System for Reconstituting a Dry Reagent Composition and Method for Using the Same

Abstract

A system for reconstituting a dry reagent composition is provided. Aspects of the system include a liquid container having an inner wall with the dry reagent composition, and a solid volume displacer positioned inside the liquid container and configured to occupy a majority of the liquid container volume below the upper portion of the dry reagent composition. Aspects of the invention further include a method of using the system and a kit comprising the system.

Description

Background Art

[0001] Assays for determining the presence and concentration of an analyte in a biological sample fluid often rely on the specific binding of a detectable label to the target analyte. The detectable label can be a marker that can be visualized by the naked eye or detected by spectroscopic methods such as fluorescence or UV-Vis spectroscopy. Typically, a fluorescent dye can be used as the detectable label that contains a specific fluorophore.

[0002] A fluorophore can have certain specific properties such as its absorption spectrum, extinction coefficient at an excitation-favorable wavelength, its emission spectrum, and its quantum efficiency. The quantum efficiency is the number of photons emitted per absorbed photon.

[0003] A multiplexed assay enables the simultaneous detection of multiple analytes in a single assay. In the case of immunoassays, a multiplexed assay involves a cocktail of antibodies each labeled with a different dye. Each antibody binds to a specific analyte or antigen in the sample. In this way, different analytes or antigens can be distinguished and quantified based on different dyes.

[0004] For convenience, a multiplex assay reagent can be supplied as a pre-mixed cocktail of individual binding molecules such as antibodies. Other reaction components such as buffers, salts, and surfactants can be included in the cocktail. Most preferably, it is a cocktail (single assay reagent) containing all the necessary components. In this case, the assay can be performed by simply adding the sample to a reaction vessel containing the cocktail.

[0005] It is advantageous to provide stable assay reagents, particularly reagents that are stable at room temperature. This allows for the transportation and storage of reagents without refrigeration, which is particularly important for assays conducted in rural areas where resources, including electricity, may be limited. For this purpose, stable reagents can be provided by drying an aqueous solution of the reagent or by lyophilization. However, there are problems with some labeled reagents. These reagents crosslink when they physically contact each other. They remain crosslinked even after resuspension in a liquid solution, so that the label no longer binds to the individual antibodies and causes false results. This can be particularly problematic in fields such as flow cytometry, which uses multiple polymer dyes to create multiple, e.g., ten or more, fluorescence channels by using only one wavelength for excitation. However, the chemical properties of these polymer dyes prevent the storage of multiple dyes in a pooled cocktail. Polymer entities have been observed to bind to each other in solution, which results in either a false or abnormal cell population that does not actually exist.

[0006] U.S. Patent No. 10,545,137 discloses a reagent device having a solid support and first and second dried polymer dye compositions that are precisely positioned relative to the surface of the solid support, whereby dye-dye interactions are reduced compared to reagent devices in which two or more dye compositions are provided but not precisely positioned relative to each other. When in use, the precisely positioned dye compositions can be reconstituted by introducing an aqueous liquid to contact the dye compositions. To achieve good assay performance, it is necessary to maintain the reagent concentration. Thus, the aqueous liquid introduced to contact the dye compositions can be a biological sample fluid to maintain the reagents and analytes of the sample at a desired concentration. SUMMARY OF THE INVENTION

[0007] However, in the device of U.S. Patent No. 10,545,137, although dye-dye interactions are reduced, the inventors recognize that the large amount of liquid required to reconstitute the precisely positioned dye composition can result in a reduction in dye concentration. This problem expands as the number of precisely positioned dye compositions increases, effectively limiting the number of dye compositions that can be used in the assays performed by the device.

[0008] For this reason, the inventors recognize that it is desirable to reconstitute the dye composition with a small amount of liquid.

[0009] The present disclosure is made in view of such circumstances, and its object is to provide a system for reconstituting a dry reagent composition that can reduce the volume of liquid required to reconstitute the dry reagent composition. Accordingly, embodiments of the present invention reduce the volume of liquid required to reconstitute or rehydrate a dry reagent composition.

[0010] A system for reconstituting a dry reagent composition is provided. Aspects of the system include a liquid container having an inner wall with a dry reagent composition, and a solid volume displacer positioned inside the liquid container and configured to occupy a majority of the liquid container volume below the top of the dry reagent composition. Aspects of the present invention further include a method of using the system and a kit including the system.

Brief Description of the Drawings

[0011]

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DETAILED DESCRIPTION OF THE INVENTION

[0012] A system for reconstituting a dry reagent composition is provided. Aspects of the system include a liquid container having an inner wall with the dry reagent composition, and a solid volume displacer positioned inside the liquid container and configured to occupy a majority of the liquid container volume below the upper portion of the dry reagent composition. Aspects of the present invention further include a method of using the system and a kit including the system.

[0013] Before the present invention is described in more detail, it is to be understood that the present invention is not limited to the specific embodiments described, and accordingly, of course, can vary. It is also to be understood that the terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting, as the scope of the present invention is defined only by the appended claims.

[0014] When a range of values is provided, unless the context clearly dictates otherwise, each intervening value, to the tenth of the unit of the lower limit, is understood to be included between the upper and lower limits of the range and any other stated value or intervening value within the stated range. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also included within the invention, subject to any specific excluded limitations within the scope of the description. When the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0015] A particular range is presented herein with the term "about" preceding the numerical value. As used herein, the term "about" is used to provide literal support for the exact number preceding it and for numbers that are close to or approximate the number preceding the term. In determining whether a number is close to or substantially the same as a specifically recited number, a number that is close to or substantially the same as an unrecited number may be a number that provides substantial equivalence to the specifically recited number in the context presented.

[0016] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, representative, exemplary methods and materials are described herein.

[0017] All publications and patents cited in this specification are hereby incorporated by reference as if each individual publication or patent was specifically and individually indicated to be incorporated by reference, and incorporated by reference herein to disclose and describe the methods and / or materials to which the publication is cited. Any citation of a publication is for its disclosure prior to the filing date of this application 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 the publications provided may be different from the actual publication dates which may need to be independently confirmed.

[0018] It should be noted that, as used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. It should further be noted that the claims may be drafted to exclude any optional element. Accordingly, this description is intended to serve as a basis for use of exclusive terms such as "solely" and "only" in connection with the recitation of claim elements, or for use of "negative" limitations.

[0019] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein can be readily separated from, or combined with, any of the features of some other embodiments without departing from the scope or spirit of the present invention. Any recited method can be performed in the order of recited events or in any other order that is logically possible.

[0020] The apparatus and method are described or are to be described in functional terms for grammatical fluidity, but the claims should not be construed as necessarily limited by a "means" or "step" limitation syntax unless expressly recited under 35 U.S.C. § 112, and should be accorded the full scope of the meaning ascribed by the claim and the equivalents thereof under the doctrine of judicial estoppel, and it should be clearly understood that where the claims are expressly recited under 35 U.S.C. § 112, full statutory equivalents under 35 U.S.C. § 112 should be accorded.

[0021] As summarized above, a system for reconstituting a dry reagent composition is provided. In further describing various embodiments of the present invention, the subject system will first be described in more detail. Next, methods of using the subject system, as well as kits containing the same, will be described.

[0022] System for Reconstituting a Dry Reagent Composition Embodiments of the present invention include a system for reconstituting a dry reagent composition that is optimized for the purpose of reconstituting or rehydrating a dry reagent composition that is ultimately used in an assay, such as an assay of a liquid sample. Aspects of the system include a liquid container having an inner wall with a dry reagent composition, and a solid volume displacer positioned inside the liquid container and configured to occupy a majority of the liquid container volume below the upper portion of the dry reagent composition. The system enables a controlled displacement of the liquid volume in a manner sufficient to reconstitute the dry reagent composition, thereby providing a number of advantages including, but not limited to, improved usability, cost, and performance.

[0023] Liquid Container As summarized above, the system of the present invention includes a liquid container. In certain embodiments, the liquid container is useful, for example, in an assay for the presence of one or more analytes in a sample, such as an assay of a liquid sample, such as a biological sample. A liquid container according to certain embodiments of the present disclosure includes an open end and a bottom separated by an intervening wall, the inner surface of the wall including a dry reagent composition.

[0024] The liquid container can be any convenient container that is compatible with a liquid sample and / or reagent or analyte that can contact the container. For example, the liquid container can be a liquid-compatible container configured to contain a liquid sample. In some cases, the liquid sample can be an aqueous liquid sample, and in these cases, the liquid container can be compatible with the aqueous sample. "Compatible" means that the liquid container (e.g., the material from which the container is made) is substantially inert (e.g., does not significantly react) with respect to the liquid and / or reagent or analyte in contact with the container.

[0025] The liquid container can include an open end and a bottom opposite the open end. In these cases, the open end of the liquid container exposes the interior of the liquid container to the ambient environment (e.g., through an opening) such that the contents of the liquid container are at the same atmospheric pressure as the ambient environment. The longest cross-sectional dimension of the liquid container, e.g., the diameter, can vary and in some cases is in the range of 0.5 cm to 5 cm, e.g., 0.5 cm to 4.5 cm, or 0.5 cm to 4 cm, or 0.5 cm to 3.5 cm, or 0.5 cm to 3 cm, or 0.5 cm to 2.5 cm, or 0.5 cm to 2 cm, or 0.5 cm to 1.5 cm, or 0.5 cm to 1.2 cm, or 0.9 cm to 1.0 cm. In some embodiments, the open end of the liquid container can include a protrusion such that, for example, a snap-on cap can be applied to the open end. In other embodiments, the open end of the liquid container can include a thread such that, for example, a threaded cap can be screwed onto the opening of the open end.

[0026] In some cases, the bottom of the liquid container facing the open end may include a closed end from which a wall protrudes. Thus, the liquid container may be configured to hold a specific volume of fluid (e.g., gas or liquid). In certain embodiments, the liquid container is configured to hold a specific volume of liquid. The size of the liquid container may depend on the volume of liquid to be held in the liquid container. For example, the liquid container may be configured to hold a volume (e.g., liquid volume) in the range of 0.1 nl to 1000 ml, such as 0.1 ml to 900 ml, or 0.1 ml to 800 ml, or 0.1 ml to 700 ml, or 0.1 ml to 600 ml, or 0.1 ml to 500 ml, or 0.1 ml to 400 ml, or 0.1 ml to 300 ml, or 0.1 ml to 200 ml, or 0.1 ml to 100 ml, or 0.1 ml to 50 ml, or 0.1 ml to 25 ml, or 0.1 ml to 10 ml, or 0.1 ml to 5 ml, or 0.1 ml to 1.5 ml, or 0.1 ml to 1 ml, or 0.1 ml to 0.5 ml. In certain cases, the liquid container is configured to hold a volume (e.g., liquid volume) in the range of 0.1 ml to 200 ml, such as 0.5 ml to 100 ml, such as 1.0 ml to 50 ml.

