Dispensing reagent containers and methods of dispensing

Abstract

The present disclosure provides a dispensing reagent container that is capable of precise small volume reagent dispensing. The dispensing reagent container comprises a proximal end and a distal end; a first storage region at the proximal end having: a first volume, and a first end at a proximal end having an opening for filling with reagent; a second storage region in fluid communication with the first storage region having a second volume, wherein the second volume is less than the first volume, a dispenser positioned at the distal end of the second storage region; and a plunger having a proximal end and a distal end, wherein: the plunger is positioned coaxially within the reagent container spanning the first storage region and the second storage region, and the distal end of the plunger is capable of being in contact with the dispenser at the distal end of the reagent container.

Description

DISPENSING REAGENT CONTAINERS AND ME HODS OF DISPENSINGCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 613,508, filed December 21, 2023, which application is incorporated herein by reference in its entirety.INTRODUCTION

[0002] Reagent dispensing using automated sample processing and analysis devices commonly uses aspiration of the reagent into a probe, transport of the reagents to a reaction vessel, followed by dispensing of the reagents into the reaction vessel. Following reagent transfer, the probe is washed to clean the probe and to avoid cross-contamination with reagents aspirated and dispensed in the following test. The usage of a probe runs the risk of ineffective washing of ‘sticky’ reagents which can result in cross-contamination. The washing of probes additionally places limits on the timing of the washing to avoid impacting the throughput of assays. The usage of wash solutions requires the collection and disposal of the wash solutions in addition to the need for pumps and reservoirs which is associated with failure modes that result in leaks and spills of potentially hazardous fluids. The inclusion of additional components to an automated sample processing and analysis device creates increased costs and complexity that serves as a barrier for low-cost and low- volume applications.

[0003] Alternative strategies for reagent dispensing include self-dispensing containers and syringes. Self-dispensing containers make use of squeeze bottles to avoid the need for washing, however, they lack the precision and accuracy needed for low-volume applications. The accuracy of dispensing using squeeze bottles highly depends on the container geometry, fill volume, and mechanical tolerance which change based on the size of the container. Syringes allow for more precise and accurate dispensing of reagents which are generally associated with a smaller diameter syringe. The smaller diameter syringe lacks the capacity necessary for reagent dispensing in automated sample processing and analysis devices and increasing the diameter of the syringe creates the problem that a very small displacement is needed to dispense a small volume.

[0004] Dispensing reagent containers that can dispense small and precise volumes without greater mechanical complexity are highly desirable in the field of automated sample processingand analysis devices. Such dispensing reagent containers would allow for the development of lower cost and lower complexity automated sample processing and analysis devices.SUMMARY

[0005] The present disclosure provides dispensing reagent containers that are capable of precise small volume reagent dispensing. The dispensing reagent container comprises a proximal end and a distal end; a first storage region at the proximal end having: a first volume, and a first end at a proximal end having an opening for filling with reagent; a second storage region in fluid communication with the first storage region having a second volume, wherein the second volume is less than the first volume, a dispenser positioned at the distal end of the second storage region; and a plunger having a proximal end and a distal end, wherein: the plunger is positioned coaxially within the reagent container spanning the first storage region and the second storage region, and the distal end of the plunger is capable of being in contact with the dispenser at the distal end of the reagent container.

[0006] Also provided are methods for using the dispensing reagent container using the dispensing reagent container.BRIEF DESCRIPTION OF THE DRAWINGS

[0007] FIG. 1 illustrates a dispensing reagent container according to an embodiment.

[0008] FIG. 2 illustrates a dispensing reagent container according to an embodiment.

[0009] FIG. 3 illustrates a deconstructed dispensing reagent container according to an embodiment.

[0010] FIG. 4 illustrates a cross-section of a dispensing reagent container according to an embodiment.

[0011] FIG. 5 illustrates a cross-section of a dispensing reagent container according to an embodiment.

[0012] FIG. 6 illustrates a cross-section of a dispensing reagent container according to an embodiment.

[0013] FIG. 7 illustrates a cross-section of a dispensing reagent container according to an embodiment.

[0014] FIG. 8 illustrates a second storage region and dispenser according to an embodiment.

[0015] FIG. 9 illustrates a collection of dispensing reagent containers according to an embodiment.

[0016] FIG. 10 illustrates a collection of dispensing reagent containers of various sizes according to an embodiment.DETAILED DESCRIPTION OF THE INVENTION

[0017] Embodiments of the present disclosure relate to dispensing reagent containers and methods for dispensing reagents.

[0018] Before the present invention is described in greater detail, it is to be understood that this invention is not limited to a particular embodiment described, as such may, of course, 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, since the scope of the present invention will be limited only by the appended claims.

[0019] It must be 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. Thus, for example, refer to “an elastomeric gasket” includes plurality of such elastomeric gaskets and reference to “the reagent” includes reference to one or more reagents and equivalents thereof known to those skilled in the art, and so forth.

[0020] All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The present disclosure is controlling to the extent there is a contradiction between the present disclosure and a publication incorporated by reference.Definitions

[0021] Before the embodiments of the present disclosure are described, it is to be understood that this invention is not limited to particular embodiments described, as such may, of course, 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.

[0022] 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 absolutevalues 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.

[0023] 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.

[0024] 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, a first vertical structure and a 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.

[0025] “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,” “and” 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.

[0026] 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.

[0027] “Microbead” and “microparticle” are used herein interchangeably and refer to a substantially spherical solid support. The microbead or microparticle is a substantially sphericalsolid support that is influenced by a magnetic field such that the magnetic field can attract or repulse the microparticle or magnetic particle. The microparticle and microbead may contain at least one specific binding member that binds to an analyte of interest and at least one detectable label. Alternatively, the microparticle and microbead may contain a first specific binding member that binds to the analyte and a second specific binding member that also binds to the analyte and contains at least one detectable label.

[0028] " Non-functional bead," "helper bead," and "assisting particle" are used interchangeably and refers to a substantially spherical assisting solid support, that is larger in diameter than a microparticle, which is configured to be chemically inert with respect to other components of an assay. As used herein, an assisting particle refers to a spherical particle which generally does not chemically interact with other particles (including a microparticle, conjugate, and / or reagent), but which is magnetic or paramagnetic. In certain exemplary embodiments, assisting particle may be coated so as to chemically interact with interferents, that is, any materials which would interfere with assay or analysis of an analyte of interest within the targeted sample. In such embodiments, the assisting particles can also improve binding efficiency of the microparticles including, for the purpose of illustration and not limitation, by binding with interferents. Additionally and alternatively, the shape of a solid support can be roughly spherical, though not limited to such shapes.

[0029] The assisting solid supports can be larger in diameter than the other support mediums within the storage region and configured so as to not chemically interact with any other components within the mixing region. Specifically, the diameter of the assisting solid supports (e.g. helper beads) can be at least 1 %, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least about 11 %, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 50%, at least about 75%, at least about 100%, at least about 150%, at least about 200%, at least about 250%, at least about 300%, at least about 350%, at least about 400%, at least about 450%, at least about 500%, at least about 550%, at least about 600%, at least about 650%, at least about 700%, at least about 750%, at least about 800%, at least about 850%, at least about 900%, at least about 950%, at least about 1000% greater or larger than the diameter of other support mediums (e.g., microparticles).

