Cartridges for multiplex analysis of analytes and methods of using them

Abstract

Aspects of the present disclosure provide an analysis cartridge that allows analyzing multiple chemical components of a sample, such as blood. In certain embodiments, the analysis cartridge comprises a chemical analysis module for analyzing multiple analytes. A chemical analysis module comprises a top panel and a bottom panel and a plurality of channels disposed between the top panel and the bottom panel, wherein one or more channels of the plurality of channels comprise one or more reagents that produce, in each such channel, a detectable signal indicative of the concentration of an analyte. The chemical analysis module can have a sample port that delivers a sample to the plurality of channels and one or more sensors that detect the signals generated in the channels. Also provided herein are methods of analyzing a sample, such as a plasma, serum, or blood sample, in the analysis cartridges provided herein.

Description

[0001] CARTRIDGES FOR MULTIPLEX ANALYSIS OF ANALYTES AND METHODS OF USING THEM

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 612,216, filed December 19, 2023, which application is incorporated herein by reference in its entirety

[0004] INTRODUCTION

[0005] Analyses of biological samples, such as blood samples, often involve testing multiple chemical components, for example, metabolites and proteins. The analysis of these biological samples is often limited by the availability of the samples. Therefore, multiplex analyses of biological samples, particularly for their chemical components, while using minute amounts of the sample is desirable.

[0006] SUMMARY

[0007] In certain aspects, this disclosure provides an analysis cartridge comprising a plurality of channels that allows multiplex analysis of chemical components in a sample, particularly, a biological sample, such as blood, serum, or plasma.

[0008] In certain aspects, this disclosure provides an analysis cartridge comprising a chemical analysis module for multiplex analysis of analytes in a sample. In some cases, the chemical analysis module comprises a top panel and a bottom panel and a plurality of channels disposed between the top panel and the bottom panel. In some cases, the top panel and the bottom panel have a plurality of vertical walls disposed between the top panel and the bottom panel thereby producing the plurality of channels. The plurality of channels can run from a proximal end to the distal end of the top panel and the bottom panel.

[0009] In some cases, one or more channels from the plurality of channels contain one or more reagents that produce, in each such channel, a detectable signal indicative of the concentration of an analyte. The chemical analysis module can have at a proximal end, a sample port that delivers a sample to the plurality of channels. The chemical analysis module can have one or more sensors that detect a detectable signal generated in one or more of the plurality of channels. In certain embodiments, the sample is a serum or plasma. Separation of plasma from blood can be performed using a plasma separation membrane or by digital microfluidics, for example, by using agglutination or magnetic beads to separate blood cells from plasma. In some cases, the sample can be whole blood.

[0010] In some cases, the analysis cartridge further comprises a sample delivery module configured to deliver a sample to a sample port thereby allowing the sample to fill the channels of the chemical analysis module.

[0011] In certain embodiments, the chemical analysis module comprises one or more channels within the plurality of channels, each configured to detect an analyte, such as, glucose, calcium, blood urea nitrogen (BUN), creatinine, sodium, potassium, chloride, carbon dioxide (CO2), serum total protein (TP), serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

[0012] In certain aspects, this disclosure also provides a chemical analysis module that allows multiplex analysis of chemical components in a sample, particularly, a biological sample, such as blood, serum, or plasma. In some cases, the chemical analysis module comprises a top panel and a bottom panel and a plurality of channels disposed between the top panel and the bottom panel. In some cases, the top panel and the bottom panel have a plurality of vertical walls disposed between the top panel and the bottom panel thereby producing the plurality of channels. The plurality of channels can run from a proximal end to the distal end of the top panel and the bottom panel.

[0013] Also provided herein are methods of analyzing a sample, such as a plasma, serum, or blood sample, in the analysis cartridges provided herein.

[0014] BRIEF DESCRIPTION OF THE FIGURES

[0015] FIG. 1 depicts an example of a chemical analysis module having multiple amperometric sensors.

[0016] FIG. 2 depicts an example of a chemical analysis module having a combination of amperometric sensors and optical sensors.

[0017] FIG. 3 depicts an example of a sample delivery module comprising a sample drain.

[0018] FIG. 4 depicts an example of a sample delivery module.

[0019] FIG. 5 depicts an example of a sample delivery module comprising a digital microfluidics (DMF) chip. DETAILED DESCRIPTION

[0020] This disclosure relates to the United States provisional patent application no. 63 / 605,046 filed on December 1 , 2023, the contents of which are herein incorporated by reference in their entirety.

[0021] Certain aspects of the present disclosure provide analysis cartridges that allow the analysis of chemical components of samples, such as blood samples. An analysis cartridge described herein can allow a comprehensive metabolic panel (CMP) analysis of a blood sample.

[0022] In some cases, a chemical analysis module comprises a top panel and a bottom panel and a plurality of channels disposed between the top panel and the bottom panel. The plurality of channels can run from a proximal end to a distal end of the top panel and the bottom panel.

[0023] The “proximal end” of the chemical analysis module is where the sample port is located. The “distal end” of the chemical analysis module is opposite to the proximal end. In some cases, in one or more channels of the plurality of channels, one or more sensors can be located at or near the distal end.

[0024] One or more of the plurality of channels may be open to air at the distal ends thereby allowing a sample to load into the plurality of channels, for example, via capillary forces. Moreover, the dimensions of the top panel, the bottom panel, the distance between the top panel and the bottom panel, and the dimensions of the channels within the plurality of channels are such that the surface tension of a sample does not allow the sample to spill over from the distal end, thus, retaining the sample within the channels.

[0025] In some cases, the top panel and the bottom panel have a plurality of vertical walls disposed between the top panel and the bottom panel thereby producing the plurality of channels. Alternatively, the top panel and / or the bottom panel have indents or grooves that engage with the other panel to form the plurality of channels. For example, the top panel and the bottom panel may have semicircular indents, which, when the top panel and the bottom panel are joined together, form channels having circular cross sections.

[0026] In some cases, indents or grooves can be only on the top panel or only on the bottom panel. In such embodiments, the non-indented or non-grooved panel can be substantially flat. A plurality of channels can then be formed when the top panel and the bottom panel are joined together with or without an adhesive. The channels thus formed would have the shape of the indents or grooves. Indents or grooves can be in any other suitable shapes so as to produce channels having rectangular, square, oval, circular, elliptical, irregular cross sections, or a combination thereof. Channels having any other suitable cross section could be used and such embodiments are within the purview of the disclosure.

[0027] In some cases, one or more channels within the plurality of channels contain one or more reagents that produce, in each such channel, a detectable signal indicative of the concentration of an analyte.

[0028] In some cases, one or more channels contain one or more reagents that produce, in each such channel, a detectable signal indicative of the concentration of an analyte, while one or more channels of the plurality of channels may have one or more reagents for detection of an analyte, except for a reagent that generates a detectable signal. Such channels can be used as references or blanks for the corresponding analytes.

[0029] The chemical analysis module can have at the proximal end a sample port that delivers a sample to the plurality of channels.

[0030] The chemical analysis module can have one or more sensors that detect the detectable signals generated in the one or more channels of the plurality of channels.

[0031] In some cases, the analysis cartridge further comprises a sample delivery module configured to deliver the sample to a sample port thereby allowing the sample to fill the plurality of channels of the chemical analysis module.

[0032] Also provided herein are methods of analyzing a sample, such as a plasma, serum, or blood sample, in the analysis cartridges provided herein.

[0033] Before the present devices and methods 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.

[0034] 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 devices and methods. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the devices 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 devices and methods. 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.

[0035] 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.

[0036] 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.

[0037] 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 or not. The singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.

[0038] The term “comprising” is used herein as requiring the 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 not change 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). 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.

[0039] For the recitation of numeric ranges herein, each intervening number there between 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.

[0040] All ranges disclosed herein are inclusive of the recited endpoint and independently combinable (for example, the range of “from 2 grams to 10 grams” is inclusive of the endpoints, 2 grams and 10 grams, 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.

[0041] 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 .

[0042] 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.

[0043] The terms “horizontal” and “vertical” are used to indicate direction relative to an absolute reference, i.e., 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.

[0044] 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.

[0045] 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.

[0046] It is noted that, as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. It is further noted that 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.

[0047] 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 devices and methods. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.

[0048] DEVICES

[0049] As summarized above, certain aspects of the present disclosure provide analysis cartridges for analyzing chemical components of biological samples, such as blood, plasma, or serum samples. Also, certain aspects of the disclosure provide analysis cartridges that allow analysis of small volumes of samples. A suitable 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 sample also includes extracts from soaking a swab in an appropriate buffer. For example, a nasal or throat swab could be soaked in a buffer, such as saline to prepare a sample that could be analyzed in the analysis cartridges disclosed herein. A sample can also be a lysate of a tissue produced in an appropriate buffer. Even additional 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.

[0050] Certain biological samples to be analyzed in the analysis cartridges disclosed herein include venous blood, capillary blood, serum, or plasma.