[0027] The shape of the liquid container may vary and may depend on the use of the container. For example, as described herein, the liquid container may find use in an assay such as an assay of a liquid sample (e.g., a biological sample). In these cases, the liquid container may be configured in a shape that is compatible with the assay and / or with the method or other device used to perform the assay. For example, the liquid container may be in the shape of typical laboratory equipment used to perform the assay or in a shape that is compatible with other devices used to perform the assay. In certain embodiments, the walls of the liquid container are cylindrical in shape, the cylinder having an opening at a first end and a closed bottom joined to the cylinder at a second end opposite the first end. In some cases, the liquid container has a circular cross-section. The shape of the liquid container may vary and may depend on the use of a solid volume displacer. For example, the liquid container may be configured in a shape that is compatible with the methods of the present disclosure. In some embodiments, the walls of the liquid container are tapered. In some cases, the liquid container may be conical or may include a conical section. For example, the liquid container may include a substantially cylindrical wall section adjacent to the opening at the first end and a conical wall section adjacent to the second closed end opposite the first end. In some embodiments, the bottom of the liquid container is rounded. In some embodiments, the liquid container may be a vial, a test tube, or a centrifuge tube (e.g., a microcentrifuge tube). In certain cases, the liquid container is a vial. In certain cases, the liquid container is a test tube. As previously described, the liquid container may be configured to hold a volume (e.g., a volume of liquid). In embodiments where the liquid container is a vial or a test tube, the liquid container is 1×10 -7It can be configured to hold a volume (e.g., the volume of a liquid) in the range of ml to 1000 ml, for example, 0.5 ml to 900 ml, or 0.5 ml to 800 ml, or 0.5 ml to 700 ml, or 0.5 ml to 600 ml, or 0.5 ml to 500 ml, or 0.5 ml to 400 ml, or 0.5 ml to 300 ml, or 0.5 ml to 200 ml, or 0.5 ml to 100 ml, or 0.5 ml to 50 ml, or 0.5 ml to 25 ml, or 0.5 ml to 10 ml, or 0.5 ml to 5 ml, or 1 ml to 5 ml. In certain cases, the vial or test tube is configured to hold a volume (e.g., the volume of a liquid) in the range of 0.5 ml to 5 ml.

[0028] In certain embodiments, the liquid container has a width (which can also be referred to as the diameter of a cylindrical liquid container) in the range of 0.5 cm to 5 cm, for example, 0.5 cm to 4.5 cm, or 0.5 cm to 4 cm, or 0.5 cm to 3.5 cm, or 0.5 cm to 3 cm, or 0.5 cm to 2.5 cm, or 0.5 cm to 2 cm, or 0.5 cm to 1.5 cm, or 0.5 cm to 1.2 cm, or 0.9 cm to 1.0 cm. In some cases, the liquid container has a width (or diameter) in the range of 0.5 cm to 2.5 cm. In some cases, the liquid container has a width (or diameter) in the range of 0.5 cm to 1.2 cm, for example, 0.9 cm to 1.0 cm. In certain embodiments, the liquid container has a length in the range of 1 cm to 20 cm, for example, 1 cm to 15 cm, or 1 cm to 10 cm, or 1 cm to 5 cm, or 1 cm to 2.5 cm. In some cases, the liquid container has a length in the range of 1 cm to 10 cm. In some cases, the liquid container has a length in the range of 1 cm to 5 cm. In some cases, the liquid container has a length in the range of 1 cm to 2.5 cm.

[0029] As described above, embodiments of the liquid container may be compatible with liquid samples and / or reagents or analytes that contact the container. Examples of suitable container materials include, but are not limited to, ceramics, metals (e.g., stainless steel, aluminum, anodized aluminum, titanium, copper, bronze, nickel, etc.), resins, paper, glass, and plastics. For example, the container may be composed of, but is not limited to, glasses such as silicate glass, borosilicate glass, sodium borosilicate glass (e.g., PYREX (trademark)), fused quartz glass, fused silica glass, etc. Other examples of suitable container materials include polymeric materials. The polymeric materials can be hydrophobic polymeric materials. Hydrophobic polymeric materials can be plastics such as polystyrene, polypropylene, polymethylpentene, polytetrafluoroethylene (PTFE), perfluoroether (PFE), fluorinated ethylene propylene (FEP), perfluoroalkoxy alkane (PFA), polyethylene terephthalate (PET), polyethylene (PE), low density polyethylene (LDPE), polyether ether ketone (PEEK), etc., but are not limited thereto.

[0030] In some embodiments, as described above, the liquid container is configured to hold a specific volume of fluid (e.g., gas or liquid). In some cases, the liquid container is configured to hold a specific volume of liquid. In some embodiments, the liquid container may be sealed. That is, the liquid container may include a seal that substantially prevents the contents of the liquid container (e.g., the liquid within the liquid container) from exiting the liquid container. The seal of the liquid container may also substantially prevent other substances from entering the liquid container. For example, the seal may be a watertight seal that substantially prevents liquid from entering or exiting the container, or it may be an airtight seal that substantially prevents gas from entering or exiting the container. In some cases, the seal is a removable or breakable seal, so that the contents of the liquid container can be exposed to the surrounding environment as needed, for example, when it is desirable to remove a portion of the contents of the liquid container. In some cases, the seal is made of an elastic material to provide a barrier (e.g., a watertight and / or airtight seal) for holding a sample within the container. Specific types of seals include, but are not limited to, films such as polymer films, caps, etc., depending on the type of container. Suitable materials for the seal include, but are not limited to, rubber or polymer seals such as silicone rubber, natural rubber, styrene-butadiene rubber, ethylene-propylene copolymer, polychloroprene, polyacrylate, polybutadiene, polyurethane, styrene-butadiene, and combinations thereof. For example, in certain embodiments, the seal is a septum that can be penetrated by a needle, syringe, or cannula. The seal can also provide for convenient access to the sample within the liquid container and a protective barrier covering the opening of the container. In some cases, the seal is a removable seal such as a threaded or snap-on cap or other suitable sealing element that can be applied to the opening of the liquid container. For example, a threaded cap can be screwed onto the opening before or after adding a sample to the liquid container.

[0031] As described above, the liquid container can be configured to hold a fluid (e.g., gas or liquid) of a certain specific volume. In some cases, the bottom of the liquid container has an inner surface and an outer surface. In some cases, the walls of the liquid container have an inner surface and an outer surface. In these embodiments, the inner surface of the liquid container is the surface of the liquid container that faces the inside of the container. The inner surface can contact the contents of the liquid container. The outer surface of the liquid container is the surface of the container that faces away from the inside of the container. The outer surface does not contact the contents of the liquid container.

[0032] As discussed previously, the liquid container may contain a dry reagent composition. In certain embodiments, the dry reagent composition is positioned on the surface (e.g., the inner surface) of the liquid container. A reagent means a substance or compound that interacts with other substances or compounds, for example, to facilitate a chemical reaction or to test whether a reaction has occurred. For example, the substance or compound may include, but is not limited to, beads, preservatives, surfactants, defoamers, buffers (e.g., pH buffers), reactants, specific binding members (e.g., antibodies), dyes, and the like. Dry means that the reagent composition contains a small amount of solvent (i.e., substantially no solvent, for example), or that the reagent composition can reduce its viscosity by combining with the volume of the solvent. For example, in some embodiments, the dry reagent composition can be a liquid having a relatively high viscosity (e.g., syrup), and by adding a volume of solvent (e.g., water), under standard conditions, the viscosity of the reagent composition can be reduced by 10 cP or more, or 100 cP or more, or 500 cP or more, or 1,000 cP or more, or 10,000 cP or more, or 100,000 cP or more, or 250,000 cP or more, or 1,000,000 cP or more. In certain embodiments, the dry reagent composition contains 25 wt% or less of solvent, for example, 20 wt% or less, or 15 wt% or less, or 10 wt% or less, or 5 wt% or less, or 3 wt% or less, or 1 wt% or less, or 0.5 wt% or less of solvent. In some cases, the dry reagent composition is not a fluid. In some cases, the dry reagent composition is substantially solid. For example, the dry reagent composition can have a high viscosity such as 10,000 cP or more, or 25,000 cP or more, or 50,000 cP or more, or 75,000 cP or more, or 100,000 cP or more, or 150,000 cP or more, or 200,000 cP or more, or 250,000 cP or more under standard conditions. In some embodiments, the dry reagent can be dissolved, either partially or completely, in the volume of the solvent. For example, the dry reagent composition can be rehydrated or reconstituted in water.In some embodiments, the dry reagent composition can maintain its shape and position (e.g., on the inner surface of the liquid container to which the composition is attached, as discussed in more detail below) before it is contacted by a solvent.

[0033] In some cases, the dry reagent composition is positioned on the inner surface of the wall of the liquid container. The liquid container may contain one or more dry reagent compositions (e.g., dye compositions) on the container surface such as the inner surface of the wall of the liquid container. For example, on the inner surface of the wall of the liquid container, there may be two or more dry reagent compositions, or three or more, or four or more, or five or more, or six or more, or seven or more, or eight or more, or nine or more, or ten or more, or eleven or more, or twelve or more, or thirteen or more, or fourteen or more, or fifteen or more, sixteen or more, or seventeen or more, or eighteen or more, or nineteen or more, or twenty or more, or twenty-five or more, or thirty or more, or thirty-five or more, or forty or more, or forty-five or more, or fifty or more dry reagent compositions. In some embodiments, the container contains from 2 to 50 dry reagent compositions on its inner surface. For example, on the inner surface of the wall, it contains from 2 to 40, or 2 to 30, or 2 to 20, or 2 to 15, or 2 to 10, or 2 to 7, or 2 to 5 dry reagent compositions. For example, the container may contain 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or 10, or 11, or 12, or 13, or 14, or 15, or 16, or 17, or 18, or 19, or 20 dry reagent compositions on the inner surface of the wall of the liquid container. In certain cases, the liquid container contains 2 dry reagent compositions on the inner surface of the wall of the liquid container. In certain cases, the liquid container contains 5 dry reagent compositions on the inner surface of the wall of the liquid container. In certain cases, the liquid container contains 7 dry reagent compositions on the inner surface of the wall of the liquid container. In certain cases, the liquid container contains 10 dry reagent compositions on the inner surface of the wall of the liquid container. In certain cases, the liquid container contains 15 dry reagent compositions on the inner surface of the wall of the liquid container.