[0030] The assisting particles can be configured such that they do not chemically bond or pair with other components of the targeted solution, such as the microparticles, target conjugates, and / or the target analyte. In certain exemplary embodiments, both microparticles and assistingparticles can be magnetic, paramagnetic, or superparamagnetic particles (or any combination therein). In such exemplary embodiments, both microparticles and assisting particles, under the influence of a magnetic field or force, can form into chains of connected particles which facilitate mixing within the targeted solution.

[0031] “Specific binding partner” or “specific binding member” as used interchangeably herein refer to one of two different molecules that specifically recognizes the other molecule compared to substantially less recognition of other molecules. The one of two different molecules has an area on the surface or in a cavity, which specifically binds to and is thereby defined as complementary with a particular spatial and polar organization of the other molecule. The molecules may be members of a specific binding pair. For example, a specific binding member may include, but not limited to, a protein, such as a receptor, an enzyme, an antibody and an aptamer, a peptide, a nucleotide, oligonucleotide, a nucleic acid, a polynucleotide and combinations thereof.

[0032] “Specific binding” or “specifically binding” as used herein may refer to the interaction of an antibody, a protein, or a peptide with a second chemical species, wherein the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody is specific for epitope “A”, the presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled “A” and the antibody, will reduce the amount of labeled A bound to the antibody.

[0033] The terms “polynucleotide” and “nucleic acid,” used interchangeably herein, refer to a polymeric form of nucleotides of any length, either ribonucleotides or deoxy nucleotides. Thus, this term includes, but is not limited to, single-, double-, or multi-stranded DNA or RNA, genomic DNA, cDNA, DNA-RNA hybrids, or a polymer including purine and pyrimidine bases or other natural, chemically or biochemically modified, non-natural, or derivatized nucleotide bases. The terms “polynucleotide” and “nucleic acid” should be understood to include, as applicable to the embodiment being described, single-stranded (such as sense or antisense) and double-stranded polynucleotides.

[0034] The terms "peptide," "polypeptide," and "protein" are used interchangeably herein, and refer to a polymeric form of amino acids of any length, which can include coded and noncoded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides having modified peptide backbones.

[0035] "Analyte", "target analyte", "analyte of interest" as used interchangeably herein, refers to a substance, material or chemical constituent the presence, absence and / or amount of which is being analyzed in a biological sample obtained from a subject. In some aspects, the analyte is a biomolecule. Non-limiting examples of biomolecules include macromolecules such as, proteins, lipids, and carbohydrates. In certain instances, the analyte may be hormones, antibodies, growth factors, cytokines, enzymes, receptors (e.g., neural, hormonal, nutrient, and cell surface receptors) or their ligands, cancer markers (e.g., PSA, TNF-alpha), markers of myocardial infarction (e.g., troponin, creatine kinase, Creatinine kinase-cardiac muscle biomarker (CK- MB), B-type natriuretic peptide (also known as brain natriuretic peptide; BNP), N-terminal prohormone of brain natriuretic peptide (NT-proBNP) and the like), toxins, drugs (e.g., drugs of addiction), metabolic agents (e.g., including vitamins), and the like. Non-limiting examples of protein analytes include peptides, polypeptides, protein fragments, protein complexes, fusion proteins, recombinant proteins, phosphoproteins, glycoproteins, lipoproteins, or the like.

[0036] “Antibody” and “antibodies” as used herein refers to monoclonal antibodies, multispecific antibodies, human antibodies, humanized antibodies (fully or partially humanized), animal antibodies such as, but not limited to, a bird (for example, a duck or a goose), a shark, a whale, and a mammal, including a non-primate (for example, a cow, a pig, a camel, a llama, a horse, a goat, a rabbit, a sheep, a hamster, a guinea pig, a cat, a dog, a rat, a mouse, etc.) or a non-human primate (for example, a monkey, a chimpanzee, etc.), recombinant antibodies, chimeric antibodies, single-chain Fvs (“scFv”), single chain antibodies, single domain antibodies, Fab fragments, F(ab’) fragments, F(ab')2 fragments, disulfide-linked Fvs (“sdFv”), and anti-idiotypic (“anti-Id”) antibodies, dual-domain antibodies, dual variable domain (DVD) or triple variable domain (TVD) antibodies (dual -variable domain immunoglobulins and methods for making them are described in Wu, C., et al., Nature Biotechnology, 25(11): 1290- 1297 (2007) and PCT International Application WO 2001 / 058956, the contents of each of which are herein incorporated by reference), and functionally active epitope-binding fragments of any of the above. Antibodies include immunoglobulin molecules and immunologically active fragments of immunoglobulin molecules, namely, molecules that contain an analyte-binding site. Immunoglobulin molecules can be of any type (for example, IgG, IgE, IgM, IgD, IgA, and IgY), class (for example, IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2), or subclass. For simplicity sake, an antibody against an analyte is frequently referred to herein as being either an “antianalyte antibody” or merely an “analyte antibody”.

[0037] “Antibody fragment” as used herein refers to a portion of an intact antibody comprising the antigen-binding site or variable region. The portion does not include the constant heavy chain domains (i.e., CH2, CH3, or CH4, depending on the antibody isotype) of the Fc region of the intact antibody. Examples of antibody fragments include, but are not limited to, Fab fragments, Fab' fragments, Fab'-SH fragments, F(ab')2 fragments, Fd fragments, Fv fragments, diabodies, single-chain Fv (scFv) molecules, single-chain polypeptides containing only one light chain variable domain, single-chain polypeptides containing the three CDRs of the light-chain variable domain, single-chain polypeptides containing only one heavy chain variable region, and single-chain polypeptides containing the three CDRs of the heavy chain variable region.

[0038] 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 invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety to disclose and describe the methods and / or materials in connection with which the publications are cited. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.1. OVERVIEW

[0039] Provided herein are dispensing reagent containers that may be used with automated sample processing and analysis devices. To provide structure to the description of the dispensing reagent containers, first, the overall dispensing reagent container design will be disclosed. Second, the benefits of the dispensing reagent container will be disclosed. Following an overview of the basic design of the dispensing reagent container, a detailed disclosure of the dispensing reagent container’s design will be disclosed. Following the detailed disclosure of the dispensing reagent container’s design, methods of dispensing reagents using the dispensing reagent container will be disclosed.

[0040] I. Dispensing reagent container design

[0041] The dispensing reagent container comprises a proximal and a distal end and a first storage region having at the proximal end having a first volume and a first end at a proximal end having an opening for filling with reagents. The dispensing reagent container comprises asecond storage region in fluid communication with the first storage region having a second volume, wherein the second volume is less than the first volume. The dispensing reagent container comprises a dispenser positioned at the distal end of the second storage region and a plunger having a proximal end and a distal end wherein the plunger is positioned coaxially within the reagent bottle spanning the first storage region and the second storage region and the distal end of the plunger of the plunger is capable of being in contact with the dispenser at the distal end of the reagent container. In some embodiments, the dispensing reagent container comprises an external cap that covers the dispenser when the dispensing reagent bottle is not in use. The term “external cap” as used herein refers to a cap that is on the outside of the dispensing reagent container, e.g., not within the first or second volume or within a region that stores or houses reagents.

