrifugation force on the diluted sample in the mixing chamber to move it from the mixing chamber to the one or more detection chambers. The third centrifugation then exerts enough centrifugation force on the diluted sample in the mixing chamber to move it from the mixing chamber to the one or more detection chambers.

[0138] Accordingly, the analysis cartridge can be centrifuged at different speeds in a step-wise manner to cause different fluids to move from certain specific compartments to certain other specific compartments.

[0139] In some cases, the sample metering chamber further comprises a sample overflow conduit. The sample overflow conduit can be fluidically connected to a sample overflow chamber. Thus, the sample beyond the volume of the sample metering chamber flows to the sample overflow chamber. With this arrangement, a predetermined volume of the sample is collected in the sample metering chamber. This volume can be used in calculating concentrations of one or more analytes in the sample.

[0140] In some embodiments, the sample metering chamber further comprises a cellular components chamber. The cellular component chamber can be used to separate cellular components of the sample. For example, when the sample is blood, blood cells can be collected in the cellular components chamber and blood plasma can be collected in the sample metering chamber. In some cases, centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components chamber and leaves the non-cellular components of the sample into the sample metering chamber.

[0141] In some cases, the one or more detection chambers of an analysis cartridge comprise optical detection chambers. In addition, an analysis cartridge can further comprise one or more electrochemical sensors to electrochemically detect one or more analytes in the sample. For electrochemical detection of analytes, cellular components need not be separated from the non- cellular components.

[0142] Accordingly, one or more electrochemical sensors can be fluidically connected to a sample chamber. For example, a first set of one or more electrochemical sensors that detect a first set of one or more analytes in the sample can be located in the sample overflow chamber and / or the sample overflow conduit.

[0143] Alternatively, a sample chamber is fluidically connected to an electrochemical analysis module. In some cases, a fifth centrifugation of the analysis cartridge moves a portion of the sample from the sample chamber into the electrochemical analysis module. In some cases, the fifth centrifugation can exert on a sample in the sample chamber sufficient centrifugal force to move the sample from the sample chamber into the electrochemical analysis module. The fifth centrifugal force can be below the fourth threshold, discussed above.

[0144] The one or more electrochemical sensors, for example, the electrochemical sensors present in the sample overflow chamber, the sample overflow conduit, and / or the electrochemical analysis module can comprise amperometric sensors, coulometric sensors, potentiometric sensors, voltametric sensors, impedance sensors, or a combination thereof.

[0145] In some cases, the one or more electrochemical sensors, for example, the electrochemical sensors present in the sample overflow chamber, the sample overflow conduit, and / orthe electrochemical analysis module can comprise sensors to detect one or more analytes in a sample, such as one or more analytes in a blood sample. Such analytes include tests that are routinely ordered to determine a subject's general health status. In some cases, the clinical chemistry panels include metabolic panels. The clinical chemistry panels help evaluate, for example, the body's electrolyte balance and / or the status of several major body organs. Examples of clinical chemistry panels that may be detected by assays of the present disclosure include, but are not limited to, basic metabolic panel (BMP), comprehensive metabolic panel (CMP), electrolyte panel, lipid panel, liver panel, renal panel, and thyroid function panel. The basic metabolic panel (BMP) includes 8 tests, all of which are found in the CMP. The BMP provides information about the current health of kidneys and respiratory system as well as electrolyte and acid / base balance and level of blood glucose. The CMP measurement is used for liver and kidney health, level of blood glucose, acid / base balance in blood, fluid and electrolytebalance, and important blood proteins. In some cases, the CMP measures glucose, calcium, total amount of albumin and globulins, bilirubin, BUN (blood urea nitrogen), creatinine, albumin, sodium, potassium, bicarbonate, chloride, alkaline phosphatase (ALP), alanine transaminase (ALT), and aspartate aminotransferase (AST). The electrolyte panel is used to detect a problem with the body's fluid and electrolyte balance. For example, the electrolyte panel measures the blood levels of carbon dioxide, chloride, potassium, and sodium. The lipid panel is used to assess a subject's risk of developing cardiovascular disease. For example, the lipid panel measures the amount of cholesterol and other fats in blood, such as total cholesterol, LDL (low-density lipoprotein), HDL (high-density lipoprotein), and triglycerides. The liver panel (hepatic function panel) is used to screen for, detect, evaluate, and monitor acute and chronic liver inflammation (hepatitis), liver disease and / or damage. The liver panel measures different enzymes, proteins, and other substances made by liver. For example, the liver panel includes albumin, total protein, ALP, ALT, AST, gamma-glutamyl transferase (GGT), bilirubin, Lactate dehydrogenase (LD), Prothrombin time (PT). The renal panel (kidney function panel) includes tests such as albumin, creatinine, BUN, eGFR to evaluate kidney function. The thyroid Function Panel is used to evaluate thyroid gland function and to help diagnose thyroid disorders. The thyroid function panel measure thyroid hormone such as thyroxine (T4), triiodothyronine (T3), and thyroid stimulating hormone (TSH). In some cases, a high TSH level indicates that the thyroid gland is not making enough thyroid hormone (primary hypothyroidism). The opposite situation, in which the TSH level is low, usually indicates that the thyroid is producing too much thyroid hormone (hyperthyroidism). In other cases, the finding of an elevated TSH and low free T4 (FT4) or free T4 index (FTI) indicates primary hypothyroidism due to disease in the thyroid gland. A low TSH and low FT4 or FTI indicate hypothyroidism due to a problem involving the pituitary gland. A low TSH with an elevated FT4 or FTI is found in individuals who have hyperthyroidism. These clinical chemistry panels are well known in the art and are further described in the assay portion of the present disclosure.

[0146] In some cases, such as where an analysis cartridge is employed for a complete metabolic panel (CMP), reagents comprise one or more of the following: 2,4,6-Tribromo-3-hydroxybenzoic acid (TBHBA), 2-Chloro-4-nitrophenyl-a-maltotrioside (CNPG3), 2-Methyl-4-isothizolin-3-one hydrochloride (MIT), 4,7,13,16,21-Pentaoxa-l,10-diazabicyclo[8.8.5]tricosane (Kryptofix 221), 4- Aminoantipyrine hydrochloride, Adenosine 5'-diphosphate, Adenosine 5'-triphosphate, a- ketoglutaric acid, Amylase, Arsenazo III, sodium salt, Ascorbate oxidase (Cucurbita spp.), Bilirubin oxidase, Bromcresol purple, Calcium acetate, Creatine amidinohydrolase (Actinobacillus spp.),Creatinine amidohydrolase (Pseudomonas spp.), Cupric sulfate, Ethylene glycol-bis(B- aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), Ethylenediaminetetraacetic acid (EDTA), R- Galactosidase, Glucose-6-phosphate dehydrogenase (yeast), Glutamate Dehydrogenase (bovine liver), Glutamine synthetase, Hexokinase (yeast), Imidazole, Lactate dehydrogenase, L-alanine, L-aspartic acid, L-glutamic acid, Magnesium chloride, Magnesium sulfate, Malate dehydrogenase (porcine heart), Manganese chloride, N-Acetyl cysteine, R-Nicotinamide adenine dinucleotide (NAD), R-Nicotinamide adenine dinucleotide, reduced (NADH), o-Nitrophenyl-15-D- galactopyranoside (ONPG), Peroxidase (horseradish), Phosphoenol pyruvate, Phosphoenol pyruvate carboxylase, p-NPP, Potassium ferrocyanide, Potassium iodide, Pyruvate kinase, Sarcosine oxidase (microorganism), Sodium potassium tartrate, Urease (jack bean), Zinc sulfate, as well as other buffers, surfactants, excipients and preservatives.

[0147] In some cases, the one or more electrochemical sensors are also fluidically connected to a calibration fluid chamber. A calibration fluid can be contacted to the one or more electrochemical sensors via centrifugation of an analysis cartridge. The calibration fluid can be used to provide a blank reading or a reference reading for one or more analytes analyzed via the one or more electrochemical sensors.

[0148] As noted above, different chambers in an analysis cartridge described herein can be connected to each other via valves. Any suitable microfluidic valve can be implemented in an analysis cartridge.

[0149] In some cases, the valves used in the analysis cartridges disclosed herein are siphon valves. For example, a diluent valve is a diluent siphon, a sample valve is a sample siphon, and / or a diluted sample valve is a diluted sample siphon.

[0150] In some cases, when the diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon, the siphon is operated by priming. Operation of an exemplary siphon valve in a microfluidic centrifugation based cartridge is described by Zehnle et al. (2015), Microfluidics and Nanofluidics; 19(6):1259-1269, which is incorporated herein by reference in its entirety.

[0151] Briefly, a siphon comprises a crest, a critical fill level, and a first and second chambers connected by the siphon. Certain such arrangement is shown and described in Zehnle et al. When a siphon valve is in a closed state, the liquid is retained in a first chamber, which, as compared to a second chamber, is closer to the rotational axis of an analysis cartridge, i.e., the first chamber is proximate and the second chamber is distal. In an open state the siphon is primed by filling the siphon to a critical fill level for siphon priming. When a siphon is primed,the liquid is transferred under centrifugal force from the first chamber to a second chamber, the second chamber, as compared to the first chamber, being farther to the rotational axis of an analysis cartridge. To prime a siphon, an analysis cartridge is centrifuged to exert a centrifugal force on a liquid in a first chamber such that the liquid crosses the siphon crest, which then allows the liquid to flow to a second chamber. For example, the diluent siphon and the sample siphon are primed by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold. Similarly, the diluted sample siphon is primed by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

[0152] Once a diluted sample is provided to the one or more detection chambers, the reagents in the detection chambers produce signals, such as optical signals, that indicate concentrations of one or more analytes in the sample. In some cases, distribution of the diluted sample to the one or more detection chambers can be via a distribution channel that fluidically connects the mixing chamber to the one or more detection chambers.

[0153] Exemplary detection chambers include microcuvettes. The number of detection chambers may vary. In some cases, the number of detection chambers ranges from 2 to 100, such as 5 to 50, such as 10 to 25 and including 14 to 20. In some cases, cartridges include 10 or more detection chambers, such as 11 or more detection chambers, such as 12 or more detection chambers, such as 13 or more detection chambers, such as 14 or more detection chambers, such as 15 or more detection chambers, such as 16 or more detection chambers, such as 17 or more detection chambers, such as 18 or more detection chambers, such as 19 or more detection chambers, and including 20 or more detection chambers. The shape and size of the detection chambers in the plurality may vary, as desired.

[0154] The shape and size of the detection chambers may vary, as desired. In certain cases, the detection chambers have an elongated structure (e.g., having a length greater than width). The elongate structure may have any convenient cross-sectional shape, where cross-sectional shapes of interest include, but are not limited to rectilinear cross-sectional shapes, e.g., squares, rectangles, trapezoids, triangles, hexagons, etc., curvilinear cross-sectional shapes, e.g., circles, ovals, as well as irregular shapes, e.g., a parabolic bottom portion coupled to a planar top portion. In certain cases, the detection chambers of the plurality have a circular cross section. In other embodiments, the detection chambers have a square cross section. In still other embodiments, detection chambers have a rectangular cross section. The volume of the detectionchambers may also vary. In some cases, detection chambers of the plurality have a volume ranging from 0.3 pl to 500 pl, such as 2 pl to 300 pl, such as 3 pl to 200 pl, such as 4 pl to 100 pl and including 5 pl to 10 pl. In select versions, detection chambers of the plurality have a volume ranging from 0.3 pl to 50 pl. In certain cases, detection chambers of a volume of 5 pl or more, such as 6 pl or more, such as 7 pl or more, such as 8 pl or more, such as 9 pl or more, and including 10 pl or more. In some instances, detection chambers have a diameter ranging from 0.1 mm to 20 mm, such as 0.5 mm to 15 mm, such as 1 mm to 10 mm, and including 1.5 mm to 2 mm. In certain cases, detection chambers have a diameter of 1.5 mm or more, such as 1.6 mm or more, such as 1.7 mm or more, such as 1.8 mm or more, such as 1.9 mm or more, and including 2 mm or more. Adjacent detection chambers may be separated by a distance ranging from 1 mm to 10 mm, such as 2 mm to 8 mm, and including 4 mm to 5 mm. Space between detection chambers may in some cases be sufficient to ensure that each one can be interrogated by a beam of light from the illuminator without interfering with its neighbor detection chambers.

[0155] The detection chambers in the subject cartridge may be constructed from any suitable material. In some cases, the detection chambers are comprised of a polymeric material that is transparent in a detection wavelength band. In some such cases, the detection chambers of the plurality are comprised of polystyrene (PS), PMMA, CoC, or CoP.

[0156] Detection chambers of the subject analysis cartridges may include an inlet for receiving a diluted sample, and an outlet where air and / or excess diluted sample fluid may escape as the detection chambers are being filled. In some embodiments where the detection chambers are comprised of an elongate structure, the detection chambers include an inlet at a proximal end of the elongate structure, and an outlet at the distal end of the elongate structure. In certain instances, the cartridge is configured such that the detection chambers are arranged upright (i.e., vertically). In some such embodiments, the inlets may be arranged at the bottom such that the detection chambers fill with diluted sample from the bottom and ascend via capillary action. In some cases, this arrangement is sufficient to minimize the generation of air bubbles when sample fluid fills the detection chambers. In some alternate embodiments, detection chambers fill from the top-down. In certain cases, the detection chambers are light-accessible at certain windows. In some such cases, each detection chamber comprises a first light-accessible window configured to permit entry of light, and a second light-accessible window configured to permit an exit of the light from the light source. The remainder of the detection chambers may or may not also be light-accessible. In select versions, aside from the first and second light-accessible windows, the detection chambers are opaque.

[0157] In embodiments, the outlets of the detection chambers have edges that are not perpendicular to the edges of the detection chamber. In some such embodiments, the outlets are beveled. Put another way, at least a portion of the outlet may be chamfered. Beveling the interface between the detection chamber and additional components (e.g., outlet vent, overflow reservoir, etc.) may be helpful to reduce bubble formation.

[0158] In some embodiments, cartridges include one or more capillary stops (i.e., capillary stop valves). As is understood in the art, capillary stops halt the flow of liquid in microchannels without external intervention using an abrupt change in microchannel geometry. In some cases, the capillary stop constitutes an abrupt expansion of detection chamber geometry. In other cases, the capillary stop constitutes an abrupt narrowing of detection chamber geometry. In certain instances, microfluidic channels are molded between detection chambers for transferring sample fluid and waste. In select instances, cartridges include an overflow reservoir fluidical ly connected to each detection chamber. Said overflow reservoir may constitute a void in the cartridge into which diluted sample fluid may flow if / when said fluid surpasses the capillary stop. Additionally or alternatively, cartridges may include an overflow outlet vent gaseously connected to each capillary stop. Said overflow outlet vent may constitute a microfluidic channel gaseously connected to each capillary stop that vents to the surrounding environment.

[0159] The one or more detection chambers can contain reagents, for example, dried reagents, that are configured to analyze specific compounds within the sample. In some cases, the cartridge is pre-filled with reagents and is ready to use. In some embodiments, all or a portion of the reagents may be present. Depending on the type of reagents present in the detection chambers and the resultant signal produced for the analyzed non-cellular constituents, appropriate detectors can be used to detect the signals.

[0160] The analysis cartridges may need to be stored in appropriate conditions to preserve the reactivity of the reagents. For example, depending on the reagents present, the analysis cartridges may need to be stored in a refrigerator or a freezer before use. When the reagents are not sensitive to room temperature, the analysis cartridges may be stored at room temperature.

[0161] The one or more dry reagents may in certain embodiments comprise a one or more non- fluorescent or fluorescent dyes such as Eosin, Methylene Blue, Acridine Orange (also referred to as "Basic Orange 15" or "ACO"), or Astrazon Orange (also referred to as "AO" or Basic Orange 21), a component to bind to nucleic acids, an anticoagulant, an antibody, an antibody fragment, an ionophore, an enzyme, a set of enzymes, a peptide with a cleavable detectable moiety, asubstrate, an optical marker dye identifying a type of assay bead, and / or combinations thereof. In another case, when dry reagents are mixed with sample, for example, plasma, a colorimetric reaction occurs which may be optically detectable.

[0162] The one or more detection chambers can contain reagents for analysis of one or more analytes within a sample. The analytes described above with respect to electrochemical sensors could also be analyzed in one or more detection chambers using optical detection techniques. Suitable optical detection techniques for various analytes are known in the art and use of such techniques are within the purview of the disclosure.ANALYSIS CARTRIDGES HAVING METERING FINGERS

[0163] As noted above, in some cases, the disclosure provides analysis cartridges comprising one or more metering fingers. Such one or more metering fingers are fluidically connected to one or more detection chambers that carry out detection and analysis of one or more analytes. Certain details of the analysis cartridges describes above under "Analysis Cartridges Without Metering Fingers" are also applicable to analysis cartridges having metering fingers and such embodiments are within the purview of the disclosure. Such details include the various fluidic connections between different chambers, use and functionality of siphons, types of analytes analyzed, types of reagents used, shapes and geometries of various chambers, shapes and geometries of the analysis cartridges, volumes of various chambers, material used for fabrication of the analysis cartridges, a first and a second set of electrochemical sensors and their locations, and the like.

[0164] Accordingly, certain embodiments of the disclosure provide an analysis cartridge for analyzing a sample, the analysis cartridge configured to be centrifuged around a vertical axis, the analysis cartridge comprising:

[0165] a d iluent chamber fluidically connected to a diluent metering chamber, wherein a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0166] a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve and to a sample metering chamber via a sample valve such that, when the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent meteringchamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0167] one or more metering fingers f luidica I ly connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0168] one or more detection chambers, each fluidical ly connected to one of the one or more metering fingers via a capillary channel such that, a fourth centrifugation of the analysis cartridge moves the diluted sample from the one or more metering fingers into the one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold.

[0169] In some cases, the fourth threshold is higherthan the third threshold, the third threshold is higher than the second threshold and the second threshold is higher than the first threshold.

[0170] For example, the first centrifugation only allows a diluent from the diluent chamber to flow to the diluent metering chamber. However, the first centrifugation does not exert enough centrifugation force on the diluent in the diluent metering chamber to move it from the diluent metering chamber to the mixing chamber.

[0171] Si milarly, the second centrifugation then exerts enough centrifugation force on the diluent in the diluent metering chamber to move it from the diluent metering chamber to the mixing chamber. The second centrifugation also exerts enough centrifugation force on a sample in the sample metering chamber to move it from the sample metering chamber to the mixing chamber. However, the second centrifugation does not exert enough centrifugation force on the diluted sample in the mixing chamber to move it from the mixing chamber to the one or more detection chambers.

[0172] The third centrifugation then exerts enough centrifugation force on the diluted sample in the mixing chamber to move it from the mixing chamber to the one or more metering fingers. The third centrifugation does not exert enough centrifugation force on the diluted sample in the one or more metering fingers to move it to the one or more detection chambers. The fourth centrifugation then exerts enough centrifugation force on the diluted sample in the metering fingers to move it from the metering fingers to the one or more detection chambers.

[0173] In some cases, the analysis cartridge comprises a diluent chamber fluidical ly connected to a diluent metering chamber. The connection can be microfluidic channel with or without avalve. A diluent chamber is fluidically connected to a diluent metering chamber such that a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber. The first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold. For example, a centrifugation force above a first threshold can overcome surface tension of the diluent in a microfluidic channel or force open a valve to allow the diluent from the diluent chamber to flow to the diluent metering chamber.

[0174] A diluent metering chamber has a pre-determined volume and is fluidically connected to a diluent overflow chamber. Thus, upon centrifugation of the analysis cartridge, a diluent from a diluent chamber fills a predetermined volume of the diluent in the diluent metering chamber, with the excess diluent flowing into the diluent overflow chamber. Thus, a first centrifugation of an analysis cartridge loads a predetermined volume of the diluent in the diluent metering chamber. This volume can be used in calculating concentrations of the one or more analytes in the sample.

[0175] The diluent chamber can have a volume sufficient to accommodate a diluent having a volume selected from: between 500 pL and 600 pL, about 500 pL, about 510 pL, about 520 pL, about 530 pL, about 540 pL, about 550 pL, about 560 pL, about 570 pL, about 580 pL, about 580 pL, and about 600 pL. The diluent metering chamber can have a volume selected from: between 450 pL and 500 pL, about 450 pL, about 455 pL, about 460 pL, about 465 pL, about 470 pL, about 475 pL, about 480 pL, about 485 pL, about 490 pL, about 495 pL, and about 500 pL. In some cases, the total volume of the diluent chamber is between 20% and 100% higherthan the volume of the diluent collected therein.

[0176] In some cases, the sample metering chamber is fluidically connected to a sample chamber. A sample can be applied to the sample chamber through a fluidic connection to a sample reservoir. A sample can also be applied to the sample chamber by a user, for example, using a pipette.

[0177] A sample loaded into the sample chamber can be moved to the sample metering chamber by subjecting the analysis cartridge to a fifth centrifugation. The fifth centrifugation exerts on the sample in the sample chamber a fifth centrifugation force above a fifth threshold. For example, a centrifugation force above the fifth threshold can overcome surface tension of the sample in a microfluidic channel or force open a valve to allow the sample from the sample chamber to flow to the sample metering chamber.