[0051] “Subject” as used herein refers to any vertebrate, 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. The subject or patient may be undergoing other forms of treatment.

[0052] In certain embodiments, the analysis cartridge comprises a chemical analysis module. In some cases, a chemical analysis module allows multiplex analysis of analytes in a sample.

[0053] In certain embodiments, the chemical analysis module comprises a top panel and a bottom panel and a plurality of channels disposed between the top panel and the bottom panel. The plurality of channels can run from a proximal end to a distal end of the top panel and the bottom panel.

[0054] In some cases, the top panel and the bottom panel have a plurality of vertical walls disposed between the top panel and the bottom panel thereby producing the plurality of channels. Alternatively, the top panel and / or the bottom panel have indents or grooves that engage with other to form a plurality of channels. For example, the top panel and the bottom panel may have semicircular indents or grooves, which, when the top panel and the bottom panel are joined together form channels having circular cross sections. Indents or grooves can be in any other suitable shapes so as to produce channels having rectangular, square, oval, circular, or elliptical cross sections. Channels having any suitable cross section could be used and such embodiments are within the purview of the disclosure. In some cases, indents or grooves can be only on the top panel or only on the bottom panel. In such embodiments, the non-indented or non-grooved panel can be substantially flat. A plurality of channels can then be formed when the top panel and the bottom panel are joined together with or without an adhesive. The channels thus formed would have the shape of the indents or grooves.

[0055] The top panel and the bottom panel can be made of any suitable material, for example, polyethylene terephthalate (PET). Alternatively, the top and / or the bottom panel can comprise laser ablated gold or platinum electrodes on a PET sheet. The top panel and the bottom panel can be made from a flexible or rigid plastic. The top panel and the bottom panel may be, independent of each other, transparent, translucent, or opaque. In some cases, the top panel has a different level of transparency than the bottom panel.

[0056] In some cases, the plurality of vertical walls disposed between the top panel and the bottom panel are made from an adhesive. For example, lines of an adhesive in an appropriate pattern can be deposited onto the top panel and / or the bottom panel. The top panel and the bottom panel can then be contacted with each other, for example, pressed on each other, so that the adhesive lines facilitate joining of the top panel and the bottom panel thereby also forming channels comprising the top panel and the bottom panel and the vertical walls of the adhesive.

[0057] In some cases, the adhesive deposited on the top panel and / or the bottom panel is a pressure adhesive. Certain examples of such adhesives include acrylate adhesives or silicone adhesives. In some cases, the adhesive is a medical grade adhesive.

[0058] In some cases, roll to roll manufacturing is used to contact the top panel and the bottom panel comprising the adhesive lines to produce chemical analysis modules comprising a plurality of channels.

[0059] In some cases, the top panel comprises pillars extending down into the channels. Such pillars facilitate filling of the channels with a sample as the pillars induce capillary action. For example, a sample introduced in a sample port can load into the plurality of channels of a chemical analysis module via capillary or wicking force, which can be facilitated by the pillars extending down into from the top panel into the plurality of channels.

[0060] In some cases, the top panel and the bottom panel of a chemical analysis module are separated by a distance between 10 pm and 100 pm, such as 10 pm, 15 pm, 20 pm, 15 pm, 30 pm, 35 pm, 40 pm, 45 pm, 50 pm, 55 pm, 60 pm, 65 pm, 70 pm, 75 pm, 80 pm, 85 pm, 90 pm, 95 pm, or 100 pm. In some cases, one or more channels of the plurality of channels contain one or more reagents that produce, in each such channel, a detectable signal indicative of a concentration of an analyte. In some cases, one or more channels have one or more reagents for detection of an analyte, except for a reagent that generates a detectable signal. Such channels can be used as references or blanks for the corresponding analytes.

[0061] The chemical analysis module can comprise, at the proximal end, a sample port that delivers the sample to the plurality of channels.

[0062] In some cases, the chemical analysis module can have one or more sensors that detect a detectable signal generated in one or more of the plurality of channels.

[0063] In some cases, a sensor is located within or adjacent to a channel within the plurality of channels and detects a detectable signal generated in the channel. A sensor can also be located at any suitable position within a channel, for example, in the middle of the channel or at a distal end of a channel.

[0064] The term “located within the channel” indicates that a sensor is fully or partially enclosed within a channel, for example, at any suitable position, such as the middle or the distal end of a channel.

[0065] The term “located adjacent to a channel” indicates that a sensor is at a distance from a channel but is capable of detecting a signal generated in the channel.

[0066] A sensor that is located within or adjacent to a channel can detect an electrochemical signal, such as current or charge generated in the channel. For example, an amperometric sensor can detect a current generated in a channel. Similarly, a potentiometric sensor can detect a charge generated in a channel.

[0067] A sensor that is located within or adjacent to a channel can also detect a chemical signal, such as oxygen generated in a channel. A coulometric sensor can detect oxygen generated in a channel.

[0068] In some cases, a sensor is directed at a channel and detects a detectable signal generated in the channel.

[0069] The term “a sensor directed at a channel” refers to a sensor that may or may not be physically within or adjacent to the channel but is in an appropriate position to detect a detectable signal generated in the channel. For example, an optical sensor can be at an appropriate position to detect an optical signal generated in a channel. Accordingly, in some cases, a chemical analysis module comprises one or more optical sensors that are directed at one or more channels to detect optical signals generated in the channels. In one or more channels within a plurality of channels of a chemical analysis module, depending on the generated signal, appropriate sensors detect the generated signals. For example, to detect a current generated in a channel, a channel can contain an amperometric sensor. To detect oxygen, a channel can contain a coulometric sensor. To detect a charge, a sensor can contain a potentiometric sensor. To detect an optical signal, an optical sensor can be directed at a channel. An optical sensor can be a CCD (charge coupled device) sensor or a CMOS (complementary metal oxide semiconductor) sensor.

[0070] A chemical analysis module can have only amperometric sensors, only potentiometric sensors, only coulometric sensors, only optical sensors, or a combination thereof.

[0071] For example, detectable signals generated in a first set of one or more channels of the plurality of channels can be electrochemical signals and the corresponding sensors can be amperometric or potentiometric sensors, the detectable signals generated in a second set of one or more channels of the plurality of channels can be optical signals and the corresponding sensors can be optical sensors, and the detectable signals generated in a third set of one or more channels of the plurality of channels can be oxygen and the corresponding sensors can be coulometric sensors. Thus, in a plurality of channels, a combination of amperometric sensors, potentiometric sensors, coulometric sensors, and optical sensors can be employed to detect the signals generated in the corresponding channels.

[0072] The electrochemical signals generated in two or more channels of the plurality of channels can be different from each other, for example, different ions as detectable electrochemical signals or different chemical as detectable signals. Accordingly, different sensors can be implemented in different channels of the plurality of channels.

[0073] Similarly, optical signals generated in two or more channels of the plurality of channels can be different from each other, for example, different wavelengths of detectable optical signals. Accordingly, different optical sensors can be implemented in detecting signals from different channels of the plurality of channels.

[0074] Additional detectable signals that can be generated to determine a concentration of a chemical as well as corresponding sensors are known in the art and such embodiments are within the purview of the disclosure. In certain exemplary embodiments, an analysis cartridge comprises two or more channels of the plurality of channels comprising the following combinations of sensors: one or more electrochemical sensors and one or more optical sensors; one or more electrochemical sensors and one or more chemical sensors; one or more optical sensors and one or more chemical sensors; one or more amperometric sensors and one or more potentiometric sensors; one or more amperometric sensors and one or more coulometric sensors; one or more amperometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more coulometric sensors.

[0075] In some cases, the width of a channel is between 0.1 mm to 1 mm, such as 0.1 mm, 0.2 mm, 0.3 mm, 0.4 mm, 0.5 mm, 0.6 mm, 0.7 mm, 0.8 mm, 0.9 mm, or 1 mm. The width of different channels of the plurality of channels within a chemical analysis module can be different from each other.

[0076] In certain embodiments, the length of a channel is between 5 mm and 100 mm, such as, 5 mm, 10 mm, 15 mm, 20 mm, 25 mm, 30 mm, 35 mm, 40 mm, 45 mm, 50 mm, 55 mm, 60 mm, 65 mm, 70 mm, 75 mm, 80 mm, 85 mm, 90 mm, 95 mm, or 100 mm. The length of different channels of the plurality of channels within a chemical analysis module can be different from each other.

[0077] Depending upon the distance between the top panel and the bottom panel, the width of a channel, and the length of a channel, the volume of a channel can be between 5 nl to 10 pl. In certain embodiments, the distance between the top panel and the bottom panel, the width of a channel, and the length of a channel are selected such that the volume of a channel is between 0.01 pl and 1 pl, such as 0.01 pl, 0.05 pl, 0.1 pl, 0.2 pl, 0.3 pl, 0.4 pl, 0.5 pl, 0.6 pl, 0.7 pl, 0.8 pl, 0.9 pl, or 1 .0 pl.