[0042] In some embodiments, the dispensing reagent container further comprises a base region proximal to the first storage region having a stanchion positioned coaxially in the base region wherein the base region is in fluid communication with the first storage region, the stanchion has a proximal end and a distal end having an opening with a diameter positioned coaxially spanning the height of the stanchion, and the plunger is positioned within the opening of the stanchion thereby spanning the base region, the first storage region, and the second storage region. When the dispensing reagent container comprises a base region, a first storage region, a second storage region, and a dispenser, the base region is at the proximal end of the dispensing reagent container, the first storage region is distal to the base region, the second storage region is distal to the first storage region, and the dispenser is distal to the second storage region.

[0043] FIG. 1 discloses an illustration of the exterior of an embodiment of an upside-down dispensing reagent container. In this embodiment, the dispensing reagent container has a distal end 105 and a proximal end 106. The distal end of the dispensing reagent container 105 has a cap 101 covering the dispenser (not shown). The cap 101 is bound to the second storage region 102. The cap may be bound to the second storage region by any method deemed necessary including, without limitation, welding, treaded connections, press fitting, adhesive bonding, solvent bonding, snap fitting, etc. The second storage region 102 is proximal to the cap 101. The first storage region 103 is proximal to the second storage region 102. The volume of the first storage region 103 is larger than the volume of the second storage region 102. The base region 104 is proximal to the first storage region 103. The base region is bound to the first storage region at the interface 108 between the base region 104 and the first storage region 103.The base region may be bound to the first storage region by any method deemed necessary including, without limitation, welding, treaded connections, press fitting, adhesive bonding, solvent bonding, snap fitting, etc. The base region 104 comprises a plunger (not shown). In some embodiments, the cap is only present prior to the first use of the dispensing reagent container.

[0044] FIG. 4 discloses an illustration of a cross-section of an embodiment of the dispensing reagent bottle where the plunger is fully depressed. In this embodiment, the dispensing reagent container 400 comprises a proximal end 406 and a distal end 405. The dispensing reagent container comprises a base region 404, a plunger 407, a first storage region 403, a second storage region 402, and a dispenser 401. The base region 404 is at the proximal end 406 of the dispensing reagent container 400. The base region 404 is proximal to the first storage region 403. The base region is bound to the first storage region at the interface 408 between the based region 404 and the first storage region 403. The base region may be bound to the first storage region by any method deemed necessary including, without limitation, welding, treaded connections, press fitting, adhesive bonding, solvent bonding, snap fitting, etc. The second storage region 402 is distal to the first storage region 403. The first storage region 403 has a first volume and the second storage region has a second volume that is smaller than the first volume. A dispenser 401 is positioned at the distal end 405 of the dispensing reagent bottle. The dispenser 401 is distal to the second storage region 402. The dispensing reagent bottle 400 comprises a plunger 407 having a proximal end 412 and a distal end 413. The plunger 407 is positioned coaxially within the dispensing reagent container 400 spanning the base region, the first storage region, and the second storage region. The distal end of the plunger 413 is in contact with the base of the second storage region 409 thereby sealing the dispenser 401 and preventing the reagent contained within the first storage region 403 from leaking or evaporating from the dispensing reagent container.

[0045] FIG. 8 discloses an illustration of a close-up view of a cross-section of the second storage region. The dispensing reagent container 800 comprises a second storage region 802 that comprises a small volume for dispensing precise small volumes. In some embodiments, the total volume of the second storage region comprises the total volume dispensed for a single assay. In some embodiments, the second storage region comprises more than the total volume dispensed for a single assay. The volume of the second storage region may be a range of different including without limitation, about 5- lOpL, about 10- 15pL, about 15-20glL, about 20-25pL, about 25-30ftL, about 30-35|lL, about 35-40|lL or greater than about 4O|1L. In someembodiments, the volume of the second storage region is about 5-20pL. The second storage region comprises a base 809. The base 809 of the second storage region 802 may have a range of different shapes. In some embodiments, the base of the second storage region is conical. In some embodiments, the base of the second storage region is flat. In some embodiments, the base of the second storage region is spherical. In some embodiments, the base of the second storage region is pyramidal. The second storage region is fluidically connected to dispenser 801. The dispenser 801 discharges the reagent contained in the second storage region when the plunger 807 is depressed into the second storage region. The diameter of the dispenser is sufficiently large to allow the passage of microparticles in the absence of clogging and small enough to create a jet for dispensing reagents with a precise cutoff that does not leave a droplet hanging on the tip of the dispenser. The diameter of the dispenser may be a range of different diameters including about 300-800|im. For instance, the diameter of the dispenser may be about 300-350p.m, about 350-400|im, about 400-450|im, about 45O-5OO|om, about 550-600|J.m, about 650-70|im, about 700-750|J.m, about 750-800|J.m, about 800-850pm, about 850-900|Jm, about 900-950 .m, about 950-1000|J.m, about 1000-1 lOO m, about 1100-1200|J.m, about 1200-1300|Jm, about 1300- 1400|im, about 1400-15 OOftm or greater than about 1500|am.

[0046] FIG. 9 discloses an illustration of a collection of dispensing reagent containers connected to each other according to an embodiment. In this embodiment, the dispensing reagent containers 900a, 900b, and 900c comprise external clipping features 901a and 901b on two opposite sides of the perimeter of the dispensing reagent container 900c that are capable of joining two or more dispensing reagent containers together. Any number of dispensing reagent containers may be joined together including, without limitation, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more.

[0047] The base region, the first storage region, the second storage region, and the plunger of the present disclosure may be made of a range of different materials such that the materials facilitate the methods and designs disclosed herein. The material may be rigid or flexible. The rigidity and flexibility may be controlled both by the material used and the thickness of the material of the base region, the first storage region, the second storage region, and the plunger. In some embodiments, the base region, the first storage region, the second storage region, and the plunger are made of the same material. In some embodiments, the base region, the first storage region, the second storage region, and the plunger are made from different materials. In some embodiments, the first storage region and the second storage region are made of the same material. In some embodiments, the base region, the first storage region and the second storageregion are made of the same material. In some embodiments, the base region, the first storage region, the second storage region, and the dispenser are made of optically transparent materials. In some embodiments, the base region, the first storage region and the second storage region are made of the same material. In some embodiments, the base region, the first storage region, the second storage region, and the dispenser are made of optically semi-transparent materials. In some embodiments, the base region, the first storage region and the second storage region are made of the same material. In some embodiments, the base region, the first storage region, the second storage region, and the dispenser are made of optically opaque materials. In general the materials are chemically inert and do not react with an of the reagents contained within the dispensing reagent container. For instance, materials that find use in the present disclosure include, without limitation, polyethylene, high density polyethylene, polypropylene, etc.