[0034] As described above, the container can include two or more dry reagent compositions positioned relative to the surface of the container (e.g., the inner surface of the wall of the liquid container). In some cases, the wall of the liquid container can include a second dry reagent composition positioned on its inner surface. In some cases, the first and second, e.g., two or more, dry reagent compositions are adhered to the inner surface of the wall of the liquid container. By "adhered" is meant that the dry reagent compositions are stably bound to the location on the inner surface where they are positioned such that they do not move from that location when they are in the dry state. Upon reconstitution, the reagents of the dry reagent compositions, e.g., dyes, are dissociated from the location on the surface where they are positioned and thus are present in the liquid used to reconstitute the dry reagent compositions. The dry reagent compositions can be disposed at different locations on the inner surface of the wall of the liquid container. For example, the first and second dry reagent compositions can be clearly positioned on the inner surface of the wall of the liquid container. By "clear position" or "clearly positioned" is meant that the dry reagent composition is disposed at a position different from the position of another dry reagent composition. The position of the dry reagent composition can refer to the location of the dry reagent composition on the surface of the liquid container and / or to the position of the dry reagent composition relative to the surface of the liquid container. In some cases, the dry reagent composition occupies a defined volume of space. For example, the dry reagent composition can occupy the volume of space on the surface of the liquid container. A clearly positioned dry reagent composition can occupy a volume of space that does not significantly coincide or overlap with the volume of space occupied by another dry reagent composition, and in some cases, this is a volume of space that does not coincide or overlap at all with the volume of space occupied by another dry reagent composition. Embodiments in which the dry reagent compositions are clearly positioned can provide for minimization of the interaction between each of the dry reagent compositions, e.g., dye-dye interaction.

[0035] Put another way, a precisely positioned dry reagent composition is not significantly mixed with another dry reagent composition (e.g., a polymer dye composition). For example, substantially no part of the precisely positioned dry reagent composition is mixed with a part of another dry reagent composition (e.g., a polymer dye composition). In some cases, the precisely positioned dry reagent composition is not mixed with another dry reagent composition (e.g., a polymer dye composition). For example, no part of the precisely positioned dry reagent composition is mixed with a part of another dry reagent composition (e.g., a polymer dye composition). In certain embodiments, the precisely positioned dry reagent composition contains a single reagent. For example, the precisely positioned dry reagent composition may be substantially composed of a single reagent and contain no significant amount of other reagents. The precisely positioned dry reagent composition may contain an excess of reagent relative to other reagents that may be present in the dry reagent composition. For example, it may contain 75 wt% or more, such as 80 wt% or more, or 85 wt% or more, or 90 wt% or more, or 95 wt% or more, or 97 wt% or more, or 99 wt% or more, or 100 wt% of the reagent relative to other reagents that may be present in the dry reagent composition. In certain embodiments, the precisely positioned dry reagent composition contains a single dye. For example, the precisely positioned dry reagent composition may be substantially composed of a single dye and contain no significant amount of other dyes. The precisely positioned dry reagent composition may contain an excess of dye relative to other dyes that may be present in the dry reagent composition. For example, it may contain 75 wt% or more, such as 80 wt% or more, or 85 wt% or more, or 90 wt% or more, or 95 wt% or more, or 97 wt% or more, or 99 wt% or more, or 100 wt% of the dye relative to other dyes that may be present in the dry reagent composition. In some cases, the dry reagent composition contains two or more reagents. In some cases, the first and / or second dry reagent composition contains two or more reagents, such as three or more reagents, four or more reagents, five or more reagents, six or more reagents, seven or more reagents, eight or more reagents, nine or more reagents, or ten or more reagents. In some cases, the dry reagent composition contains two or more dyes.In some cases, the first and / or second dry reagent composition comprises two or more dyes, for example, three or more dyes, four or more dyes, five or more dyes, six or more dyes, seven or more dyes, eight or more dyes, nine or more dyes, or ten or more dyes.

[0036] In some cases, the clearly positioned dry reagent composition is spaced from the bottom of the liquid container. The dry reagent composition spaced from the bottom of the liquid container can be physically separated from the bottom of the liquid container. For example, the clearly positioned dry reagent composition is at a distance from the bottom of the liquid container such that there is a specific distance between the edge of the dry reagent composition closest to the bottom of the liquid container and the edge of the bottom of the liquid container closest to the dry reagent composition, and can be positioned on the inner surface of the wall of the liquid container. The edge of the bottom of the liquid container can be any point where the bottom of the liquid container contacts the wall of the liquid container. In some embodiments, the bottom of the liquid container is considered to contact the wall of the liquid container, and the wall of the liquid container no longer has a specific shape or is no longer continuous at a specific angle. In an embodiment where the wall of the liquid container is in the shape of a cylinder (e.g., a tapered cylinder) and the bottom of the liquid container is round, the bottom of the liquid container can be considered to contact the wall of the liquid container at this location, and the opposing horizontal ends of the wall of the liquid container are no longer extending parallel to each other or are no longer extending substantially parallel to each other in an embodiment where the cylinder is tapered (i.e., the liquid container is no longer a cylinder and instead resembles, for example, a hemisphere). In some embodiments, the distance between the dry reagent composition on the inner surface of the wall of the liquid container and the bottom of the liquid container is 0.1 mm or more, for example, 0.5 mm or more, or 1 mm or more, or 2 mm or more, or 3 mm or more, or 4 mm or more, or 5 mm or more, or 6 mm or more, or 7 mm or more, or 8 mm or more, or 9 mm or more, or 10 mm or more, or 12 mm or more, or 14 mm or more, or 16 mm or more, or 18 mm or more, or 20 mm or more, or 25 mm or more, or 30 mm or more, or 35 mm or more, or 40 mm or more, or 50 mm or more, or 60 mm or more, or 70 mm or more, or 80 mm or more, or 90 mm or more, or 100 mm or more, or 110 mm or more, or 120 mm or more, or 130 mm or more, or 140 mm or more, or 150 mm or more, or 160 mm or more, or 170 mm or more, or 180 mm or more, or 190 mm or more, or 200 mm or more.For example, the distance between the dry reagent composition on the inner surface of the wall of the liquid container and the bottom of the liquid container can range from 0.1 mm to 200 mm, for example, from 0.1 mm to 190 mm, or from 0.1 mm to 180 mm, or from 0.1 mm to 170 mm, or from 0.1 mm to 160 mm, or from 0.1 mm to 150 mm, or from 0.1 mm to 140 mm, or from 0.1 mm to 130 mm, or from 0.1 mm to 120 mm, or from 0.1 mm to 110 mm, or from 0.1 mm to 100 mm, or from 0.1 mm to 90 mm, or from 0.1 mm to 80 mm, or from 0.1 mm to 70 mm, or from 0.1 mm to 60 mm, or from 0.1 mm to 50 mm, or from 0.1 mm to 40 mm, or from 0.1 mm to 30 mm, or from 0.1 mm to 20 mm, or from 0.1 mm to 10 mm, or from 0.1 mm to 9 mm, or from 0.1 mm to 8 mm, or from 0.1 mm to 7 mm, or from 0.1 mm to 6 mm, or from 0.1 mm to 5 mm, or from 0.1 mm to 4 mm, or from 0.1 mm to 3 mm, or from 0.1 mm to 2 mm, or from 0.1 mm to 1 mm, or from 0.1 mm to 0.5 mm. In certain cases, the distance between the dry reagent composition on the inner surface of the wall of the liquid container and the bottom of the liquid container ranges from 0.1 mm to 200 mm. In some cases, the distance between the dry reagent composition on the inner surface of the wall of the liquid container and the bottom of the liquid container ranges from 0.1 mm to 10 mm.

[0037] In some cases, the clearly positioned dry reagent compositions are separated from each other at separate locations on the surface of the liquid container. A dry reagent composition separated from another dry reagent composition can be physically separated from the adjacent dry reagent composition. For example, the clearly positioned dry reagent compositions can be positioned on the surface of the liquid container at separate locations such that there is a specific distance between the edge of the dry reagent composition and the edge of the adjacent dry reagent composition. In some embodiments, the distance between separate locations of the dry reagent compositions on the surface of the liquid container is 0.1 mm or more, for example, 0.5 mm or more, or 1 mm or more, or 2 mm or more, or 3 mm or more, or 4 mm or more, or 5 mm or more, or 6 mm or more, or 7 mm or more, or 8 mm or more, or 9 mm or more, or 10 mm or more, or 12 mm or more, or 14 mm or more, or 16 mm or more, or 18 mm or more, or 20 mm or more, or 25 mm or more, or 30 mm or more, or 35 mm or more, or 40 mm or more, or 50 mm or more, or 60 mm or more, or 70 mm or more, or 80 mm or more, or 90 mm or more, or 100 mm or more, or 110 mm or more, or 120 mm or more, or 130 mm or more, or 140 mm or more, or 150 mm or more, or 160 mm or more, or 170 mm or more, or 180 mm or more, or 190 mm or more, or 200 mm or more.For example, the distance between separate locations of the dry reagent composition on the surface of the liquid container can range from 0.1 mm to 200 mm, such as from 0.1 mm to 190 mm, or from 0.1 mm to 180 mm, or from 0.1 mm to 170 mm, or from 0.1 mm to 160 mm, or from 0.1 mm to 150 mm, or from 0.1 mm to 140 mm, or from 0.1 mm to 130 mm, or from 0.1 mm to 120 mm, or from 0.1 mm to 110 mm, or from 0.1 mm to 100 mm, or from 0.1 mm to 90 mm, or from 0.1 mm to 80 mm, or from 0.1 mm to 70 mm, or from 0.1 mm to 60 mm, or from 0.1 mm to 50 mm, or from 0.1 mm to 40 mm, or from 0.1 mm to 30 mm, or from 0.1 mm to 20 mm, or from 0.1 mm to 10 mm, or from 0.1 mm to 9 mm, or from 0.1 mm to 8 mm, or from 0.1 mm to 7 mm, or from 0.1 mm to 6 mm, or from 0.1 mm to 5 mm, or from 0.1 mm to 4 mm, or from 0.1 mm to 3 mm, or from 0.1 mm to 2 mm, or from 0.1 mm to 1 mm, or from 0.1 mm to 0.5 mm. In certain cases, the distance between separate locations of the dry reagent composition on the surface of the liquid container ranges from 0.1 mm to 200 mm. In some cases, the distance between separate locations of the dry reagent composition on the surface of the liquid container ranges from 0.1 mm to 10 mm.

[0038] In certain embodiments, the clearly positioned dry reagent compositions are positioned adjacent to each other on the inner surface of the wall of the liquid container but are not separated from each other. In these cases, the edges of the dry reagent compositions can contact the edges of adjacent dry reagent compositions. For example, the volume of the space occupied by a dry reagent composition can contact the volume of the space occupied by another (adjacent) dry reagent composition but does not significantly overlap. In these embodiments, adjacent dry reagent compositions can contact each other but are not significantly mixed together. For example, substantially no part of a clearly positioned dry reagent composition is mixed with a part of another (adjacent) dry reagent composition.

[0039] The clearly positioned dry reagent composition may be present on the same surface of the liquid container (e.g., the inner surface of the wall of the liquid container), but may be disposed at a different position on or relative to the surface of the liquid container (e.g., the inner surface of the wall of the liquid container). For example, as described above, the liquid container may include an inner surface and an outer surface. In certain instances, the dry reagent composition is positioned on the surface of the liquid container (e.g., the inner surface of the wall of the liquid container). In some cases, the dry reagent composition is clearly positioned on the inner surface of the liquid container (e.g., the inner surface of the wall of the liquid container).