[0078] The reagents provided in a channel within the plurality of channels are designed to detect an analyte and provide a detectable signal indicating the concentration of the analyte. The analytes detected in different channels of the plurality of channels can be glucose, calcium, BUN, creatinine, sodium, potassium, chloride, CO2, TP, serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

[0079] Various methods are known in the art to detect each of the abovementioned analytes. Various reagents used to detect each of the abovementioned analytes are also known in the art. Accordingly, channels containing specific reagents designed to detect one or more of the abovementioned analytes are within the purview of the disclosure. Moreover, reagents and methods of detecting additional analytes are known in the art and devices containing such reagents for detecting such additional analytes are within the purview of the disclosure.

[0080] In some cases, a channel in a plurality of channels is configured to analyze an analyte selected from: glucose, calcium, BUN, creatinine, sodium, potassium, chloride, CO2, TP, serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

[0081] In certain embodiments, two or more channels in a plurality of channels are configured to analyze two or more analytes selected from: glucose, calcium, BUN, creatinine, sodium, potassium, chloride, CO2, TP, serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

[0082] For example, a chemical analysis module can have 17 channels, each configured to analyze one analyte selected from: glucose, calcium, BUN, creatinine, sodium, potassium, chloride, CO2, TP, serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase. Thus, a chemical analysis module can analyze all of the following analytes: glucose, calcium, BUN, creatinine, sodium, potassium, chloride, CO2, TP, serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase. Within the plurality of channels, additional control / reference / blank channels may be included beyond the 17 channels.

[0083] In some cases, the plurality of channels of the chemical analysis module are designed to provide a CMP of a blood sample. In such cases, the chemical analysis module can have a plurality of channels configured to analyze the combination of the following analytes: glucose, calcium, sodium, potassium, carbon dioxide, chloride, albumin, TP, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, BUN, and creatinine.

[0084] In some cases, the cartridge comprises a module for separating a plasma from blood cells of a blood sample. The separated plasma can then be delivered to the sample port.

[0085] A plasma separation module can comprise a filter for separating the plasma from blood cells in the blood sample. A plasma separation module can also comprise a DMF chip for separating the plasma from blood cells in the blood sample. Typically, a DMF chip comprises a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets comprising the blood and one or more reagents to transport the droplets between the first and the second substrates. The one or more reagents in a DMF for separating plasma from blood can comprise one or more agglutination agents or magnetic beads. The agglutination agents or magnetic beads can adhere to blood cells, which can be selectively moved in through the DMF electrodes away from the liquid portion of the blood, i.e., plasma.

[0086] An exemplary DMF based plasma separation microfluidic device is described in Dixon et al. (2020), Lab on a Chip, 20,1845, which is incorporated by reference in its entirety.

[0087] An exemplary embodiment of an analysis cartridge comprising a chemical analysis module is provided in FIGS. 1 -2.

[0088] An exemplary embodiment shown in FIG. 1 comprises the analysis cartridge 100 comprising the chemical analysis module 101 , which comprises at the proximal end, the sample port 103. The sample port 103 can receive the sample from a sample delivery module. The sample can be a plasma, serum, or blood. Any other suitable sample can also be loaded into the chemical analysis module and such embodiments are within the purview of the disclosure.

[0089] The chemical analysis module 101 can comprise a top panel and a bottom panel, which are not apparent in the perspective view shown in FIG. 1. The top panel can be transparent, opaque, or translucent. In FIG. 1 , the top panel is transparent, opaque, or translucent but not shown as such to make visible the underlying channels in the chemical analysis module. The bottom panel is below the channels and can be transparent, opaque, or translucent.

[0090] The different channels 102 are formed by the top panel, the bottom panel, and the vertical walls 105. The channels 102 are open to air at the distal ends thereby facilitating loading of the channels via capillary force.

[0091] At the distal end of the chemical analysis module are a plurality of amperometric sensors 104. In this embodiment, all the channels comprise an amperometric sensor.

[0092] Another exemplary embodiment of an analysis cartridge is shown in FIG. 2. In this figure, the analysis cartridge 200 comprises the chemical analysis module 201 , which comprises at a proximal end, the sample port 203. A sample delivery module (not shown) can deliver a sample to the sample port 203. The sample can be a plasma, serum, or blood. Any other suitable sample can also be loaded into the chemical analysis module and such embodiments are within the purview of the invention.

[0093] The chemical analysis module 201 can comprise a top panel and a bottom panel, which are not apparent in the perspective view shown in FIG. 2. The top panel can be transparent, opaque, or translucent. In FIG. 2, the top panel is transparent, opaque, or translucent but not shown as such to make visible the underlying channels in the chemical analysis module. The bottom panel is below the channels and can also be transparent, opaque, or translucent. The top panel may have a different level of transparency than the bottom panel.

[0094] The plurality of channels 202 are formed by the top panel, the bottom panel, and the vertical walls 206. The distal end of the channels 202 are open to air thereby facilitating loading of the channels via capillary force.

[0095] The Channels 204 are designed to produce electrochemical current signals. These electrochemical current signals are detected by amperometric sensors 207. While FIG. 2 shows together numerous channels 204 generating electrochemical current signals and having amperometric sensors 207, such channels and the sensors can be present in any one or more of the channels.

[0096] In FIG. 2, the channels 205 are designed to produce optical signals indicated by the ovals 208. One of more optical sensors (not shown) can be directed towards channels 205 to detect the optical signals 208.

[0097] Thus, in the embodiment exemplified in FIG. 2, some of the channels generate electrochemical signals and some of the channels generate optical signals. The corresponding sensors are accordingly placed or configured.

[0098] In some cases, a sample delivery module is configured to deliver a sample to the sample port. In some cases, between 100 nl and 200 pl of a sample is delivered. In certain embodiments, the sample delivery module is configured to deliver to the sample port a sample in a volume between 2 pl and 50 pl, such as 2 pl, 5 pl, 10 pl, 15 pl, 20 pl, 25 pl, 30 pl, 35 pl, 40 pl, 45 pl, 50 pl, 55 pl, 60 pl, 65 pl, 70 pl, 75 pl, 80 pl, 85 pl, 90 pl, or 100 pl. A sample delivered to the sample port is loaded in different channels. Depending on the geometry of different channels, different volumes of a sample can be loaded in different channels. In some cases, a sample delivery module comprises a sample chamber, a sample drain, and a sample deposition member.

[0099] In some cases, a sample deposition member can deposit a sample into a sample chamber. Any suitable fluid movement mechanism can be used to deposit a sample into a sample chamber. For example, appropriate positive or negative pressure can be applied to a sample from other parts of the analysis cartridge into a sample chamber. For example, a sample may be moved into a sample chamber via diffusion, convection, pumping, applied pressure, gravity-driven flow, density gradients, temperature gradients, chemical gradients, pressure gradients (positive or negative), pneumatic pressure, gasproducing chemical reactions, centrifugal flow, capillary pressure, wicking, electric field- mediated, electrode-mediated, electrophoresis, dielectrophoresis, magnetophoresis, magnetic fields, magnetically driven flow, optical force, chemotaxis, phototaxis, surface tension gradient driven flow, Marangoni stresses, hermos-capillary convection, surface energy gradients, acoustophoresis, surface acoustic waves, electroosmotic flow, thermophoresis, electrowetting, opto-electrowetting, a pipette, a peristaltic pump, syringe pump, a pressure-drive flow control pump, or the like.

[0100] A sample chamber is fluidically connected to the sample port and a sample deposited into the sample chamber is delivered to the sample port. At the sample port, the delivered sample loads into the channels of the chemical analysis module and fills the channels.

[0101] In some cases, a sample chamber is also fluidically connected to a sample drain. When a sample reaches the sample port, the sample fills the channels of the chemical analysis module. Capillary action can facilitate such filling of the channels. In some cases, when the chemical analysis module are substantially filled with the sample, the back-pressure from the sample avoids further movement of the sample into the channels. The excess fluid then drains from the sample chamber into the fluidically connected sample drains.

[0102] Various parameters of the sample drain can be adjusted. For example, the depth of the sample drain can be between 0.1 mm and 1 mm, such as 0.1 , 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1 .0 mm. Also, the length of the sample drain can be between 10 mm and 100 mm, such as 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 100 mm. The width of the sample drain can be between 0.5 and 1.5 mm, such as 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1.0, 1 .05, 1.1 , 1 .15, 1.2, 1.25, 1.3, 1 .35, 1 .4, 1 .45, or 1 .5 mm. FIG. 3 shows an embodiment of an analysis cartridge comprising a sample delivery module that delivers a sample to the sample port and comprises a sample drain. The analysis cartridge 300 comprises the chemical analysis module 301 and the sample delivery module 303.

[0103] The sample delivery module 303 comprises a sample chamber 307. A sample deposition member (not shown) delivers a sample into the sample chamber 307, which is fluidically connected to the sample port 302 and, thus, delivers the sample to the sample port 302. The sample delivers to the sample port can load into the channels 306 via capillary action. At the distal end of the chemical analysis module, the channels have sensors 305 that detect the detectable signals generated in the corresponding channels. While the sensors in FIG. 3 are shown as amperometric sensors, any other sensors or combination of sensors can be used.