[0178] In some cases, a sample chamber is fluidically connected to a sample metering chamber via a channel having a cross section area that does not cause clogging of the channel, forexample, because of the surface tension of the sample. To that end, in some cases, a sample chamber is fluidically connected to a sample metering chamber via a channel having a cross section area of: between 0.2 mm2and 0.5 mm2, about 0.2 mm2, about 0.25 mm2, about 0.3 mm2, about 0.35 mm2, about 0.4 mm2, about 0.45 mm2, and about 0.5 mm2.

[0179] In some cases, the fifth threshold is lower than the first and / or the second threshold so that the fifth threshold does not cause the sample from the sample metering chamber to move into the mixing chamber. Thus, in some cases, the fifth centrifugation only allows a sample from the sample chamber to flow to the sample metering chamber. However, the fifth centrifugation does not exert enough centrifugation force on the sample in the sample metering chamber to move it from the sample metering chamber to the mixing chamber.

[0180] In some cases, the sample metering chamber has fluidically connected to it a cellular components chamber. In certain such cases, a first or a fifth centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components chamber. In some cases, when a fifth centrifugation is used to move the cellular components of the sample into the cellular components chamber, the fifth threshold is lower than the first and / or the second threshold so that the fifth centrifugation does not allow the sample from the sample metering chamber to enter the mixing chamber during the fifth centrifugation.

[0181] To facilitate movement of sample, particularly, cellular components, from a sample metering chamber into a cellular components chamber, a fluidic connection between the sample metering chamber and the cellular components chamber comprises two channels with different depths. Thus, in some cases, a fluidic connection between the sample metering chamber and the cellular components chamber has a deeper channel and a shallower channel. An example of such connecting channel is shown in FIGS. 16A-16B. Such an arrangement of a connecting channels reduces the risk of air lock during the initial transport of the sample from the sample metering chamber into the cellular components chamber. In some cases, the deeper channel is between 0.2 and 0.5 mm deeper than the shallower channel.

[0182] As noted above, in the context of the location on an analysis cartridge refers to, a proximate location that is closer to the vertical axis around which the analysis cartridge is centrifuged or configured to be centrifuged. In some cases, as shown in FIG. 16B, the proximate wall of the sample metering chamber can be angled such that the fluidic connection between the sample metering chamber and the sample overflow chamber is proximate compared to the other end of the proximate wall of the sample metering chamber. In some cases, this angle is about 2 degrees. Such a design makes the connection to the sample overflow chamber moreproximate than the other end of the proximate wall of the sample metering chamber, which also facilitates venting of any bubbles into the sample overflow chamber.

[0183] In some cases, a sample metering chamber is fluidically connected to a sample overflow chamber such that the first centrifugation or the fifth centrifugation of the analysis cartridge moves into the sample overflow chamber excess sample that does not fit in the sample metering chamber.

[0184] In certain embodiments of analysis cartridges, the fluidic connection between the sample metering chamber and the sample overflow chamber is towards the top of the proximate wall of the sample metering chamber. Thus, a sample loaded in the sample metering chamber flows to the sample overflow chamber only when the sample metering chamber is fully filled. Also, in some cases, a fluidic connection between a sample metering chamber and a sample overflow chamber covers at the most 20% of the height and at the most 30% of the width of the proximate wall of the sample metering chamber. This location and dimension of such connection ensures that a sample that has flowed to a sample overflow chamber cannot return to a sample metering chamber. This location and dimension of such connection also improves the accuracy of the sample metering.

[0185] Moreover, to facilitate that the sample metering chamber is first filled at the proximate end and a sample does not easily flow into the cellular component chamber, in some cases, the floor of the sample metering chamber has an upward ramp relative to the bottom of the chamber, wherein the upward ramp is in the proximate to the distal direction, i.e., the ramp is slanted from the proximal end to the distal end of the sample metering chamber. An example of such ramp is described is described in FIG. 16A. In some cases, the ramp can comprise one or more stages, for example, one, two, three, four, or five stages.

[0186] In certain such cases, the sample overflow chamber comprises a sample overflow vent that vents the gas from the sample metering chamber as the sample moves into the sample metering chamber.

[0187] As noted above, the first centrifugation or the fifth centrifugation of an analysis cartridge separates cellular components in a sample in the cellular components chamber and collects a liquid portion of the sample in the sample metering chamber. After centrifugation and separation of cellular components of the sample in the cellular components chamber, it is desirable to keep the separated cellular components within the cellular component chamber. Therefore, cellular components chamber is designed to reduce the risk of cellular components flowing from the cellular components chamber back into the sample metering chamber. Toachieve that, in some cases, a cellular components chamber has a substantially horizontal floor and substantially perpendicular walls. An example of such design is shown in FIGS. 17B and 17C. As shown in FIG. 17B, the cellular components chamber can be designed to collect and pack cells, e.g., RBCs, at the widest and the most distal area. Also, as shown in FIG. 17C, during deceleration, the packed cells, e.g., RBC's may resuspend, for example, because of acceleration vortices. During resuspension, the steep perpendicular walls of the cellular components chamber prevents the cells from traveling towards and into the sample metering chamber.

[0188] In some cases, to facilitate the movement of sample between various chambers, the connections are designed to be in line with the vertical axis. For example, in some cases, the fluidic connection between the sample chamber and the sample metering chamber is in line with the vertical axis. Similarly, in some cases, the fluidic connection between the sample metering chamber and the cellular components chamber is in line with the vertical axis.

[0189] In some cases, the sample chamber has a volume sufficient to accommodate a sample having a volume selected from: between 50 pL and 70 pL, about 50 pL, about 52 pL, about 54 pL, about 56 pL, about 58 pL, about 60 pL, about 62 pL, about 64 pL, about 66 pL, about 68 pL, and about 70 pL. In some cases, the total volume of the sample chamber is between 20% and 100% higher than the volume of the sample loaded therein.

[0190] In some cases, the sample metering chamber has a volume selected from: between 8 pL and 15 pL, about 8 pL, about 9 pL, about 10 pL, about 11 pL, about 12 pL, about 13 pL, about 14 pL, and about 15 pL.

[0191] When the sample is blood, a blood sample loaded in the sample chamber can be centrifuged to separate the blood cells in the cellular components chamber thereby allowing blood plasma to collect in the sample metering chamber.

[0192] In some cases, a blood sample is a human blood sample. In some cases, the human blood sample a hematocrit range between 30% and 70%.

[0193] In some cases, a fluidic connection between a diluent metering chamber and a diluent overflow chamber is towards the top of the proximate wall of the diluent metering chamber. In some cases, such fluidic connection covers at the most 20% of the height and at the most 10% of the width of the proximate wall of the diluent metering chamber. This arrangement facilitates accuracy of diluent metering and reduces the risk that a diluent that has left the diluent metering chamber towards the diluent overflow chamber does not return to the diluent metering chamber.

[0194] In some cases, to facilitate the accuracy of diluent metering, a ramp is provided in the diluent metering chamber. Particularly, the distal wall of the diluent metering chamber comprises a ramp in one or more stages, for example, one, two, three, or four stages. The ramp in one or more stages in the diluent metering chamber is from the proximate to the distal direction, i.e., the one or more stages of the ramp are slanted up from the proximal end to the distal end of the diluent metering chamber. As shown in FIG. 19A, in an exemplary diluent metering chamber, the distal wall has a ramp with the first stage 1901 and the second stage 1902.

[0195] The ramp with one or more stages in the diluent metering chamber allows keeping the local volume of diluent fluid in excess compared to where the entry to the diluent siphon is from the diluent metering chamber. As the analysis cartridge is centrifuged, the ramp in the diluent metering chamber facilitates the movement of the diluent distally and upwards towards the diluent siphon, e.g., towards the diluent metering outlet 1903 as shown in FIG. 19B.

[0196] Without the one or more ramps in the diluent metering chamber, the last diluent volume to leave the diluent metering chamber could get shallow and be prone to breaking apart or creating bubbles in the diluent siphon. This may impede fil l / d raw of the diluent siphon. This may also affect complete transfer of the metered diluent and affect the intended dilution factor.

[0197] Thus, the ramp with the one or more stages in the diluent metering chamber facilitates accurate metering of the diluent and transfer of the entirety of the metered diluent from the diluent metering chamber to the mixing chamber. Accurate metering of the diluent also provides for accurate dilution factor of the diluted sample and, consequently, accurate analysis results.

[0198] In some cases, a fluidic connection between the diluent metering chamber and a diluent siphon is not concentric to the vertical axis. An exemplary arrangement is shown in FIG. 19C. Such arrangement encourages a diluent to flow outward into the diluent siphon.

[0199] In some cases, a fluidic connection between the mixing chamber and the diluted sample siphon is in line with the mixing chamber. This encourages any fluid in the diluted sample siphon to re-enter the mixing chamber during mixing. An example of such arrangement is show in FIGS. 20A-20B.

[0200] An analysis cartridge disclosed herein also comprises a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve. The mixing chamber is also fluidically connected to a sample metering chamber via a sample valve. When the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into themixing chamber. The second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold. For example, a centrifugation force above the second threshold can overcome surface tension of the diluent in a microfluidic channel or force open a valve to allow the diluent from the diluent metering chamber to flow into the mixing chamber. Similarly, a centrifugation force above the second threshold can overcome surface tension of the sample in a microfluidic channel or force open a valve to allow the sample from the sample metering chamber to flow to the mixing chamber.

[0201] While the second centrifugation can move both the diluent and the sample into the mixing chamber, in some cases, the analysis cartridge can be configured so that the diluent is moved into the mixing chamber in a centrifugation step and the sample is moved into the mixing chamber in a separate centrifugation step.

[0202] Mixing of a sample and a diluent is achieved in the mixing chamber. In some cases, mixing is achieved by rapidly changing the centrifugation speed, for example, from ~4000 RPM to ~750RPM, while driving in the same direction.

[0203] Without being bound by any theory, the change in centrifugation speed may causes macro- and microscopic mixing and intermingling of the two fluids thereby homogenizing them. For example, the change in centrifugation speed may cause micro vortices between the fluid layers due to the change in force experienced by each layer radius.

[0204] In some cases, a mixing chamber comprises a mixing chamber vent that vents the gas from the mixing chamber as the diluted sample moves into the mixing chamber.

[0205] An analysis cartridge disclosed herein can also further comprise one or more metering fingers. Metering fingers have a known volume and is used to separate a diluted sample of known volume before the diluted sample is loaded into the one or more detection chambers. The one or more metering fingers are fluidical ly connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel. The third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold. For example, a centrifugation force above the third threshold can overcome surface tension of the diluted sample in a microfluidic channel or force open a valve to allow the diluted sample from the mixing chamber to flow to the one or more metering fingers.

[0206] An analysis cartridge disclosed herein can also further comprise one or more detection chambers. The one or more detection chambers are fluidically connected to the one or more metering fingers via a capillary channel such that, a fourth centrifugation of the analysis cartridge moves the diluted sample from the one or more metering fingers into the one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold. The fourth centrifugation exerts on the diluted sample in a metering finger a fourth centrifugal force above a fourth threshold. For example, a centrifugation force above the fourth threshold can overcome surface tension of the diluted sample in the capillary channels or overcome the air pressure in the detection chambers to allow the diluted sample from the one or more metering fingers to flow to the one or more detection chambers.

[0207] As noted above, a third centrifugation of the analysis cartridge moves a diluted sample to one or more metering fingers via a distribution channel. Filling of the metering fingers from one end to the other of the analysis cartridge can occur in a substantially sequential manner, with one metering finger is filled at a time before the next one is filled. Once mixing of the sample and the diluent is complete, the centrifugation of the analysis cartridge slows down and allows the diluted sample to prime the diluted sample siphon 1016. Once primed, the centrifuge speeds up to begin distribution.

[0208] As the centrifugation of the analysis cartridge continues, the dilute sample enters the distribution channel 1017 and begins to fill up the first metering finger without jetting into the corresponding first detection chamber. This step is depicted in FIGS. 21B and 24C-24D.

[0209] As the first channel is filled, overfill occurs and starting the filling sequence of the next metering finger. This is depicted in FIGS. 21C and 24E. The second metering finger is filled next, as depicted in FIG. 21D.

[0210] This sequential process is repeated until all 20 metering fingers are filled as shown in FIGS. 21E and 25. As the last metering finger fills up the centrifugation speed is increased to spin off any excess diluted sample into the diluted sample overflow chamber 1019. The diluted sample overflow chamber comprises the diluted sample overflow chamber vent 1020, which allows the air to go out when the diluted sample fills in the metering fingers.

[0211] In some cases, the proximate walls of the one or more metering fingers are sloped away from the proximate towards the distal direction. This allows appropriate filling of the metering chambers as the sample trickles down towards the distal end of the metering fingers.

[0212] In some cases, to facilitate movement of a diluted sample from a metering finger to the capillary channel, the distal edges of the metering fingers are rounded. An example is shown in FIG. 24A, where the distal edges 2401 are rounded.

[0213] In some cases, the capillary channels between the one or more metering fingers and the corresponding detection chambers are on the top surface of the analysis cartridge. To provide appropriate capillary stop pressure, in some cases, the capillary channels have a depth of between 0.1 mm and 0.3 mm. In some cases, the capillary channels have a width of between 0.4 mm and 0.6 mm.

[0214] Once all the metering fingers are filled and all excess has been spun off to the waste chamber, distribution of the diluted sample is complete, as shown in FIG. 21F.

[0215] To improve timings and reliability, the distribution channel can be designed to be non- concentric to the axis of rotation. This allows the start of the channel and the end of the channel to be at different radial heights. This is shown in FIG. 22, where the distribution channel is not concentric to the axis of rotation. For example, the end of the distribution channel near the fluidic connection with the diluted sample siphon is proximate than the end of the distribution channel near the fluidic connection with the diluted sample overflow chamber. This feature would allow the last detection chamber to be filled despite the mixing chamber being emptied. In an exemplary embodiment, a 2 mm step down is used. However, by increasing this parameter, the last wells can be filled faster.

[0216] Accordingly, the analysis cartridge can be centrifuged at different speeds in a step-wise manner to cause different fluids to move from certain specific compartments to certain other specific compartments.

[0217] In some cases, the mixing chamber has a volume that is sufficient to accommodate a diluted sample of: between 450 pL and 500 pL, about 450 pL, about 455 pL, about 460 pL, about 465 pL, about 470 pL, about 475 pL, about 480 pL, about 485 pL, about 490 pL, about 495 pL, and about 500 pL. In some cases, the total volume of the mixing chamber is between 20% and 100% higher than the volume of the diluted sample collected therein.

[0218] In some cases, the one or more metering fingers have a volume of: between 15 pL and 30 pL, about 15 pL, about 16 pL, about 17 pL, about 18 pL, about 19 pL, about 20 pL, about 21 pL, about 22 pL, about 23 pL, about 24 pL, about 25 pL, about 26 pL, about 27 pL, about 28 pL, about 29 pL, about 30 pL. In certain embodiments, different metering fingers have different volumes. In certain embodiments, all the one or more metering fingers have substantiallyidentical volumes. In some cases, some of the one or more metering fingers have substantially identical volumes and some of the one or more metering fingers have different volumes.

[0219] In some cases, the one or more detection chambers have a volume of: between 15 inland 30 pL, about 15 pL, about 16 iL, about 17 pL, about 18 pL, about 19 pL, about 20 pL, about 21 pL, about 22 pL, about 23 pL, about 24 pL, about 25 pL, about 26 pL, about 27 pL, about 28 |j.L, about 29 pL, about 30 pL. In certain embodiments, different detection chambers have different volumes. In certain embodiments, all the one or more detection chambers have substantially identical volumes. In some cases, some of the one or more detection chambers have substantially identical volumes and some of the one or more detection chambers have different volumes.

[0220] As noted above, a capillary channel between a metering finger and a detection chamber provides capillary stop pressure. The third centrifugation does not exert enough centrifugation force on the diluted sample in the one or more metering fingers to move it to the one or more detection chambers. However, the fourth centrifugal force above the fourth threshold exerts sufficient centrifugal force on the diluted sample in the one or more metering fingers to overcome the capillary stop pressure of the one or more capillary channels. To that end, in some cases, a capillary channel connecting a metering finger and a detection chamber has the dimensions of 0.2 - 0.3 mm x 0.4 to 0.6 mm, which allows consistent filling of the diluted sample in the metering fingers without jetting the diluted sample into the corresponding detection chambers.

[0221] In some cases, the analysis cartridges having metering fingers described above also have a first and a second set of electrochemical sensors. Certain details of such sets of electrochemical sensors are provided above under "Analysis Cartridges Without Metering Fingers" and such details are also applicable to analysis cartridges having one or more metering fingers.

[0222] The term "valve" as used herein refers to a connection that is opened and closed. For example, a valve can be a siphon, where a hydrophilic draw of fluid into a siphon that connect reservoirs is used to allow a fluid to flow from one chamber to another. A valve can also be a capillary stop, where the capillary stop prevents a fluid from going into a subsequent chamber until the centrifugal force exerted on the fluid is high enough, e.g., beyond a threshold, to overcome the fluid / material / opening resistance of the capillary stop.

[0223] In some cases, a diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon. Operation of a siphon, for example, as used in a sample siphon, a diluent siphon, or a diluted sample siphon is provided above under"Analysis Cartridges Without Metering Fingers" and such details are also applicable to the siphons in analysis cartridges having one or more metering fingers. For example, wherein the diluent siphon, the sample siphon, and / or the diluted sample siphon can be opened by priming. For example, the diluent siphon and the sample siphon can be primed by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold. Similarly, the diluted sample siphon can be primed by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

[0224] In some cases, a diluent metering chamber is fluidically connected to a diluent overflow chamber such that the first centrifugation of the analysis cartridge moves into the diluent overflow chamber excess diluent that does not fit in the diluent metering chamber. In some cases, the diluent overflow chamber comprises a diluent overflow vent that vents the gas from the diluent metering chamber as the diluent moves into the diluent metering chamber.

[0225] In some cases, the one or more metering fingers are fluidically connected to a diluted sample overflow chamber such that the third centrifugation of the analysis cartridge moves into the diluted sample overflow chamber excess diluted sample that does not fit in the one or more metering fingers. In certain such cases, the diluted sample overflow chamber comprises a diluted sample overflow chamber vent that vents the gas from the one or more metering fingers as the diluted sample moves into the one or more metering fingers.

[0226] In some cases, a fluidic connection between the mixing chamber and the diluted sample siphon is not concentric to the vertical axis. An exemplary embodiment is shown in FIG. 20A where the connection 2001 is not concentric to the vertical axis. Such arrangement encourages a diluted sample to flow outward into the diluted sample siphon.

[0227] The filling of the detection chamber is driven by the combination of capillary stop force, back pressure from compressing enclosed air, and the burst frequency where instability occurs. By adjusting the volume of compressed air and the plug length, the centrifugation speed at which instability occurs can be modified. The capillary stop is strengthened due to the back pressure generated by the compression of the enclosed air volume in the detection chamber, as shown in FIG. 25. This allows the capillary stop to function at higher forces than traditional capillary stops.

[0228] The diluted sample siphon outlet may act as a bottle neck for dispense speeds. To address this potential issue, a trumpet design can be implemented in diluted sample siphon. Thisis shown in FIG. 23, where a trumpet shaped outlet of the diluted sample siphon connects to the distribution channel 2302.

[0229] As noted above, the connecting channels between the metering fingers and the corresponding detection chambers have dimensions that allow the channels to be consistently filled without jetting the diluted sample into the detection chamber.

[0230] Certain features of the metering fingers shown in FIG. 24A facilitate proper filling and metering of the diluted sample. For example, the rounded ends 2401 of the metering fingers guide the fluid flow towards the capillary channel during initial filling. Also, ramped inlets 2402 ensure that air gets pushed out during filling of the metering chamber. Moreover, the channels 2403, with the dimensions of 0.2 mm x 0.5 mm consistently filled with the diluted sample without jetting the diluted sample into the corresponding detection chambers 2404.

[0231] In some cases, the one or more detection chambers comprise one or more reagents. These reagents are configured to detect one or more analytes. In some cases, the one or more reagents are dried reagents. For example, one or more reagents can be dried in the one or more detection chambers, which dissolve when a diluted sample is introduced into the one or more detection chambers. Alternatively, one or more reagents can be provided in a dried bead, for example, a lyophilized bead, which releases the one or more reagents when a diluted sample wets the bead in a detection chamber.

[0232] As noted above, some of the detection chambers can have two chambers. One such exemplary detection chamber is shown in FIGS. 26A-26B. The 2-step reaction being the last step in the filling process, provides two main benefits: as the last cuvette, there is no cross contamination with previous cuvettes and the slower flow rate increases the amount of time the first Lyo bead (bead containing lyophilized reagents) has to dissolve so the bead doesn't get carried to the waste chamber.

[0233] As shown in FIG. 26A, the detection chamber having two chambers 2602 and 2603 is connected to the corresponding metering finger via channel 2601. Compared to the 2 mm width of the channels for a single chambered detection chambers, the width for this channel is reduced to 1.5 mm. This feature minimizes the backflow of dissolved Lyo beads.