[0040] Examples of clearly positioned dry reagent compositions include embodiments where the dry reagent composition is disposed at a particular location on the surface of the liquid container (e.g., the inner surface of the wall of the liquid container) and another dry reagent composition is also disposed at a different location on the surface of the container. Thus, the clearly positioned dry reagent compositions can be positioned at separate locations on the surface of the liquid container. For example, an embodiment of the container may include first and second dry reagent compositions, where the first dry reagent composition is positioned at a particular location on the inner surface of the wall of the liquid container and the second dry reagent composition is positioned at a different location on the inner surface of the wall of the liquid container than the first dry reagent composition. As described above, the first and second dry reagent compositions can be spaced apart from each other such that there is a distance between the separate locations of the first and second dry reagent compositions on the surface of the liquid container (e.g., the inner surface of the wall of the liquid container). The distance between the first dry reagent composition and the second dry reagent composition can be according to the ranges and values described above.

[0041] The dry reagent composition can have any suitable dimensions. In some cases, the first and second dry reagent compositions have the same dimensions. In certain embodiments, the first and second dry reagent compositions include different dimensions. The dry reagent composition can have any suitable cross-sectional shape, including, for example, a linear cross-sectional shape such as a square, rectangle, trapezoid, triangle, hexagon, etc., a curved cross-sectional shape such as a circle, ellipse, and an irregular shape such as a parabolic shape. In some cases, the first and second dry reagent compositions have a diameter in the range of 0.01 mm to 5 mm, including, for example, 0.01 mm to 4 mm, 0.01 mm to 3 mm, 0.01 to 2 mm, 0.01 to 1 mm, 0.1 mm to 5 mm, 0.1 mm to 4 mm, 0.1 mm to 3 mm, 0.1 to 2 mm, 0.1 to 1 mm, 0.5 mm to 5 mm, 0.5 mm to 4 mm, 0.5 mm to 3 mm, 0.5 to 2 mm, 0.5 to 1 mm. In some cases, the first and second dry reagent compositions have a surface area in the range of 5×10 -5 mm 2 ~20 mm 2 、5×10 -3 mm 2 ~20 mm 2 、0.1 mm 2 ~20 mm 2 、5×10 -5 mm 2 ~0.5 mm 2 、5×10 -3 mm 2 ~0.5 mm 2 、0.1 mm 2 ~0.5 mm 2 including 5×10 -5 mm 2 ~20 mm 2 .

[0042] Additional drying reagent compositions can be provided on the inner surface of the liquid container (e.g., the inner surface of the wall of the liquid container). For example, the container may include a third drying reagent composition (e.g., a dye composition) that is clearly positioned on the inner surface of the wall of the liquid container. The third drying reagent composition can be clearly positioned relative to the first drying reagent composition and can also be clearly positioned relative to the second drying reagent composition. Thus, as described herein, each of the drying reagent compositions (e.g., the first, second, and third drying reagent compositions) can be clearly positioned relative to each other on the inner surface of the wall of the liquid container. As previously described, additional clearly positioned drying reagent compositions, e.g., four or more, or five or more, seven or more, ten or more, etc., clearly positioned drying reagent compositions can be provided on the surface of the liquid container. In some instances, the drying reagent composition is adhered to the inner surface of the wall of the liquid container.

[0043] Additional examples of the precisely positioned dry reagent compositions include embodiments in which the dry reagent composition is disposed on the surface of the liquid container (e.g., the inner surface of the wall of the liquid container) at a particular location and another dry reagent composition is disposed at the same location. Thus, the precisely positioned dry reagent compositions can be co-located at the same location on the surface of the liquid container (e.g., the inner surface of the wall of the liquid container). The dry reagent compositions may be co-located at the same location but can still be precisely positioned. For example, the dry reagent compositions can be separated from each other by a non-dye material. In some cases, the non-dye material is sandwiched between the precisely positioned dry reagent compositions. The non-dye material can substantially cover the surface of the dry reagent composition such that adjacent dry reagent compositions are separated from the dry reagent composition. For example, a dry reagent composition can have a non-dye material disposed on its surface and another dry reagent composition can be disposed on the surface of the non-dye material. In these instances, the dry reagent compositions can be physically separated from other dry reagent compositions by the non-dye material. In some cases, the precisely positioned dry reagent compositions can be provided as alternating layers of dry reagent composition and non-dye material on the surface of the container. Thus, in a particular embodiment, two or more dry reagent compositions are precisely positioned relative to each other and co-located at the same location on the surface of the liquid container. In some embodiments, the non-dye material (e.g., non-reagent material) can be positioned within the liquid container and spaced apart from the precisely positioned dry reagent compositions such that there is a distance between the non-dye material and the one or more dry reagent compositions.

[0044] In certain embodiments, the non-dye material is a material that is compatible with other assay components (e.g., reagents, buffers, analytes, etc.) that may be present in the liquid container during use. The non-dye material can be substantially inert with respect to other assay components (e.g., reagents, buffers, analytes, etc.) that may be present in the container during use so that there is no significant reaction between the non-dye material and the other assay components. Examples of non-dye materials include, but are not limited to, any of the reagents, non-reagent materials, stabilizers, buffers, soluble inert materials (e.g., water-soluble inert materials), beads, etc. discussed above. Exemplary stabilizers include, but are not limited to, sugars and polyalcohols. Sugars and polyalcohols suitable for use in lyophilized dye compositions include sugars that are compatible with other reagents, buffers, dyes, and sample components used. Examples of suitable sugars include, but are not limited to, sucrose, maltose, trehalose, 2-hydroxypropyl-β-cyclodextrin (β-HPCD), lactose, glucose, fructose, galactose, glucosamine, etc., and combinations thereof. In certain instances, the sugar is a disaccharide. For example, the disaccharide can be sucrose. Examples of suitable polyalcohols include, but are not limited to, mannitol, glycerol, erythritol, threitol, xylitol, sorbitol, etc., and combinations thereof. The non-dye material can include, for example, bovine serum albumin (BSA), sodium azide, glycerol, phenylmethylsulfonyl fluoride (PMSF), ethylenediaminetetraacetic acid (EDTA), buffered citrate, phosphate buffered saline (PBS), sodium chloride, paraformaldehyde, etc., and combinations thereof.

[0045] For example, embodiments of the liquid container may include first and second dry reagent compositions, where the first dry reagent composition is positioned at a particular location on the surface of the liquid container (e.g., the inner surface of the wall of the liquid container), and the second dry reagent composition is co-located with the first dry reagent composition at the same location. As described above, the first dry reagent composition and the second dry reagent composition may be spaced apart from each other such that there is a distance between the first dry reagent composition and the second dry reagent composition. For example, as described above, the first and second dry reagent compositions may be separated from each other by a non-dye material. The distance between the first dry reagent composition and the second dry reagent composition may be according to the ranges and values described above. For example, the non-dye material may be sandwiched between the clearly positioned first and second dry reagent compositions. In these embodiments, the first dry reagent composition may be positioned on the surface of the liquid container, the non-dye material may be disposed as a layer on the surface of the first dry reagent composition, and the second dry reagent composition may be disposed on the surface of the non-dye composition. In these cases, the first dry reagent composition may be physically separated from the second dry reagent composition by the non-dye material. Thus, in certain embodiments, the first and second dry reagent compositions are clearly positioned relative to each other and are co-located at the same location on the surface of the liquid container. For example, the layer of non-dye material on the surface of the first dry reagent composition may substantially cover the entire surface of the first dry reagent composition. In these cases, the second dry reagent composition disposed on the surface of the non-dye composition may not be in significant contact with the first dry reagent composition. In some cases, the non-dye material is a substantially continuous layer of non-dye material on the surface of the first dry reagent composition. For example, the non-dye material may cover a significant portion of the surface of the first dry reagent composition, such as 75% or more of the surface of the first dry reagent composition, or 80% or more, or 85% or more, or 90% or more, or 95% or more, or 97% or more, or 99% or more of the surface of the first dry reagent composition. Embodiments in which the surface of the first dry reagent composition is substantially covered by the non-dye material may provide for minimization of dye-dye interactions between the first dry reagent composition and the second dry reagent composition.

[0046] In certain embodiments, the non-dye material has a thickness in the range of 0.01 mm to 5 mm, such as 0.05 mm to 5 mm, or 0.1 mm to 5 mm, or 0.1 mm to 4 mm, or 0.1 mm to 3 mm, or 0.1 mm to 2 mm, or 0.1 mm to 1 mm, or 0.1 mm to 0.9 mm, or 0.1 mm to 0.8 mm, or 0.1 mm to 0.7 mm, or 0.1 mm to 0.6 mm, or 0.1 mm to 0.5 mm. In certain cases, the non-dye material has a thickness of 0.1 mm to 1 mm. In some cases, the non-dye material has a thickness of 0.1 mm to 0.05 mm.

[0047] Additional dry reagent compositions may also be provided. For example, the liquid container may include a third dry reagent composition that is clearly positioned relative to the first and second dry reagent compositions. Thus, the third dry reagent composition can be clearly positioned relative to the first dry reagent composition and also clearly positioned relative to the second dry reagent composition. Thus, as described herein, each of the dry reagent compositions (e.g., the first, second, and third dry reagent compositions) can be clearly positioned relative to each other. In some cases, each of the dry reagent compositions can be separated from each other by a non-dye material. For example, each of the dry reagent compositions can be separated from each other by a non-dye material. In some cases, the non-dye material is sandwiched between each of the clearly positioned dry reagent compositions. In certain cases, each of the clearly positioned dry reagent compositions is provided as a layer having a layer of non-dye material between each of the clearly positioned dry reagent compositions. As described above, additional layers of clearly positioned dry reagent compositions may be provided, such as four or more, or five or more, seven or more, ten or more, etc. of clearly positioned dry reagent compositions. Thus, the plurality of dry reagent compositions can be clearly positioned relative to each other and co-located at the same location on the surface of the liquid container (e.g., the inner surface of the wall of the liquid container).

[0048] In certain embodiments, the dry reagent composition on the surface of the liquid container is a dry dye composition that includes, for example, a dye. The dry dye composition is a dye composition that includes a small amount of a solvent. For example, the dry dye composition may include a low amount of a liquid such as water. In some cases, the dry dye composition is substantially free of a solvent. For example, the dry dye composition may be substantially free of a liquid such as water. In certain embodiments, the dry dye composition includes 25 wt% or less, for example, 20 wt% or less, or 15 wt% or less, or 10 wt% or less, or 5 wt% or less, or 3 wt% or less, or 1 wt% or less, or 0.5 wt% or less of a solvent. In some cases, the dry dye composition is not a fluid. In some cases, the dry dye composition is substantially solid. For example, the dry dye composition may have a high viscosity such as 10,000 cP or more, or 25,000 cP or more, or 50,000 cP or more, or 75,000 cP or more, or 100,000 cP or more, or 150,000 cP or more, or 200,000 cP or more, or 250,000 cP or more under standard conditions.