[0048] II. Benefits of the device

[0049] The dispensing reagent container of the present disclosure provides certain benefits over reagent containers known in the art. The benefits of the dispensing reagent containers of the present disclosure include the ability to dispense consistent small volumes regardless of the volume of the container, the ability to dispense directly from the dispensing reagent container without the need for a reservoir, the lack of a need to wash the dispenser in between dispenses, the ability to connect multiple dispensing reagent containers together, and increased shelf-life of the reagents contained within the dispensing reagent container. In terms of the ability to dispense consistent small volumes, the multiple exemplary embodiments of the dispensing reagent containers disclosed herein combine the degree of precision provided by a syringe by having a small volume second storage region and the capacity of a larger reagent container by having a larger volume first storage region that is fluidically connected to the smaller volume second storage region. The ability to dispense without the need for washes allows the usage of reagents that are “stickier” or have a tendency to adhere to dispensing tips. Overall, the design of the exemplary dispensing reagent containers disclosed herein allows for the elimination of many expensive components, failure modes, and the space allocation in automated sample processing and analysis devices.2. DEVICE DESIGN

[0050] The dispensing reagent containers of the present disclosure have a range of different volumes and features. Exemplary embodiments of the dispensing reagent containers are disclosed in FIG. 1-7.

[0051] FIG. 1 discloses an illustration of the exterior of an embodiment of an upside-down dispensing reagent container. In this embodiment, the dispensing reagent container has a distal end 105 and a proximal end 106. The distal end of the dispensing reagent container 105 has a cap 101 covering the dispenser (not shown). The cap 101 is bound to the second storage region 102. The cap may be bound to the second storage region by any method deemed necessary including, without limitation, welding, treaded connections, press fitting, adhesive bonding, solvent bonding, snap fitting, etc. The second storage region 102 is proximal to the cap 101. The first storage region 103 is proximal to the second storage region 102. The volume of the first storage region 103 is larger than the volume of the second storage region 102. The base region 104 is proximal to the first storage region 103. The base region is bound to the first storage region at the interface 108 between the base region 104 and the first storage region 103. The base region may be bound to the first storage region by any method deemed necessary including, without limitation, welding, treaded connections, press fitting, adhesive bonding, solvent bonding, snap fitting, etc. The base region 104 comprises a plunger (not shown). In some embodiments, the cap is only present prior to the first use of the dispensing reagent container. This embodiment is a representation of the dispensing reagent container when the dispensing reagent container is not in use or in storage. This orientation allows the reagents contained in the dispensing reagent container to settle away from the dispenser.

[0052] FIG. 2 discloses an illustration of the exterior of an embodiment of the dispensing reagent container. In this embodiment, the dispensing reagent container 200 comprises a proximal end 206 and a distal end 205. The dispensing reagent container comprises a base region 204, a plunger 207, a first storage region 203, a second storage region 202, and a dispenser 201 . The base region 204 is at the proximal end 206 of the dispensing reagent container 200. The base region 204 is proximal to the first storage region 203. The base region is bound to the first storage region at the interface 208 between the base region 204 and the first storage region 203. The base region may be bound to the first storage region by any method deemed necessary including, without limitation, welding, treaded connections, press fitting, adhesive bonding, solvent bonding, snap fitting, etc. The second storage region 202 is distal to the first storage region 203. The first storage region 203 has a first volume and the second storage region 202 has a second volume that is smaller than the first volume. A dispenser 201 is positioned at the distal end 205 of the dispensing reagent bottle. The dispenser 201 is distal to the second storage region 202. This embodiment is a representation of the orientation of the dispensing reagent container when the dispensing reagent container is in use.

[0053] FIG. 3 discloses an illustration of a cross-section of an embodiment of a deconstruction of the dispensing reagent bottle. In this embodiment, the first storage region 303 has a first end 316 at a proximal end having an opening for filling with one or more reagents. The first storage region 303 has a first volume 317 (region between white dotted lines). The first storage region 303 is proximal to the second storage region 302. The second storage region has an entrance 308 and a base 309. The second storage region 302 has a second volume 318 defined by the space between the entrance 308 and the base 309. The second volume 318 is smaller than the first volume 317. The second storage region 302 is in fluid communication with the first storage region 303. In some embodiments, the first storage region comprises one or more internal baffles along the perimeter of the first storage region. In some embodiments, the one or more internal baffles promote the mixing of one or more reagents present in the first storage region and / or second storage region. In some embodiments, the internal baffles promote the mixing of one or more reagents present in the first storage region and / or second storage region when the plunger is depressed. The base region 304 has a stanchion 310 positioned coaxially in the base region 304. The stanchion has a proximal end with an opening 311a and a distal end with an opening 311b with a diameter positioned coaxially within the base region spanning the height of the stanchion (the distance between 31 la to 311b). The plunger 307 has a proximal end 312 and a distal end 313 and the plunger 307 is positioned within the opening 311a and 311b thereby spanning the base region 304. In some embodiments, the distal end of the plunger 313 has comprises an elastomeric gasket 314. In some embodiments, the proximal end of the plunger comprises one or more elastomeric gaskets 315a, 315b, and / or 315c. In some embodiment, the one or more elastomeric gaskets are integrated into the plunger. In some embodiments, the one or more elastomeric gaskets are added to the exterior of the plunger. This embodiment is a representation of the dispensing reagent container when the first storage region and the second storage region are being filled with reagent.

[0054] FIG. 4 discloses an illustration of a cross-section of an embodiment of the dispensing reagent bottle where the plunger is fully depressed. In this embodiment, the dispensing reagent container 400 comprises a proximal end 406 and a distal end 405. The dispensing reagent container comprises a base region 404, a plunger 407, a first storage region 403, a second storage region 402, and a dispenser 401. The base region 404 is at the proximal end 406 of the dispensing reagent container 400. The base region 404 is proximal to the first storage region 403. The base region is bound to the first storage region at the interface 408 between the based region 404 and the first storage region 403. The base region may be bound to the first storage region by any method deemed necessary including, without limitation, welding, treadedconnections, press fitting, adhesive bonding, solvent bonding, snap fitting, etc. The second storage region 402 is distal to the first storage region 403. The first storage region 403 has a first volume and the second storage region has a second volume that is smaller than the first volume. A dispenser 401 is positioned at the distal end 405 of the dispensing reagent bottle. The dispenser 401 is distal to the second storage region 402. The dispensing reagent bottle 400 comprises a plunger 407 having a proximal end 412 and a distal end 413. The plunger 407 is positioned coaxially within the dispensing reagent container 400 spanning the base region, the first storage region, and the second storage region. The distal end of the plunger 413 is in contact with the base of the second storage region 409 thereby sealing the dispenser 401 and preventing the reagent contained within the first storage region 403 from leaking or evaporating from the dispensing reagent container. In some embodiments, the distal end of the plunger is the same diameter as the second storage reagent. In some embodiments, the distal end of the plunger comprises an elastomeric gasket. In some embodiments, the elastomeric gasket is equal to or greater than the diameter of the second storage region. This embodiment is a representation of the dispensing reagent container just prior to dispensing reagent or after completing the dispensing of the reagent.