[0234] Moreover, in some cases, the two-step metering uses a deep 4.8 mm circular geometry to inhibit the Lyo bead in the detection chamber from traveling upstream when it is wetted. This is shown in FIG. 26B.

[0235] When all the metering fingers are filled, a further centrifugation of the analysis cartridge at a higher speed moves the diluted sample from the metering fingers into the detection chambers 1021. This is shown in FIG. 14H.

[0236] Detection chambers comprise one or more reactants that are configured to detect one or more analytes. The reagents can be provided in the form of beads that contain lyophilized reagents. Thus, when a diluted sample is loaded into a reaction mixture, the lyophilized reagents in the beads slowly dissolve in the diluted sample.

[0237] The analyte detection can be based on electrochemical or optical detection. Each detection chamber is a well of about 2.35 mm diameter (with a minimum of 2.32 mm diameter) having a draft angle of about 0.5 degrees and depth of about 5 mm. Thus, each well is configured to accommodate about 22 to 24 pL with about 1% variance in the loaded volume.

[0238] An exemplary combination of analytes in a centrifugation cartridge comprises: glucose, albumin, ALP, calcium, total protein, CO2, total bilirubin, blood urea nitrogen, ALT, AST, creatinine, sodium, chloride, and potassium. Additional detection chambers can be used for certain duplicates and reference readings.Methods

[0239] The analysis cartridges disclosed herein can be used for analyzing a biological sample, for example, a blood sample for its non-cellular constituents.

[0240] The term "biological sample" as used herein includes any sample comprising non-cellular constituents. In some embodiments, a biological sample is a biological fluid sample. A suitable biological sample analyzed in the analysis cartridges disclosed herein can be blood, urine, saliva, sweat, sputum, semen, mucus, lacrimal fluid, lymph fluid, amniotic fluid, interstitial fluid, lung lavage, cerebrospinal fluid, feces, or the like. A biological sample can also be a slurry of an otherwise solid tissue, such as a liver or a kidney, homogenized in a buffer to prepare a slurry comprising cells and non-cellular constituents. A biological sample can also comprise cultured cells in a suitable buffer. Additional examples of biological samples that could be analyzed in the analysis cartridges disclosed herein can be readily identified by a person of ordinary skill in the art and such embodiments are within the purview of the disclosure.

[0241] A portion of the biological sample from which the cells are removed is referenced herein as "a non-cellular portion of the biological sample." A non-cellular portion of the biological sample comprises one or more non-cellular constituents, including metabolites and small molecules, such as glucose and other biochemicals as well as large biomolecules, such asproteins. An analysis cartridge disclosed herein can analyze one or more non-cellular constituents.

[0242] A biological sample can be obtained from any suitable subject. "Subject" as used herein refers to any animal, including, but not limited to, a mammal (e.g., cow, pig, camel, llama, horse, goat, rabbit, sheep, hamsters, guinea pig, cat, dog, rat, and mouse, a non-human primate (for example, a monkey, such as a cynomolgus or rhesus monkey, chimpanzee, etc.) and a human). In some embodiments, the subject may be a human or a non-human. In some embodiments, the subject is a human.

[0243] Depending on the biological sample, different non-cellular constituents can be analyzed in the chemical analysis module. For example, if the biological sample is blood, the cellular analysis module is configured to analyze blood plasma for CMP. Additional embodiments of non- cellular constituents that could be analyzed in the chemical analysis module are well known to a person of ordinary skill in the art and such embodiments are within the purview of this disclosure.

[0244] Any structural elements of the analysis cartridges described elsewhere in this disclosure, for example, those described under "Analysis Cartridges" above, are applicable to the methods disclosed herein.

[0245] Certain aspects of the disclosure provide a method of analyzing a biological sample, for example, a blood sample, for its non-cellular constituents, such as plasma, in the analysis cartridges disclosed herein.METHODS OF USING ANALYSIS CARTRIDGES WITHOUT METERING FINGERS

[0246] In some cases, the disclosure provides a method of analyzing a sample in an analysis cartridge without metering fingers. In certain such cases, the method comprises:

[0247] conducting a first centrifugation of the analysis cartridge to move a diluent from a diluent chamber to a f luidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0248] conducting a second centrifugation of the analysis cartridge to move into a mixing chamber the diluent from the diluent metering chamber and the sample from the sample metering chamber; wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0249] conducting a third centrifugation to move into a one or more detection chambers the diluted sample from the mixing chamber, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0250] In some cases, the method comprises conducting a fourth centrifugation of the analysis cartridge before conducting the first centrifugation of the analysis cartridge, wherein the fourth centrifugation moves the sample from a sample chamber into the sample metering chamber, and wherein the fourth centrifugation exerts on the sample in the sample chamber a fourth centrifugation force above a fourth threshold.

[0251] In some cases, the third threshold is higher than the second threshold and the second threshold is higher than the first threshold. In certain embodiments, the fourth threshold is lower than the first and / or the second threshold. Thus, the fourth, the first, the second, and the third centrifugations can be carried out sequentially to move the sample and / or reagents in a step-wise manner through different chambers of the analysis cartridge.

[0252] After the third centrifugation, a diluted sample is loaded into the one or more detection chambers. Accordingly, in some cases, the method comprises detecting in the one or more detection chambers one or more signals that indicate the concentration of one or more analytes in the sample. The detected one or more signals can be used to determine the concentrations of the one or more analytes in the sample.

[0253] As noted above, in some cases, the sample metering chamber of the analysis cartridge further comprises a sample overflow conduit. The sample overflow conduit can be fluidically connected to a sample overflow chamber. Thus, the sample beyond the volume of the sample metering chamber flows to the sample overflow chamber. With this arrangement, a specific and predetermined volume of the sample is collected in the sample metering chamber. This volume can be used in calculating concentrations of one or more analytes in the sample.

[0254] Accordingly, in some cases of the methods described herein, conducting the fourth centrifugation of the analysis cartridge comprises receiving into a sample overflow chamber via a sample overflow conduit a portion of the sample that does not fit in the sample metering chamber.

[0255] Also, as noted above, in some embodiments, the sample metering chamber further comprises a cellular components chamber, which can be fluidically connected to the sample metering chamber. Accordingly, in some cases of the methods disclosed herein, conducting the fourth centrifugation of the analysis cartridge comprises receiving the cellular components of the sample into the cellular components chamber.

[0256] Moreover, as noted above, in some cases, the one or more detection chambers of an analysis cartridge comprise optical detection chambers. In addition, an analysis cartridge can further comprise one or more electrochemical sensors to detect one or more analytes in the sample. For electrochemical detection of analytes, cellular components need not be separated from the non-cellular components.

[0257] Accordingly, some cases of the methods disclosed herein further comprises detecting a first set of one or more analytes in the sample in one or more electrochemical sensors located in the sample overflow chamber and / or the sample overflow conduit.

[0258] Further, as noted above, in some embodiments of the analysis cartridges disclosed herein, a sample chamber is fluidically connected to an electrochemical analysis module. Accordingly, in some cases, methods disclosed herein comprises conducting a fifth centrifugation of the analysis cartridge to move a portion of the sample from the sample chamber into the electrochemical analysis module. In some cases, the fifth centrifugation can exert on a sample in the sample chamber sufficient centrifugal force to move the sample from the sample chamber into the electrochemical analysis module. The fifth centrifugal force can be below the fourth threshold, discussed above.

[0259] In some cases, the methods further comprise detecting a second set of one or more analytes in the sample using the second set of one or more electrochemical sensors.

[0260] Furthermore, as noted above, in some cases, in the analysis cartridges disclosed herein, the diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon. Accordingly, in some cases, the methods disclosed herein comprise opening by priming the diluent siphon, the sample siphon, and / or the diluted sample siphon. For example, priming the diluent siphon and the sample siphon comprises centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold. Similarly, priming the diluted sample siphon comprises centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

[0261] In some embodiments, methods disclosed herein comprise assaying clinical chemistry panels in a blood sample. The clinical chemistry panels referto groups of tests that are routinely ordered to determine a subject's general health status. In some cases, the clinical chemistry panels include metabolic panels. The clinical chemistry panels help evaluate, for example, the body's electrolyte balance and / or the status of several major body organs. In some cases, theassays are performed on a blood sample, usually drawn from a vein. Examples of clinical chemistry panels that may be detected by assays of the present disclosure include, but are not limited to, basic metabolic panel (BMP), comprehensive metabolic panel (CMP), electrolyte panel, lipid panel, liver panel, renal panel, and thyroid function panel. The basic metabolic panel (BMP) includes 8 tests, all of which are found in the CMP. The BMP provides information about the current health of kidneys and respiratory system as well as electrolyte and acid / base balance and level of blood glucose. The CMP measurement is used for liver and kidney health, level of blood glucose, acid / base balance in blood, fluid and electrolyte balance, and important blood proteins. In some cases, the CMP measures glucose, calcium, total amount of albumin and globulins, bilirubin, BUN (blood urea nitrogen), creatinine, albumin, sodium, potassium, bicarbonate, chloride, alkaline phosphatase (ALP), alanine transaminase (ALT), and aspartate aminotransferase (AST). The electrolyte panel is used to detect a problem with the body's fluid and electrolyte balance. For example, the electrolyte panel measures the blood levels of carbon dioxide, chloride, potassium, and sodium. The lipid panel is used to assess a subject's risk of developing cardiovascular disease. For example, the lipid panel measures the amount of cholesterol and other fats in blood, such as total cholesterol, LDL (low-density lipoprotein), HDL (high-density lipoprotein), and triglycerides. The liver panel (hepatic function panel) is used to screen for, detect, evaluate, and monitor acute and chronic liver inflammation (hepatitis), liver disease and / or damage. The liver panel measures different enzymes, proteins, and other substances made by liver. For example, the liver panel includes albumin, total protein, ALP, ALT, AST, gamma-glutamyl transferase (GGT), bilirubin, Lactate dehydrogenase (LD), Prothrombin time (PT). The renal panel (kidney function panel) includes tests such as albumin, creatinine, BUN, eGFR to evaluate kidney function. The thyroid Function Panel is used to evaluate thyroid gland function and to help diagnose thyroid disorders. The thyroid function panel measure thyroid hormone such as thyroxine (T4), triiodothyronine (T3), and thyroid stimulating hormone (TSH). In some cases, a high TSH level indicates that the thyroid gland is not making enough thyroid hormone (primary hypothyroidism). The opposite situation, in which the TSH level is low, usually indicates that the thyroid is producing too much thyroid hormone (hyperthyroidism). In other cases, the finding of an elevated TSH and low free T4 (FT4) or free T4 index (FTI) indicates primary hypothyroidism due to disease in the thyroid gland. A low TSH and low FT4 or FTI indicate hypothyroidism due to a problem involving the pituitary gland. A low TSH with an elevated FT4 or FTI is found in individuals who have hyperthyroidism. These clinicalchemistry panels are well known in the art and are further described in the assay portion of the present disclosure.

[0262] In some cases, such as where the cartridge is employed for a complete metabolic panel (CMP), reagents comprise one or more of the following: 2,4,6-Tribromo-3-hydroxybenzoic acid (TBHBA), 2-Chloro-4-nitrophenyl-a-maltotrioside (CNPG3), 2-Methyl-4-isothizolin-3-one hydrochloride (MIT), 4,7,13,16,21-Pentaoxa-l,10-diazabicyclo[8.8.5]tricosane (Kryptofix 221), 4- Aminoantipyrine hydrochloride, Adenosine 5'-diphosphate, Adenosine 5'-triphosphate, a- ketoglutaric acid, Amylase, Arsenazo III, sodium salt, Ascorbate oxidase (Cucurbita spp.), Bilirubin oxidase, Bromcresol purple, Calcium acetate, Creatine amidinohydrolase (Actinobacillus spp.), Creatinine amidohydrolase (Pseudomonas spp.), Cupric sulfate, Ethylene glycol-bis(R-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), Ethylenediaminetetraacetic acid (EDTA), R- Galactosidase, Glucose-6-phosphate dehydrogenase (yeast), Glutamate Dehydrogenase (bovine liver), Glutamine synthetase, Hexokinase (yeast), Imidazole, Lactate dehydrogenase, L-alanine, L-aspartic acid, L-glutamic acid, Magnesium chloride, Magnesium sulfate, Malate dehydrogenase (porcine heart), Manganese chloride, N-Acetyl cysteine, ^-Nicotinamide adenine dinucleotide (NAD), R-Nicotinamide adenine dinucleotide, reduced (NADH), o-Nitrophenyl-iS-D- galactopyranoside (ONPG), Peroxidase (horseradish), Phosphoenol pyruvate, Phosphoenol pyruvate carboxylase, p-NPP, Potassium ferrocyanide, Potassium iodide, Pyruvate kinase, Sarcosine oxidase (microorganism), Sodium potassium tartrate, Urease (jack bean), Zinc sulfate, as well as other buffers, surfactants, excipients and preservatives.

[0263] In some cases, it may be desirable to analyze a control sample. The control sample may be analyzed concurrently with the sample from the subject as described above. The results obtained from the subject sample can be compared to the results obtained from the control sample. Standard curves may be provided, with which assay results for the sample may be compared. Using samples taken from multiple donors, standard curves can be provided for reference levels in normal healthy subjects.

[0264] Thus, in view of the above, a method for determining the presence and / or amount of non-cellular constituents in a sample is provided. In some cases, the method comprises assaying the sample for non-cellular constituents and comparing it to a control. In some cases, a calibrator is optionally, and is preferably, part of a series of calibrators in which each of the calibrators differs from the other calibrators in the series by the concentration of the analyte.METHODS OF USING ANALYSIS CARTRIDGES HAVING METERING FINGERS

[0265] As described above under "Analysis Cartridges Having Metering Fingers," some embodiments of the disclosure provide analysis cartridges comprising one or more metering fingers. Such one or more metering fingers are fluidica lly connected to one or more detection chambers that carry out detection and analysis of one or more analytes. Certain embodiments of the disclosure provides methods of analyzing a sample in an analysis cartridges describes above under "Analysis Cartridges Having Metering Fingers."

[0266] As noted above, in some cases, the disclosure provides analysis cartridges comprising one or more metering fingers. Such one or more metering fingers are fluidically connected to one or more detection chambers that carry out detection and analysis of one or more reactants. Certain details of the analysis cartridges describes above under "Analysis Cartridges Without Metering Fingers" are also applicable to analysis cartridges having metering fingers. Such details include the various fluidic connections between different chambers, use and functionality of siphons, types of analytes analyzed, types of reagents used, shapes and geometries of various chambers, shapes and geometries of the analysis cartridges, volumes of various chambers, material used for fabrication of the analysis cartridges, and the like and implementation of such details in the methods of using analysis cartridges having metering fingers are within the purview of the disclosure.

[0267] In certain such embodiments, the disclosure provides a method of analyzing a sample in an analysis cartridge, the method comprising:

[0268] conducting a first centrifugation of the analysis cartridge to move a diluent from a diluent chamber to a fluidically connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0269] conducting a second centrifugation of the analysis cartridge to move into a mixing chamberthe diluent from the diluent metering chamberand the sample from a sample metering chamber; wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0270] conducting a third centrifugation of the analysis cartridge to move the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0271] conducting a fourth centrifugation to move into a one or more detection chambers the diluted sample from the one or more metering fingers, wherein the fourth centrifugation exerts on the diluted sample in the one or more metering fingers a fourth centrifugal force above a third threshold.

[0272] Certain details of the analysis cartridges having one or more metering fingers described above are also applicable to the methods of using such analysis cartridges described herein.

[0273] For example, in some embodiments of such methods, the fourth threshold is higher than the third threshold, the third threshold is higher than the second threshold, and the second threshold is higher than the first threshold.

[0274] In some cases, the first centrifugation of the analysis cartridge also moves the sample from a sample chamber into the sample metering chamber. In some cases, the sample metering chamber is fluidica lly connected to a cellular components chamber and the first centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components chamber.

[0275] In certain embodiments, a method disclosed herein comprises conducting a fifth centrifugation to move a sample from a sample chamber into a sample metering chamber. Accordingly, the method comprises conducting a fifth centrifugation of the analysis cartridge to move the sample from the sample chamber into the sample metering chamber, and wherein the fifth centrifugation exerts on the sample in the sample chamber a fifth centrifugation force above a fifth threshold. In some cases, the fifth centrifugation of the analysis cartridge also moves the cellular components of the sample into the cellular components chamber. In some cases, the fifth threshold is lower than the first and / or the second threshold so that the fifth threshold does not cause the sample from the sample metering chamber to move into the mixing chamber during the fifth centrifugation.

[0276] In some cases, the sample metering chamber further comprises a sample overflow conduit fluidical ly connected to a sample overflow chamber that receives a portion of the sample that does not fit in the sample metering chamber, wherein the sample overflow chamber and / or the sample overflow conduit comprises a first set of one or more electrochemical sensors that detect a first set of one or more analytes in the sample. In certain such cases, conducting the first centrifugation or the fifth centrifugation of the analysis cartridge comprises detecting the first set of one or more analytes in the sample in the first set of one or more electrochemical sensors. For example, in certain such cases, the method comprises detecting in the one or moredetection chambers one or more signals that indicate the concentration of one or more analytes in the sample and determining the concentrations of the one or more analytes in the sample.

[0277] During the first centrifugation or the fifth centrifugation of the analysis cartridge, the method comprises receiving into a sample overflow chamber a portion of the sample that does not fit in the sample metering chamber. Also, during the first centrifugation or the fifth centrifugation, the method comprises receiving the cellular components of the sample into the cellular components chamber that is fluid ica lly connected to the sample metering chamber.

[0278] In some cases, the sample chamber is fu rther fluidical ly connected to an electrochemical analysis module comprising a second set of one or more electrochemical sensors. In certain such cases, a method disclosed herein comprises conducting a sixth centrifugation of the analysis cartridge to move a portion of the sample from the sample chamber into the electrochemical analysis module. In certain such cases, the method further comprises detecting a second set of one or more analytes in the sample using the second set of one or more electrochemical sensors.

[0279] In some cases, the diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon. Accordingly, in some cases, the method comprises opening by priming the diluent siphon, the sample siphon, and / or the diluted sample siphon. In some cases, priming the diluent siphon and the sample siphon is done by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold. Similarly, priming the diluted sample siphon can be done by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

[0280] In some cases, the method comprises detecting one or more detection chambers one or more signals that indicate the concentration of one or more analytes in the sample and determining the concentrations of the one or more analytes in the sample. Certain details of such detection are described above under the "Methods of Using an Analysis Cartridge without Metering Fingers" section and such details are also applicable to methods of detecting one or more analytes using an analysis cartridge having one or more metering fingers and such embodiments are within the purview of the disclosure. Certain such details include, the type of analytes detected, the types of reagents used, the detection methods used, volumes of various chambers and fluids, centrifugation speeds, etc.Analyzers for analysis cartridges

[0281] In some aspects, the analysis cartridges are processed in an analyzer that contains software to execute one or more tasks, including the performance of the methods described herein. In some aspects, the analyzer contains software to automatically determine the next appropriate step in methods as described herein. For example, the instrument may contain software that determines amount or presence of an analyte of interest. The software may display this determination, such as on a graphical user interface.

[0282] In some aspects, the analyzer stores software that instructs a processor to execute a given task. In some aspects, the software stores machine readable instructions that instruct a processor to execute a given task. The machine-readable instructions may be one or more executable programs or portion(s) of an executable program for execution by a computer. The programs may be embodied in software stored on a non-transitory computer readable storage medium such as a CD-ROM, a floppy disk, a hard drive, a DVD, a Blu-ray disk, or a memory associated with the processors. Alternatively, the entire programs and / or parts thereof could alternatively be executed by a device otherthan the processors and / or embodied in firmware or dedicated hardware. Additionally or alternatively, processes may be implemented by one or more hardware circuits (e.g., discrete and / or integrated analog and / or digital circuitry, an FPGA, an ASIC, a comparator, an operational-amplifier (op-amp), a logic circuit, etc.) structured to perform the corresponding operation without executing software or firmware.

[0283] The machine-readable instructions may be stored in one or more of a compressed format, an encrypted format, a fragmented format, a compiled format, an executable format, a packaged format, etc. Machine readable instructions as described herein may be stored as data (e.g., portions of instructions, code, representations of code, etc.) that may be utilized to create, manufacture, and / or produce machine executable instructions. For example, the machine- readable instructions may be fragmented and stored on one or more storage devices and / or computing devices (e.g., servers). The machine-readable instructions may require one or more of installation, modification, adaptation, updating, combining, supplementing, configuring, decryption, decompression, unpacking, distribution, reassignment, compilation, etc. in order to make them directly readable, interpretable, and / or executable by a computing device and / or other machine. For example, the machine-readable instructions may be stored in multiple parts, which are individually compressed, encrypted, and stored on separate computing devices, wherein the parts when decrypted, decompressed, and combined form a set of executable instructions that implement a program such as that described herein.