[0049] In some cases, the dye composition is a lyophilized dye composition. In certain cases, the lyophilized dye composition is a dye composition from which water has been removed by sublimation, and the water in the dye composition undergoes a phase transition from solid to gas. For example, the lyophilized dye composition can be a dye composition in which the dye composition is frozen (e.g., the water in the dye composition is frozen), and then the pressure around the dye composition is reduced so that the water in the dye composition undergoes sublimation and is removed from the composition. In certain cases, the lyophilized dye composition contains a low amount of water, such as 25% or less, or 20% or less, or 15% or less, or 10% or less, or 9% or less, or 8% or less, or 7% or less, or 6% or less, or 5% or less, or 4% or less, or 3% or less, or 2% or less, or 1% or less, or 0.5% or less, or 0.25% or less, or 0.1% or less, as measured by Karl Fischer (KF) titration. In some cases, the lyophilized dye composition has 3% or less water as measured by Karl Fischer titration. In some cases, the lyophilized dye composition has 1% or less water as measured by Karl Fischer titration. In some cases, the lyophilized dye composition has 0.5% or less water as measured by Karl Fischer titration. The lyophilized dye composition can contain additives such as stabilizers and / or excipients. In some cases, the lyophilized dye composition contains a stabilizer such as a sugar or a polyalcohol. Sugars and polyalcohols suitable for use in the lyophilized dye composition include sugars that are compatible with other reagents, buffers, dyes, and sample components used. Examples of suitable sugars include sucrose, maltose, trehalose, 2-hydroxypropyl-beta-cyclodextrin (β-HPCD), lactose, glucose, fructose, galactose, glucosamine, etc., and combinations thereof, but are not limited thereto. In certain cases, the sugar is a disaccharide. For example, the disaccharide can be sucrose. Examples of suitable polyalcohols include mannitol, glycerol, erythritol, threitol, xylitol, sorbitol, etc., and combinations thereof, but are not limited thereto.

[0050] The dyes in the dye composition can be used as detectable labels. In certain cases, the dye includes a detectable moiety or marker that is detectable based on, for example, fluorescence emission maximum, fluorescence polarization, fluorescence lifetime, light scattering, mass, molecular weight, or a combination thereof. In certain embodiments, the detectable label is a fluorophore (i.e., a fluorescent label, a fluorescent dye, etc.). The fluorophores of interest can include, but are not limited to, dyes suitable for use in analytical applications (e.g., flow cytometry, imaging, etc.).

[0051] In some cases, the fluorophore is a polymeric dye. In some cases of the method, the polymeric dye includes a conjugated polymer. A conjugated polymer (CP) is characterized by a delocalized electronic structure that includes a backbone of alternating unsaturated bonds (e.g., double bonds and / or triple bonds) and saturated (e.g., single bonds) bonds, and the π electrons can move from one bond to the other. Thus, the conjugated backbone can restrict the bond angles between the repeating units of the polymer, imparting an extended linear structure to the polymeric dye. For example, proteins and nucleic acids are also macromolecules, but in some cases, they do not form an extended rod structure but rather fold into a higher-order three-dimensional shape. In addition to this, the CP can form a "rigid rod" polymer backbone, with the torsional (e.g., twist) angle between monomer repeating units along the polymer backbone chain being restricted. In some cases, the polymeric dye includes a CP having a rigid rod structure. The structural features of the polymeric dye can affect the fluorescence properties of the molecule.

[0052] The polymer dyes of interest include, but are not limited to, U.S. Patent Publications 20040142344, 20080293164, 20080064042, 20100136702, 20110256549, 20110257374, 20120028828, 20120252986, 20130190193, 20160264737, 20160266131, 20180231530, 20180009990, 20180009989, and 20180163054, the entire disclosures of which are incorporated herein by reference, and Gaylord et al., J. Am. Chem. Soc., 2001, 123(26), pp 6417-6418; Feng et al., Chem. Soc. Rev., 2010, 39, 2411-2419; and Traina et al., J. Am. Chem. Soc., 2011, 133(32), pp12600-12607, the entire disclosures of which are incorporated herein by reference.

[0053] Polymer dyes can have one or more desirable spectroscopic properties, such as a specific absorption maximum wavelength, a specific emission maximum wavelength, an extinction coefficient, a quantum yield, etc. (see, for example, Chattopadhyay et al., "Brilliant violet fluorophore: A new class of ultrabright fluorescent compounds for immunofluorescence experiments", Cytometry Part A, 81A(6) 456 - 466, 2012). In some embodiments, the polymer dye has an absorption curve between 280 nm and 475 nm. In certain embodiments, the polymer dye has an absorption maximum (excitation maximum) in the range of 280 nm to 475 nm. In some embodiments, the polymer dye absorbs incident light having a wavelength between 280 nm and 475 nm. In some embodiments, the polymer dye has an emission maximum wavelength in the range of 400 nm to 850 nm, such as 415 nm to 800 nm. Specific examples of the maximum emission values of interest include, but are not limited to, 421 nm, 510 nm, 570 nm, 602 nm, 650 nm, 711 nm, and 786 nm. In some cases, the polymer dye has an emission maximum wavelength in a range selected from the group consisting of 410 nm to 430 nm, 500 nm to 520 nm, 560 nm to 580 nm, 590 nm to 610 nm, 640 nm to 660 nm, 700 nm to 720 nm, and 775 nm to 795 nm. In certain embodiments, the polymer dye has an emission maximum wavelength of 421 nm. In some cases, the polymer dye has an emission maximum wavelength of 510 nm. In some instances, the polymer dye has an emission maximum wavelength of 570 nm. In certain embodiments, the polymer dye has an emission maximum wavelength of 602 nm. In some cases, the polymer dye has an emission maximum wavelength of 650 nm. In certain cases, the polymer dye has an emission maximum wavelength of 711 nm. In some embodiments, the polymer dye has an emission maximum wavelength of 786 nm. In certain cases, the polymer dye has an emission maximum wavelength of 421 nm ± 5 nm. In some embodiments, the polymer dye has an emission maximum wavelength of 510 nm ± 5 nm.In certain cases, the polymeric dye has a maximum emission wavelength of 570 nm ± 5 nm. In some cases, the polymeric dye has a maximum emission wavelength of 602 nm ± 5 nm. In some embodiments, the polymeric dye has a maximum emission wavelength of 650 nm ± 5 nm. In certain cases, the polymeric dye has a maximum emission wavelength of 711 nm ± 5 nm. In some cases, the polymeric dye has a maximum emission wavelength of 786 nm ± 5 nm. In certain embodiments, the polymeric dye has a maximum emission value selected from the group consisting of 421 nm, 510 nm, 570 nm, 602 nm, 650 nm, 711 nm, and 786 nm.

[0054] Specific polymeric dyes that can be used include, but are not limited to, BD Horizon Brilliant™ Dyes, such as BD Horizon Brilliant™ Violet Dyes (e.g., BV421, BV480, BV510, BV570, BV605, BV650, BV711, BV750, BV786); BD Horizon Brilliant™ Ultraviolet Dyes (e.g., BUV395, BUV496, BUV563, BUV615, BUV661, BUV737, BUV805); and BD Horizon Brilliant™ Blue Dyes (e.g., BB515, BB630-P2, BB660-P2, BB700, BB755-P, BB-790P) (BD Biosciences, San Jose, CA).

[0055] In certain embodiments, as described above, the liquid container (e.g., the inner surface of the wall of the liquid container) contains more than one dye composition, such as, for example, two dye compositions (e.g., a first and a second dye composition). In these embodiments, the dye composition can be a polymer dye composition, as described above. For example, the liquid container can contain a first and a second polymer dye composition. As described above, the first and second polymer dyes can be conjugated polymers (CPs). In certain cases, the first and second polymer dyes are water-soluble conjugated polymers, as described above. In some instances, the dye compositions contained in the liquid container can be different dye compositions, such as different polymer dye compositions. The different dye compositions can be different from each other in terms of their chemical composition and / or in terms of one or more properties of the dyes. For example, the different dye compositions can have at least one of their excitation maxima and emission maxima different from each other. In some cases, the different dye compositions have different excitation maxima from each other. In some cases, the different dye compositions have different emission maxima from each other. In some cases, the different dye compositions have both their excitation maxima and emission maxima different from each other. Thus, in embodiments containing a first and a second dye, the first and second dyes can have at least one of their excitation maxima and emission maxima different from each other. For example, the first and second dyes can have different excitation maxima, different emission maxima, or both different excitation maxima and emission maxima from each other. Additional dye compositions may be contained in the liquid container, and each of the dye compositions within the container is different from each other, as described above.

[0056] In certain embodiments, the liquid container (e.g., the inner surface of the wall of the liquid container) also contains other types of dye compositions, such as one or more non-polymeric dye compositions. As discussed above, the dye may include a detectable moiety or marker that is detectable based on, for example, the fluorescence emission maximum, fluorescence polarization, fluorescence lifetime, light scattering, mass, molecular weight, or combinations thereof. In certain embodiments, the non-polymeric dye includes a fluorophore (i.e., a fluorescent label, fluorescent dye, etc.). The fluorophores of interest can include, but are not limited to, dyes suitable for use in analytical applications (e.g., flow cytometry, imaging, etc.). A number of non-polymeric dyes are commercially available from various sources, such as Molecular Probes (Eugene, OR) and Exciton (Dayton, OH). For example, fluorophores of non-polymeric dyes include 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid; acridine, and derivatives thereof, such as acridine, acridine orange, acridine yellow, acridine red, and acridine isothiocyanate; 5-(2'-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS); 4-amino-N-[3-(vinylsulfonyl)phenyl]naphthalimide-3,5-disulfonate (Lucifer Yellow VS); N-(4-anilino-1-naphthyl)maleimide; anthranilamide; brilliant yellow; coumarin, and derivatives thereof, such as coumarin, 7-amino-4-methylcoumarin (AMC, Coumarin 120), 7-amino-4-trifluoromethylcoumarin (Coumaran 151); cyanine, and derivatives thereof, such as cyanocine, Cy3, Cy3.5, Cy5, Cy5.5, and Cy7; 4',6-diamidino-2-phenylindole (DAPI); 5',5”-dibromopyrogallol-sulfonphthalein (bromopyrogallol red); 7-diethylamino-3-(4'-isothiocyanatophenyl)-4-methylcoumarin; diethylaminocoumarin; diethylenetriaminepentaacetate; 4,4'-diisothiocyanatodihydro-stilbene-2,2'-disulfonic acid; 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid; 5-[dimethylamino]naphthalene-1-sulfonyl chloride (DNS, dansyl chloride);4-(4'-dimethylaminophenylazo)benzoic acid (DABCYL); 4-dimethylaminophenylazophenyl-4'-isothiocyanate (DABITC); eosin, and derivatives such as eosin and eosin isothiocyanate; erythrosin, and derivatives such as erythrosin B and erythrosin isothiocyanate; ethidium; fluorescein and derivatives such as 5-carboxyfluorescein (FAM), 5-(4,6-dichlorotriazin-2-yl)aminofluorescein (DTAF), 2'7'-dimethoxy-4'5'-dichloro-6-carboxyfluorescein (JOE), fluorescein isothiocyanate (FITC), fluorescein chlorotriazinyl, naphthofluorescein, and QFITC (XRITC); fluorescamine; IR144; IR1446; green fluorescent protein (GFP); reef coral fluorescent protein (RCFP); Lissamine™; Lissamine rhodamine, lucifer yellow; malachite green isothiocyanate; 4-methylumbelliferone; orthocresolphthalein; nitrotyrosine; pararosaniline; Nile red; Oregon green; phenol red; B-phycoerythrin (PE); o-phthalaldehyde; pyrene, and derivatives such as pyrene, pyrene butyrate and succinimidyl 1-pyrenebutyrate; reactive red 4 (Cibacron™ brilliant red 3B-A); rhodamine, and derivatives such as 6-carboxy-X-rhodamine (ROX), 6-carboxyrhodamine (R6G), 4,7-dichlororhodamine lissamine, rhodamine B sulfonyl chloride, rhodamine (Rhod), rhodamine B, rhodamine 123, rhodamine X isothiocyanate, sulforhodamine B, sulforhodamine 101, sulforhodamine 101 sulfonyl chloride derivative (Texas red), N,N,N',N'-tetramethyl-6-carboxyrhodamine (TAMRA), tetramethylrhodamine, and tetramethylrhodamine isothiocyanate (TRITC); riboflavin; rose bengal and terbium chelate derivatives; xanthene; carotenoid-protein complexes such as peridinin-chlorophyll protein (PerCP); allophycocyanin (APC);or combinations thereof;