[0104] The sample chamber 307 is fluidically connected to the sample drain 304. As shown in FIG. 3, the sample drain 304 comprises two channels, each fluidically connected to the sample chamber.

[0105] In some cases, a sample delivery module comprises a sample chamber fluidically connected to the sample port of the chemical analysis module. The fluidic connection with a sample port can be used to deliver a sample to the chemical analysis module. An exemplary analysis cartridge for such embodiments is provided in FIG. 4. In FIG. 4, the sample chamber 403 can be filled with a sample using a sample deposition member (not shown).

[0106] The chemical analysis module 401 comprises the plurality of channels 406 and sensors 407. The plurality of channels 406 can be open to air. A sample once deposited into the sample chamber 403 in a sufficient amount would deliver the sample into the sample port 404. The sample can then enter the channels of the chemical analysis module, for example, via capillary force. As the sample fills the chemical analysis module it pushes out air until it completely fills the channels. The dimensions of the chemical analysis module / channels are designed so that the surface tension of the sample at the edge of the open channels does not allow spilling of the sample from the distal ends of the channels. The surface tension also provides sufficient back pressure such that no more sample enters the channels.

[0107] In some cases, a sample delivery module comprises a DMF chip that delivers the sample to the sample port 404. As well-known in the art, a DMF chip comprises: a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on sample droplets between the first and the second substrates to transport the sample droplets.

[0108] In certain embodiments, a DMF chip includes a first substrate and a second substrate, where the second substrate is positioned over the first substrate and separated from the first substrate by a gap. The first or the second substrate may include a plurality of DMF electrodes. The plurality of DMF electrodes may be an array or a series of electrodes that are individually controllable for activation and deactivation. The plurality of DMF electrodes may be overlayed with an insulating material to electrically isolate the DMF electrodes. In certain embodiments, the space / gap between the first and second substrates may be filled with air or with an inert fluid, such as oil. In exemplary embodiments, a series of DMF electrodes may be disposed on the first substrate and a single electrode disposed on the second substrate in a facing configuration with the series of electrodes on the first substrate. The series of electrodes and the single electrode may be covered with an insulating layer. In other cases, the series or plurality of electrodes on the first substrate may be configured as co-planar electrodes and the second substrate may not include an electrode. Various configurations of DMF electrodes are known in the art and are described, for example, in United States Patent No. 1 1 ,016,053, which is incorporated herein in its entirety. Any of these configurations of DMF electrodes can be present in the analysis cartridges disclosed here.

[0109] FIG. 5 provides an exemplary analysis cartridge 500 comprising the DMF chip 501 for moving the sample into the chemical analysis module 504. The sample may be provided to the DMF chip from sample chamber 502. The DMF chip moves the sample from the sample chamber to the sample port 505 of the chemical analysis module 404. The optional wicking pads 503 may absorb excess sample that does not enter the chemical analysis module 504.

[0110] In some cases, the DMF chip comprises one or more dried reagents that facilitates movement of a sample over the DMF electrodes. The DMF electrodes can be coated with a surfactant, namely, ethylenediamine tetrakis(ethoxylate-block-propoxylate) tetrol (90r4 or Tetronic 904™). Tetronic 904™ facilitates better movement of the droplets compared to electrodes without it. Any suitable surfactant can be used to facilitate such fluid movement. In some cases, the electrodes comprise EDTA, which avoids coagulation of a blood sample when manipulated in the DMF chip.

[0111] Chemical analysis module Certain aspects of the disclosure provide a chemical analysis module for multiplex analysis of analytes in a sample.

[0112] In some cases, the chemical analysis module comprises: a top panel and a bottom panel and a plurality of channels disposed between the top panel and the bottom panel, wherein one or more channels of the plurality of channels contain one or more reagents that produce, in each such channel, a detectable signal indicative of a concentration of an analyte, a sample port that delivers the sample to the plurality of channels, and one or more sensors that detect the detectable signals generated in the one or more channels of the plurality of channels.

[0113] Certain details of the chemical analysis modules described elsewhere in this disclosure in relation to the analysis cartridges of the disclosure also apply to the chemical analysis modules disclosed herein. Such details are incorporated herein by reference in their entirety.

[0114] The plurality of channels can run from a proximal end to a distal end of the top panel and the bottom panel.

[0115] The plurality of channels can be formed using the top panel and / or the bottom panel having indents or grooves that form the plurality of channels. The channels of the plurality of channels can be rectangular, square, oval, circular, elliptical, or irregular in the cross section.

[0116] In some cases, the plurality of channels are formed via a plurality of vertical walls disposed between the top panel and the bottom panel thereby producing the plurality of channels. In some cases, the plurality of vertical walls disposed between the top panel and the bottom panel comprise an adhesive.

[0117] In certain cases, in the chemical analysis module a sensor of the one or more sensors is located within or adjacent to a channel and detects the detectable signal generated in the channel. For example, the sensor can be located in the middle of the channel or at the distal end of the channel.

[0118] In some cases, the sensor can be an amperometric sensor that detects a current generated in the channel. The sensor can also be a potentiometric sensor that detects a charge generated in the channel. The sensor is a coulometric sensor that detects oxygen generated in the channel.

[0119] In some cases, a sensor from the one or more sensors is directed at a channel and detects the detectable signal generated in the channel. Such sensor can be an optical sensor that detects an optical signal generated in the channel. An optical sensor can be a CCD sensor or a CMOS sensor.

[0120] In some cases, within the plurality of channels of the chemical analysis module, the detectable signals generated in a first set of one or more channels are electrochemical signals and the corresponding sensors are amperometric and / or potentiometric sensors. Similarly, within the plurality of channels of the chemical analysis module, the detectable signals generated in a second set of one or more channels are optical signals and the corresponding sensors are optical sensors. Further, within the plurality of channels of the chemical analysis module, the detectable signals generated in a third set of one or more channels are oxygen and the corresponding sensors are coulometric sensors.

[0121] In some cases, two or more channels of the plurality of channels comprise the following combinations of sensors: one or more electrochemical sensors and one or more optical sensors; one or more electrochemical sensors and one or more chemical sensors; one or more optical sensors and one or more chemical sensors; one or more amperometric sensors and one or more potentiometric sensors; one or more amperometric sensors and one or more coulometric sensors; one or more amperometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more coulometric sensors

[0122] For example, the detectable signals generated in the first set of one or more channels of the plurality of channels are electrochemical signals and the corresponding sensors are amperometric and / or potentiometric sensors; the detectable signals generated in the second set of one or more channels of the plurality of channels are optical signals and the corresponding sensors are optical sensors; and the detectable signals generated in the third set of one or more channels of the plurality of channels are oxygen and the corresponding sensors are coulometric sensors.

[0123] In some cases, the top panel comprises pillars extending down into the channels. Such pillars would facilitate loading of a sample into the channels via capillary action.

[0124] In some cases, the top panel and the bottom panel are separated by a distance between 10 pm and 100 pm. The longest dimension in the cross section of a channel can be between 0.1 mm and 1 mm. Also, the length of a channel can be between 5 mm and 100 mm. In some cases, the volume of one or more channels within the plurality of channels is between 0.01 pl and 1 pl.

[0125] In some cases, a channel within the plurality of channels of the chemical analysis module disclosed herein is configured to detect an analyte selected from: glucose, calcium, blood urea nitrogen (BUN), creatinine, sodium, potassium, chloride, carbon dioxide (CO2), serum total protein (TP), serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

[0126] In some cases, the plurality of channels are configured to detect two or more analytes selected from: glucose, calcium, BUN, creatinine, sodium, potassium, chloride, carbon dioxide CO2, TP, serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

[0127] In even further cases, the plurality of channels are configured to detect two or more analytes selected from: glucose, calcium, sodium, potassium, carbon dioxide, chloride, albumin, TP, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, BUN, and creatinine.

[0128] Certain aspects of the disclosure provide an analysis cartridge comprising a chemical analysis module disclosed herein.

[0129] In addition to the chemical analysis module, the analysis cartridge can further a plasma separation module for separating a plasma from blood cells in a blood sample.

[0130] The plasma separation module can comprise a filter for separating the plasma from blood cells in the blood sample.

[0131] The plasma separation module can also comprise a first digital microfluidic (DMF) chip for separating the plasma from blood cells in the blood sample, the first DMF chip comprising a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets comprising the blood and one or more reagents to transport the droplets between the first and the second substrates.

[0132] In some cases, un addition to the chemical analysis module, the analysis cartridge further comprises a sample delivery module that delivers the sample to a sample port that delivers the sample to the chemical analysis module.

[0133] In some cases, the sample delivery module comprises a sample chamber, a sample deposition member that delivers the sample into the sample chamber, and a sample drain fluidically connected to the sample chamber, wherein the sample drain receives excess sample beyond the sample filled into the chemical analysis module. The sample drain comprises two or more channels fluidically connected to the sample chamber.