[0284] In another example, the machine-readable instructions may be stored in a state in which they may be read by a computer, but require addition of a library (e.g., a dynamic link library (DLL)), a software development kit (SDK), an application programming interface (API), etc. in order to execute the instructions on a particular computing device or other device. In another example, the machine-readable instructions may need to be configured (e.g., settings stored, data input, network addresses recorded, etc.) before the machine-readable instructions and / or the corresponding program(s) can be executed in whole or in part. Thus, the disclosed machine- readable instructions and / or corresponding program(s) are intended to encompass such machine-readable instructions and / or program(s) regardless of the particular format or state of the machine-readable instructions and / or program(s) when stored or otherwise at rest or in transit.

[0285] The machine-readable instructions described herein can be represented by any past, present, or future instruction language, scripting language, programming language, etc. For example, the machine-readable instructions may be represented using any of the following languages: C, C++, Java, C#, Perl, Python, JavaScript, HyperText Markup Language (HTML), Structured Query Language (SQL), Swift, etc.

[0286] The machine readable instructions may be stored on a non-transitory computer and / or machine readable medium such as a hard disk drive, a flash memory, a read-only memory, a compact disk, a digital versatile disk, a cache, a random-access memory and / or any other storage device or storage disk in which information is stored for any duration (e.g., for extended time periods, permanently, for brief instances, for temporary buffering, and / or for caching of the information). As used herein, the term non-transitory computer readable medium is expressly defined to include any type of computer readable storage device and / or storage disk and to exclude propagating signals and to exclude transmission media.

[0287] Thus, in some aspects, the disclosure provides an analyzer for analyzing a sample in an analysis cartridges, the analyzer comprising:

[0288] a rotor comprising a slot that holds the analysis cartridge such that, when the rotor rotates, the analysis cartridge is centrifuged around a vertical axis,

[0289] a processor, and

[0290] a non-transitory computer readable medium comprising instructions, which when executed by the processor, cause the analyzer to perform:

[0291] a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into a fluidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0292] when a diluent valve and a sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from a sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0293] a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more detection chambers, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0294] In some cases, the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform the first centrifugation followed by the second centrifugation followed by the third centrifugation. In some embodiments, the third threshold is higherthan the second threshold and the second threshold is higher than the first threshold.

[0295] In certain embodiments, the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform a fourth centrifugation of the analysis cartridge before conducting the first centrifugation of the analysis cartridge. The fourth centrifugation moves the sample from a sample chamber into the sample metering chamber. The fourth centrifugation exerts on the sample in the sample chamber a fourth centrifugation force above a fourth threshold. In some cases, the fourth threshold is lower than the first and / or the second threshold.

[0296] In some cases, an analyzer disclosed herein further comprises a detectorthat detects one or more signals from the one or more detection chambers. The one or more signals indicate the concentration of one or more analytes in the sample. Forexample, the detector can be an optical detector that detects one or more optical signals from the one or more detection chambers.

[0297] As noted above, in some cases, an analysis disclosed herein comprises a first set of one or more electrochemical sensors that detect one or more analytes in a sample. Accordingly, in some cases, the analyzer disclosed herein comprises a first set of one or more electrochemical detectors that detect a first set of one or more electrochemical signals from a first set of one or more electrochemical sensors.

[0298] Moreover, as noted above, in some cases, an analysis disclosed herein comprises a second set of one or more electrochemical sensors that detect one or more analytes in a sample. Accordingly, in some cases, the analyzer disclosed herein comprises a second set of one or more electrochemical detectors that detect a second set of one or more electrochemical signals from a second set of one or more electrochemical sensors.

[0299] The detectors in the first set and / or the second set of one or more electrochemical detectors can comprise an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof. In some cases, the first set of one or more electrochemical detectors and / or the second set of one or more electrochemical are parts of an electrical detection unit comprising electrical circuits that detect electrical signals.ANALYZERS FOR ANALYSIS CARTRIDGES HAVING METERING FINGERS

[0300] As described above under "Analysis Cartridges Having Metering Fingers," some embodiments of the disclosure provide analysis cartridges comprising one or more metering fingers. Such one or more metering fingers are fluidica lly connected to one or more detection chambers that carry out detection and analysis of one or more analytes. Certain embodiments of the disclosure provide analyzers that contains software to execute one or more tasks, including the performance of the methods described herein using the analysis cartridges having metering fingers. In some aspects, the analyzer contains software to automatically determine the next appropriate step in methods as described herein. For example, the instrument may contain software that determines amount or presence of an analyte of interest. The software may display this determination, such as on a graphical user interface.

[0301] Certain aspects of analyzers of the disclosure are described above under "Analyzers for Analysis Cartridges" and such aspects are also applicable to the analyzers of the disclosure that analyze samples using analysis cartridges having metering fingers as described herein. Certain such aspects include software that instructs processors to execute tasks, machine-readable instructions executable by programs, non-transitory computer readable storage media, hardware circuits structured to perform operations without executing software or firmware, machine-readable instructions, appropriate instruction languages. Application of these aspects to analyzers for analysis cartridges having metering fingers are within the purview of the disclosure.

[0302] Certain embodiments of the disclosure provide an analyzer for analyzing a sample in an analysis cartridges, the analyzer comprising:

[0303] a rotor comprising a slot that holds the analysis cartridge such that, when the rotor rotates, the analysis cartridge is centrifuged around a vertical axis,

[0304] a processor, and

[0305] a non-transitory computer readable medium comprising instructions, which when executed by the processor, cause the analyzer to perform:

[0306] a first centrifugation of the analysis cartridge that moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0307] when a diluent valve and a sample valve are open, a second centrifugation of the analysis cartridge that moves the diluent from the diluent metering chamber and a sample from a sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0308] when a diluted sample valve is open, a third centrifugation of the analysis cartridge that moves the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0309] a fourth centrifugation of the analysis cartridge that moves the diluted sample from the one or more metering fingers into the corresponding one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold.

[0310] In some cases, non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform the first centrifugation followed by the second centrifugation, which is followed by the third centrifugation, which is followed by the fourth centrifugation. In certain cases, the fourth threshold is higher than the third threshold, the third threshold is higher than the second threshold, and the second threshold is higher than the first threshold.

[0311] In some cases, the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform a fifth centrifugation of the analysis cartridge before conducting the first centrifugation of the analysis cartridge, wherein the fifth centrifugation moves the sample from a sample chamber into the sample meteringchamber, and wherein the fifth centrifugation exerts on the sample in the sample chamber a fifth centrifugation force above a fifth threshold. In some cases, fifth threshold is lower than the first and / or the second threshold, so that the fifth threshold does not cause the sample from the sample metering chamber to move into the mixing chamber.

[0312] In some cases, the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform the fifth centrifugation followed by the first centrifugation, which is followed by the second centrifugation, which is followed by the third centrifugation, which is followed by the fourth centrifugation.

[0313] In some cases, the analyzers comprise a detector that detects one or more signals from the one or more detection chambers, the one or more signals indicate the concentration of one or more analytes in the sample. In some cases, the detector is an optical detector that detects one or more optical signals from the one or more detection chambers.

[0314] In some cases, one or more reactions in the one or more detection chambers produce an electrical or electrochemical signal. In such cases, other detectors, such as one or more detectors amperometric sensors, coulometric sensors, potentiometric sensors, voltametric sensors, impedance sensors, or a combination thereof.

[0315] When an analysis cartridge having the one or more metering fingers comprises a first set and / or a second set of electrochemical sensors, the analyzer configured to manipulate such analysis cartridge further comprises a first set of one or more electrochemical detectors that detect a first set of one or more electrochemical signals from a first set of one or more electrochemical sensors located in a sample overflow chamber and / or the sample overflow conduit of the analysis cartridge. The first set of one or more electrochemical detectors can comprise an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof.

[0316] Si milarly, when an analysis cartridge having the one or more metering fingers comprises a second set and / or a second set of electrochemical sensors, the analyzer configured to manipulate such analysis cartridge further comprises a second set of one or more electrochemical detectors that detect a second set of one or more electrochemical signals from a second set of one or more electrochemical sensors located in a sample overflow chamber and / or the sample overflow conduit of the analysis cartridge. The second set of one or more electrochemical detectors can comprise an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof.

[0317] In some cases, the first set of one or more electrochemical detectors and / or the second set of one or more electrochemical are parts of an electrical detection unit comprising electrical circuits that detect electrical signals.

[0318] For purposes of completeness, various aspects of the present disclosure are set out in the following numbered clauses.

[0319] Clause 1. An analysis cartridge for analyzing a sample, the analysis cartridge configured to be centrifuged around a vertical axis, the analysis cartridge comprising:

[0320] a d iluent chamber fluidical ly connected to a diluent metering chamber, wherein a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0321] a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve and to a sample metering chamber via a sample valve such that, when the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0322] one or more detection chambers fluidically connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more detection chambers, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0323] Clause 2. The analysis cartridge of clause 1, wherein the third threshold is higher than the second threshold and the second threshold is higher than the first threshold.

[0324] Clause 3. The analysis cartridge of clause 1 or 2, wherein the sample metering chamber is fluidically connected to a sample chamber such that a fourth centrifugation of the analysis cartridge moves the sample from the sample chamber into the sample metering chamber, wherein the fourth centrifugation exerts on the sample in the sample chamber a fourth centrifugation force above a fourth threshold.

[0325] Clause 4. The analysis cartridge of clause 3, wherein the fourth threshold is lower than the first and / or the second threshold.

[0326] Clause 5. The analysis cartridge of any one of clauses 1 to 4, wherein the sample metering chamber further comprises a sample overflow conduit fluidically connected to a sample overflow chamber that receives a portion of the sample that does not fit in the sample metering chamber.

[0327] Clause 6. The analysis cartridge of clause 5, wherein the sample metering chamber further comprises a cellular components sub-chamber and the fourth centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components sub-chamber.

[0328] Clause 7. The analysis cartridge of clause 5 or 6, wherein the sample overflow chamber and / or the sample overflow conduit comprises a first set of one or more electrochemical sensors that detect a first set of one or more analytes in the sample.

[0329] Clause 8. The analysis cartridge of clause 7, wherein the sample overflow conduit is fluidically connected to a calibration fluid chamber.

[0330] Clause 9. The analysis cartridge of any one of clauses 1 to 8, wherein the sample chamber is further fluidically connected to an electrochemical analysis module, wherein a fifth centrifugation of the analysis cartridge moves a portion the sample from the sample chamber into the electrochemical analysis module.

[0331] Clause 10. The analysis cartridge of clause 9, wherein the electrochemical analysis module comprises a second set of one or more electrochemical sensors that detect a second set of one or more analytes in the sample.

[0332] Clause 11. The analysis cartridge of clause 10, wherein the electrochemical analysis module comprises a calibration fluid chamber fluidically connected to the second set of one or more electrochemical sensors.

[0333] Clause 12. The analysis cartridge of any one of clauses 1 to 11, wherein the diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon.

[0334] Clause 13. The analysis cartridge of clause 12, wherein the diluent siphon, the sample siphon, and / or the diluted sample siphon is / are opened by priming.

[0335] Clause 14. The analysis cartridge of clause 13, wherein the diluent siphon and the sample siphon are primed by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold.

[0336] Clause 15. The analysis cartridge of clause 13 or 14, wherein the diluted sample siphon is primed by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

[0337] Clause 16. A method of analyzing a sample in an analysis cartridge, the method comprising:

[0338] conducting a first centrifugation of the analysis cartridge to move a diluent from a diluent chamber to a f luidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0339] conducting a second centrifugation of the analysis cartridge to move into a mixing chamber the diluent from the diluent metering chamber and the sample from the sample metering chamber; wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0340] conducting a third centrifugation to move into one or more detection chambers the diluted sample from the mixing chamber, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0341] Clause 17. The method of clause 16, wherein the third threshold is higher than the second threshold and the second threshold is higher than the first threshold.

[0342] Clause 18. The method of clause 16 or 17, further comprising conducting a fourth centrifugation of the analysis cartridge before conducting the first centrifugation of the analysis cartridge, wherein the fourth centrifugation moves the sample from a sample chamber into the sample metering chamber, and wherein the fourth centrifugation exerts on the sample in the sample chamber a fourth centrifugation force above a fourth threshold.

[0343] Clause 19. The method of clause 18, wherein the fourth threshold is lower than the first and / or the second threshold.

[0344] Clause 20. The method of any one of clauses 16 to 19, comprising detecting in the one or more detection chambers one or more signals that indicate the concentrations of one or more analytes in the sample and determining the concentrations of the one or more analytes in the sample.

[0345] Clause 21. The method of any one of clauses 16 to 20, wherein conducting the fourth centrifugation of the analysis cartridge comprises receiving into a sample overflow chamber via a sample overflow conduit a portion of the sample that does not fit in the sample metering chamber.

[0346] Clause 22. The method of any one of clauses 18 to 21, wherein conducting the fourth centrifugation of the analysis cartridge comprises receiving the cellular components of the sample into the cellular components sub-chamber that is fluidically connected to the sample metering chamber.

[0347] Clause 23. The method of clause 21 or 22, further comprising detecting a first set of one or more analytes in the sample in one or more electrochemical sensors located in the sample overflow chamber and / or the sample overflow conduit.

[0348] Clause 24. The method of any one of clauses 16 to 23, wherein the sample chamber is further fluidically connected to an electrochemical analysis module comprising a second set of one or more electrochemical sensors, and wherein conducting a fifth centrifugation of the analysis cartridge moves a portion of the sample from the sample chamber into the electrochemical analysis module.

[0349] Clause 25. The method of clause 24, further comprising detecting a second set of one or more analytes in the sample using the second set of one or more electrochemical sensors.

[0350] Clause 26. The method of any one of clauses 16 to 25, wherein the diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon.

[0351] Clause 27. The method of clause 26, comprising opening by priming the diluent siphon, the sample siphon, and / or the diluted sample siphon.

[0352] Clause 28. The method of clause 27, comprising priming the diluent siphon and the sample siphon by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold.

[0353] Clause 29. The method of clause 26 or 27, comprising priming the diluted sample siphon by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

[0354] Clause 30. An analyzer for analyzing a sample in an analysis cartridges, the analyzer comprising:

[0355] a rotor comprising a slot that holds the analysis cartridge such that, when the rotor rotates, the analysis cartridge is centrifuged around a vertical axis,

[0356] a processor, and

[0357] a non-transitory computer readable medium comprising instructions, which when executed by the processor, cause the analyzer to perform:

[0358] a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into a fluidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0359] when a diluent valve and a sample valve are open, a second centrifugation of the analysis cartridge that moves the diluent from the diluent metering chamber and a sample from a sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0360] a third centrifugation of the analysis cartridge that moves the diluted sample from the mixing chamber into the one or more detection chambers, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

[0361] Clause 31. The analyzer of clause 30, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform the first centrifugation followed by the second centrifugation followed by the third centrifugation.

[0362] Clause 32. The analyzer of clause 30 or 31, wherein the third threshold is higher than the second threshold and the second threshold is higher than the first threshold.

[0363] Clause 33. The analyzer of any one of clauses 30 to 32, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzerto perform a fourth centrifugation of the analysis cartridge before conducting the first centrifugation of the analysis cartridge, wherein the fourth centrifugation moves the sample from a sample chamber into the sample metering chamber, and wherein the fourth centrifugation exerts on the sample in the sample chamber a fourth centrifugation force above a fourth threshold.

[0364] Clause 34. The analyzer of clause 33, wherein the fourth threshold is lower than the first and / or the second threshold.

[0365] Clause 35. The analyzer of clause 33 or 34, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform the fourth centrifugation followed by the first centrifugation followed by the second centrifugation followed by the third centrifugation.

[0366] Clause 36. The analyzer of any one of clauses 30 to 35, further comprising a detector that detects one or more signals from the one or more detection chambers, wherein the one or more signals indicate the concentration of one or more analytes in the sample.

[0367] Clause 37. The analyzer of clause 36, wherein the detector is an optical detector that detects one or more optical signals from the one or more detection chambers.

[0368] Clause 38. The analyzer of any one of clauses 30 to 37, wherein the analyzer further comprises a first set of one or more electrochemical detectors that detect a first set of one or more electrochemical signals from a first set of one or more electrochemical sensors located in a sample overflow chamber and / or the sample overflow conduit of the analysis cartridge.

[0369] Clause 39. The analyzer of clause 38, wherein the first set of one or more electrochemical detectors comprises an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof.

[0370] Clause 40. The analyzer of any one of clauses 30 to 39, wherein the analyzer further comprises a second set of one or more electrochemical detectors that detect a second set of one or more electrochemical signals from a second set of one or more electrochemical sensors located in an electrochemical analysis module of the analysis cartridge.

[0371] Clause 41. The analyzer of clause 40, wherein the second set of one or more electrochemical detectors comprises an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof.

[0372] Clause 42. The analyzer of any one of clauses 38 to 41, wherein the first set of one or more electrochemical detectors and / or the second set of one or more electrochemical are parts of an electrical detection unit comprising electrical circuits that detect electrical signals.

[0373] Clause 43. An analysis cartridge for analyzing a sample, the analysis cartridge configured to be centrifuged around a vertical axis, the analysis cartridge comprising:

[0374] a d iluent chamber fluidical ly connected to a diluent metering chamber, wherein a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0375] a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve and to a sample metering chamber via a sample valve such that, when the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluentfrom the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0376] one or more metering fingers fluidica I ly connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0377] one or more detection chambers fluidically connected to the one or more metering fingers via capillary channels such that, a fourth centrifugation of the analysis cartridge moves the diluted sample from the one or more metering fingers into the one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold.

[0378] Clause 44. The analysis cartridge of clause 43, wherein the fourth threshold is higher than the third threshold, the third threshold is higherthan the second threshold, and the second threshold is higher than the first threshold.

[0379] Clause 45. The analysis cartridge of clause 43 or 44, wherein the sample metering chamber is fluidically connected to a sample chamber such that the first centrifugation of the analysis cartridge moves the sample from the sample chamber into the sample metering chamber.

[0380] Clause 46. The analysis cartridge of clause 45, wherein the sample metering chamber is fluidically connected to a cellular components chamber and the first centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components chamber.

[0381] Clause 47. The analysis cartridge of clause 43 or 44, wherein the sample metering chamber is fluidically connected to a sample chamber such that a fifth centrifugation of the analysis cartridge moves the sample from the sample chamber into the sample metering chamber, and wherein the fifth centrifugation exerts on the sample in the sample chamber a fifth centrifugation force above a fifth threshold.

[0382] Clause 48. The analysis cartridge of clause 47, wherein the sample metering chamber is fluidically connected to a cellular components chamber and the fifth centrifugation of theanalysis cartridge moves the cellular components of the sample into the cellular components chamber.

[0383] Clause 49. The analysis cartridge of clause 48, wherein the fifth threshold is lower than the first and / or the second threshold.

[0384] Clause 50. The analysis cartridge of any one of clauses 43 to 49, wherein the sample metering chamber further comprises a sample overflow conduit fluidically connected to a sample overflow chamber that receives a portion of the sample that does not fit in the sample metering chamber.

[0385] Clause 51. The analysis cartridge of clause 50, wherein the sample overflow chamber and / or the sample overflow conduit comprises a first set of one or more electrochemical sensors that detect a first set of one or more analytes in the sample.

[0386] Clause 52. The analysis cartridge of clause 51, wherein the sample overflow conduit is fluidically connected to a calibration fluid chamber.

[0387] Clause 53. The analysis cartridge of any one of clauses 43 to 52, wherein the sample chamber is further fluidically connected to an electrochemical analysis module, wherein a sixth centrifugation of the analysis cartridge moves a portion the sample from the sample chamber into the electrochemical analysis module.

[0388] Clause 54. The analysis cartridge of clause 53, wherein the electrochemical analysis module comprises a second set of one or more electrochemical sensors that detect a second set of one or more analytes in the sample.

[0389] Clause 55. The analysis cartridge of clause 54, wherein the electrochemical analysis module comprises a calibration fluid chamber fluidically connected to the second set of one or more electrochemical sensors.

[0390] Clause 56. The analysis cartridge of any one of clauses 43 to 55, wherein the diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon.

[0391] Clause 57. The analysis cartridge of clause 56, wherein the diluent siphon, the sample siphon, and / or the diluted sample siphon is / are opened by priming.

[0392] Clause 58. The analysis cartridge of clause 57, wherein the diluent siphon and the sample siphon are primed by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold.

[0393] Clause 59. The analysis cartridge of clause 57 or 58, wherein the diluted sample siphon is primed by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

[0394] Clause 60. The analysis cartridge of clause 59, wherein the diluted sample from the one or more metering fingers is moved into the one or more detection chambers by centrifuging the analysis cartridge to exert on the diluted sample in the one or more metering fingers the fourth centrifugal force above the fourth threshold such that the diluted sample in the one or more metering fingers overcomes the capillary stop pressure of the one or more capillary channels.

[0395] Clause 61. The analysis cartridge of any one of clauses 43 to 60, wherein the diluent metering chamber is fluidically connected to a diluent overflow chamber such that the first centrifugation of the analysis cartridge moves into the diluent overflow chamber excess diluent that does not fit in the diluent metering chamber.