[0057] In certain embodiments, the dye composition contained in the liquid container (e.g., the inner surface of the wall of the liquid container) comprises a polymeric dye composition, as described above. In some cases, the dye composition contained in the liquid container comprises a non-polymeric dye composition, as described above. In some instances, the dye composition contained in the liquid container comprises both a polymeric dye composition and a non-polymeric dye composition. As discussed above, each of the dye compositions (e.g., polymeric and non-polymeric dye compositions) can be clearly positioned on the surface of the liquid container. In some cases, the liquid container comprises a plurality of dye compositions, as described above. For example, the liquid container can comprise two or more, such as three or more, different polymeric dye compositions and two or more, such as three or more, or four or more, or five or more, different non-polymeric dye compositions. In some cases, the liquid container comprises three or more different polymeric dye compositions and five or more different non-polymeric dye compositions.

[0058] As described above, the liquid container can contain both the polymeric dye composition and the non-polymeric dye composition. In some cases, the polymeric dye composition is mixed with the non-polymeric dye composition. In certain embodiments, the mixture of the polymeric dye composition and the non-polymeric dye composition does not undergo significant dye-dye interactions between the polymeric dye composition and the non-polymeric dye composition. For example, the fluorescence emission energy of the polymeric dye composition is not significantly quenched by interaction with the non-polymeric dye composition. In some cases, the fluorescence emission energy of the polymeric dye composition is not significantly dissipated by non-radiative transitions. In these embodiments, the detectable fluorescence of the polymeric dye composition is not significantly less than expected compared to the fluorescence of the polymeric dye composition in the absence of the non-polymeric dye composition. Similarly, in some embodiments, the fluorescence emission energy of the non-polymeric dye composition is not significantly quenched by interaction with the polymeric dye composition. For example, the fluorescence emission energy of the non-polymeric dye composition may not be significantly dissipated by non-radiative transitions. In these embodiments, the detectable fluorescence of the non-polymeric dye composition is not significantly less than expected compared to the fluorescence of the non-polymeric dye composition in the absence of the polymeric dye composition.

[0059] In certain embodiments, the dye composition includes dyes such as polymeric and / or non-polymeric dyes, as described above. The dye composition may also include other components, such as solvents, buffers, stabilizers, etc., but is not limited thereto. For example, the dye composition may include a stabilizer that reduces and / or substantially prevents the decomposition of the dyes in the dye composition. In some cases, the presence of the stabilizer in the dye composition is sufficient to reduce and / or substantially prevent the decomposition of the dyes in the dye composition over a certain period, such as 24 hours or more, or 48 hours or more, or 72 hours or more, or 4 days or more, or 5 days or more, or 6 days or more, or 1 week or more, or 2 weeks or more, or 3 weeks or more, or 4 weeks or more, or 2 months or more, or 3 months or more, or 4 months or more, or 5 months or more, or 6 months or more, or 9 months or more, or 1 year or more. Examples of stabilizers include, but are not limited to, bovine serum albumin (BSA), sodium azide, glycerol, phenylmethanesulfonyl fluoride (PMSF), etc. Additional additives, such as additives that maintain the cells present in whole blood, such as platelet stabilizing factors, may also be present in the composition. Examples of additives that may be included in the composition are anticoagulants such as ethylenediaminetetraacetic acid (EDTA), buffered citrate, heparin, etc. The composition may include these additives in liquid or dry form.

[0060] In some cases, the dye component of a given dry dye composition is a conjugate of a dye moiety and a specific binding member. The specific binding member and the dye moiety can be conjugated (e.g., covalently) to each other at any convenient location of the two molecules via an optional linker.

[0061] As used herein, the term "specific binding member" refers to a member of a pair of molecules that have binding specificity for each other. One member of the pair of molecules may have a surface region or depression that specifically binds to a surface region or depression of the other member of the pair of molecules. Thus, the pair of members has the property of specifically binding to each other to form a binding complex. In some embodiments, the affinity between the specific binding members in the binding complex is 10 -6 M or less, for example, 10-8 10 including below M -7 Below M, for example, 10 -15 10 including below M -9 Below M, 10 -10 Below M, 10 -11 Below M, 10 -12 Below M, 10 -13 Below M, 10 -14 K below M d (Dissociation constant). In some embodiments, the specific binding member binds specifically with high affinity. High affinity means that the binding member binds specifically at 10×10 -9 Below M, for example, 1×10 -9 Below M, 3×10 -10 Below M, 1×10 -10 Below M, 3×10 -11 Below M, 1×10 -11 Below M, 3×10 -12 Below M, 1×10 -12 Apparent K below M d Means binding specifically with an apparent affinity characterized by

[0062] The specific binding member can be proteinaceous. As used herein, the term "proteinaceous" refers to a moiety composed of amino acid residues. The proteinaceous moiety can be a polypeptide. In certain cases, the protein specific binding member is an antibody. In certain embodiments, the proteinaceous specific binding member is an antibody fragment, e.g., a binding fragment of an antibody that specifically binds to a polymeric dye. As used herein, the terms "antibody" and "antibody molecule" are used interchangeably and refer to a protein consisting of one or more polypeptides substantially encoded by all or part of a recognized immunoglobulin gene. For example, in humans, the recognized immunoglobulin genes include the kappa (κ), lambda (λ), and heavy chain gene loci, which together include numerous variable region genes and constant region genes mu (μ), delta (δ), gamma (γ), epsilon (ε), and alpha (α) that encode the IgM, IgD, IgG, IgE, and IgA isotypes, respectively. The immunoglobulin light or heavy chain variable region consists of framework regions (FRs) interrupted by three hypervariable regions also called "complementary determining regions" or "CDRs". The ranges of the framework regions and CDRs are precisely defined (see "Sequences of Proteins of Immunological Interest," E. Kabat et al., U.S. Department of Health and Human Services, (1991)). The numbering of all antibody amino acid sequences described herein conforms to the Kabat system. The sequences of the framework regions of different light or heavy chains are relatively conserved within a species. The framework region of an antibody, which is the combined framework region of the constituent light and heavy chains, serves to position and align the CDRs. The CDRs mainly contribute to the binding to the epitope of the antigen. The term "antibody" is meant to include full-length antibodies and can refer to natural antibodies from any organism, engineered antibodies, or antibodies recombinantly produced for experimental, therapeutic, or other purposes, as further defined below.

[0063] The antibody fragments of interest include, but are not limited to, Fab, Fab’, F(ab’)2, Fv, scFv, or other antigen-binding subsequences of an antibody, those produced by modification of a whole antibody, or those newly synthesized using recombinant DNA techniques. The antibody may be monoclonal or polyclonal and may have other specific activities against cells (e.g., antagonist, agonist, neutralizing, inhibitory, or stimulatory antibodies). It is understood that the antibody may have additional conservative amino acid substitutions that do not substantially affect antigen binding or other antibody functions.

[0064] In certain embodiments, the specific binding member is a Fab fragment, an F(ab’)2 fragment, an scFv, a diabody, or a triabody. In certain embodiments, the specific binding member is an antibody. In some cases, the specific binding member is a mouse antibody or a binding fragment thereof. In certain instances, the specific binding member is a recombinant antibody or a binding fragment thereof.

[0065] In certain embodiments, the liquid container also includes a calibration standard. The calibration standard can be useful for determining the accuracy of the assay and ensuring consistency between subsequent assays. In some cases, the calibration standard includes labeled beads such as fluorescently labeled beads. The fluorescently labeled beads can be standard fluorescently labeled beads typically used as calibration standards. Examples of standard fluorescently labeled beads include, but are not limited to, fluorescently labeled microparticles or nanoparticles. In some cases, the fluorescently labeled beads are configured to remain suspended in the assay mixture and not substantially sediment or aggregate. In some embodiments, the fluorescently labeled beads include, but are not limited to, fluorescently labeled polystyrene beads, fluorescein beads, rhodamine beads, and other beads tagged with fluorescent dyes. Additional examples of fluorescently labeled beads are described in U.S. Patent Nos. 6,350,619, 7,738,094, and 8,248,597, the disclosures of each of which are incorporated herein by reference in their entirety.