[0055] FIG. 5 discloses a cross-section of an embodiment of the dispensing reagent bottle where the plunger is not fully depressed. In this embodiment, the dispensing reagent container 500 comprises a proximal end 506 and a distal end 505. The dispensing reagent container comprises a base region 504, a plunger 507, a first storage region 503, a second storage region 502, and a dispenser 501. The base region 504 is at the proximal end 506 of the dispensing reagent container 500. The base region 504 is proximal to the first storage region 503. The first storage region 503 has a first volume and the second storage region 502 has a second volume that is smaller than the first volume. A dispenser 501 is positioned at the distal end 505 of the dispensing reagent bottle. The dispensing reagent bottle 500 comprises a plunger 507 having a proximal end 512 and a distal end 513. The plunger 507 is positioned coaxially within the dispensing reagent container 500 spanning the base region, the first storage region, and the second storage region. The plunger 507 comprises one or more elastomeric gaskets 515a, 515b, and / or 515c. In some embodiments, the one or more elastomeric gaskets guide the plunger into the second storage region. In some embodiments, the elastomeric gaskets seal the opening spanning the stanchion of the base region. The distal end of the plunger 513 is not in contact with the entrance 508 to the second storage region thereby enabling the filling of the second storage region with reagent. This embodiment is a representation of the dispensing reagent container prior to depressing the plunger to dispense reagent.

[0056] FIG. 6 discloses a cross-section of an embodiment of the dispensing reagent bottle where the plunger is not fully depressed. In this embodiment, the dispensing reagent container 600 comprises a proximal end 606 and a distal end 605. The dispensing reagent container comprises a base region 604, a plunger 607, a first storage region 603, a second storage region 602, and a dispenser 601. The base region 604 is at the proximal end 606 of the dispensing reagent container 600. The base region 604 is proximal to the first storage region 603. The first storage region 603 has a first volume and the second storage region 602 has a second volume that is smaller than the first volume. A dispenser 601 is positioned at the distal end 605 of the dispensing reagent bottle. The dispensing reagent bottle 600 comprises a plunger 607 having a proximal end 612 and a distal end 613. The plunger 607 is positioned coaxially within the dispensing reagent container 600 spanning the base region, the first storage region, and the second storage region. The distal end of the plunger 613 is in contact with the entrance 608 to the second storage region thereby sealing the second storage region prior to the commencing of the dispensing. In some embodiments, the distal end of the plunger is the same diameter as the second storage region. In some embodiments, the distal end of the plunger comprises an elastomeric gasket. In some embodiments, the elastomeric gasket is equal to or greater than the diameter of the second storage region. This embodiment is a representation of the dispensing reagent container prior to depressing the plunger to dispense reagent.

[0057] FIG. 7 discloses a cross-section of an embodiment of the dispensing reagent bottle where the plunger is not fully depressed. In this embodiment, the dispensing reagent container 700 comprises a proximal end 706 and a distal end 705. The dispensing reagent container comprises a base region 704, a plunger 707, a first storage region 703, a second storage region 702, and a dispenser 701 . The base region 704 is at the proximal end 707 of the dispensing reagent container 700. The base region 704 is proximal to the first storage region 703. The first storage region 703 has a first volume and the second storage region 702 has a second volume that is smaller than the first volume. A dispenser 701 is positioned at the distal end 707 of the dispensing reagent bottle. The dispensing reagent bottle 700 comprises a plunger 707 having a proximal end 712 and a distal end 713. The plunger 707 is positioned coaxially within the dispensing reagent container 700 spanning the base region, the first storage region, and the second storage region. The distal end of the plunger 713 is within the second storage reagent thereby beginning the dispensing. In some embodiments, the distal end of the plunger is the same diameter as the second storage region. In some embodiments, the distal end of the plunger comprises an elastomeric gasket. In some embodiments, the elastomeric gasket is equal to orgreater than the diameter of the second storage region. This embodiment is a representation of the dispensing reagent container during dispensing of reagent.

[0058] The dispensing reagent containers of the present disclosed may comprise one or more reagents. The one or more reagents may be any reagent deemed useful for performing an assay. In some embodiments, the assay is an assay that detects a target analyte. The analyte may be macromolecules such as proteins, lipids, or carbohydrates. The analyte may be hormones, antibodies, growth factors, cytokines, enzymes, receptors (e.g., neural, hormonal, nutrient, and cell surface receptors) or their ligands, cancer markers (e.g., PSA, TNF-alpha), markers of myocardial infarction (e.g., troponin, creatine kinase, Creatinine kinase-cardiac muscle biomarker (CK-MB), B-type natriuretic peptide (also known as brain natriuretic peptide; BNP), N-terminal prohormone of brain natriuretic peptide (NT-proBNP) and the like), toxins, drugs (e.g., drugs of addiction), metabolic agents (e.g., including vitamins), and the like. Non-limiting examples of protein analytes include peptides, polypeptides, protein fragments, protein complexes, fusion proteins, recombinant proteins, phosphoproteins, glycoproteins, lipoproteins, or the like.

[0059] The one or more reagents may include a range of different reagents including without limitations, first specific binding members, second specific binding members, buffers containing microparticles, buffers containing assisting microparticles, buffers containing microparticles and assisting microparticles, wash buffers, lysis buffers, elution buffers, hydrophobic solutions, hydrophilic solutions, inert solutions, solutions that react with enzymes to produce detectable signals, buffers for nucleic acid amplification buffers, buffers containing primers for nucleic acid amplification, buffers containing probes for the detection of amplification products, buffers containing enzymes such as polymerases, transcriptases, reverse transcriptases, phosphatases, kinases, transferases, ligases, etc., or any combination thereof. The one or more reagents may be two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more reagents. In some embodiments, the one or more reagents are a combination of any of the reagents described above.

[0060] The dispensing reagent containers of the present disclosure have first storage regions having first volumes and second storage regions having second volumes. The second storage region has a second volume that is less than the first volume. The first storage region may have a range of different first volumes that are larger than the second volumes. For instance, the first volume may be at least about 10 times, at least about 15 times, at least about 20 times, at least about 25 times, at least about 30 times, at least about 35 times, at least about 40 times, at leastabout 45 times, at least about 50 times, at least about 75 times, at least about 100 times, at least about 125 times, at least about 150 times, at least about 175 times, at least about 200 times, at least about 300 times, at least about 400 times, at least about 500 times, at least about 600 times, at least about 700 times, at least about 800 times, at least about 900 times, at least about 1000 times, at least about 2000 times, at least about 3000 times, at least about 4000 times, at least about 5000 times, at least about 6000 times, at least about 7000 times, at least about 8000 times, at least about 9000 times, at least about 10000 times, or more than about 10000 times the second volume. The first volume of the first storage region may be a range of different volumes. For instance, the first volume of the first storage region may be about 100p.L to about lOOmL. The first volume may be at least about 100|J.L, at least about 200p.L, at least about 300|iL, at least about 400pL, at least about 500| L, at least about 6OO| L, at least about 700pL, at least about 800|J.L, at least about 900pL, at least about ImL, at least about 2mL, at least about 3mL, at least about 4mL, at least about 5mL, at least about 6mL, at least about 7mL, at least about 8mL, at least about 9mL, at least about lOmL, at least about llmL, at least about 12mL, at least about 13mL, at least about 14mL, at least about 15mL, at least about 16mL, at least about 17mL, at least about 18mL, at least about 19mL, at least about 20mL, at least about 30mL, at least about 40mL, at least about 50mL, at least about 60mL, at least about 70mL, at least about 80mL, at least about 90mL, at least about lOOmL, or greater than about lOOmL. In some embodiments, the first volume is about 100|J.L to about 50mL.