[0396] Clause 62. The analysis cartridge of clause 61, wherein the diluent overflow chamber comprises a diluent overflow vent that vents the gas from the diluent metering chamber as the diluent moves into the diluent metering chamber.

[0397] Clause 63. The analysis cartridge of any one of clauses 43 to 62, wherein the sample metering chamber is fluidically connected to a sample overflow chamber such that the first centrifugation or the fifth centrifugation of the analysis cartridge moves into the sample overflow chamber excess sample that does not fit in the sample metering chamber.

[0398] Clause 64. The analysis cartridge of clause 63, wherein the sample overflow chamber comprises a sample overflow vent that vents the gas from the sample metering chamber as the sample moves into the sample metering chamber.

[0399] Clause 65. The analysis cartridge of any one of clauses 43 to 64, wherein the mixing chamber comprises a mixing chamber vent that vents the gas from the mixing chamber as the diluted sample moves into the mixing chamber.

[0400] Clause 66. The analysis cartridge of any one of clauses 43 to 65, wherein the one or more metering fingers are fluidically connected to a diluted sample overflow chamber such that the third centrifugation of the analysis cartridge moves into the diluted sample overflow chamber excess diluted sample that does not fit in the one or more metering fingers.

[0401] Clause 67. The analysis cartridge of clause 66, wherein the diluted sample overflow chamber comprises a diluted sample overflow chamber vent that vents the gas from the one or more metering fingers as the diluted sample moves into the one or more metering fingers.

[0402] Clause 68. The analysis cartridge of any one of clauses 43 to 67 , wherein the diluent chamber has a volume sufficient to accommodate a diluent having a volume selected from: between 500 pL and 600 pL, about 500 p.L, about 510 pL, about 520 p.L, about 530 |al_, about 540 pL, about 550 pL, about 560 pL, about 570 pL, about 580 pL, about 580 pL, and about 600 pL.

[0403] Clause 69. The analysis cartridge of any one of clauses 43 to 68, wherein the diluent metering chamber has a volume selected from: between 450 pL and 500 pL, about 450 pL, about 455 pL, about 460 pL, about 465 pL, about 470 pL, about 475 pL, about 480 pL, about 485 pL, about 490 pL, about 495 pL, and about 500 pL.

[0404] Clause 70. The analysis cartridge of any one of clauses 43 to 69, wherein the sample chamber has a volume sufficient to accommodate a sample having a volume selected from: between 50 pL and 70 pL, about 50 pL, about 52 pL, about 54 pL, about 56 pL, about 58 pL, about 60 pL, about 62 pL, about 64 pL, about 66 pL, about 68 pL, and about 70 pL.

[0405] Clause 71. The analysis cartridge of any one of clauses 43 to 70, wherein the sample metering chamber has a volume selected from: between 8 pL and 15 pL, about 8 pL, about 9 pL, about 10 pL, about 11 pL, about 12 pL, about 13 pL, about 14 pL, and about 15 pL.

[0406] Clause 72. The analysis cartridge of any one of clauses 43 to 71, wherein the mixing chamber has a volume that is sufficient to accommodate a diluted sample of: between 450 pL and 500 pL, about 450 pL, about 455 pL, about 460 pL, about 465 pL, about 470 pL, about 475 pL, about 480 pL, about 485 pL, about 490 pL, about 495 pL, and about 500 pL.

[0407] Clause 73. The analysis cartridge of any one of clauses 43 to 72, wherein the one or more metering fingers have a volume of: between 15 pL and 30 pL, about 15 pL, about 16 pL, about 17 pL, about 18 pL, about 19 pL, about 20 pL, about 21 pL, about 22 pL, about 23 pL, about 24 pL, about 25 pL, about 26 pL, about 27 pL, about 28 pL, about 29 pL, and about 30 pL.

[0408] Clause 74. The analysis cartridge of any one of clauses 43 to 73, wherein the one or more detection chambers have a volume of: between 15 pL and 30 pL, about 15 pL, about 16 pL, about 17 pL, about 18 pL, about 19 pL, about 20 pL, about 21 pL, about 22 pL, about 23 pL, about 24 pL, about 25 pL, about 26 pL, about 27 pL, about 28 pL, about 29 pL, and about 30 pL.

[0409] Clause 75. The analysis cartridge of any one of clauses 43 to 74, wherein the sample chamber is fluidically connected to the sample metering chamber via a channel having a cross section area of: between 0.2 mm2 and 0.5 mm2, about 0.2 mm2, about 0.25 mm2, about 0.3 mm2, about 0.35 mm2, about 0.4 mm2, about 0.45 mm2, and about 0.5 mm2.

[0410] Clause 76. The analysis cartridge of any one of clauses 43 to 75, wherein the sample metering chamber comprises a proximate wall closer to the vertical axis, and wherein the fluidicconnection between the sample metering chamber and the sample overflow chamber is towards the top of the proximate wall of the sample metering chamber, and wherein the fluidic connection covers at the most 20% of the height and at the most 30% of the width of the proximate wall of the sample metering chamber.

[0411] Clause 77. The analysis cartridge of any one of clauses 43 to 76, wherein the floor of the sample metering chamber has an upward ramp relative to the bottom of the chamber, wherein the upward ramp is in the proximate to the distal direction.

[0412] Clause 78. The analysis cartridge of any one of clauses 43 to 77, wherein a connecting channel between the sample metering chamber and the cellular components chamber has a deeper channel and a shallower channel, wherein the deeper channel is between 0.2 and 0.5 mm deeper than the shallower channel.

[0413] Clause 79. The analysis cartridge of any one of clauses 42 to 78, wherein the cellular component chamber has a substantially horizontal floor and substantially perpendicular walls.

[0414] Clause 80. The analysis cartridge of any one of clauses 42 to 79, wherein the proximate wall of the sample metering chamber is angled such that the fluidic connection between the sample metering chamber and the sample overflow chamber is proximate compared to the other end of the proximate wall of the sample metering chamber.

[0415] Clause 81. The analysis cartridge of clause 75, wherein the channel between the sample chamber and the sample metering chamber is in line with the vertical axis.

[0416] Clause 82. The analysis cartridge of clause 78, wherein the connecting channel between the sample metering chamber and the cellular components chamber is in line with the vertical axis.

[0417] Clause 83. The analysis cartridge of clause 56, wherein the sample siphon has a siphon crest proximate to the sample metering chamber.

[0418] Clause 84. The analysis cartridge of clause 83, wherein the sample siphon has a siphon exit that connects to the mixing chamber, and wherein the siphon exit is in line with the vertical axis.

[0419] Clause 85. The analysis cartridge of any one of clauses 43 to 84, wherein, the distal wall of the diluent metering chamber comprises a ramp in one or more stages, wherein the one or more stages of the ramp are slanted from the proximal end to the distal end of the diluent metering chamber.

[0420] Clause 86. The analysis cartridge of clause 85, wherein the ramp in the diluent metering chamber comprises two stages.

[0421] Clause 87. The analysis cartridge of clause 56, wherein a fluidic connection between the diluent metering chamber and the diluent siphon is not concentric to the vertical axis.

[0422] Clause 88. The analysis cartridge of any one of clauses 43 to 87, wherein the diluent metering chamber has a proximate wall closer to the vertical axis, and wherein the fluidic connection between the diluent metering chamber and the diluent overflow chamber is towards the top of the proximate wall of the diluent metering chamber, and wherein the fluidic connection covers at the most 20% of the height and at the most 10% of the width of the proximate wall of the diluent metering chamber.

[0423] Clause 89. The analysis cartridge of clause 56, wherein a fluidic connection between the mixing chamber and the diluted sample siphon is not concentric to the vertical axis.

[0424] Clause 90. The analysis cartridge of any one of clauses 43 to 89, wherein the fluidic connection between the mixing chamber and the diluted sample siphon is in line with the mixing chamber.

[0425] Clause 91. The analysis cartridge of any one of clauses 43 to 90, wherein the distribution channel is not concentric to the vertical axis.

[0426] Clause 92. The analysis cartridge of clause 91, wherein the end of the distribution channel near the fluidic connection with the diluted sample siphon is proximate than the end of the distribution channel near the fluidic connection with the diluted sample overflow chamber.

[0427] Clause 93. The analysis cartridge of clause 92, wherein the end of the distribution channel nearthe fluidic connection with the diluted sample siphon is proximate by at least 2 mm than the end of the distribution channel near the fluidic connection with the diluted sample overflow chamber.

[0428] Clause 94. The analysis cartridge of any one of clauses 43 to 93, wherein the fluidic connection between the diluted sample siphon and the distribution channel is conical with the broader end connected to the distribution channel.

[0429] Clause 95. The analysis cartridge of any one of clauses 43 to 94, wherein the one or more metering fingers comprise proximate walls towards the vertical axis, and the proximate walls of the one or more metering fingers are sloped away from the proximate and towards the distal end of the analysis cartridge.

[0430] Clause 96. The analysis cartridge of any one of clauses 43 to 95, wherein the distal edges of the one or more metering fingers are rounded.

[0431] Clause 97. The analysis cartridge of any one of clauses 43 to 96, wherein, when in operation, the capillary channels between the one or more metering fingers and thecorresponding detection chambers are on the top surface of the analysis cartridge and have a depth of between 0.1 mm and 0.3 mm.

[0432] Clause 98. The analysis cartridge of clause 97, wherein, when in operation, the capillary channels between the one or more metering fingers and the corresponding detection chambers are on the top surface of the analysis cartridge and have a width of between 0.4 mm and 0.6 mm.

[0433] Clause 99. The analysis cartridge of any one of clauses 43 to 98, wherein one or more of the one or more detection chambers have two chambers.

[0434] Clause 100. The analysis cartridge of any one of clauses 43 to 99, wherein the one or more detection chambers comprise one or more reagents.

[0435] Clause 101. The analysis cartridge of clause 100, wherein the one or more reagents are dried reagents.

[0436] Clause 102. The analysis cartridge of clause 101, wherein the dried reagents are in a lyophilized bead.

[0437] Clause 103. A method of analyzing a sample in an analysis cartridge, the method comprising:

[0438] conducting a first centrifugation of the analysis cartridge to move a diluent from a diluent chamber to a f luidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,

[0439] conducting a second centrifugation of the analysis cartridge to move into a mixing chamberthe diluent from the diluent metering chamberand the sample from a sample metering chamber; wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0440] conducting a third centrifugation of the analysis cartridge to move the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0441] conducting a fourth centrifugation to move into a one or more detection chambers the diluted sample from the one or more metering fingers, wherein the fourth centrifugation exerts on the diluted sample in the one or more metering fingers a fourth centrifugal force above a third threshold.

[0442] Clause 104. The method of clause 103, wherein the fourth threshold is higher than the third threshold, the third threshold is higher than the second threshold, and the second threshold is higher than the first threshold.

[0443] Clause 105. The method of clause 103 or 104, wherein the first centrifugation of the analysis cartridge also moves the sample from a sample chamber into the sample metering chamber.

[0444] Clause 106. The method of any one of clauses 103 to 105, wherein the sample metering chamber is fluidically connected to a cellular components chamber and wherein the first centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components chamber.

[0445] Clause 107. The method of any one of clauses 103 to 105, wherein the sample metering chamber is fluidically connected to a sample chamber, and the method comprises conducting a fifth centrifugation of the analysis cartridge to move the sample from the sample chamber into the sample metering chamber, and wherein the fifth centrifugation exerts on the sample in the sample chamber a fifth centrifugation force above a fifth threshold.

[0446] Clause 108. The method of clause 107, wherein the sample metering chamber is fluidically connected to a cellular components chamber and the fifth centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components chamber.

[0447] Clause 109. The method of clause 107 or 108, wherein the fifth threshold is lower than the first and / or the second threshold.

[0448] Clause 110. The method of any one of clauses 103 to 109, wherein the sample metering chamber further comprises a sample overflow conduit fluidically connected to a sample overflow chamber that receives a portion of the sample that does not fit in the sample metering chamber, wherein the sample overflow chamber and / or the sample overflow conduit comprises a first set of one or more electrochemical sensors that detect a first set of one or more analytes in the sample, and wherein conducting the first centrifugation or the fifth centrifugation of the analysis cartridge comprises detecting the first set of one or more analytes in the sample in the first set of one or more electrochemical sensors.

[0449] Clause 111. The method of clause 110, comprising detecting in the one or more detection chambers one or more signals that indicate the concentration of one or more analytes in the sample and determining the concentrations of the one or more analytes in the sample.

[0450] Clause 112. The method of any one of clauses 103 to 111, wherein conducting the first centrifugation or the fifth centrifugation of the analysis cartridge comprises receiving into a sample overflow chamber a portion of the sample that does not fit in the sample metering chamber.

[0451] Clause 113. The method of any one of clauses 103 to 112, wherein conducting the first centrifugation or the fifth centrifugation comprises receiving the cellular components of the sample into the cellular components chamber that is fluidically connected to the sample metering chamber.

[0452] Clause 114. The method of any one of clauses 103 to 113, wherein the sample chamber is further fluidically connected to an electrochemical analysis module comprising a second set of one or more electrochemical sensors, and wherein conducting a sixth centrifugation of the analysis cartridge moves a portion of the sample from the sample chamber into the electrochemical analysis module.

[0453] Clause 115. The method of clause 114, further comprising detecting a second set of one or more analytes in the sample using the second set of one or more electrochemical sensors.

[0454] Clause 116. The method of any one of clauses 103 to 115, wherein the diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon.

[0455] Clause 117. The method of clause 116, comprising opening by priming the diluent siphon, the sample siphon, and / or the diluted sample siphon.

[0456] Clause 118. The method of clause 116 or 117, comprising priming the diluent siphon and the sample siphon by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold.

[0457] Clause 119. The method of any one of clauses 116 to 118, comprising priming the diluted sample siphon by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

[0458] Clause 120. The method of any one of clauses 103 to 119, further comprising detecting in the one or more detection chambers one or more signals that indicate the concentration of one or more analytes in the sample and determining the concentrations of the one or more analytes in the sample.

[0459] Clause 121. An analyzer for analyzing a sample in an analysis cartridges, the analyzer comprising:

[0460] a rotor comprising a slot that holds the analysis cartridge such that, when the rotor rotates, the analysis cartridge is centrifuged around a vertical axis,

[0461] a processor, and

[0462] a non-transitory computer readable medium comprising instructions, which when executed by the processor, cause the analyzer to perform:

[0463] a first centrifugation of the analysis cartridge that moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold

[0464] when a diluent valve and a sample valve are open, a second centrifugation of the analysis cartridge that moves the diluent from the diluent metering chamber and a sample from a sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold,

[0465] when a diluted sample valve is open, a third centrifugation of the analysis cartridge that moves the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and

[0466] a fourth centrifugation of the analysis cartridge that moves the diluted sample from the one or more metering fingers into the corresponding one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold.

[0467] Clause 122. The analyzer of clause 121, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform the first centrifugation followed by the second centrifugation, which is followed by the third centrifugation, which is followed by the fourth centrifugation.

[0468] Clause 123. The analyzer of clause 121 or 122, wherein the fourth threshold is higher than the third threshold, the third threshold is higherthan the second threshold, and the second threshold is higher than the first threshold.

[0469] Clause 124. The analyzer of any one of clauses 121 to 123, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform a fifth centrifugation of the analysis cartridge before conducting the first centrifugation of the analysis cartridge, wherein the fifth centrifugation moves the sample from a sample chamber into the sample metering chamber, and wherein the fifthcentrifugation exerts on the sample in the sample chamber a fifth centrifugation force above a fifth threshold.

[0470] Clause 125. The analyzer of clause 124, wherein the fifth threshold is lower than the second threshold.

[0471] Clause 126. The analyzer of clause 124 or 125, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform the fifth centrifugation followed by the first centrifugation, which is followed by the second centrifugation, which is followed by the third centrifugation, which is followed by the fourth centrifugation.

[0472] Clause 127. The analyzer of any one of clauses 121 to 126, further comprising a detectorthat detects one or more signals from the one or more detection chambers, the one or more signals indicate the concentration of one or more analytes in the sample.

[0473] Clause 128. The analyzer of clause 127, wherein the detector is an optical detector that detects one or more optical signals from the one or more detection chambers.

[0474] Clause 129. The analyzer of any one of clauses 121 to 128, wherein the analyzer further comprises a first set of one or more electrochemical detectors that detect a first set of one or more electrochemical signals from a first set of one or more electrochemical sensors located in a sample overflow chamber and / or the sample overflow conduit of the analysis cartridge.

[0475] Clause 130. The analyzer of clause 129, wherein the first set of one or more electrochemical detectors comprises an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof.

[0476] Clause 131. The analyzer of any one of clauses 121 to 130, wherein the analyzer further comprises a second set of one or more electrochemical detectors that detect a second set of one or more electrochemical signals from a second set of one or more electrochemical sensors located in an electrochemical analysis module of the analysis cartridge.

[0477] Clause 132. The analyzer of clause 131, wherein the second set of one or more electrochemical detectors comprises an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof.

[0478] Clause 133. The analyzer of any one of clauses 129 to 132, wherein the first set of one or more electrochemical detectors and / or the second set of one or more electrochemical are parts of an electrical detection unit comprising electrical circuits that detect electrical signals.EXAMPLESExample 1: An exemplary analysis cartridge

[0479] FIGS. 1A-1B depict an exemplary analysis cartridge according to this disclosure. In this example, the analysis cartridge is in a pie shape and can be centrifuged around an axis that passes through the center of the circle of which the pie is a part. Any suitable number of analysis cartridges could be centrifuged at a time depending on the angle of the pie. Thus, if the angle of the pie is about 60°, six analysis cartridges can be loaded onto a rotor of a centrifuge. If the angle of the pie is about 72°, five analysis cartridges can be loaded onto a rotor of a centrifuge. If the angle of the pie is about 90°, four analysis cartridges can be loaded onto a rotor of a centrifuge.

[0480] The analysis cartridge 100 of FIG. 1A comprises a diluent chamber 101, which is fluidically connected to diluent metering chamber 103. A centrifugation of the analysis cartridge at a certain speed exerts on a diluent in the diluent chamber sufficient centrifugal force to move the diluent from the diluent chamber 101 to diluent metering chamber 103. The volume of the diluent metering chamber is fixed and known. Any excess diluent beyond this volume is transported to the diluent overflow chamber 104.

[0481] The analysis cartridge of FIG. 1A also comprises a sample metering chamber 105. The sample metering chamber 105 is fluidically connected to a sample chamber 102. Centrifugation of the analysis chamber at a certain speed exerts on a sample in the sample chamber sufficient centrifugal force to move the sample from the sample chamberto the sample metering chamber. The volume of the sample metering chamber is predetermined and known. Any excess sample beyond this volume is transported to the sample overflow chamber 106.

[0482] The analysis cartridge of FIG. 1A further comprises a cellular components chamber 107. Upon centrifugation of the analysis cartridge, if any cells are present in the sample, the cells get collected in this cellular components chamber.

[0483] The analysis cartridge of FIG. 1A further comprises a diluent siphon 109 and a sample siphon 108. The diluent siphon and the sample siphon are primed by centrifuging the analysis cartridge at a certain speed. Such centrifugation opens the diluent siphon 109 and allows the metered diluent from the diluent metering chamber 103 to flow to the mixing chamber 111. Similarly, such centrifugation opens the sample siphon 108 and allows the metered sample fromthe sample metering chamber to flow to the mixing chamber 111. In the mixing chamber 111, the diluent and the sample are mixed to produce a diluted sample.

[0484] Furthermore, the analysis cartridge of FIG. 1A comprises a diluted sample siphon 110, which connects the mixing chamber to the one or more detection chambers 113. The diluted sample siphon is primed by centrifuging the analysis cartridge at a certain speed. Such centrifugation opens the diluted sample siphon 111 and allows the diluted sample from the mixing chamber 111 to flow to the one or more detection chambers 113. Distribution of the diluted sample to the one or more detection chambers 113 can be via the distribution channel 112. In the one or more detection chambers 113, the diluted sample is analyzed to detect one or more analytes. The one or more detection chambers comprise reagents for producing an optical signal that indicates the concentration of an analyte in the sample. The optical signals so produced are detected and quantified by one or more optical detectors that could be a part of an analyzer that processes the analysis cartridge 100 to identify concentrations of one or more analytes in the sample.

[0485] FIG. IB provides an alternate view of the analysis cartridge 100 of FIG. 1A.

[0486] FIGS. 2A to 2G describes certain features of an exemplary analysis cartridge 200, as shown in FIG. 2A. The analysis cartridge comprises a diluent chamber 201 and a sample chamber 202, as shown in FIG 2B. Further, as shown in FIG. 2C, particularly, in the enlarged view of a portion of the analysis cartridge, the sample chamber 202 is fluidically connected to the sample metering chamber 205. The sample metering chamber comprises a cellular components chamber 207, where cellular components of a sample are collected upon centrifugation of the analysis cartridge. The sample metering chamber is also fluidically connected to sample overflow chamber 206, which collects the sample beyond the volume of the sample metering chamber 205.

[0487] The analysis cartridge of FIG. 2C also shows the diluent chamber 201 fluidically connected to diluent metering chamber 203. The diluent metering chamber is fluidically connected to the diluent overflow chamber 204, which collects the diluent beyond the volume of the diluent metering chamber 203.