[0134] The sample delivery module can also comprises a DMF chip that delivers the sample to the sample port, wherein the DMF chip comprises: a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets of the sample between the first and the second substrates to transport the droplets.

[0135] In some cases, the sample delivery module comprises a sample chamber and a sample deposition member that deposits the sample into the sample chamber.

[0136] METHODS

[0137] The analysis cartridges or the chemical analysis modules disclosed herein can be used for analyzing a sample, for example, a blood, serum, or plasma sample for its chemical components.

[0138] Any suitable sample analyzed in the methods 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. In some cases, the methods comprise providing a CMP of a subject’s blood. Additional samples that could be analyzed in the methods 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.

[0139] Any structural elements of the analysis cartridges or the chemical analysis modules described elsewhere in this disclosure, for example, those described under “Devices” above, are applicable to the methods disclosed herein. For example, the structure of the chemical analysis module as discussed above is also applicable to the methods disclosed herein.

[0140] Certain aspects of the disclosure provide a method of analyzing a sample, for example, a blood, plasma, or serum sample, in the analysis cartridges disclosed herein. In some cases, the methods comprise delivering to a chemical analysis module a sample in a small volume, for example, a sample between 100 nl to 200 pl. In certain embodiments, the methods comprise delivering to the sample port a sample in a volume between 2 pl and 50 pl, such as 2 pl, 5 pl, 10 pl, 15 pl, 20 pl, 25 pl, 30 pl, 35 pl, 40 pl, 45 pl, 50 pl, 55 pl, 60 pl, 65 pl, 70 pl, 75 pl, 80 pl, 85 pl, 90 pl, or 100 pl. The sample, such as a blood, serum, or plasma sample loaded into the chemical analysis module can be analyzed to obtain, store, and process concentrations of one or more analytes in the sample. Such information can be used to determine chemical composition of the sample. For example, depending on the detection of certain types and concentration of chemicals detected in the chemical analysis module, the methods comprise providing CMP values for the blood sample.

[0141] In some cases, the methods comprise using a DMF chip to load a sample into the chemical analysis module of an analysis cartridge.

[0142] In some cases, the methods comprising using a sample chamber and a sample deposition member to load a sample into the chemical analysis module of an analysis cartridge. In some cases, using a sample chamber and a sample deposition member to load a sample into the chemical analysis module of an analysis cartridge further comprises using a sample drain to receive excess sample beyond the sample filled into the chemical analysis module.

[0143] Thus, in certain cases, the disclosure provides a method of analyzing a sample in an analysis cartridge that comprises a sample delivery module comprising a sample chamber, a sample deposition member that delivers the sample into the sample chamber, and a sample drain fluidically connected to the sample chamber, wherein excess sample beyond the sample filled into the chemical analysis module is received in a sample drain fluidically connected to the sample chamber. In some cases, the excess sample beyond the sample filled into the chemical analysis module is received in two or more channels of the sample drain, such as, one, two, three, four, five, or six channels fluidically connected to the sample chamber.

[0144] Similarly, in some cases, the disclosure provides a method of analyzing a sample in an analysis cartridge that comprises a sample delivery module comprising a DMF chip that delivers the sample to the sample port, wherein the DMF chip comprises: a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets of sample between the first and the second substrates to transport the liquid droplet.

[0145] Moreover, in some cases, the disclosure provides a method of analyzing a sample in an analysis cartridge that comprises a sample delivery module comprising a sample chamber and a sample deposition member that deposits the sample into the sample chamber. 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.

[0146] Thus, in view of the above, a method for determining the presence, amount, or amount of analyte in a test sample is provided. The method comprises assaying the test sample for a target analyte and comparing it to a control. The 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.

[0147] For reasons of completeness, various aspects of the invention are set out in the following numbered clauses:

[0148] Clause 1 . An analysis cartridge comprising a chemical analysis module for multiplex analysis of analytes in a sample, the chemical analysis module comprising: a top panel and a bottom panel and a plurality of channels disposed between the top panel and the bottom panel, wherein one or more channels of the plurality of channels contain one or more reagents that produce, in each such channel, a detectable signal indicative of a concentration of an analyte, a sample port that delivers the sample to the plurality of channels, and one or more sensors that detect the detectable signals generated in the one or more channels of the plurality of channels.

[0149] Clause 2. The analysis cartridge of Clause 1 , wherein the plurality of channels run from a proximal end to a distal end of the top panel and the bottom panel.

[0150] Clause 3. The analysis cartridge of Clause 1 or 2, wherein the top panel and / or the bottom panel have indents or grooves that form the plurality of channels.

[0151] Clause 4. The analysis cartridge of any one of Clauses 1 to 3, wherein the channels are rectangular, square, oval, circular, elliptical, or irregular in the cross section. Clause 5. The analysis cartridge of Clause 1 or 2, wherein the top panel and the bottom panel have a plurality of vertical walls disposed between the top panel and the bottom panel thereby producing the plurality of channels.

[0152] Clause 6. The analysis cartridge of Clause 5, wherein the plurality of vertical walls disposed between the top panel and the bottom panel comprise an adhesive.

[0153] Clause 7. The analysis cartridge of any one of Clauses 1 to 6, wherein a sensor from the one or more sensors is located within or adjacent to a channel and detects the detectable signal generated in the channel.

[0154] Clause 8. The analysis cartridge of Clause 7, wherein the sensor is located in the middle of the channel or at the distal end of the channel.

[0155] Clause 9. The analysis cartridge of Clause 6 or 7, wherein the sensor is an amperometric sensor that detects a current generated in the channel.

[0156] Clause 10. The analysis cartridge of Clause 6 or 7, wherein the sensor is a potentiometric sensor that detects a charge generated in the channel.

[0157] Clause 1 1 . The analysis cartridge of Clause 6 or 7, wherein the sensor is a coulometric sensor that detects oxygen generated in the channel.

[0158] Clause 12. The analysis cartridge of any one of Clauses 1 to 6, wherein a sensor from the one or more sensors is directed at a channel and detects the detectable signal generated in the channel.

[0159] Clause 13. The analysis cartridge of Clause 12, wherein the sensor is an optical sensor that detects an optical signal generated in the channel.

[0160] Clause 14. The analysis cartridge of any one of Clauses 1 to 13, wherein the detectable signals generated in a first set of one or more channels are electrochemical signals and the corresponding sensors are amperometric and / or potentiometric sensors. Clause 15. The analysis cartridge of any one of Clauses 1 to 14, wherein the detectable signals generated in a second set of one or more channels are optical signals and the corresponding sensors are optical sensors.

[0161] Clause 16. The analysis cartridge of any one of Clauses 1 to 15, wherein the detectable signals generated in a third set of one or more channels are oxygen and the corresponding sensors are coulometric sensors.

[0162] Clause 17. The analysis cartridge of any one of Clauses 1 to 8, wherein two or more channels of the plurality of channels comprise the following combinations of sensors: one or more electrochemical sensors and one or more optical sensors; one or more electrochemical sensors and one or more chemical sensors; one or more optical sensors and one or more chemical sensors; one or more amperometric sensors and one or more potentiometric sensors; one or more amperometric sensors and one or more coulometric sensors; one or more amperometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more coulometric sensors

[0163] Clause 18. The analysis cartridge of any one of Clauses 14 to 17, wherein the detectable signals generated in the first set of one or more channels are electrochemical signals and the corresponding sensors are amperometric and / or potentiometric sensors; the detectable signals generated in the second set of one or more channels are optical signals and the corresponding sensors are optical sensors; and the detectable signals generated in the third set of one or more channels are oxygen and the corresponding sensors are coulometric sensors.

[0164] Clause 19. The analysis cartridge of any one of Clauses 1 to 18, wherein the top panel comprises pillars extending down into the channels.

[0165] Clause 20. The analysis cartridge of any one of Clauses 1 to 19, wherein the top panel and the bottom panel are separated by a distance between 10 pm and 100 pm. Clause 21 . The analysis cartridge of any one of Clauses 1 to 20, wherein the longest dimension in the cross section of a channel is between 0.1 mm and 1 mm.

[0166] Clause 22. The analysis cartridge of any one of Clauses 1 to 21 , wherein the length of a channel is between 5 mm and 100 mm.

[0167] Clause 23. The analysis cartridge of any one of Clauses 1 to 22, wherein the volume of one or more channels within the plurality of channels is between 0.01 pl and 1 pl.

[0168] Clause 24. The analysis cartridge of any one of Clauses 1 to 23, wherein a channel from the plurality of channels is configured to detect an analyte selected from: glucose, calcium, blood urea nitrogen (BUN), creatinine, sodium, potassium, chloride, carbon dioxide (CO2), serum total protein (TP), serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

[0169] Clause 25. The analysis cartridge of any one of Clauses 1 to 24, wherein the plurality of channels are configured to detect two or more analytes selected from: glucose, calcium, BUN, creatinine, sodium, potassium, chloride, carbon dioxide CO2, TP, serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

[0170] Clause 26. The analysis cartridge of any one of Clauses 1 to 25, wherein the plurality of channels are configured to detect two or more analytes selected from: glucose, calcium, sodium, potassium, carbon dioxide, chloride, albumin, TP, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, BUN, and creatinine.