[0061] The second volume of the second storage region may be a range of different volumes. For instance, the second volume of the second storage region may be about 5-5O| L. The second volume of the second storage region may be about 5- I OpL, about 10- 15|1L, about 15-20pL, about 2O-25|1L, about 25-30|J.L, about 30-35|lL, about 35-40|lL, about 4O-45|1L, about 45-50|J.L, or greater than about 50|lL. The second volume of the second storage region may be at least about 5pL. at least about 10|TL, at about least 15|1L, at least about 20|J.L, at least about 25|1L, at least about 30|lL, at least about 35|1L, at least about 40|_tL, at least about 45|1L, or at least about 50|J.L. In some embodiments, the volume of the second storage region is about 5-30|J.L.

[0062] The second storage region has an entrance and a base. The base of the second storage has a variety of different shapes. For instance, the base of the storage region may be flat, conical, spherical, pyramidal, a non-regular shape etc. In some embodiments, the base of the second storage region is conical. In some embodiments, the base of the second storage region is flat. In some embodiments, the base of the second storage region is spherical. In some embodiments,the base of the second storage region is pyramidal. In some embodiments, the base of the second storage region is a non-regular shape.

[0063] FIG. 8 discloses an illustration of a close-up view of a cross-section of the second storage region. The dispensing reagent container 800 comprises a second storage region 802 that comprises a small volume for dispensing precise small volumes. In some embodiments, the total volume of the second storage region comprises the total volume dispensed for a single assay. In some embodiments, the second storage region comprises more than the total volume dispensed for a single assay. The volume of the second storage region may be a range of different including without limitation, about 5- lOpL, about 10-15pL, about 15-20pL, about 20-25pL, about 25-30pL, about 30-35pL, about 35-40pL or about 40-45pL, about 45-50pL, or greater than about 50pL. In some embodiments, the volume of the second storage region is about 5- 30pI .. The second storage region comprises a base 809. The base 809 of the second storage region 802 may have a range of different shapes. In some embodiments, the base of the second storage region is conical. In some embodiments, the base of the second storage region is flat. In some embodiments, the base of the second storage region is spherical. In some embodiments, the base of the second storage region is pyramidal. The second storage region is flui dically connected to dispenser 801. The dispenser 801 discharges the reagent contained in the second storage region when the plunger 807 is depressed into the second storage region. The diameter of the dispenser is sufficiently large to allow the passage of microparticles in the absence of clogging and small enough to create a jet for dispensing reagents with a precise cutoff that does not leave a droplet hanging on the tip of the dispenser. The diameter of the dispenser may be a range of different diameters including about 3OO-8OOpm. For instance, the diameter of the dispenser may be about 3OO-35O|jm, about 350-400pm, about 400-450pm, about 450-500pm, about 550-600pm, about 650-70pm, about 700-750pm, about 750-800pm, or greater than about 800pm.

[0064] The first volume and second volume are determined by the shape of the dispensing reagent containers or the shape of the first storage region or second storage region. The shape may be a variety of different shapes. For instance, the shape may include a cylinder, a cube, a cuboid, a triangular prism, a pentagonal prism, a hexagonal prism, a heptagonal prism, an octagonal prism, a nonagonal prism, a decagonal prism, etc.

[0065] The first storage region may have certain features. In some embodiments, the first storage region has one or more internal baffles. In some embodiments, the one or more internalbaffles are along the perimeter of the first storage region. By “internal baffle” it is meant that the baffle is on the interior of the first storage region and may be in contact with the one or more reagents contained therein. In some embodiments, the one or more internal baffles promote the mixing of the one or more reagents when the plunger is depressed. In some embodiments, the one or more internal baffles promote the mixing of the one or more reagents when the dispensing reagent container is turned right-side-up from an upside-down orientation or is turned upside-down from a right-side-up orientation. In some embodiments, the dispensing reagent container is moved to mixed the one or more reagents contained therein. In some embodiments, the moving is spinning the dispensing reagent container on its axis. In some embodiments, the moving is moving the dispensing reagent container in a planar manner. In some embodiments, the first storage region has one or more internal fins. In some embodiments, the one or more internal fins are along the perimeter of the first storage region. By “internal fin” it is meant that the fin is on the interior of the first storage region and may be in contact with the one or more reagents contained therein. In some embodiments, the one or more internal fins promote the mixing of the one or more reagents when the plunger is depressed. In some embodiments, the one or more internal fins promote the mixing of the one or more reagents when the dispensing reagent container is turned right-side-up from an upside-down orientation or is turned upsidedown from a right-side-up orientation.

[0066] The exterior of the dispensing reagent containers may have certain features. In some embodiments, the dispensing reagent container comprises two or more external clipping features on opposite sides on the perimeter of the dispensing reagent container that are capable of joining two or more dispensing reagent containers together. By “external clipping features” it is meant that the clipping features are on the exterior of the dispensing reagent container and are never in contact with the one or more reagents contained within the dispensing reagent bottle. In some embodiments, the external clipping features are on the exterior of the first storage region. In some embodiments, the external clipping features are on the exterior of the base region.

[0067] FIG. 9 discloses an illustration of a collection of dispensing reagent containers connected to each other by external clipping features according to an embodiment. In this embodiment, the dispensing reagent containers 900a, 900b, and 900c comprise external clipping features 901a and 901b on two opposite sides of the perimeter of the dispensing reagent container 900c that are capable of joining two or more dispensing reagent containers together. Any number of dispensing reagent containers may be joined together including, withoutlimitation, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more.

[0068] FIG. 10 discloses an illustration of a collection of dispensing reagent containers of various sizes connected to each other by external clipping features according to an embodiment. In this embodiment, the small dispensing reagent containers 1002a, 1002b, and 1002c comprise external clipping features 1005a and 1005b on two opposite sides of the perimeter of the dispensing reagent container 1002c that are capable of joining two or more dispensing reagent containers together. While only three small dispensing reagent containers are shown, any number of small dispensing reagent containers may be joined together including, without limitation, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more. The medium dispensing reagent containers 1001a, 1001b, and 1001c comprise external clipping features 1004a and 1004b on two opposite sides of the perimeter of the dispensing reagent container 1001c that are capable of joining two or more dispensing reagent containers together. While only three medium dispensing reagent containers are shown, any number of small dispensing reagent containers may be joined together including, without limitation, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more. The large dispensing reagent containers 1000a, 1000b, and 1000c comprise external clipping features 1003a and 1003b on two opposite sides of the perimeter of the dispensing reagent container 1000c that are capable of joining two or more dispensing reagent containers together. While only three large dispensing reagent containers are shown, any number of small dispensing reagent containers may be joined together including, without limitation, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more. In some embodiments, one or more small dispensing reagent containers are joined to one or more medium dispensing reagent containers. In some embodiments, one or more small dispensing reagent containers are joined to one or more large dispensing reagent containers. In some embodiments, one or more medium dispensing reagent containers are joined to one or more large dispensing reagent containers. In some embodiments, one or more small dispensing reagent containers are joined to one or more medium dispensing reagent containers and one or more large dispensing reagent containers.