[0488] FIG. 2D shows the priming of the diluent siphon 216 and the sample siphon 217. Centrifugation of the cartridge at a certain speed would cause the diluent from the diluent metering chamber 218 to flow to the mixing chamber 211. Similarly, centrifugation of the cartridge at a certain speed would cause the sample from the sample chamber 219 to move tothe mixing chamber 211. The diluent 214 and the sample 215 so moved are mixed in the mixing chamber to produce a diluted sample 221, as shown in FIG. 2E.

[0489] FIG. 2F shows the priming of the diluted sample siphon 210. Centrifugation of the cartridge at a certain speed would cause the diluent from the mixing chamber 211 to flow via the distribution channel 220 to the one or more detection chambers 223 as shown in FIG. 2G. The diluent sample is then analyzed in the one or more detection chambers 223 to determine concentrations of one or more analytes.

[0490] FIG. 3 shows certain exemplary analysis cartridge 300 that comprises one or more electrochemical sensors 302. The electrochemical sensors are located in the sample chamber 301. The electrochemical sensors 302 can detect one or more analytes within the sample loaded into the sample chamber 301.

[0491] FIG. 4 shows certain exemplary analysis cartridge 400 that comprises one or more electrochemical sensors 401. The one or more electrochemical sensors are located in the sample overflow chamber 403, which is connected to the sample metering chamber via the sample overflow conduit 402. The electrochemical sensors 401 can detect one or more analytes within the sample that overflows into the sample overflow chamber 403.

[0492] FIG. 5 shows certain exemplary analysis cartridge 500 that comprises one or more electrochemical sensors 502. The one or more electrochemical sensors are located in the sample overflow conduit 501 that is connected to the sample metering chamber 503. The electrochemical sensors 502 can detect one or more analytes within the sample that overflows through the sample overflow conduit 501.

[0493] FIG. 6A shows certain parts of an exemplary analysis cartridge. The analysis cartridge comprises a sample chamber 601, which is connected to a sample metering chamber 605. The sample chamber 601 is also connected to an electrochemical analysis module via the conduit602. The electrochemical analysis module comprises the one or more electrochemical sensors603. The electrochemical analysis module also comprises the calibration fluid chamber 604. The calibration fluid chamber can comprise a calibration fluid, which is provided to the one or more electrochemical sensors 603 upon centrifugation of the analysis cartridge.

[0494] FIG. 6B shows the exemplary analysis cartridge of FIG. 6A in operation. The sample chamber 601 is filled with a blood sample, which flows to the one or more electrochemical sensors 603 via the conduit 602. The calibration fluid chamber 604 comprises a calibration fluid, which is provided to the one or more electrochemical sensors 603 upon centrifugation of the analysis cartridge.

[0495] FIG. 7 shows a further exemplary analysis cartridge. The sample chamber 701 is filled with a sample, which flows to the one or more electrochemical sensors 703 via the conduit 702. The calibration fluid chamber 704 comprises a calibration fluid, which is provided to the one or more electrochemical sensors 703 upon centrifugation of the analysis cartridge.

[0496] FIG. 8 shows an exemplary analysis cartridge comprising one or more detection chambers having a capillary stop. As shown in FIG. 8, the analysis cartridge comprises detection chambers 801, which comprise two compartments 802 and 803, which are connected via a capillary stop.

[0497] FIG. 9 shows an exemplary analysis cartridge comprising one or more detection chambers having two compartments with a siphon valve in between. As shown in FIG. 9, the analysis cartridge comprises detection chambers 901, which comprise two compartments 902 and 903, which are connected via a siphon valve.

[0498] The arrangements of the two-compartments shown in the one or more detection chambers of FIGS. 8 and 9 can be used where detection reaction involves two steps. Thus, a first step can be carried out in the compartment 802 or 902 and the reagents can be moved to the compartment 803 or 903 for further analysis.Example 2 - An exemplary method of analyzing a sample using an analysis cartridge disclosed in Example 1

[0499] The sample chamber holds between 50 pl to 1 ml of a sample, for example, between 30 pl to 100 pl, such as 30 pl, 40 pl, 50 pl, 60 pl, 70 pl, 80 pl, 90 pl, or 100 pl. The sample chamber can be vented to allow a sample, such as blood or a control to enter the chamber. The sample chamber can be capped with a lid and / or a sealing element to protect a user and / or instrument from contamination.

[0500] The sample chamber may also comprise one or more channels that f luidica lly connect it to other chambers / compartments of the analysis cartridge. Exit channels can have dimensions that restrict or prevent the flow past the entry of this same channel and / or shape at end of this same channel which prevents fluid from entering the next fluidic chamber, the sample metering chamber, unintentionally and only when initiated by a first centrifugation of the analysis cartridge.

[0501] A diluent chamber may contain a rigid or flexible container filled with a desired volume of diluent fluid. Following insertion of the analysis cartridge into an analyzer, the element which seals and protects the diluent fluid from ambient conditions is compromised. The diluent chamber has an exit channel, which connects to a diluent metering chamber.

[0502] The first centrifugation: Upon the first centrifugation of the analysis cartridge, i.e., a rotation in a certain direction and within a certain rpm range, the resistance provided by the sample chamber exit channel described above is overcome and the sample is emptied from the sample chamber and delivered into the sample metering chamber and / or sample overflow chamber.

[0503] The first centrifugation may also be used to separate a whole blood sample into plasma and a packed red blood cell layer and deliver excess whole blood / sample / control into the sample excess chamber. The first centrifugation further delivers diluent from the diluent chamber into the diluent metering chamber and delivers excess diluent into the diluent overflow chamber. Further, excess diluent can also be delivered into certain calibration chambers.

[0504] Sample metering chamber: The sample metering chamber has a volumetric capacity which is less than the volume of sample chamber described above. The excess volume of whole blood / sample / control delivered from the sample chamber to the sample metering chamber is diverted to a sample overflow chamber via a conduit located at the top (nearest center of rotation) of the sample metering chamber.

[0505] The sample overflow conduit has dimensional features and shape / design that allow the excess whole blood / sample / control volume to enter this channel and flow into the sample overflow chamber once the sample metering is filled via the first centrifugation of the analysis cartridge.

[0506] The sample metering chamber has a channel located a certain distance and volume below the top (nearest the center or rotation) of the sample metering chamber. The position of the metering channel within the sample metering chamber and its channel dimensions / features are designed to precisely deliver to the mixing chamber a sample of a desired volume.

[0507] During the first centrifugation of the analysis cartridge, the sample metering channel's design prevents fluid from entering because the centrifugal force applied is lesser than the capillary force of this same channel.

[0508] Sample overflow chamber: This chamber holds excess whole blood / sample / control that was delivered to the sample metering chamber during the first centrifugation. This chamber can be interrogated for the presence of liquid and confirm that the sample metering chamber was filled: with a metered volume of whole blood / sample / control. The whole blood / sample / control diverted into the sample overflow chamber becomes contained and is isolated from all further analysis described below.

[0509] Diluent metering chamber: The diluent metering chamber has a desired volumetric capacity, which is less than the volume of the diluent contained within the diluent chamber. The excess diluent delivered from the diluent chamber to the diluent metering chamber during the first centrifugation is diverted to a diluent overflow chamber via an overflow conduit located at the very top (nearest center of rotation) of the diluent metering chamber.

[0510] The diluent overflow conduit has dimensional features and shape / design which allow the excess diluent to enter this channel and flow into the diluent overflow chamber once the diluent metering chamber has been filled via rotation of the consumable in the first centrifugation.

[0511] The diluent metering chamber also has a diluent metering channel located a certain distance and volume below the top (nearest the center or rotation) of the diluent metering chamber. The position of the diluent metering channel within the diluent metering chamber and its channel dimensions / features are designed to deliver to the mixing chamber a certain diluent volume.

[0512] Diluent overflow chamber holds excess diluent that was delivered to the diluent metering chamber during the first centrifugation. The diluent diverted into the diluent overflow chamber becomes contained and is isolated from all further analysis described below unless it is to be delivered / used within calibration chambers.

[0513] If calibration chambers are to be filled with diluent, an exit channel flows from the diluent overflow chamber to the calibration chambers. The design of the dilution overflow chamber's exit channel permits diluent to flow into all calibration chambers to which it is connected during centrifugation of the analysis cartridge.

[0514] This chamber can be interrogated for the presence of liquid and confirm that diluent metering chamber was filled.

[0515] Calibration chambers may be filled with air or diluent. If connected by diluent excess chamber exit channel, or other materials including reagents, reference surfaces etc. as required for specific purposes. In the case where diluent is delivered to these chambers it would be done so via an appropriate centrifugation step. These chambers are interrogated by the instrument for calibration and both consumable and instrument QC purposes. For example: presence of liquid confirms the diluent metering chamber was filled with a metered volume of diluent, sample quality: e.g. HIL index, instrument optics calibration: e.g. % transmission, et al.

[0516] The analysis cartridge rotation is brought to a stop. While slowing down and / or when stopped, the capillary forces of the sample metering and diluent metering channels cause these same channels to fill with sample and diluent respectively.

[0517] Upon centrifugation of the analysis cartridge, i.e., rotation in a certain direction and within a certain rpm range, the metered sample volume and the metered diluent volume are emptied into a metering chamber.

[0518] In cases where the channel features of both or eitherof the sample metering and dilution metering channels were designed to prevent unintentional flow of sample and diluent into the mixing chamber, the centrifugation at certain speed and duration overcomes such features, and the metered volumes of sample and diluent are delivered to the mixing chamber.

[0519] The Mixing chamber has a desired volumetric capacity, which is greater than any combination of the metered sample and diluent. The plasma dilution chamber is designed with a volumetric capacity to ensure all metered sample and diluent delivered via the third centrifugation fit within it and to accommodate a certain volume of air to potentially facilitate and encourage mixing and homogenization of the sample and diluent within this chamber.

[0520] The mixing chamber can be designed to allow a desired dilution of the sample. The mixing chamber has an exit channel located a certain distance and volume below the top (nearest the center or rotation) of the mixing chamber.

[0521] Once the mixing chamber's exit channel begins to fill, dimensional features and design of the end of this channel may be used to prevent fluid from entering the next fluidic elements: detection chambers and excess diluted sample chamber (if present), unintentionally and only when initiated by appropriate centrifugation of the analysis cartridge.

[0522] Upon the second centrifugation of the analysis cartridge, i.e., rotation in a certain direction and within a certain rpm range or a program or series of rotations in either direction at varying rpms, the metered sample and the metered diluent are emptied into a mixing chamber.

[0523] The analysis cartridge can be rotated or rocked back and forth to ensure the sample and diluent are fully mixed, homogenized prior to delivery to the next fluidic elements of the analysis cartridge. When the analysis cartridge rotation is brought to a stop, the slowing down and / or when stopped, the capillary forces of the mixing chamber exit channel cause this channel to fill with homogeneously mixed diluted sample.

[0524] Upon third centrifugation of the consumable, i.e., rotation in a certain direction and within a certain rpm range, the diluted sample is delivered to the one or more detection chambers via the diluted sample distribution channel. In cases where the channel features of mixing chamber's exit channel are designed to prevent unintentional flow of diluted sample into the detection chambers and excess diluted sample chamber if present, the third centrifugation overcomes such features.

[0525] Detection chambers comprise a series of chambers which may have varying length, width and height as required to yield volumes and optical pathlengths suitable for specific and individual assay reaction and detection schemes. The detection chambers contain dried reagents: usually lyophilized which are specific for each individual assay. Some detection chambers may not contain any reagents to serve calibration purposes. During the third centrifugation, the detection chambers are filled with diluted sample and any air contained within them is evacuated.

[0526] Excess diluted sample chamber mayor may not be present in analysis cartridges disclosed herein. Any diluted sample volume that is not used to fill the reaction cuvettes can be delivered to the excess diluted plasma chamber during the third centrifugation. Delivery of the excess diluted sample to this chamberfluidically isolates the contents of each detection chambers.

[0527] The diluted sample sent to the excess diluted sample chamber will not be used for any analytical purposes. This chamber can be interrogated for the presence of liquid and confirm that the reaction cuvettes were properly filled.

[0528] The analysis cartridge can be further centrifuged in a certain direction and within a certain rpm range or a sequence of rotations in possibly varying directions and rpms, to generate flows within each of the reaction cuvettes to promote mixing and homogenization of the reagents with the diluted sample.

[0529] Analysis: For all analytes not requiring 2-step detection chambers, the chambers could be optically interrogated.

[0530] Two-Step detection chambers: In some cases, all reagents necessary for detection of an analyte cannot be located within a single detection chambers. In such cases, separate detection chambers each containing a subset of the reagents required for detection of the analyte are employed. The 2-step detection chambers may have varying length, width and height as required to yield volumes and optical pathlengths suitable for specific and individual assay reaction and detection schemes. When 2-step reaction cuvettes are employed, the first reaction cuvette is as described above except that it has an exit channel leading from it into a second detection chamber.

[0531] During certain centrifugations of the analysis cartridge related to the first detection chamber, this exit channel's design prevents fluid from entering the second detection chamber either because a) the centrifugal force is greater that the capillary force of this same channel or b) the end of the exit channel leading away from the first detection chamber is designed to only be breached at a higher centrifugal force. Upon further centrifugation of the analysis cartridge,i.e., rotation in a certain direction and within a certain rpm range, the diluted sample and reagents from the first detection chamber are delivered to the second detection chamber.

[0532] Even further centrifugation of the analysis cartridge, i.e., rotation in a certain direction and within a certain rpm range or a sequence of rotations in possibly varying directions and rpms, generates flows within each of the detection chambers intended to promote mixing and homogenization of the reagents with the diluted sample in the second detection chamber.

[0533] Analysis: For all analytes requiring 2-step detection chambers, the chambers could be optically interrogated.Example 3: An exemplary analysis cartridge having metering fingers

[0534] FIGS. 10 depicts an exemplary analysis cartridge having one or more metering fingers. In this example, the analysis cartridge is in a pie shape and can be centrifuged around an axis that passes through the center of the circle of which the pie is a part. Any suitable number of analysis cartridges could be centrifuged at a time depending on the angle of the pie. Thus, if the angle of the pie is about 60°, six analysis cartridges can be loaded onto a rotor of a centrifuge. If the angle of the pie is about 72°, five analysis cartridges can be loaded onto a rotor of a centrifuge. If the angle of the pie is about 90°, four analysis cartridges can be loaded onto a rotor of a centrifuge. If the angle of the pie is about 120°, three analysis cartridges can be loaded onto a rotor of a centrifuge.

[0535] The analysis cartridge 1000 of FIG. 10 comprises the mounting hole 1001. The mounting hole can engage with a mounting pin, for example, the mounting pin of an appropriate centrifuge rotor to load the analysis cartridge for centrifugation and analysis in an analyzer.

[0536] The analysis cartridge comprises the diluent chamber 1003, which has the diluent loading port 1002. The diluent chamber is fluidically connected to the diluent metering chamber 1006. The analysis cartridge of FIG. 10 also comprises the sample chamber 1005, which has the sample loading port 1004. A diluent can be loaded into the diluent chamber and a sample can be loaded into the sample chamber via the corresponding inlets. An analysis cartridge with the sample loaded in the sample chamber and the diluent loaded into the diluent chamber is shown in FIG. 14A.

[0537] Centrifugation of the analysis cartridge at a certain speed exerts on a diluent in the diluent chamber sufficient centrifugal force to move the diluent from the diluent chamber 1003 to the diluent metering chamber 1006. The volume of the diluent metering chamber is fixed and known, for example, between 450 pL to 500 pL. Any excess diluent beyond this volume istransported to the diluent overflow chamber 1007. The diluent overflow chamber comprises the diluent overflow chamber vent 1008, which allows the air to go out when the diluent chamber and the diluent overflow chamber is filled with a diluent.

[0538] The sample chamber 1005 is fluidically connected to the sample metering chamber 1009 via the channel 1024. The sample metering chamber 1009 is fluidically connected to the cellular components chamber 1010. Centrifugation of the analysis chamber at a certain speed exerts on a sample in the sample chamber sufficient centrifugal force to move the sample from the sample chamber to the sample metering chamber and the cellular components chamber. The volume of the sample metering chamber is predetermined and known, for example, about 12 pL. Any excess sample that does not fit in the sample metering chamber and the cellular components chamber flows to the sample overflow chamber 1011. The sample overflow chamber comprises the sample overflow vent 1012, which allows the air to go out when the sample overflow chamber is filled with a sample. An analysis cartridge after the first centrifugation such that the sample is loaded in the sample chamber and the cellular components chamber (with excess in the sample overflow chamber) and the diluent is loaded into the diluent chamber (with excess diluent in the diluent overflow chamber) is shown in FIG. 14B.

[0539] A centrifugation of the analysis chamber at a certain speed exerts on a sample in the sample metering chamber sufficient centrifugal force to force the cells in the sample (e.g., blood cells in a blood sample) into the cellular components chamber. Thus, the non-cellular portion of the sample is collected in the sample metering chamber 1009. The volumes of the loaded samples, the sample metering chamber, and the cellular components chamber are chosen such that the entirety of the sample chamber is filled with the sample and the cellular components chamber is partially filled with the cellular components and partially filled with the sample. This arrangement ensures that the sample metering chamber is filled with the sample and facilitates accurate metering of the sample. An analysis cartridge after centrifugation such that the cellular components are separated in the cellular components sub-folder and the sample is collected in the sample metering chamber is shown in FIG. 14C.

[0540] Certain details of the separation of cellular components from the sample are depicted in FIGS. 15A-15E. During a specific centrifugation, the sample (e.g., a blood sample) enters the cellular components chamber 1010.

[0541] The entry of the sample into the cellular component chamber can be facilitated via the stepped feature in the connecting channel, which is depicted in FIGS. 16A-16B and 17A-17C.

[0542] As shown in FIG. 16A, the sample loaded into the sample chamber can enter the sample metering chamber 1602 through the channel 1601. The cross section area of the channel 1601 is at least 0.2 mm2. This size promotes jetting.

[0543] The fluidic connection 1608 between the metering chamber and the sample overflow chamber has a reduced cross-sectional area. This improves the resolution and accuracy of metering. The sharp cutoff 1610 to the sample overflow chamber further aids the accuracy and resolution of metering.

[0544] The floor of the sample metering chamber is the ramp 1603, which can be, for example, a 20° ramp. This feature ensures that ensures the lower section of the sample metering chamber is filled first. The metering chamber is fluid ica I ly connected to the cellular components chamber via the connection channel 1611. The connecting channel 1611 has the deeper channel 1604 and the shallower channel 1605. The deeper channel can be between 0.2 and 0.5 mm, e.g., 0.2 mm, 0.3 mm, 0.4 mm, or 0.5 mm lower than the shallower channel. For example, the deeper channel has a 0.4 mm down-step compared to the shallower channel. This reduces the risk of air lock during the initial fill of the cellular component chamber 1606. Also, the smaller cross- sectional area of the connecting channel allows for greater metering accuracy.

[0545] During centrifugation of the analysis cartridge, the sample flow path is line with the axis of rotation minimizing sheer experienced by the cellular components. The cellular component chamber has a vertical wall 1607. The cellular component chamber 1606 is designed to collect and pack the cells at the widest and lowest area of this chamber. As shown in FIGS. 17B and 17C, during deceleration, the packed cells may resuspend (acceleration vortices). During such resuspension, the steep vertical walls prevent the cells from traveling upstream into the sample metering chamber.

[0546] As depicted in FIG. 16B, an angle, e.g., a 2-degree angle, is added to the proximate face of the sample metering chamber. This ensures that the overflow exit is at a proximate compared to the other end of the wall, which also encourages venting bubbles through to the sample overflow chamber 1011.

[0547] During a specific centrifugation step of the analysis cartridge, the cellular components of the sample are collected in the cellular component chamber and a non-cellular sample is collected and metered in the sample metering chamber. An exemplary cellular separation is depicted in FIGS. 15A-15E. For example, a sample is pipetted into the sample chamber. If it is a blood sample, 60% or lower haematocrit is preferred.

[0548] During a specific centrifugation, the sample enters the cellular component chamber via the stepped feature in the connecting channel 1503. The sample runs down the side to allow for air swap. Once the metering chamber 1502 and the cellular component chamber 1504 are filled, excess blood is spun off into the sample overflow / waste chamber 1505.

[0549] Once all excess sample is spun off, the sample volume dictated by the volume of the metering chamber is metered. A specific centrifugation induces separation of the cellular components and their accumulation in the cellular component chamber 1504. The metered sample in the metering chamber 1504 is then siphoned through the sample siphon 1506.

[0550] The analysis cartridge of FIG. 10 further comprises the diluent siphon 1014 and a sample siphon 1013. The diluent siphon and the sample siphon are primed by centrifuging the analysis cartridge at a certain speed. Such centrifugation opens the diluent siphon and allows the metered diluent from the diluent metering chamber 1006 to flow to the mixing chamber 1015. Similarly, such centrifugation opens the sample siphon 1013 and allows the metered sample from the sample metering chamber to flow to the mixing chamber 1015. The mixing chamber comprises the mixing chamber vent 1025, which allows the air to go out when the mixing chamber fills with the diluted sample.