[0171] Clause 27. The analysis cartridge of any one of Clauses 1 to 26, wherein the analysis cartridge further comprises a plasma separation module for separating a plasma from blood cells in a blood sample. Clause 28. The analysis cartridge of Clause 27, wherein the plasma separation module comprises a filter for separating the plasma from blood cells in the blood sample.

[0172] Clause 29. The analysis cartridge of Clause 27, wherein the plasma separation module comprises a first digital microfluidic (DMF) chip for separating the plasma from blood cells in the blood sample, the first DMF chip comprising a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets comprising the blood and one or more reagents to transport the droplets between the first and the second substrates.

[0173] Clause 30. The analysis cartridge of any one of Clauses 1 to 29, further comprising a sample delivery module that delivers the sample to a sample port that delivers the sample to the chemical analysis module.

[0174] Clause 31 . The analysis cartridge of Clause 30, wherein the sample delivery module comprises a sample chamber, a sample deposition member that delivers the sample into the sample chamber, and a sample drain fl uidically connected to the sample chamber, wherein the sample drain receives excess sample beyond the sample filled into the chemical analysis module.

[0175] Clause 32. The analysis cartridge of Clause 31 , wherein the sample drain comprises two or more channels f luid ically connected to the sample chamber.

[0176] Clause 33. The analysis cartridge of Clause 30, wherein, the sample delivery module comprises a second DMF chip that delivers the sample to the sample port, wherein the second DMF chip comprises: a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets of the sample between the first and the second substrates to transport the droplets.

[0177] Clause 34. The analysis cartridge of Clause 30, wherein the sample delivery module comprises a sample chamber and a sample deposition member that deposits the sample into the sample chamber. Clause 35. A method of analyzing a sample, comprising loading the sample in any one of the analysis cartridges of Clauses 1 to 34 and analyzing the sample.

[0178] Clause 36. The method of Clause 35, comprising analyzing between 0.1 pl to 2 pl of the sample in one or more channels of the plurality of channels.

[0179] Clause 37. The method of Clause 35 or 36, wherein the sample is: blood, plasma urine, saliva, sweat, sputum, semen, mucus, lacrimal fluid, lymph fluid, amniotic fluid, interstitial fluid, lung lavage, cerebrospinal fluid, feces, a nasal swab soaked in a buffer, or a throat swab soaked in a buffer.

[0180] Clause 38. The method of Clause 37, wherein the blood is venous blood or capillary blood.

[0181] Clause 39. A method of fabricating an analysis cartridge of any one of Clauses 1 to 34, comprising: providing a top panel having certain features, providing a bottom panel having certain features, depositing glue lines on the top panel and / or the bottom panel; joining the two panels together so as to produce a chemical analysis module comprising a plurality of channels.

[0182] Clause 40. The method of Clause 39, wherein, the top panel and the bottom panel comprise a PET film.

[0183] Clause 41 . A chemical analysis module for multiplex analysis of analytes in a sample comprising: a top panel and a bottom panel and a plurality of channels disposed between the top panel and the bottom panel, wherein one or more channels of the plurality of channels contain one or more reagents that produce, in each such channel, a detectable signal indicative of a concentration of an analyte, a sample port that delivers the sample to the plurality of channels, and one or more sensors that detect the detectable signals generated in the one or more channels of the plurality of channels.

[0184] Clause 42. The chemical analysis module of Clause 41 , wherein the plurality of channels run from a proximal end to a distal end of the top panel and the bottom panel.

[0185] Clause 43. The chemical analysis module of Clause 41 or 42, wherein the top panel and / or the bottom panel have indents or grooves that form the plurality of channels.

[0186] Clause 44. The chemical analysis module of any one of Clauses 41 to 43, wherein the channels are rectangular, square, oval, circular, elliptical, or irregular in the cross section.

[0187] Clause 45. The chemical analysis module of Clause 41 or 42, wherein the top panel and the bottom panel have a plurality of vertical walls disposed between the top panel and the bottom panel thereby producing the plurality of channels.

[0188] Clause 46. The chemical analysis module of Clause 45, wherein the plurality of vertical walls disposed between the top panel and the bottom panel comprise an adhesive.

[0189] Clause 47. The chemical analysis module of any one of Clauses 41 to 46, wherein a sensor from the one or more sensors is located within or adjacent to a channel and detects the detectable signal generated in the channel.

[0190] Clause 48. The chemical analysis module of Clause 47, wherein the sensor is located in the middle of the channel or at the distal end of the channel.

[0191] Clause 49. The chemical analysis module of Clause 46 or 47, wherein the sensor is an amperometric sensor that detects a current generated in the channel.

[0192] Clause 50. The chemical analysis module of Clause 46 or 47, wherein the sensor is a potentiometric sensor that detects a charge generated in the channel. Clause 51 . The chemical analysis module of Clause 46 or 47, wherein the sensor is a coulometric sensor that detects oxygen generated in the channel.

[0193] Clause 52. The chemical analysis module of any one of Clauses 41 to 46, wherein a sensor from the one or more sensors is directed at a channel and detects the detectable signal generated in the channel.

[0194] Clause 53. The chemical analysis module of Clause 52, wherein the sensor is an optical sensor that detects an optical signal generated in the channel.

[0195] Clause 54. The chemical analysis module of any one of Clauses 41 to 53, wherein the detectable signals generated in a first set of one or more channels are electrochemical signals and the corresponding sensors are amperometric and / or potentiometric sensors.

[0196] Clause 55. The chemical analysis module of any one of Clauses 41 to 54, wherein the detectable signals generated in a second set of one or more channels are optical signals and the corresponding sensors are optical sensors.

[0197] Clause 56. The chemical analysis module of any one of Clauses 41 to 55, wherein the detectable signals generated in a third set of one or more channels are oxygen and the corresponding sensors are coulometric sensors.

[0198] Clause 57. The chemical analysis module of any one of Clauses 41 to 48, wherein two or more channels of the plurality of channels comprise the following combinations of sensors: one or more electrochemical sensors and one or more optical sensors; one or more electrochemical sensors and one or more chemical sensors; one or more optical sensors and one or more chemical sensors; one or more amperometric sensors and one or more potentiometric sensors; one or more amperometric sensors and one or more coulometric sensors; one or more amperometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more coulometric sensors Clause 58. The chemical analysis module of any one of Clauses 54 to 57, wherein the detectable signals generated in the first set of one or more channels are electrochemical signals and the corresponding sensors are amperometric and / or potentiometric sensors; the detectable signals generated in the second set of one or more channels are optical signals and the corresponding sensors are optical sensors; and the detectable signals generated in the third set of one or more channels are oxygen and the corresponding sensors are coulometric sensors.

[0199] Clause 59. The chemical analysis module of any one of Clauses 41 to 58, wherein the top panel comprises pillars extending down into the channels.

[0200] Clause 60. The chemical analysis module of any one of Clauses 41 to 59, wherein the top panel and the bottom panel are separated by a distance between 10 pm and 100 pm.

[0201] Clause 61 . The chemical analysis module of any one of Clauses 41 to 60, wherein the longest dimension in the cross section of a channel is between 0.1 mm and 1 mm.

[0202] Clause 62. The chemical analysis module of any one of Clauses 41 to 61 , wherein the length of a channel is between 5 mm and 100 mm.

[0203] Clause 63. The chemical analysis module of any one of Clauses 41 to 62, wherein the volume of one or more channels within the plurality of channels is between 0.01 pl and 1 pl.

[0204] Clause 64. The chemical analysis module of any one of Clauses 41 to 63, wherein a channel from the plurality of channels is configured to detect an analyte selected from: glucose, calcium, blood urea nitrogen (BUN), creatinine, sodium, potassium, chloride, carbon dioxide (CO2), serum total protein (TP), serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase. Clause 65. The chemical analysis module of any one of Clauses 41 to 64, wherein the plurality of channels are configured to detect two or more analytes selected from: glucose, calcium, BUN, creatinine, sodium, potassium, chloride, carbon dioxide CO2, TP, serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

[0205] Clause 66. The chemical analysis module of any one of Clauses 41 to 65, wherein the plurality of channels are configured to detect two or more analytes selected from: glucose, calcium, sodium, potassium, carbon dioxide, chloride, albumin, TP, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, BUN, and creatinine.

[0206] Clause 67. An analysis cartridge comprising the chemical analysis module of any one of Clauses 41 to 66.

[0207] Clause 68. The analysis cartridge of Clause 67, further comprising a plasma separation module for separating a plasma from blood cells in a blood sample

[0208] Clause 69. The analysis cartridge of Clause 68, wherein the plasma separation module comprises a filter for separating the plasma from blood cells in the blood sample.