[0069] The exterior of the dispensing reagent containers may have certain features. In some embodiments, the dispensing reagent container comprises an external handle feature. The handle feature may be a feature that allows a user or an automated system to grab and transport one or more dispensing reagent containers. In some embodiments, the external handle feature is on theexterior of the first storage region. In some embodiments, the external handle feature is on the exterior of the base region. In some embodiments, the dispensing reagent container comprises an external tab feature. The tab feature may be a feature that allows a user or an automated system to grab and transport one or more dispensing reagent containers. In some embodiments, the external tab feature is on the exterior of the first storage region. In some embodiments, the external tab feature is on the exterior of the base region.

[0070] In some embodiments, in place of the base region, a compression region may be present. In some embodiments, the compression region is connected to the plunger such that when the compression region is compressed the plunger is depressed. In some embodiments, the compression region may be in the form of a bellows structure. In some embodiments, the compression region compresses in staged manner comprising a two or more stages where in the first stage depresses the plunger at a set amount that is not the total second volume of the second storage region thereby dispensing a set volume of reagent and the second stage depresses the plunger at a set amount that it the remainder of the second volume of the second storage region thereby dispensing a set volume of reagent that is the same volume as the first stage. The compression of the compression region is reversible such that the compression region may be uncompressed to the size of the uncompressed state.

[0071] The plunger of the present disclosure may have certain features. In some embodiments, the plunger has one or more elastomeric gaskets at the proximal end of the plunger. In some embodiments, the one or more elastomeric gaskets at the proximal end align the distal end of the plunger such that misalignment or jamming is prevented. In some embodiments, the one or more elastomeric gaskets at the proximal end seal the stanchion of the base region such that reagents are not able to leak or spill from the opening in the stanchion. In some embodiments, the one or more elastomeric gaskets are equal to or greater than the diameter of the opening of the stanchion of the base region such that the elastomeric gaskets completely seal the gasket. The elastomeric gaskets of the present disclosure are compressible such that even if they are greater in diameter than the opening the in the stanchion of the base region they can be compressed to fit in the opening. In some embodiments, the plunger comprises a retention feature on the proximal end such that the plunger cannot be removed from the base region. In some embodiments, the distal end of the plunger comprises an elastomeric gasket. In some embodiments, the elastomeric gasket is added to the exterior of the plunger. In some embodiments, the elastomeric gasket is fused or apart of the plunger. In some embodiments, the diameter of the elastomeric gasket on the distal end of the plunger is equal to or greater than thediameter of the second storage region. In some embodiments, the diameter of the plunger is equal to the diameter of the second storage region.3. METHODS OF DISPENSING A REAGENT

[0072] The present disclosure provides methods of dispensing a reagent using the dispensing reagent containers of the present disclosure. The method of dispensing a reagent comprises a) contacting the proximal end of the plunger of the reagent container of any of dispensing reagent containers described above, wherein the distal end of the plunger is in contact with the dispenser and the reagent container comprises one or more reagents, b) retracting the plunger such that the distal end is not located within the second storage region thereby enabling the filling of the second storage region with the one or more reagents, and c) depressing the plunger such that the distal end is located within the second storage region thereby dispensing all or a portion of the one or more reagents contained within the second volume of the second storage region.

[0073] The contacting of the proximal end of the plunger may be performed by any means that allows for precise depression of the plunger. In some embodiments, the contacting is performed by a mechanical device. In some embodiments, when the dispensing reagent container comprises a compression region in place of a base region, the contacting is performed by compressing the compressible region.

[0074] In some embodiments, the method further comprises step d) inverting the reagent container prior to step a), wherein prior to step d) the distal end of the dispensing reagent container is facing upward and the proximal end of the reagent container is facing downward and after step d) the distal end of the dispensing reagent container is facing downward and the proximal end of the reagent container is facing upward.

[0075] In some embodiments, the method further comprises repeating step c) until all of the reagent contained within the second volume is dispensed. The repeating step may be repeated a range of different times. In some embodiments, the repeating occurs one time. In some embodiments, the repeating occurs two times. In some embodiments, the repeating occurs three times. In some embodiments, the repeating occurs four times. In some embodiments, the repeating occurs three times. In some embodiments, the repeating occurs five times. In some embodiments, the repeating occurs three times. In some embodiments, the repeating occurs six times. In some embodiments, the repeating occurs seven times. In some embodiments, the repeating occurs eight times. In some embodiments, the repeating occurs nine times. In some embodiments, the repeating occurs ten times.

[0076] In some embodiments, the method further comprises repeating step b) thereby refilling the second volume of the second storage region. The repeating step may be repeated a range of different times. For instance, the repeating may occur one to ten thousand times. The repeating may occur about 1 to about 10 times, about 10 to about 50 times, about 50 to about 100 times, about 100 to about 200 times, about 200 to about 500 times, about 500 to about 1000 times, about 1000 to about 2000 times, about 2000 to about 5000 times, about 5000 to about 10000 times. In some embodiments, the repeating occurs at least about one time. In some embodiments, the repeating occurs at least about ten times. In some embodiments, the repeating occurs at least about one hundred times. In some embodiments, the repeating occurs at least about two hundred times. In some embodiments, the repeating occurs at least about five hundred times. In some embodiments, the repeating occurs at least about one thousand times. In some embodiments, the repeating occurs at least about two thousand times. In some embodiments, the repeating occurs at least about five thousand times. In some embodiments, the repeating occurs at least about ten thousand times.

[0077] In some embodiments, the method further comprises step e) inverting the reagent container after step c), wherein prior to step e) the distal end of the dispensing reagent container is facing downward and the proximal end of the reagent container is facing upward and the distal end of the plunger is in contact with the dispenser and after step d) the distal end of the dispensing reagent container is facing upward and the proximal end of the reagent container is facing downward.

[0078] In some embodiments, the method further comprises step f) retracting the plunger after step c) wherein prior to step f) the distal end of the plunger is in contact with the dispenser, wherein the retracting does not retract the distal end of the plunger outside of the second storage region thereby drawing air into the second volume of the second storage region.

[0079] An exemplary method of dispensing of a reagent is disclosed in FIG. 4-7. FIG. 4 represents step a) wherein the distal end of the plunger 413 is in contact with dispenser or the base of the second storage region 309. FIG. 5 represents step b) wherein the plunger 507 has been retracted such that the distal end of the plunger 514 is not located within the second storage region 502 thereby enabling the filling of the second storage region 502 with one or more reagents. FIG. 6 represents just prior to step c) wherein the plunger 607 is depressed such that the distal end of the plunger 614 is at the entrance 608 of the second storage region 602. FIG. 7 represents step c) wherein the plunger 707 is depressed such that the distal end of the plunger713 is located within the second storage region 702 thereby dispensing all or a portion of the one or more reagents contained within the second volume of the second storage region 702.

Claims

CLAIMS1. A dispensing reagent container, comprising: a proximal end and a distal end; a first storage region at the proximal end having: a first volume, and a first end at a proximal end having an opening for filling with reagent; a second storage region in fluid communication with the first storage region having a second volume, wherein the second volume is less than the first volume, a dispenser positioned at the distal end of the second storage region; and a plunger having a proximal end and a distal end, wherein: the plunger is positioned coaxially within the reagent container spanning the first storage region and the second storage region, and the distal end of the plunger is capable of being in contact with the dispenser at the distal end of the reagent container.