[0551] FIG. 14D shows the analysis cartridge in the process of centrifugation such that a portion of the metered diluent from the diluent metering chamber and a portion of the metered sample from the sample metering chamber is introduced in to the mixing chamber and is getting mixed. Mixing is achieved by rapidly changing speed from ~4000 RPM to ~750RPM while driving in the same direction. This change in speed causes micro vortices between the fluid layers due to the change in force experienced by each layer radius. 14E shows the analysis cartridge after centrifugation such that the metered diluent from the diluent metering chamber and the metered sample from the sample metering chamber is introduced in to the mixing chamber and is mixed. In the mixing chamber 1015, the diluent and the sample are mixed to produce a diluted sample.

[0552] In some cases, the fluidic connection between the diluted sample siphon and the distribution channel is conical with the broader end connected to the distribution channel. Such design avoids a bottle neck for dispense speeds and allow smooth flow of diluted sample from the mixing chamber into a distribution channel.

[0553] Furthermore, the analysis cartridge of FIG. 10 comprises the diluted sample siphon 1016, which connects the mixing chamber to the one or more detection chambers 1021 via one or more metering fingers 1018. The diluted sample siphon is primed by centrifuging the analysiscartridge at a certain speed. Such centrifugation opens the diluted sample siphon 1016 and allows the diluted sample from the mixing chamber 1015 to flow to the one or more metering fingers 1018. The metering fingers are designed such that a metered volume of diluted sample is loaded into each of the metering fingers before it is introduced in the corresponding detection chambers 1021 via the corresponding capillary channels 1023. The dimensions of the channels are selected so that the channels consistently fill without jetting the diluted sample into the corresponding detection chambers. Some of the detection chambers comprise multiple chambers, as exemplified by the chambers 1022.

[0554] Distribution of the diluted sample to the one or more metering fingers 1018 can be via the distribution channel 1017. Filling of the metering fingers from one end to the other of the analysis cartridge can occur in a substantially sequential manner, with one metering finger is filled at a time before the next one is filled. FIG. 14F shows the analysis cartridge with the diluted sample filled in the first two metering fingers and in the process of continuing filling the subsequent metering fingers. FIG. 14G shows the analysis cartridge with the diluted sample filled in all of the metering fingers.

[0555] The details of filling up the metering fingers and the corresponding detection chambers are further described below and depicted in FIGS. 21A-21F, 22-23, 24A-24E, and 25.

[0556] Once mixing of the sample and the diluent is complete, the centrifugation of the analysis cartridge slows down and allows the diluted sample to prime the diluted sample siphon 1016. Once primed, the centrifuge speeds up to begin distribution. This step is depicted in FIGS. 21A- 21B and FIG. 24B.

[0557] As the centrifugation of the analysis cartridge continues, the diluted sample enters the distribution channel 1017 and begins to fill up the first metering finger without jetting into the corresponding first detection chamber. This step is depicted in FIGS. 21B and 24C-24D.

[0558] As the first channel is filled, overfill occurs and starting the filling sequence of the next metering finger. This is depicted in FIGS. 21C and 24E. The second metering finger is filled next, as depicted in FIG. 21D.

[0559] This sequential process is repeated until all 20 metering fingers are filled as shown in FIGS. 21E and 25. As the last metering finger fills up the centrifugation speed is increased to spin off any excess diluted sample into the diluted sample overflow chamber 1019. The diluted sample overflow chamber comprises the diluted sample overflow chamber vent 1020, which allows the air to go out when the diluted sample fills in the metering fingers.

[0560] Once all the metering fingers are filled and all excess has been spun off to the diluted sample overflow chamber, the distribution of the diluted sample is complete, as shown in FIG. 21F.

[0561] To improve timings and reliability, the distribution channel can be designed to be non- concentric to the axis of rotation. This allows the start of the channel and the end of the channel to be at different radial heights. This is shown in FIG. 22, where the distribution channel is not concentric to the axis of rotation. This feature would allow the last detection chamber to be filled despite the mixing chamber being emptied. In an exemplary embodiment, a 2 mm step down is used, i.e., the end of the distribution channel towards the diluted sample siphon is 2 mm proximate to the end of the distribution channel towards the diluted sample overflow chamber. By increasing the step down, the last wells can be filled faster.

[0562] The filling of the detection chamber is driven by the combination of capillary stop force, back pressure from compressing enclosed air, and the burst frequency where instability occurs. By adjusting the volume of compressed air and the plug length, the centrifugation speed at which instability occurs can be modified. The capillary stop is strengthened due to the back pressure generated by the compression of the enclosed air volume in the detection chamber, as shown in FIG. 25. This allows the capillary stop to function at higher forces than without it.

[0563] The diluted sample siphon outlet may act as a bottle neck for dispense speeds. To address this potential issue, a trumpet design can be implemented in diluted sample siphon. This is shown in FIG. 23, where a trumpet shaped outlet of the diluted sample siphon connects to the distribution channel 2302.

[0564] As noted above, the connecting channels between the metering fingers and the corresponding detection chambers have dimensions that allow the channels to be consistently filled without jetting the diluted sample into the detection chamber.

[0565] Certain features of the metering fingers shown in FIG. 24A facilitate proper filling and metering of the diluted sample. For example, the rounded distal ends 2401 of the metering fingers guide the fluid flow towards the capillary channel during initial filling. Also, ramped inlets 2402 ensure that air gets pushed out during filling of the metering chamber. Moreover, the channels 2403, with the dimensions of 0.2 mm x 0.5 mm consistently filled with the diluted sample without jetting the diluted sample into the corresponding detection chambers 2404.

[0566] As noted above, some of the detection chambers can have two chambers. One such exemplary detection chamber is shown in FIGS. 26A-26B. The 2-step reaction being the last step in the filling process, provides two main benefits: as the last cuvette, there is no crosscontamination with previous cuvettes and the slower flow rate increases the amount of time the first Lyo bead (bead containing lyophilized reagents) has to dissolve so the bead doesn't get carried to the waste chamber.

[0567] As shown in FIG. 26A, the detection chamber having two chambers 2602 and 2603 is connected to the corresponding metering finger via channel 2601. Compared to the 2 mm width of the channels for a single chambered detection chambers, the width for this channel is reduced to 1.5 mm. This feature minimizes the backflow of dissolved Lyo beads.

[0568] Moreover, the two-step metering uses a deep 4.8 mm circular geometry to inhibit the Lyo bead in the detection chamber from traveling upstream when it is wetted. This is shown in FIG. 26B.

[0569] When all the metering fingers are filled, a further centrifugation of the analysis cartridge at a higher speed moves the diluted sample from the metering fingers into the detection chambers 1021. This is shown in FIG. 14H.

[0570] Detection chambers comprise one or more reactants that are configured to detect an analyte. The reagents can be provided in the form of beads that contain lyophilized reagents. Thus, when a diluted sample is loaded into a reaction mixture, the lyophilized reagents in the beads slowly dissolve in the diluted sample.

[0571] The analyte detection can be based on electrochemical or optical detection. Each detection chamber is a well of about 2.35 mm diameter (with a minimum of 2.32 mm diameter) having a draft angle of about 0.5 degrees and depth of about 5 mm. Thus, each well is configured to accommodate about 22 to 24 pL with about 1% variance in the loaded volume.

[0572] An exemplary combination of analytes in a centrifugation cartridge comprises: glucose, albumin, ALP, calcium, total protein, CO2, total bilirubin, blood urea nitrogen, ALT, AST, creatinine, sodium, chloride, and potassium. Additional detection chambers can be used for certain duplicates and reference readings.Example 4: Work-flow details and volume profiles for an exemplary analysis cartridge

[0573] This example describes certain details of various reagent and sample volumes as well as work-flow that could be implemented in analyzing a sample using the exemplary analysis cartridge described in Example 3.

[0574] FIG. 11 provides a flow chart for analyzing a sample in the analysis cartridge described in Example 3 and depicted in FIG. 10. Briefly, a sample is introduced into the sample chamber 1005 via the sample loading port 1004. In a specific centrifugation, the sample is metered and cellularcomponents are separated. For example, when blood is the sample, blood is separated into plasma and cellular components, and measured volume of blood plasma (sample) is collected in the sample metering chamber. Excess sample is collected in the sample overflow chamber 1011.

[0575] Similarly, a diluent is introduced into the diluent chamber 1003 via the diluent loading port 1002. In a specific (the first) centrifugation, the diluent is metered and excess diluent is collected in the diluent overflow chamber 1007.

[0576] In a further centrifugation (the second centrifugation), sample from the sample metering chamber and the diluent from the diluent chamber are introduced via the sample siphon and the diluent siphon, respectively, into the mixing chamber. In the mixing chamber 1015, the diluent and the sample are mixed to produce a diluted sample.

[0577] In an even further centrifugation (the third centrifugation), the diluted sample is introduced into a number of metering fingers via the distribution channel 1017. Excess diluted sample is collected in the diluted sample overflow chamber 1019.

[0578] In a further centrifugation (the fourth centrifugation), the diluted sample from the metering fingers is introduced into the detection chambers 1021, where a plurality of analytes are analyzed to determine their amounts in the sample. Typically, an optical signal is used to detect the analytes. However, electrochemical signal or electrical signals can also be used.

[0579] Certain exemplary volumes of different fluids and chambers utilized in different steps during the operation of an analysis cartridge described herein are provided in FIG. 12.

[0580] In the exemplary embodiment depicted in FIG. 12, about 545 pL of diluent is introduced into the diluent chamber and about 70 pL of a sample, e.g., blood, is introduced into the sample chamber. The diluent metering chamber is about 468 pL, with the excess diluent collected in the diluent overflow chamber. Upon centrifugation and cellular component separation in the sample metering chamber, about 12 pL, for example, about 11.91 pL, of the non-cellular portion of the sample, e.g., plasma is collected in the sample metering chamber.

[0581] For example, as shown in FIG. 18A, about 550 pL of diluent is pipetted or otherwise introduced into the diluent chamber. Upon a specific centrifugation, the diluent enters the metering chamber, as shown in FIG. 18B, the diluent enters the metering chamber. As shown in FIG. 18C, when the diluent metering chamber is full, the excess flows into the diluent overflow chamber. Once all the excess diluent is in the waste chamber then the diluent metering is complete, as shown in FIG. 18D.

[0582] Certain design features of the diluent chamber facilitate accurate metering of a diluent. For example, as shown in FIG. 19A, the two stage ramp (1901 and 1902), aids removal of metereddiluent. Particularly, it keeps the height of the diluent level above the siphon before it breaks up in the siphon. Also, as shown in FIG. 19B, the metering outlet 1903 of the dilution chamber is shallow with a sharp drop off as shown by the wall 1904. This design improves metering precision. Further, the outlet of the dilution chamber 1905 is not concentric to encourage fluid to flow outward into the siphon.

[0583] Further, as shown in FIG. 20A, the outlet 2001 of the diluent metering chamber is not concentric. This encourages the diluent to flow outward into the diluent siphon. Also, as shown in FIG. 20B, the connection 2002 of outlet of the diluent metering chamber is in line with the mixing chamber. This arrangement allows any fluid in the siphon to re-enter the mixing chamber during mixing.

[0584] Mixing of the diluent with the sample is achieved in the mixing chamber by rapidly changing the centrifugation speed from about 4000 RPM to about 750 RPM while driving in the same direction. This change in speed causes micro vortices between the fluid layers due to the change in force experienced by each layer radius (r), which is shown in FIG. 20C. This force is shown in the formula Fc= - mw x (w x r). This mixing method may occur in a superior manner in a taller mixing chamber rather than wider mixing chamber.

[0585] Certain design features of the mixing chamber and its connection to the diluted sample siphon facilitate the flow of diluted sample to the siphon.

[0586] Mixing of the metered diluent and the metered sample into the mixing chamber produces about 480 pL of diluted sample. The diluted sample is introduced into the 20 metered fingers, each containing about 23 pL, for example, about 22.96 pL of the diluted sample. Thus, the total sample introduced into the 20 metered fingers is about 460 pL. The detection chamber with two subchambers can receive about 22.75 pL of the diluted sample. The remaining about 20 pL, for example, about 22.15 pL of the diluted sample is collected in the diluted sample overflow chamber.

[0587] Each detection chamber receives about 23 pL, for example, about 22.90 pL of the diluted sample.

[0588] Certain exemplary dimensions for the analysis cartridge described in Example 3 are provided in FIG. 13.

[0589] In some cases, ALTUGLAS™ PMMA VSUVT is the resin material used for this cartridge. This resin has a reported contact angle of 66-72 degrees. This is a high flow moulding resin suitable for injection moulding of complex parts. This resin provides exceptional UV transmissionand excellent transparency, making it a suitable choice for making the analysis cartridges described herein.

[0590] Example 5 - Desirable process features achieved by usingthe analysis cartridge described in Examples 4 and 5

[0591] Due to various features of the analysis cartridge described in Examples 3 and 4, analysis of certain non-cellular components of blood can be achieved in a fast and accurate manner. Certain such desired features and parameters of the process are described in Table 1 below:

[0592] Table 1: Desired features and parameters of analysis cartridge of Examples 3 and 4

[0593] An exemplary combination of non-cellular components analyzed using the analysis cartridge described in Examples 3 and 4 is provided in the Table 2 below:

[0594] Table 2: An exemplary combination of analytes analyzed in the analysis cartridge of Examples 3 and 4.

[0595] The preceding merely illustrates the principles of the present disclosure. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of the invention and the concepts contributed by the inventors to furthering the art and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of present invention is embodied by the appended claims.

Claims

1. CL IMSWE CLAIM:

1. An analysis cartridge for analyzing a sample, the analysis cartridge configured to be centrifuged around a vertical axis, the analysis cartridge comprising: a diluent chamber fluidical ly connected to a diluent metering chamber, wherein a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold, a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve and to a sample metering chamber via a sample valve such that, when the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold, one or more detection chambers fluidically connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more detection chambers, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

2. The analysis cartridge of claim 1, wherein the third threshold is higher than the second threshold and the second threshold is higher than the first threshold.

3. The analysis cartridge of claim 1 or 2, wherein the sample metering chamber is fluidically connected to a sample chamber such that a fourth centrifugation of the analysis cartridge moves the sample from the sample chamber into the sample metering chamber, wherein the fourth centrifugation exerts on the sample in the sample chamber a fourth centrifugation force above a fourth threshold.

4. The analysis cartridge of claim 3, wherein the fourth threshold is lower than the first and / or the second threshold.

5. The analysis cartridge of any one of claims 1 to 4, wherein the sample metering chamber further com rises a sample overflow conduit fluid ica lly connected to a sample overflow chamberthat receives a portion of the sample that does not fit in the sample metering chamber.

6. The analysis cartridge of claim 5, wherein the sample metering chamber further comprises a cellular components sub-chamber and the fourth centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components subchamber.

7. The analysis cartridge of claim 5 or 6, wherein the sample overflow chamber and / or the sample overflow conduit comprises a first set of one or more electrochemical sensors that detect a first set of one or more analytes in the sample.

8. The analysis cartridge of claim 7, wherein the sample overflow conduit is fluidical ly connected to a calibration fluid chamber.

9. The analysis cartridge of any one of claims 1 to 8, wherein the sample chamber is further fluidically connected to an electrochemical analysis module, wherein a fifth centrifugation of the analysis cartridge moves a portion the sample from the sample chamber into the electrochemical analysis module.

10. The analysis cartridge of claim 9, wherein the electrochemical analysis module comprises a second set of one or more electrochemical sensors that detect a second set of one or more analytes in the sample.

11. The analysis cartridge of claim 10, wherein the electrochemical analysis module comprises a calibration fluid chamber fluidically connected to the second set of one or more electrochemical sensors.

12. The analysis cartridge of any one of claims 1 to 11, wherein the diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon.

13. The analysis cartridge of claim 12, wherein the diluent siphon, the sample siphon, and / or the diluted sample siphon is / are opened by priming.

14. The analysis cartridge of claim 13, wherein the diluent siphon and the sample siphon are primed by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold.

15. The analysis cartridge of claim 13 or 14, wherein the diluted sample siphon is primed by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

16. A method of analyzing a sample in an analysis cartridge, the method comprising: conducting a first centrifugation of the analysis cartridge to move a diluent from a diluent chamber to a f luidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold, conducting a second centrifugation of the analysis cartridge to move into a mixing chamber the diluent from the diluent metering chamber and the sample from the sample metering chamber; wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold, conducting a third centrifugation to move into one or more detection chambers the diluted sample from the mixing chamber, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

17. The method of claim 16, wherein the third threshold is higher than the second threshold and the second threshold is higher than the first threshold.

18. The method of claim 16 or 17, further comprising conducting a fourth centrifugation of the analysis cartridge before conducting the first centrifugation of the analysis cartridge, wherein the fourth centrifugation moves the sample from a sample chamber into the sample metering chamber, and wherein the fourth centrifugation exerts on the sample in the sample chamber a fourth centrifugation force above a fourth threshold.

19. The method of claim 18, wherein the fourth threshold is lower than the first and / or the second threshold.

20. The method of any one of claims 16 to 19, comprising detecting in the one or more detection chambers one or more signals that indicate the concentrations of one or more analytes in the sample and determining the concentrations of the one or more analytes in the sample.

21. The method of any one of claims 16 to 20, wherein conducting the fourth centrifugation of the analysis cartridge comprises receiving into a sample overflow chamber via a sample overflow conduit a portion of the sample that does not fit in the sample metering chamber.

22. The method of any one of claims 18 to 21, wherein conducting the fourth centrifugation of the analysis cartridge comprises receiving the cellular components of the sample into the cellular components sub-chamber that is fluidically connected to the sample metering chamber.

23. The method of claim 21 or 22, further comprising detecting a first set of one or more analytes in the sample in one or more electrochemical sensors located in the sample overflow chamber and / or the sample overflow conduit.

24. The method of any one of claims 16 to 23, wherein the sample chamber is further fluidically connected to an electrochemical analysis module comprising a second set of one or more electrochemical sensors, and wherein conducting a fifth centrifugation of the analysiscartridge moves a portion of the sample from the sample chamber into the electrochemical analysis module.

25. The method of claim 24, further comprising detecting a second set of one or more analytes in the sample using the second set of one or more electrochemical sensors.

26. The method of any one of claims 16 to 25, wherein the diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon.

27. The method of claim 26, comprising opening by priming the diluent siphon, the sample siphon, and / or the diluted sample siphon.

28. The method of claim 27, comprising priming the diluent siphon and the sample siphon by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold.

29. The method of claim 26 or 27, comprising priming the diluted sample siphon by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamberthe third centrifugal force above the third threshold.

30. An analyzer for analyzing a sample in an analysis cartridges, the analyzer comprising: a rotor comprising a slot that holds the analysis cartridge such that, when the rotor rotates, the analysis cartridge is centrifuged around a vertical axis, a processor, and a non-transitory computer readable medium comprising instructions, which when executed by the processor, cause the analyzer to perform: a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into a fluidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold,when a diluent valve and a sample valve are open, a second centrifugation of the analysis cartridge that moves the diluent from the diluent metering chamber and a sample from a sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold, a third centrifugation of the analysis cartridge that moves the diluted sample from the mixing chamber into the one or more detection chambers, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold.

31. The analyzer of claim 30, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform the first centrifugation followed by the second centrifugation followed by the third centrifugation.

32. The analyzer of claim 30 or 31, wherein the third threshold is higher than the second threshold and the second threshold is higher than the first threshold.

33. The analyzer of any one of claims 30 to 32, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform a fourth centrifugation of the analysis cartridge before conducting the first centrifugation of the analysis cartridge, wherein the fourth centrifugation moves the sample from a sample chamber into the sample metering chamber, and wherein the fourth centrifugation exerts on the sample in the sample chamber a fourth centrifugation force above a fourth threshold.

34. The analyzer of claim 33, wherein the fourth threshold is lower than the first and / or the second threshold.

35. The analyzer of claim 33 or 34, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer toperform the fourth centrifugation followed by the first centrifugation followed by the second centrifugation followed by the third centrifugation.

36. The analyzer of any one of claims 30 to 35, further comprising a detector that detects one or more signals from the one or more detection chambers, wherein the one or more signals indicate the concentration of one or more analytes in the sample.

37. The analyzer of claim 36, wherein the detector is an optical detector that detects one or more optical signals from the one or more detection chambers.

38. The analyzer of any one of claims 30 to 37, wherein the analyzer further comprises a first set of one or more electrochemical detectors that detect a first set of one or more electrochemical signals from a first set of one or more electrochemical sensors located in a sample overflow chamber and / or the sample overflow conduit of the analysis cartridge.

39. The analyzer of claim 38, wherein the first set of one or more electrochemical detectors comprises an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof.

40. The analyzerof any one of claims 30 to 39, wherein the analyzer further comprises a second set of one or more electrochemical detectors that detect a second set of one or more electrochemical signals from a second set of one or more electrochemical sensors located in an electrochemical analysis module of the analysis cartridge.

41. The analyzer of claim 40, wherein the second set of one or more electrochemical detectors comprises an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof.