[0066] In some cases, the liquid container facilitates the storage of the dye composition over a long period. For example, the liquid container can be a storage-stable liquid container. In some cases, the dye composition contained in the liquid container is a storage-stable dye composition, and the dye composition is substantially stable over a long period. "Stability" or "storage stability" or "substantially stability" means a dye composition that does not significantly reduce and / or lose its activity over a long period. For example, a storage-stable dye composition may not have a significant loss of fluorescence activity due to the decomposition of the dye composition over a long period. For example, over a long period, it may not have a loss of fluorescence activity of 10% or less, or 9% or less, or 8% or less, or 7% or less, or 6% or less, or 5% or less, or 4% or less, or 3% or less, or 2% or less, or 1% or less. In a particular case, the storage-stable dye composition has a loss of fluorescence activity of 5% or less over a long period. In some cases, the storage-stable dye composition substantially retains its fluorescence activity over a long period. For example, over a long period, it retains 100% of its activity, or 99% or more of its activity, or 98% or more, or 97% or more, or 96% or more, or 95% or more, or 94% or more, or 93% or more, or 92% or more, or 91% or more, or 90% or more, or 85% or more, or 80% or more, or 75% or more. For example, a storage-stable dye composition can retain 90% or more of its fluorescence activity over a long period. In some cases, the storage-stable composition retains 95% or more of its fluorescence activity over a long period. The long period is, for example, a period of 1 week or more, or 2 weeks or more, or 3 weeks or more, or 1 month or more, or 2 months or more, or 3 months or more, or 4 months or more, or 6 months or more, or 9 months or more, or 1 year or more, or 1.5 years (e.g., 18 months) or more, or 2 years or more, or 2.5 years (e.g., 30 months) or more, or 3 years or more, or 3.5 years (e.g., 42 months) or more, or 4 years or more, or 4.5 years (e.g., 54 months) or more, or 5 years or more, etc. For example, the long period can be 6 months or more. In some cases, the long period is 9 months or more. In some cases, the long period is 1 year (e.g., 12 months) or more. In some cases, the long period is 1.5 years (e.g., 18 months) or more.In some cases, the long term is 2 years (e.g., 24 months) or more. In some instances, the long term is 10 years or less, such as 7.5 years or less, including 5 years or less, e.g., 2 years or less.

[0067] Various aspects of a liquid container (including a dry composition, e.g., a dry polymer-conjugated dye composition) that can be part of the system of embodiments of the present invention are further provided in U.S. Pat. Nos. 10,545,137 and 11,320,437, and U.S. Patent Application Publication Nos. 2020 / 0147615A1, and co-pending U.S. applications Ser. Nos. 17 / 580,130 and 17 / 830,070, the disclosures of which are incorporated herein by reference.

[0068] Solid volume displacer As summarized above, the system of the present invention also includes a solid volume displacer. A solid volume displacer according to certain embodiments of the present disclosure is configured, for example, as described above, to be positioned inside a liquid container and to occupy a majority of the liquid container volume below the upper portion of a dry reagent composition positioned on, e.g., the inner surface of the wall of the liquid container.

[0069] The solid volume displacer can be compatible with a liquid sample and / or reagent or analyte that can contact the liquid container (e.g., contained inside the liquid container). In some cases, the liquid sample can be an aqueous liquid sample, and in these cases, the solid volume displacer can be compatible with the aqueous sample. "Compatible" means that the solid volume displacer (e.g., the material from which the solid volume displacer is made) is substantially inert (e.g., does not significantly react) with respect to the liquid and / or reagent or analyte in contact with the liquid container.

[0070] A solid volume displacer may include a first end, a second end, and a body therebetween, and the second end and the body are configured to be positioned inside a liquid container such that, for example, the second end and the body displace a volume of liquid onto an inner surface of a wall of the liquid container to reconstitute a dry reagent composition, as described above. For example, the second end and the body may be configured to be positioned inside the liquid container, such that the solid volume displacer occupies a majority of the liquid container volume below the upper portion of the dry reagent composition. For example, in some embodiments, the second end and the body may be configured to be positioned inside the liquid container, such that the solid volume displacer occupies more than 50% of the liquid container volume below the upper portion of the dry reagent composition. In certain cases, the second end and the body may be configured to be positioned inside the liquid container, such that the solid volume displacer occupies a portion of the liquid container volume below the upper portion of the dry reagent composition, such as 55% or more, or 60% or more, or 70% or more, or 75% or more, or 80% or more, or 85% or more, or 90% or more, or 95% or more, or 99% or more of the volume. In embodiments where a plurality of dry reagent compositions are provided on the inner surface of the wall of the liquid container, the solid volume displacer may be configured to be positioned inside the liquid container, such that the solid volume displacer occupies a majority of the liquid container volume below the upper portion of the dry reagent composition positioned furthest from the bottom of the liquid container. In these cases, the solid volume displacer may be configured to displace a volume of liquid onto the inner surface of the wall of the liquid container to reconstitute any dry reagent composition present on the inner surface of the wall. In certain cases, the solid volume displacer is removable from the liquid container, such that the solid volume displacer can be removed from the liquid container after the liquid in the liquid container has been displaced in a manner sufficient to reconstitute the dry reagent composition. In these cases, the removable solid volume displacer enables utilization of the reconstituted reagent composition. The reconstituted reagent composition may be collected for use in analytical applications (e.g., flow cytometry, imaging, etc.).

[0071] As described above, the liquid container can be configured to hold a fluid (e.g., gas or liquid) of a certain volume. In certain embodiments, the liquid container is configured to hold a certain volume of liquid below the upper portion of the dry reagent composition. In these cases, there may be a volume of the liquid container below the upper portion of the dry reagent composition that is not configured to be occupied by the solid volume displacer. For example, thus, when the solid volume displacer is positioned inside the liquid container by introducing the liquid of this volume into the liquid container, the liquid container is filled up to the upper portion of the dry reagent composition. For example, in some cases, the volume of the liquid container below the upper portion of the dry reagent composition that is not configured to be occupied by the solid volume displacer (e.g., the volume of liquid required to fill the liquid container up to the upper portion of the dry reagent composition when the solid volume displacer is positioned inside the liquid container) can be 5000 μl or less, 1000 μl or less, for example 500 μl or less, or 400 μl or less, or 350 μl or less, or 300 μl or less, or 250 μl or less, or 200 μl or less, or 150 μl or less, or 100 μl or less, or 50 μl or less, or 20 μl or less, or 10 μl or less, or 1 μl or less. In some embodiments, the liquid container contains a volume of liquid (e.g., a biological liquid sample) that is greater than the volume of the liquid container below the upper portion of the dry reagent composition that is not configured to be occupied by the solid volume displacer.

[0072] As described above, for example, when the solid volume displacer is positioned inside the liquid container, there may be a volume of the liquid container below the upper part of the dry reagent composition that is not configured to be occupied by the solid volume displacer. In other words, when the solid volume displacer is positioned inside the liquid container, there may be a volume of liquid required to fill the liquid container up to the upper part of the dry reagent composition. Thus, the solid volume displacer can be configured to reduce the volume of liquid required to fill the liquid container up to the upper part of the dry reagent composition, for example, to contact the dry reagent composition. For example, the solid volume displacer can be configured to reduce the volume of liquid required to fill the liquid container up to the upper part of the dry reagent composition by 50% or more, for example, 60% or more, or 70% or more, or 80% or more, or 90% or more, or 95% or more.

[0073] In some embodiments, the solid volume displacer can be configured to reduce the volume of liquid required to contact the dry reagent composition in the liquid container as compared to other systems that move liquid on the wall of the liquid container. In some embodiments, the solid volume displacer can be configured to reduce the volume of liquid required to contact the dry reagent composition as compared to using a stirrer (e.g., a vortexer) alone (i.e., without a solid volume displacer). For example, the solid volume displacer can be configured to reduce the volume of liquid required to contact the dry reagent composition by 50% or more, for example, 60% or more, or 70% or more, or 80% or more, or 90% or more as compared to using a vortexer. In some embodiments, the solid volume displacer can be configured to be attached to a mechanical mixer. For example, the first end of the solid volume displacer can be configured to be attached to a mechanical mixer. In some embodiments, the first end of the solid volume displacer can be configured to be attached to a stirrer (e.g., a vortexer).

[0074] The shape of the solid volume displacer may vary and may depend on the use of the solid volume displacer. For example, as described herein, the solid volume displacer can be introduced into the previously described liquid container in a manner sufficient to displace the volume of liquid onto the inner surface of the wall of the liquid container to reconstitute the previously described dry reagent composition. In these cases, the solid volume displacer can be configured in a shape that is compatible with positioning the solid volume displacer inside the liquid container. For example, in an embodiment where the wall of the liquid container is cylindrical in shape, the body of the solid volume displacer can be configured in a cylindrical shape. In these embodiments, the solid volume displacer can include an outer surface that is concentric with the inner surface of the wall of the liquid container. In these cases, the outer surface of the solid volume displacer can have a diameter that is different from the diameter of the inner surface of the wall of the liquid container (e.g., a diameter smaller than the diameter of the inner surface of the wall of the liquid container). For example, the difference (i.e., clearance) between the diameter of the outer surface of the body of the solid volume displacer and the diameter of the inner surface of the wall of the liquid container can be 5 mm or less, such as 4 mm or less, or 3 mm or less, or 2 mm or less, or 1 mm or less, or 0.5 mm or less, or 0.2 mm or less, or 0.1 mm or less, or 0.05 mm or less, or 0.02 mm or less, or 0.01 mm or less. In some embodiments, the outer surface of the solid volume displacer may include a plurality of sections having different diameters, i.e., for example, the outer surface can be stepped. In an embodiment where the wall of the liquid container is tapered, the outer surface of the body of the solid volume displacer can be tapered at the same angle or a similar angle. In an embodiment where the bottom of the liquid container is round, the second end of the solid volume displacer can be round and concentric with the bottom of the liquid container. In some embodiments, the solid volume displacer can be configured as a pestle. In certain cases, the solid volume displacer is hollow.

[0075] As described above, the distance between the dry reagent composition on the inner surface of the wall of the liquid container and the bottom of the liquid container may vary. In some cases, the solid volume displacer may be configured to displace the volume of the liquid above the inner surface by a distance greater than the distance between the dry reagent composition on the inner surface of the wall of the liquid container and the bottom of the liquid container. For example, the solid volume displacer may be configured to displace the volume of the liquid by 0.1 mm or more, such as 0.5 mm or more, or 1 mm or more, or 2 mm or more, or 3 mm or more, or 4 mm or more, or 5 mm or more, or 6 mm or more, or 7 mm or more, or 8 mm or more, or 9 mm or more, or 10 mm or more, or 12 mm or more, or 14 mm or more, or 16 mm or more, or 18 mm or more, or 20 mm or more, or 25 mm or more, or 30 mm or more, or 35 mm or more, or 40 mm or more, or 50 mm or more, or 60 mm or more, or 70 mm or more, or 80 mm or more, or 90 mm or more, or 100 mm or more, or 110 mm or more, or 120 mm or more, or 130 mm or more, or 140 mm or more, or 150 mm or more, or 160 mm or more, or 170 mm or more, or 180 mm or more, or 190 mm or more, or 200 mm or more above the inner surface of the wall of the liquid container.

[0076] As described above, embodiments of the solid volume displacer can be compatible with a liquid sample and / or reagent or analyte that contacts a liquid container (e.g., contained within a liquid container). Examples of suitable solid volume displacer materials include, but are not limited to, ceramics, metals (e.g., stainless steel, aluminum, anodized aluminum, titanium, copper, bronze, nickel, etc.), paper, glass, resins, and plastics. For example, the solid volume displacer can be composed of a glass such as silicate glass, borosilicate glass, sodium borosilicate glass (e.g., PYREX (trademark)), fused quartz glass, fused silica glass, etc., but is not limited thereto. Other examples of suitable solid volume displacer materials include polymeric materials. The polymeric material can be a hydrophobic polymeric material. In some embodiments, the hydrophobic polymeric material can be a resin such as urethane methacrylate (UMA), Somos® 9120, Somos® WaterShed XC 11122, and Somos® EvoLVe, but is not limited thereto. In some cases, the hydrophobic polymeric material can be a plastic such as polystyrene, polypropylene, polymethylpentene, polytetrafluoroethylene (PTFE), perfluoroether (PFE), fluorinated ethylene propylene (FEP), perfluoroalkoxy alkane (PFA), polyethylene terephthalate (PET), polyethylene (PE), low density polyethylene (LDPE), polyether ether ketone (PEEK), etc., but is not limited thereto. The material constituting the solid volume displacer can be translucent / transparent, translucent, and / or any color, e.g., gray, black, white, yellow, etc.