2. The dispensing reagent container of claim 1, further comprising a base region proximal to the first storage region having a stanchion positioned coaxially within the base region, wherein: the base region is in fluid communication with the first storage region, the stanchion has a proximal end and a distal end having an opening with a diameter positioned coaxially within the base region spanning the height of the stanchion, and the plunger is positioned within the opening thereby spanning the base region, the first storage region, and the second storage region.

3. The dispensing reagent container of claims 1 or 2, wherein the further comprises one or more reagents.

4. The dispensing reagent container of any of claims 1-3, wherein the one or more reagents are selected from the group consisting of a first specific binding members, a second specific binding members, a buffer containing microparticles, a buffer containing assisting microparticles, a buffer containing microparticles and assisting microparticles, a wash buffer, a lysis buffer, an elution buffer, a hydrophobic solution, a hydrophilic solution, an inert solution,solutions that react with enzymes to produce detectable signals, a buffer for nucleic acid amplification, a buffer containing primers for nucleic acid amplification, a buffe containing probes for the detection of amplification products, a buffer containing enzymes such as polymerases, transcriptases, reverse transcriptases, phosphatases, kinases, transferases, ligases, and any combination thereof.

5. The dispensing reagent container of claims 3 or 4, wherein the first storage region comprises one or more internal baffles along a perimeter of the first storage region.

6. The dispensing reagent container of claim 5, wherein the internal baffles promote mixing of the one or more reagents when the plunger is depressed.

7. The dispensing reagent container of any of claims 1-6, wherein the volume of the first storage region is at least ten times the volume of the second storage region.

8. The dispensing reagent container of any of claims 1-7, wherein the volume of the first storage region is at least twenty-five times the volume of the second storage region.

9. The dispensing reagent container of any of claims 1-8, wherein the volume of the first storage region is at least fifty times the volume of the second storage region.

10. The dispensing reagent container of any of claims 1-9, wherein the first volume is at least seventy-five times the second volume.

11. The dispensing reagent container of any of claims 1-10, wherein the first volume is at least one hundred times the second volume.

12. The dispensing reagent container of any of claims 1-11, wherein the first volume is at least one hundred fifty times the second volume.

13. The dispensing reagent container of any of claims 1-12, wherein the volume of the first storage region is at least two hundred times the volume of the second storage region.

14. The dispensing reagent container of any of claims 1-13, wherein the first volume is at least two hundred fifty times the second volume.

15. The dispensing reagent container of any of claims 1-14, wherein the first volume is at least three hundred times the second volume.

16. The dispensing reagent container of any of claims 1-15, wherein the first volume is at least four hundred times the second volume.

17. The dispensing reagent container of any of claims 1-16, wherein the first volume is at least five hundred times the second volume.

18. The dispensing reagent container of any of claims 1-17, wherein the first volume is at least six hundred times the second volume.

19. The dispensing reagent container of any of claims 1-18, wherein the first volume is at least seven hundred times the second volume.

20. The dispensing reagent container of any of claims 1-19, wherein the first volume is at least eight hundred times the second volume.

21. The dispensing reagent container of any of claims 1-20, wherein the first volume is at least nine hundred times the second volume.

22. The dispensing reagent container of any of claims 1-21, wherein the first volume is at least one thousand times the second volume.

23. The dispensing reagent container of any of claims 1-22, wherein the second storage region comprises a conical bottom on the proximal end of the second storage region.

24. The dispensing reagent container of any of claims 1-23, wherein the first storage region is cylindrical.

25. The dispensing reagent container of any of claims 1-24, wherein the second storage region is cylindrical.

26. The dispensing reagent container of any of claims 1-25, wherein the distal end of the plunger has a diameter equal to or greater than the second diameter.

27. The dispensing reagent container of any of claims 1-26, wherein the distal end of the plunger comprises an elastomeric gasket.

28. The dispensing reagent container of any of claims 1-27, wherein the proximal end of the plunger comprises one or more elastomeric gaskets.

29. The dispensing reagent container of any of claims 1-28, wherein the one or more elastomeric gaskets on the distal end of the plunger are equal to or greater than the diameter of the opening.

30. The dispensing reagent container of any of claims 1-29, wherein the elastomeric gasket on the distal end of the plunger is equal to or greater than the diameter of the diameter of the second storage region.

31. The dispensing reagent container of any of claims 1-30, wherein the one or more elastomeric gaskets on the proximal end are equal to or greater than the diameter of the opening of the stanchion.

32. The dispensing reagent container of any of claims 1-31, wherein the plunger comprises retention features that prevent the plunger from being removed from the dispensing reagent container.

33. The dispensing reagent container of any of claims 1-32, wherein the opening of the stanchion comprises an elastomeric gasket.

34. The dispensing reagent container of any of claims 1-33, wherein the dispensing reagent container is optically opaque.

35. The dispensing reagent container of any of claims 1-34, wherein the dispensing reagent container is optically transparent.

36. The dispensing reagent container of any of claims 1-35, wherein the base region, first storage region, and the second storage region are made of the same material.

37. The dispensing reagent container of any of claims 1-35, wherein the base region, the first storage region, and the second storage region are made of different materials.

38. The dispensing reagent container of any of claims 1-37, wherein the dispensing reagent container further comprises external clipping features on two opposite sides of the perimeter of the dispensing reagent container that are capable of joining two or more dispensing reagent containers together.

39. The dispensing reagent container of any of claims 1-38, further comprising an external cap that covers the dispenser when the dispensing reagent container is not in use.

40. The dispensing reagent container of any of claims 1-39, wherein the second volume of the second storage region is 5-50pL.

41. The dispensing reagent container of any of claims 1-40, wherein the first volume of the first storage region is 100|lL-50mL.

42. The dispensing reagent container of any of claims 1-41, wherein the dispensing reagent container further comprises handle features.

43. A method of dispensing a reagent, the method comprising: a) contacting the proximal end of the plunger of the reagent container of any of claims 1-42, wherein the distal end of the plunger is in contact with the dispenser and the reagent container comprises one or more reagents, b) retracting the plunger such that the distal end of the plunger is not located within the second storage region thereby enabling the filling of the second storage region with the one or more reagents, andc) depressing the plunger such that the distal end of the plunger is located within the second storage region thereby dispensing all or a portion of the one or more reagents contained within the second volume of the second storage region.

44. The method of claim 43, wherein the contacting is performed with a mechanical device.

45. The method of claims 43 or 44, further comprising step d) inverting the reagent container prior to step a), wherein prior to step d) the distal end of the dispensing reagent container is facing upward and the proximal end of the reagent container is facing downward and after step d) the distal end of the dispensing reagent container is facing downward and the proximal end of the reagent container is facing upward.

46. The method of any of claims 43-45, further comprising repeating step c) until all of the reagent contained within the second volume is dispensed.

47. The method of any of claims 43-46, further comprising step e) inverting the reagent container after step c), wherein prior to step e) the distal end of the dispensing reagent container is facing downward and the proximal end of the reagent container is facing upward and the distal end of the plunger is in contact with the dispenser and after step d) the distal end of the dispensing reagent container is facing upward and the proximal end of the reagent container is facing downward.

48. The method of any of claims 43-47, further comprises step f) retracting the plunger after step c) wherein prior to step f) the distal end of the plunger is in contact with the dispenser, wherein the retracting does not retract the distal end of the plunger outside of the second storage region thereby drawing air into the second volume of the second storage region.

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