[0209] Clause 70. The analysis cartridge of Clause 68, wherein the plasma separation module comprises a first digital microfluidic (DMF) chip for separating the plasma from blood cells in the blood sample, the first DMF chip comprising a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets comprising the blood and one or more reagents to transport the droplets between the first and the second substrates.

[0210] Clause 71. The analysis cartridge of any one of Clauses 67 to 70, further comprising a sample delivery module that delivers the sample to a sample port that delivers the sample to the chemical analysis module. Clause 72. The analysis cartridge of Clause 71 , wherein the sample delivery module comprises a sample chamber, a sample deposition member that delivers the sample into the sample chamber, and a sample drain fl uidically connected to the sample chamber, wherein the sample drain receives excess sample beyond the sample filled into the chemical analysis module.

[0211] Clause 73. The analysis cartridge of Clause 72, wherein the sample drain comprises two or more channels fluidically connected to the sample chamber.

[0212] Clause 74. The analysis cartridge of Clause 71 , wherein, the sample delivery module comprises a second DMF chip that delivers the sample to the sample port, wherein the second DMF chip comprises: a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets of the sample between the first and the second substrates to transport the droplets.

[0213] Clause 75. The analysis cartridge of Clause 71 , wherein the sample delivery module comprises a sample chamber and a sample deposition member that deposits the sample into the sample chamber.

[0214] Clause 76. A method of analyzing a sample, comprising loading the sample in any one of the chemical analysis module of Clauses 31 to 66 or the analysis cartridge of any one of Clauses 67 to 75 and analyzing the sample.

[0215] Clause 77. The method of Clause 76, comprising analyzing between 0.1 pl to 2 pl of the sample in one or more channels of the plurality of channels.

[0216] Clause 78. The method of Clause 76 or 77, wherein, the top panel and the bottom panel comprise a PET film.

[0217] Clause 79. The method of any one of Clauses 76 to 78, wherein the sample is: blood, urine, saliva, sweat, sputum, semen, mucus, lacrimal fluid, lymph fluid, amniotic fluid, interstitial fluid, lung lavage, cerebrospinal fluid, feces, a nasal swab soaked in a buffer, or a throat swab soaked in a buffer. Clause 80. The method of Clause 79, wherein the blood is venous blood or capillary blood.

[0218] Clause 81 . A method of fabricating the chemical analysis module of any one of Clauses 41 to 66, comprising: providing a top panel having certain features, providing a bottom panel having certain features, depositing glue lines on the top panel and / or the bottom panel; joining the two panels together so as to produce a chemical analysis module comprising a plurality of channels.

[0219] 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

WE CLAIM:1 . An analysis cartridge comprising a chemical analysis module for multiplex analysis of analytes in a sample, the chemical analysis module comprising: a top panel and a bottom panel and a plurality of channels disposed between the top panel and the bottom panel, wherein one or more channels of the plurality of channels contain one or more reagents that produce, in each such channel, a detectable signal indicative of a concentration of an analyte, a sample port that delivers the sample to the plurality of channels, and one or more sensors that detect the detectable signals generated in the one or more channels of the plurality of channels.

2. The analysis cartridge of claim 1 , wherein the plurality of channels run from a proximal end to a distal end of the top panel and the bottom panel.

3. The analysis cartridge of claim 1 or 2, wherein the top panel and / or the bottom panel have indents or grooves that form the plurality of channels.

4. The analysis cartridge of any one of claims 1 to 3, wherein the channels are rectangular, square, oval, circular, elliptical, or irregular in the cross section.

5. The analysis cartridge of claim 1 or 2, wherein the top panel and the bottom panel have a plurality of vertical walls disposed between the top panel and the bottom panel thereby producing the plurality of channels.

6. The analysis cartridge of claim 5, wherein the plurality of vertical walls disposed between the top panel and the bottom panel comprise an adhesive.

7. The analysis cartridge of any one of claims 1 to 6, wherein a sensor from the one or more sensors is located within or adjacent to a channel and detects the detectable signal generated in the channel.

8. The analysis cartridge of claim 7, wherein the sensor is located in the middle of the channel or at the distal end of the channel.

9. The analysis cartridge of claim 6 or 7, wherein the sensor is an amperometric sensor that detects a current generated in the channel.

10. The analysis cartridge of claim 6 or 7, wherein the sensor is a potentiometric sensor that detects a charge generated in the channel.1 1 . The analysis cartridge of claim 6 or 7, wherein the sensor is a coulometric sensor that detects oxygen generated in the channel.

12. The analysis cartridge of any one of claims 1 to 6, wherein a sensor from the one or more sensors is directed at a channel and detects the detectable signal generated in the channel.

13. The analysis cartridge of claim 12, wherein the sensor is an optical sensor that detects an optical signal generated in the channel.

14. The analysis cartridge of any one of claims 1 to 13, wherein the detectable signals generated in a first set of one or more channels are electrochemical signals and the corresponding sensors are amperometric and / or potentiometric sensors.

15. The analysis cartridge of any one of claims 1 to 14, wherein the detectable signals generated in a second set of one or more channels are optical signals and the corresponding sensors are optical sensors.

16. The analysis cartridge of any one of claims 1 to 15, wherein the detectable signals generated in a third set of one or more channels are oxygen and the corresponding sensors are coulometric sensors.

17. The analysis cartridge of any one of claims 1 to 8, wherein two or more channels of the plurality of channels comprise the following combinations of sensors: one or more electrochemical sensors and one or more optical sensors; one or more electrochemical sensors and one or more chemical sensors; one or more optical sensors and one or more chemical sensors; one or more amperometric sensors and one or more potentiometric sensors; one or more amperometric sensors and one or more coulometric sensors; one or more amperometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more coulometric sensors.

18. The analysis cartridge of any one of claims 14 to 17, wherein the detectable signals generated in the first set of one or more channels are electrochemical signals and the corresponding sensors are amperometric and / or potentiometric sensors; the detectable signals generated in the second set of one or more channels are optical signals and the corresponding sensors are optical sensors; and the detectable signals generated in the third set of one or more channels are oxygen and the corresponding sensors are coulometric sensors.

19. The analysis cartridge of any one of claims 1 to 18, wherein the top panel comprises pillars extending down into the channels.

20. The analysis cartridge of any one of claims 1 to 19, wherein the top panel and the bottom panel are separated by a distance between 10 pm and 100 pm.21 . The analysis cartridge of any one of claims 1 to 20, wherein the longest dimension in the cross section of a channel is between 0.1 mm and 1 mm.

22. The analysis cartridge of any one of claims 1 to 21 , wherein the length of a channel is between 5 mm and 100 mm.

23. The analysis cartridge of any one of claims 1 to 22, wherein the volume of one or more channels within the plurality of channels is between 0.01 pl and 1 pl.

24. The analysis cartridge of any one of claims 1 to 23, wherein a channel from the plurality of channels is configured to detect an analyte selected from: glucose, calcium, blood urea nitrogen (BUN), creatinine, sodium, potassium, chloride, carbon dioxide (CO2), serum total protein (TP), serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

25. The analysis cartridge of any one of claims 1 to 24, wherein the plurality of channels are configured to detect two or more analytes selected from: glucose, calcium, BUN, creatinine, sodium, potassium, chloride, carbon dioxide CO2, TP, serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

26. The analysis cartridge of any one of claims 1 to 25, wherein the plurality of channels are configured to detect two or more analytes selected from: glucose, calcium, sodium, potassium, carbon dioxide, chloride, albumin, TP, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, BUN, and creatinine.

27. The analysis cartridge of any one of claims 1 to 26, wherein the analysis cartridge further comprises a plasma separation module for separating a plasma from blood cells in a blood sample.

28. The analysis cartridge of claim 27, wherein the plasma separation module comprises a filter for separating the plasma from blood cells in the blood sample.

29. The analysis cartridge of claim 27, wherein the plasma separation module comprises a first digital microfluidic (DMF) chip for separating the plasma from blood cells in the blood sample, the first DMF chip comprising a first substrate; a second substrate;a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets comprising the blood and one or more reagents to transport the droplets between the first and the second substrates.

30. The analysis cartridge of any one of claims 1 to 29, further comprising a sample delivery module that delivers the sample to a sample port that delivers the sample to the chemical analysis module.

31. The analysis cartridge of claim 30, wherein the sample delivery module comprises a sample chamber, a sample deposition member that delivers the sample into the sample chamber, and a sample drain fluidically connected to the sample chamber, wherein the sample drain receives excess sample beyond the sample filled into the chemical analysis module.

32. The analysis cartridge of claim 31 , wherein the sample drain comprises two or more channels fluidically connected to the sample chamber.

33. The analysis cartridge of claim 30, wherein, the sample delivery module comprises a second DMF chip that delivers the sample to the sample port, wherein the second DMF chip comprises: a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets of the sample between the first and the second substrates to transport the droplets.