42. The analyzer of any one of claims 38 to 41, wherein the first set of one or more electrochemical detectors and / or the second set of one or more electrochemical are parts of an electrical detection unit comprising electrical circuits that detect electrical signals.

43. An analysis cartridge for analyzing a sample, the analysis cartridge configured to be centrifuged around a vertical axis, the analysis cartridge comprising: a diluent chamber fluidical ly connected to a diluent metering chamber, wherein a first centrifugation of the analysis cartridge moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold, a mixing chamber fluidically connected to the diluent metering chamber via a diluent valve and to a sample metering chamber via a sample valve such that, when the diluent valve and sample valve are open, a second centrifugation of the analysis cartridge moves the diluent from the diluent metering chamber and a sample from the sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold, one or more metering fingers fluidically connected to the mixing chamber via a diluted sample valve such that, when the diluted sample valve is open, a third centrifugation of the analysis cartridge moves the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and one or more detection chambers fluidically connected to the one or more metering fingers via capillary channels such that, a fourth centrifugation of the analysis cartridge moves the diluted sample from the one or more metering fingers into the one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold.

44. The analysis cartridge of claim 43, wherein the fourth threshold is higherthan the third threshold, the third threshold is higher than the second threshold, and the second threshold is higher than the first threshold.

45. The analysis cartridge of claim 43 or44, wherein the sample metering chamber is fluidically connected to a sample chamber such that the first centrifugation of the analysis cartridge moves the sample from the sample chamber into the sample metering chamber.

46. The analysis cartridge of claim 45, wherein the sample metering chamber is fluidically connected to a cellular components chamber and the first centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components chamber.

47. The analysis cartridge of claim 43 or44, wherein the sample metering chamber is fluidically connected to a sample chamber such that a fifth centrifugation of the analysis cartridge moves the sample from the sample chamber into the sample metering chamber, and wherein the fifth centrifugation exerts on the sample in the sample chamber a fifth centrifugation force above a fifth threshold.

48. The analysis cartridge of claim 47, wherein the sample metering chamber is fluidically connected to a cellular components chamber and the fifth centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components chamber.

49. The analysis cartridge of claim 48, wherein the fifth threshold is lower than the first and / or the second threshold.

50. The analysis cartridge of any one of claims 43 to 49, wherein the sample metering chamber further comprises a sample overflow conduit fluidically connected to a sample overflow chamberthat receives a portion of the sample that does not fit in the sample metering chamber.

51. The analysis cartridge of claim 50, wherein the sample overflow chamber and / or the sample overflow conduit comprises a first set of one or more electrochemical sensors that detect a first set of one or more analytes in the sample.

52. The analysis cartridge of claim 51, wherein the sample overflow conduit is fluidically connected to a calibration fluid chamber.

53. The analysis cartridge of any one of claims 43 to 52, wherein the sample chamber is further fluidically connected to an electrochemical analysis module, wherein a sixthcentrifugation of the analysis cartridge moves a portion the sample from the sample chamber into the electrochemical analysis module.

54. The analysis cartridge of claim 53, wherein the electrochemical analysis module comprises a second set of one or more electrochemical sensors that detect a second set of one or more analytes in the sample.

55. The analysis cartridge of claim 54, wherein the electrochemical analysis module comprises a calibration fluid chamber fluidically connected to the second set of one or more electrochemical sensors.

56. The analysis cartridge of any one of claims 43 to 55, wherein the diluent valve is a diluent siphon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon.

57. The analysis cartridge of claim 56, wherein the diluent siphon, the sample siphon, and / or the diluted sample siphon is / are opened by priming.

58. The analysis cartridge of claim 57, wherein the diluent siphon and the sample siphon are primed by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold.

59. The analysis cartridge of claim 57 or 58, wherein the diluted sample siphon is primed by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

60. The analysis cartridge of claim 59, wherein the diluted sample from the one or more metering fingers is moved into the one or more detection chambers by centrifuging the analysis cartridge to exert on the diluted sample in the one or more metering fingers the fourth centrifugal force above the fourth threshold such that the diluted sample in the one or more metering fingers overcomes the capillary stop pressure of the one or more capillary channels.

61. The analysis cartridge of any one of claims 43 to 60, wherein the diluent metering chamber is fluidically connected to a diluent overflow chamber such that the first centrifugation of the analysis cartridge moves into the diluent overflow chamber excess diluent that does not fit in the diluent metering chamber.

62. The analysis cartridge of claim 61, wherein the diluent overflow chamber comprises a diluent overflow vent that vents the gas from the diluent metering chamber as the diluent moves into the diluent metering chamber.

63. The analysis cartridge of any one of claims 43 to 62, wherein the sample metering chamber is fluidically connected to a sample overflow chamber such that the first centrifugation or the fifth centrifugation of the analysis cartridge moves into the sample overflow chamber excess sample that does not fit in the sample metering chamber.

64. The analysis cartridge of claim 63, wherein the sample overflow chamber comprises a sample overflow vent that vents the gas from the sample metering chamber as the sample moves into the sample metering chamber.

65. The analysis cartridge of any one of claims 43 to 64, wherein the mixing chamber comprises a mixing chamber vent that vents the gas from the mixing chamber as the diluted sample moves into the mixing chamber.

66. The analysis cartridge of any one of claims 43 to 65, wherein the one or more metering fingers are fluidically connected to a diluted sample overflow chamber such that the third centrifugation of the analysis cartridge moves into the diluted sample overflow chamber excess diluted sample that does not fit in the one or more metering fingers.

67. The analysis cartridge of claim 66, wherein the diluted sample overflow chamber comprises a diluted sample overflow chamber vent that vents the gas from the one or more metering fingers as the diluted sample moves into the one or more metering fingers.

68. The analysis cartridge of any one of claims 43 to 67, wherein the diluent chamber has a volume sufficient to accommodate a diluent having a volume selected from: between 500 pL and 600 |aL about 500 p.L, about 510 pL, about 520 pL, about 530 pL, about 540 pL, about 550 pL, about 560 pL, about 570 pL, about 580 pL, about 580 pL, and about 600 pL.

69. The analysis cartridge of any one of claims 43 to 68, wherein the diluent metering chamber has a volume selected from: between 450 pL and 500 pL, about 450 pL, about 455 pL, about 460 pL, about 465 pL, about 470 pL, about 475 pL, about 480 pL, about 485 pL, about 490 pL, about 495 pL, and about 500 pL.

70. The analysis cartridge of any one of claims 43 to 69, wherein the sample chamber has a volume sufficient to accommodate a sample having a volume selected from: between 50 pL and 70 pL, about 50 pL, about 52 pL, about 54 pL, about 56 pL, about 58 pL, about 60 pL, about 62 pL, about 64 pL, about 66 pL, about 68 pL, and about 70 pL.

71. The analysis cartridge of any one of claims 43 to 70, wherein the sample metering chamber has a volume selected from: between 8 pL and 15 pL, about 8 pL, about 9 pL, about 10 pL, about 11 pL, about 12 pL, about 13 pL, about 14 pL, and about 15 pL.

72. The analysis cartridge of any one of claims 43 to 71, wherein the mixing chamber has a volume that is sufficient to accommodate a diluted sample of: between 450 pL and 500 pL, about 450 pL, about 455 pL, about 460 pL, about 465 pL, about 470 pL, about 475 pL, about 480 pL, about 485 pL, about 490 pL, about 495 pL, and about 500 pL.

73. The analysis cartridge of any one of claims 43 to 72, wherein the one or more metering fingers have a volume of: between 15 pL and 30 pL, about 15 pL, about 16 pL, about 17 pL, about 18 pL, about 19 pL, about 20 pL, about 21 pL, about 22 pL, about 23 pL, about 24 pL, about 25 pL, about 26 pL, about T1 pL, about 28 pL, about 29 pL, and about 30 pL.

74. The analysis cartridge of any one of claims 43 to 73, wherein the one or more detection chambers have a volume of: between 15 pL and 30 pL, about 15 pL, about 16 pL, about17 .L about 18 |J.L, about 19 pL, about 20 pL, about 21 pL, about 22 pL, about 23 pL, about 24 pL, about 25 pL, about 26 pL, about 27 pL, about 28 .L, about 29 pL, and about 30 pL.

75. The analysis cartridge of any one of claims 43 to 74, wherein the sample chamber is fluidically connected to the sample metering chamber via a channel having a cross section area of: between 0.2 mm2and 0.5 mm2, about 0.2 mm2, about 0.25 mm2, about 0.3 mm2, about 0.35 mm2, about 0.4 mm2, about 0.45 mm2, and about 0.5 mm2.

76. The analysis cartridge of any one of claims 43 to 75, wherein the sample metering chamber comprises a proximate wall closer to the vertical axis, and wherein the fluidic connection between the sample metering chamber and the sample overflow chamber is towards the top of the proximate wall of the sample metering chamber, and wherein the fluidic connection covers at the most 20% of the height and at the most 30% of the width of the proximate wall of the sample metering chamber.

77. The analysis cartridge of any one of claims 43 to 76, wherein the floor of the sample metering chamber has an upward ramp relative to the bottom of the chamber, wherein the upward ramp is in the proximate to the distal direction.

78. The analysis cartridge of any one of claims 43 to 77, wherein a connecting channel between the sample metering chamber and the cellular components chamber has a deeper channel and a shallower channel, wherein the deeper channel is between 0.2 and 0.5 mm deeper than the shallower channel.

79. The analysis cartridge of any one of claims 42 to 78, wherein the cellular component chamber has a substantially horizontal floor and substantially perpendicular walls.

80. The analysis cartridge of any one of claims 42 to 79, wherein the proximate wall of the sample metering chamber is angled such that the fluidic connection between the sample metering chamber and the sample overflow chamber is proximate compared to the other end of the proximate wall of the sample metering chamber.

81. The analysis cartridge of claim 75, wherein the channel between the sample chamber and the sample metering chamber is in line with the vertical axis.

82. The analysis cartridge of claim 78, wherein the connecting channel between the sample metering chamber and the cellular components chamber is in line with the vertical axis.

83. The analysis cartridge of claim 56, wherein the sample siphon has a siphon crest proximate to the sample metering chamber.

84. The analysis cartridge of claim 83, wherein the sample siphon has a siphon exit that connects to the mixing chamber, and wherein the siphon exit is in line with the vertical axis.

85. The analysis cartridge of any one of claims 43 to 84, wherein, the distal wall of the diluent metering chamber comprises a ramp in one or more stages, wherein the one or more stages of the ramp are slanted from the proximal end to the distal end of the diluent metering chamber.

86. The analysis cartridge of claim 85, wherein the ramp in the diluent metering chamber comprises two stages.

87. The analysis cartridge of claim 56, wherein a fluidic connection between the diluent metering chamber and the diluent siphon is not concentric to the vertical axis.

88. The analysis cartridge of any one of claims 43 to 87, wherein the diluent metering chamber has a proximate wall closer to the vertical axis, and wherein the fluidic connection between the diluent metering chamber and the diluent overflow chamber is towards the top of the proximate wall of the diluent metering chamber, and wherein the fluidic connection covers at the most 20% of the height and at the most 10% of the width of the proximate wall of the diluent metering chamber.

89. The analysis cartridge of claim 56, wherein a fluidic connection between the mixing chamber and the diluted sample siphon is not concentric to the vertical axis.

90. The analysis cartridge of any one of claims 43 to 89, wherein the fluidic connection between the mixing chamber and the diluted sample siphon is in line with the mixing chamber.

91. The analysis cartridge of any one of claims 43 to 90, wherein the distribution channel is not concentric to the vertical axis.

92. The analysis cartridge of claim 91, wherein the end of the distribution channel near the fluidic connection with the diluted sample siphon is proximate than the end of the distribution channel near the fluidic connection with the diluted sample overflow chamber.

93. The analysis cartridge of claim 92, wherein the end of the distribution channel near the fluidic connection with the diluted sample siphon is proximate by at least 2 mm than the end of the distribution channel near the fluidic connection with the diluted sample overflow chamber.

94. The analysis cartridge of any one of claims 43 to 93, wherein the fluidic connection between the diluted sample siphon and the distribution channel is conical with the broader end connected to the distribution channel.

95. The analysis cartridge of any one of claims 43 to 94, wherein the one or more metering fingers comprise proximate walls towards the vertical axis, and the proximate walls of the one or more metering fingers are sloped away from the proximate and towards the distal end of the analysis cartridge.

96. The analysis cartridge of any one of claims 43 to 95, wherein the distal edges of the one or more metering fingers are rounded.

97. The analysis cartridge of any one of claims 43 to 96, wherein, when in operation, the capillary channels between the one or more metering fingers and the corresponding detection chambers are on the top surface of the analysis cartridge and have a depth of between 0.1 mm and 0.3 mm.

98. The analysis cartridge of claim 97, wherein, when in operation, the capillary channels between the one or more metering fingers and the corresponding detection chambers are on the top surface of the analysis cartridge and have a width of between 0.4 mm and 0.6 mm.

99. The analysis cartridge of any one of claims 43 to 98, wherein one or more of the one or more detection chambers have two chambers.

100. The analysis cartridge of any one of claims 43 to 99, wherein the one or more detection chambers comprise one or more reagents.

101. The analysis cartridge of claim 100, wherein the one or more reagents are dried reagents.

102. The analysis cartridge of claim 101, wherein the dried reagents are in a lyophilized bead.

103. A method of analyzing a sample in an analysis cartridge, the method comprising: conducting a first centrifugation of the analysis cartridge to move a diluent from a diluent chamber to a f luidica lly connected diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first threshold, conducting a second centrifugation of the analysis cartridge to move into a mixing chamberthe diluent from the diluent metering chamberand the sample from a sample metering chamber; wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold, conducting a third centrifugation of the analysis cartridge to move the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, andconducting a fourth centrifugation to move into a one or more detection chambers the diluted sample from the one or more metering fingers, wherein the fourth centrifugation exerts on the diluted sample in the one or more metering fingers a fourth centrifugal force above a third threshold.

104. The method of claim 103, wherein the fourth threshold is higher than the third threshold, the third threshold is higher than the second threshold, and the second threshold is higher than the first threshold.

105. The method of claim 103 or 104, wherein the first centrifugation of the analysis cartridge also moves the sample from a sample chamber into the sample metering chamber.

106. The method of any one of claims 103 to 105, wherein the sample metering chamber is fluidically connected to a cellular components chamber and wherein the first centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components chamber.

107. The method of any one of claims 103 to 105, wherein the sample metering chamber is fluidically connected to a sample chamber, and the method comprises conducting a fifth centrifugation of the analysis cartridge to move the sample from the sample chamber into the sample metering chamber, and wherein the fifth centrifugation exerts on the sample in the sample chamber a fifth centrifugation force above a fifth threshold.

108. The method of claim 107, wherein the sample metering chamber is fluidically connected to a cellular components chamber and the fifth centrifugation of the analysis cartridge moves the cellular components of the sample into the cellular components chamber.

109. The method of claim 107 or 108, wherein the fifth threshold is lower than the first and / or the second threshold.

110. The method of any one of claims 103 to 109, wherein the sample metering chamber further comprises a sample overflow conduit fluidically connected to a sample overflowchamberthat receives a portion of the sample that does not fit in the sample metering chamber, wherein the sample overflow chamber and / or the sample overflow conduit comprises a first set of one or more electrochemical sensors that detect a first set of one or more analytes in the sample, and wherein conducting the first centrifugation or the fifth centrifugation of the analysis cartridge comprises detecting the first set of one or more analytes in the sample in the first set of one or more electrochemical sensors.

111. The method of claim 110, comprising detecting in the one or more detection chambers one or more signals that indicate the concentration of one or more analytes in the sample and determining the concentrations of the one or more analytes in the sample.

112. The method of any one of claims 103 to 111, wherein conducting the first centrifugation or the fifth centrifugation of the analysis cartridge comprises receiving into a sample overflow chamber a portion of the sample that does not fit in the sample metering chamber.

113. The method of any one of claims 103 to 112, wherein conducting the first centrifugation or the fifth centrifugation comprises receiving the cellular components of the sample into the cellular components chamber that is fluidically connected to the sample metering chamber.

114. The method of any one of claims 103 to 113, wherein the sample chamber is further fluidically connected to an electrochemical analysis module comprising a second set of one or more electrochemical sensors, and wherein conducting a sixth centrifugation of the analysis cartridge moves a portion of the sample from the sample chamber into the electrochemical analysis module.

115. The method of claim 114, further comprising detecting a second set of one or more analytes in the sample using the second set of one or more electrochemical sensors.

116. The method of any one of claims 103 to 115, wherein the diluent valve is a diluent si hon, the sample valve is a sample siphon, and / or the diluted sample valve is a diluted sample siphon.

117. The method of claim 116, comprising opening by priming the diluent siphon, the sample siphon, and / or the diluted sample siphon.

118. The method of claim 116 or 117, comprising priming the diluent siphon and the sample siphon by centrifuging the analysis cartridge to exert on the diluent in the diluent metering chamber and the sample in the sample metering chamber the second centrifugal force above the second threshold.

119. The method of any one of claims 116 to 118, comprising priming the diluted sample siphon by centrifuging the analysis cartridge to exert on the diluted sample in the mixing chamber the third centrifugal force above the third threshold.

120. The method of any one of claims 103 to 119, further comprising detecting in the one or more detection chambers one or more signals that indicate the concentration of one or more analytes in the sample and determining the concentrations of the one or more analytes in the sample.

121. An analyzer for analyzing a sample in an analysis cartridges, the analyzer comprising: a rotor comprising a slot that holds the analysis cartridge such that, when the rotor rotates, the analysis cartridge is centrifuged around a vertical axis, a processor, and a non-transitory computer readable medium comprising instructions, which when executed by the processor, cause the analyzer to perform: a first centrifugation of the analysis cartridge that moves a diluent from the diluent chamber into the diluent metering chamber, wherein the first centrifugation exerts on the diluent within the diluent chamber a first centrifugal force above a first thresholdwhen a diluent valve and a sample valve are open, a second centrifugation of the analysis cartridge that moves the diluent from the diluent metering chamber and a sample from a sample metering chamber into the mixing chamber, wherein the second centrifugation exerts on the diluent in the diluent metering chamber and the sample in the sample metering chamber a second centrifugal force above a second threshold, when a diluted sample valve is open, a third centrifugation of the analysis cartridge that moves the diluted sample from the mixing chamber into the one or more metering fingers via a distribution channel, wherein the third centrifugation exerts on the diluted sample in the mixing chamber a third centrifugal force above a third threshold, and a fourth centrifugation of the analysis cartridge that moves the diluted sample from the one or more metering fingers into the corresponding one or more detection chambers, wherein the fourth centrifugation exerts on the diluted sample in the metering fingers a fourth centrifugal force above a fourth threshold.

122. The analyzer of claim 121, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform the first centrifugation followed by the second centrifugation, which is followed by the third centrifugation, which is followed by the fourth centrifugation.

123. The analyzer of claim 121 or 122, wherein the fourth threshold is higher than the third threshold, the third threshold is higher than the second threshold, and the second threshold is higher than the first threshold.

124. The analyzer of any one of claims 121 to 123, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform a fifth centrifugation of the analysis cartridge before conducting the first centrifugation of the analysis cartridge, wherein the fifth centrifugation moves the sample from a sample chamber into the sample metering chamber, and wherein the fifth centrifugation exerts on the sample in the sample chamber a fifth centrifugation force above a fifth threshold.Ill125. The analyzer of claim 124, wherein the fifth threshold is lower than the second threshold.

126. The analyzer of claim 124 or 125, wherein the non-transitory computer readable medium comprises instructions, which when executed by the processor, cause the analyzer to perform the fifth centrifugation followed by the first centrifugation, which is followed by the second centrifugation, which is followed by the third centrifugation, which is followed by the fourth centrifugation.

127. The analyzer of any one of claims 121 to 126, further comprising a detector that detects one or more signals from the one or more detection chambers, the one or more signals indicate the concentration of one or more analytes in the sample.

128. The analyzer of claim 127, wherein the detector is an optical detectorthat detects one or more optical signals from the one or more detection chambers.

129. The analyzer of any one of claims 121 to 128, wherein the analyzer further comprises a first set of one or more electrochemical detectors that detect a first set of one or more electrochemical signals from a first set of one or more electrochemical sensors located in a sample overflow chamber and / or the sample overflow conduit of the analysis cartridge.

130. The analyzer of claim 129, wherein the first set of one or more electrochemical detectors comprises an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof.

131. The analyzer of any one of claims 121 to 130, wherein the analyzer further comprises a second set of one or more electrochemical detectors that detect a second set of one or more electrochemical signals from a second set of one or more electrochemical sensors located in an electrochemical analysis module of the analysis cartridge.

132. The analyzer of claim 131, wherein the second set of one or more electrochemical detectors comprises an amperometric detector, a coulometric detector, a potentiometric detector, a voltametric detector, an impedance detector, or a combination thereof.

133. The analyzer of any one of claims 129 to 132, wherein the first set of one or more electrochemical detectors and / or the second set of one or more electrochemical are parts of an electrical detection unit comprising electrical circuits that detect electrical signals.

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