34. The analysis cartridge of claim 30, wherein the sample delivery module comprises a sample chamber and a sample deposition member that deposits the sample into the sample chamber.

35. A method of analyzing a sample, comprising loading the sample in any one of the analysis cartridges of claims 1 to 34 and analyzing the sample.

36. The method of claim 35, comprising analyzing between 0.1 pl to 2 pl of the sample in one or more channels of the plurality of channels.

37. The method of claim 35 or 36, wherein the sample is: blood, plasma urine, saliva, sweat, sputum, semen, mucus, lacrimal fluid, lymph fluid, amniotic fluid, interstitial fluid, lung lavage, cerebrospinal fluid, feces, a nasal swab soaked in a buffer, or a throat swab soaked in a buffer.

38. The method of claim 37, wherein the blood is venous blood or capillary blood.

39. A method of fabricating an analysis cartridge of any one of claims 1 to 34, comprising: providing a top panel having certain features, providing a bottom panel having certain features, depositing glue lines on the top panel and / or the bottom panel; joining the two panels together so as to produce a chemical analysis module comprising a plurality of channels.

40. The method of claim 39, wherein, the top panel and the bottom panel comprise a PET film.41 . A chemical analysis module for multiplex analysis of analytes in a sample comprising: a top panel and a bottom panel and a plurality of channels disposed between the top panel and the bottom panel, wherein one or more channels of the plurality of channels contain one or more reagents that produce, in each such channel, a detectable signal indicative of a concentration of an analyte, a sample port that delivers the sample to the plurality of channels, and one or more sensors that detect the detectable signals generated in the one or more channels of the plurality of channels.

42. The chemical analysis module of claim 41 , wherein the plurality of channels run from a proximal end to a distal end of the top panel and the bottom panel.

43. The chemical analysis module of claim 41 or 42, wherein the top panel and / or the bottom panel have indents or grooves that form the plurality of channels.

44. The chemical analysis module of any one of claims 41 to 43, wherein the channels are rectangular, square, oval, circular, elliptical, or irregular in the cross section.

45. The chemical analysis module of claim 41 or 42, wherein the top panel and the bottom panel have a plurality of vertical walls disposed between the top panel and the bottom panel thereby producing the plurality of channels.

46. The chemical analysis module of claim 45, wherein the plurality of vertical walls disposed between the top panel and the bottom panel comprise an adhesive.

47. The chemical analysis module of any one of claims 41 to 46, wherein a sensor from the one or more sensors is located within or adjacent to a channel and detects the detectable signal generated in the channel.

48. The chemical analysis module of claim 47, wherein the sensor is located in the middle of the channel or at the distal end of the channel.

49. The chemical analysis module of claim 46 or 47, wherein the sensor is an amperometric sensor that detects a current generated in the channel.

50. The chemical analysis module of claim 46 or 47, wherein the sensor is a potentiometric sensor that detects a charge generated in the channel.51 . The chemical analysis module of claim 46 or 47, wherein the sensor is a coulometric sensor that detects oxygen generated in the channel.

52. The chemical analysis module of any one of claims 41 to 46, wherein a sensor from the one or more sensors is directed at a channel and detects the detectable signal generated in the channel.

53. The chemical analysis module of claim 52, wherein the sensor is an optical sensor that detects an optical signal generated in the channel.

54. The chemical analysis module of any one of claims 41 to 53, wherein the detectable signals generated in a first set of one or more channels are electrochemical signals and the corresponding sensors are amperometric and / or potentiometric sensors.

55. The chemical analysis module of any one of claims 41 to 54, wherein the detectable signals generated in a second set of one or more channels are optical signals and the corresponding sensors are optical sensors.

56. The chemical analysis module of any one of claims 41 to 55, wherein the detectable signals generated in a third set of one or more channels are oxygen and the corresponding sensors are coulometric sensors.

57. The chemical analysis module of any one of claims 41 to 48, wherein two or more channels of the plurality of channels comprise the following combinations of sensors: one or more electrochemical sensors and one or more optical sensors; one or more electrochemical sensors and one or more chemical sensors; one or more optical sensors and one or more chemical sensors; one or more amperometric sensors and one or more potentiometric sensors; one or more amperometric sensors and one or more coulometric sensors; one or more amperometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more optical sensors; one or more potentiometric sensors and one or more coulometric sensors58. The chemical analysis module of any one of claims 54 to 57, wherein the detectable signals generated in the first set of one or more channels are electrochemical signals and the corresponding sensors are amperometric and / or potentiometric sensors; the detectable signals generated in the second set of one or more channels are optical signals and the corresponding sensors are optical sensors; and the detectable signalsgenerated in the third set of one or more channels are oxygen and the corresponding sensors are coulometric sensors.

59. The chemical analysis module of any one of claims 41 to 58, wherein the top panel comprises pillars extending down into the channels.

60. The chemical analysis module of any one of claims 41 to 59, wherein the top panel and the bottom panel are separated by a distance between 10 pm and 100 pm.61 . The chemical analysis module of any one of claims 41 to 60, wherein the longest dimension in the cross section of a channel is between 0.1 mm and 1 mm.

62. The chemical analysis module of any one of claims 41 to 61 , wherein the length of a channel is between 5 mm and 100 mm.

63. The chemical analysis module of any one of claims 41 to 62, wherein the volume of one or more channels within the plurality of channels is between 0.01 pl and 1 pl.

64. The chemical analysis module of any one of claims 41 to 63, wherein a channel from the plurality of channels is configured to detect an analyte selected from: glucose, calcium, blood urea nitrogen (BUN), creatinine, sodium, potassium, chloride, carbon dioxide (CO2), serum total protein (TP), serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

65. The chemical analysis module of any one of claims 41 to 64, wherein the plurality of channels are configured to detect two or more analytes selected from: glucose, calcium, BUN, creatinine, sodium, potassium, chloride, carbon dioxide CO2, TP, serum albumin, lactate, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, lipids, and lipase.

66. The chemical analysis module of any one of claims 41 to 65, wherein the plurality of channels are configured to detect two or more analytes selected from: glucose, calcium, sodium, potassium, carbon dioxide, chloride, albumin, TP, alkaline phosphatase, alanine transaminase, aspartate aminotransferase, bilirubin, BUN, and creatinine.

67. An analysis cartridge comprising the chemical analysis module of any one of claims 41 to 66.

68. The analysis cartridge of claim 67, further comprising a plasma separation module for separating a plasma from blood cells in a blood sample69. The analysis cartridge of claim 68, wherein the plasma separation module comprises a filter for separating the plasma from blood cells in the blood sample.

70. The analysis cartridge of claim 68, wherein the plasma separation module comprises a first digital microfluidic (DMF) chip for separating the plasma from blood cells in the blood sample, the first DMF chip comprising a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets comprising the blood and one or more reagents to transport the droplets between the first and the second substrates.71 . The analysis cartridge of any one of claims 67 to 70, further comprising a sample delivery module that delivers the sample to a sample port that delivers the sample to the chemical analysis module.

72. The analysis cartridge of claim 71 , wherein the sample delivery module comprises a sample chamber, a sample deposition member that delivers the sample into the sample chamber, and a sample drain fluidically connected to the sample chamber, wherein the sample drain receives excess sample beyond the sample filled into the chemical analysis module.

73. The analysis cartridge of claim 72, wherein the sample drain comprises two or more channels fluidically connected to the sample chamber.

74. The analysis cartridge of claim 71 , wherein, the sample delivery module comprises a second DMF chip that delivers the sample to the sample port, wherein the second DMF chip comprises: a first substrate; a second substrate; a gap separating the first substrate from the second substrate; a plurality of electrodes that generate electrical actuation forces on droplets of the sample between the first and the second substrates to transport the droplets.

75. The analysis cartridge of claim 71 , wherein the sample delivery module comprises a sample chamber and a sample deposition member that deposits the sample into the sample chamber.

76. A method of analyzing a sample, comprising loading the sample in any one of the chemical analysis module of claims 31 to 66 or the analysis cartridge of any one of claims 67 to 75 and analyzing the sample.

77. The method of claim 76, comprising analyzing between 0.1 pl to 2 pl of the sample in one or more channels of the plurality of channels.

78. The method of claim 76 or 77, wherein, the top panel and the bottom panel comprise a PET film.

79. The method of any one of claims 76 to 78, wherein the sample is: blood, urine, saliva, sweat, sputum, semen, mucus, lacrimal fluid, lymph fluid, amniotic fluid, interstitial fluid, lung lavage, cerebrospinal fluid, feces, a nasal swab soaked in a buffer, or a throat swab soaked in a buffer.

80. The method of claim 79, wherein the blood is venous blood or capillary blood.81 . A method of fabricating the chemical analysis module of any one of claims 41 to 66, comprising: providing a top panel having certain features,providing a bottom panel having certain features, depositing glue lines on the top panel and / or the bottom panel; joining the two panels together so as to produce a chemical analysis module comprising a plurality of channels.-M-

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