[0077] In some embodiments, as described above, the solid volume displacer includes a first end, a second end, and a body therebetween, and the second end and the body are configured to be positioned inside the liquid container. In some embodiments, the first end of the solid volume displacer can be configured to be positioned outside the liquid container when the body and the second end of the solid volume displacer are positioned inside the liquid container. In some cases, the first end of the solid volume displacer can include a flange. In some embodiments, the flange is configured such that, for example, by contacting the open end on the liquid container, the second end of the solid volume displacer does not contact the bottom of the liquid container. In other embodiments, the flange is configured to allow the second end of the solid volume displacer to contact the bottom of the liquid container. In some embodiments, the solid volume displacer can include a ring of filter media positioned around the outside of the body near the first end. In these cases, the ring of filter media can be configured to prevent liquid from splashing out of the liquid container when the body of the solid volume displacer is positioned inside the liquid container. For example, the ring of filter media can prevent liquid from splashing out of the container by eliminating the open space between the outer surface of the body of the solid volume displacer and the inner surface of the wall of the liquid container near the open end of the liquid container when the body of the solid volume displacer is positioned inside the liquid container. In other cases, the ring of filter media can be configured to prevent liquid from splashing out of the container by contacting or sliding over and outside the open end of the liquid container (while maintaining contact with the open end, for example) when the body of the solid volume displacer is positioned inside the liquid container.

[0078] In some embodiments, the solid volume dispenser is configured to prevent the generation of an aerosol or liquid spray during use (e.g., when the body of the solid volume dispenser is inserted inside a liquid container and then removed from inside the liquid container). In some cases, a ring of filter media of the dispenser may allow air pressure to equalize between the inside and outside of the container when the body of the solid volume dispenser is positioned inside the liquid container. For example, the ring of filter media can be composed of a material that allows air to pass through but impedes or prevents the passage of liquid, such as a fibrous or porous material. In some cases, the fibrous or porous material can be configured to selectively allow the passage of certain components of the liquid while preventing the passage of other components of the liquid. For example, the fibrous or porous material can be configured to allow the passage of liquid and various substances dissolved therein while preventing the passage of cells (e.g., mammalian cells and / or bacterial cells) and virions. In some embodiments, the flange of the dispenser includes surface features, such as protrusions or recesses, on the surface of the flange that allow air pressure to equalize between the inside and outside of the container, and the flange is configured to contact the liquid container when the body of the solid volume dispenser is positioned inside the liquid container. In some embodiments, the second end of the solid volume dispenser includes protrusions (e.g., a series of ribs) and / or recesses (e.g., concentric sections with a reduced diameter) configured to prevent liquid from being discharged from the opening of the liquid container during use of the dispenser, such as for reconstituting a dry reagent composition, as described in more detail below.

[0079] In some embodiments, the first end of the solid volume displacer may include a gripping portion. In some cases, the gripping portion may include protrusions (e.g., ribs or ridges), or may include a non-slip material for improving user grip and / or control. In some embodiments, the solid volume displacer may include a gripping portion and a flange (e.g., as discussed above). In other cases, the solid volume displacer may include only one of the gripping portion or the flange. In some embodiments, the first end of the solid volume displacer is configured to seal the open end of the liquid container when the second end and the body of the solid volume displacer are positioned inside the liquid container. That is, the first end of the solid volume displacer may substantially prevent the contents of the liquid container (e.g., the liquid within the liquid container) from exiting the liquid container. The first end of the solid volume displacer may also substantially prevent other substances from entering the liquid container. For example, the first end of the solid volume displacer may form a watertight seal that substantially prevents liquid from entering or exiting the container, or may form an airtight seal that substantially prevents gas from entering or exiting the container. In some embodiments, the first end of the solid volume displacer forms a watertight seal that substantially prevents liquid from entering or exiting the liquid container, yet allows air to enter or exit the container, for example, as discussed above, to allow air pressure to equalize and prevent the formation of aerosols. In some cases, the first end of the solid volume displacer may form a removable seal. In some embodiments, the first end of the solid volume displacer includes a threaded or snap-on cap or other suitable sealing element that can be applied to the opening of the liquid container. For example, the first end of the solid volume displacer may include a threaded cap that can be screwed onto the opening of the liquid container when the second end and the body of the solid volume displacer are positioned inside the liquid container.

[0080] In some embodiments, one or more components of the solid volume displacer may be adjustable or detachable. In some cases, the flange may be adjustable or detachable. For example, the flange may include a latch or clamp configured to allow a user to attach or detach the flange depending on, for example, whether contact between the second end of the solid volume displacer and the bottom of the liquid container is desired. In some cases, the latch or clamp may allow a user to adjust the position of the flange on the solid volume displacer depending on, for example, the desired clearance between the second end of the solid volume displacer and the bottom of the liquid container. In some cases, the ring of filter media is removable. In some embodiments, the solid volume displacer may be washed and / or sterilized (e.g., in an autoclave) or discarded for later use. Thus, in some embodiments, the solid volume displacer described herein is disposable, such as after single use.

[0081] As described above, the solid volume displacer may be positioned inside the previously described liquid container and configured to occupy a majority of the liquid container volume below the top. In some instances, the second end of the solid volume displacer has an outer surface. In some instances, the body of the solid volume displacer has an outer surface. In these embodiments, the outer surface of the body of the solid volume displacer faces the inner surface of the wall of the liquid container when the body of the solid volume displacer is positioned inside the liquid container. The outer surface of the body of the solid volume displacer may contact the contents of the liquid container.

[0082] FIG. 1 provides a depiction of a system 100 according to an embodiment of the present invention. The system 100 depicted in FIG. 1 is composed of two components, a liquid container 110 and a solid volume displacer 120. The liquid container 110 includes an open end 111 and a bottom 112 separated by an intervening wall 113. The inner surface of the wall 113 includes a dry reagent composition 114 which can be a polymer dye composition (e.g., a conjugated polymer dye composition) or other dye or non-dye reagent composition. The solid volume displacer 120 includes a first end 121, a second end 122, and a body 123 therebetween.

[0083] The liquid container 110 is configured as a test tube or centrifuge tube having a cylindrical-shaped wall 113 and a rounded bottom 112. Of the dry reagent composition 114, the dry reagent composition positioned farthest from the bottom 112 of the liquid container is about 20 mm from the bottom 112 of the liquid container. The first end of the solid volume displacer 121 includes a flange that prevents the second end of the solid volume displacer 122 from contacting the bottom 112 of the liquid container, and a gripping portion 124 for increased usability / improved user grip. The body of the solid volume displacer 123 is configured as a cylinder having an outer surface concentric with the inner surface of the wall 113 of the liquid container. The second end of the solid volume displacer 122 is rounded and concentric with the bottom 112 of the liquid container. Assembly 130 provides a depiction of the system 100 with the solid volume displacer positioned inside the liquid container.

[0084] Figures 4A and 4B provide a depiction of a stepped solid volume displacer according to an embodiment of the present invention. In Figure 4A, a side view of a solid volume displacer 400 having cylindrical sections or steps 410, 420, and 430 of different diameters is provided. Step 410, which is a step adjacent to the first end of the solid volume displacer 400, is 60 mm in length and has a diameter of 9.9 mm. Step 410 is configured to function as a gripping portion when positioning the displacer 400 within a liquid container. Step 420 is 20 mm in length and has a diameter of 9.6 mm. Step 430, which is a step adjacent to the second end of the solid volume displacer 400, is 40 mm in length and has a diameter of 8.9 mm. The second end of the solid volume 400 is a spherical cap having a hemisphere with a radius of 4.6 mm, and the section of the hemisphere protruding outside step 430 is instead configured as a continuation of step 430. In Figure 4B, a three-dimensional rendering of the solid volume displacer 400 is provided adjacent to a sliced view of a liquid container 450 that is compatible with the displacer. In some cases, the liquid container 450 is approximately 75.5 mm in length and has a tapered cylindrical section and a hemispherical bottom with a thickness of 1.0 mm. Thus, when the second end of the solid volume displacer 400 contacts the bottom of the container 450, the open end of the container is approximately 69.9 mm from the center point of the spherical cap of the displacer (i.e., the tip of the second end of the displacer). In some cases, the clearance between the outer diameter of the solid volume displacer 400 and the inner diameter of the liquid container 450 can be approximately 0.2 mm near the bottom of the container 450 when the second end of the displacer is in contact with the bottom of the container. The section of the solid volume displacer 400 that extends outside the container 450 when the second end of the displacer is in contact with the bottom of the container is configured as a gripping portion.

[0085] Figures 5A and 5B provide a depiction of a linear (i.e., non-stepped) solid volume displacer according to an embodiment of the present invention. In Figure 5A, a side view of a solid volume displacer 500 is provided having a first end, a single cylindrical section of length 120 mm and diameter 9.0 mm, and a spherical cap. The spherical cap is a hemisphere with a radius of 4.6 mm, and the section of the hemisphere (i.e., of diameter 9.0 mm) that projects outside the cylindrical section is instead configured as a continuation of the cylindrical section. In Figure 5B, a 3D rendering of the solid volume displacer 500 is provided adjacent to a sliced view of a liquid container 530 that is compatible with the displacer. In some cases, the liquid container 530 has a length of about 75.5 mm and has a tapered cylindrical section and a hemispherical bottom with a thickness of 1.0 mm. Thus, when the second end of the solid volume displacer 500 contacts the bottom of the container 530, the open end of the container is about 69.9 mm from the center point of the spherical cap of the displacer (i.e., the end of the second end of the displacer). In some cases, the clearance between the outer diameter of the solid volume displacer 500 and the inner diameter of the liquid container 530 can be about 0.1 mm near the bottom of the container 530 when the second end of the displacer is in contact with the bottom of the container. The section of the solid volume displacer 500 that extends outside the container 530 when the second end of the displacer is in contact with the bottom of the container is configured as a gripping portion.

[0086] Method of Use As summarized above, aspects of the present disclosure also include methods of using the system of the invention to reconstitute a dry reagent composition. As described above, a system for reconstituting a dry reagent composition can include one or more dry reagent compositions (e.g., first and second polymer dye compositions) that are clearly positioned on a surface of a liquid container (e.g., the inner surface of the wall of the liquid container), and a solid volume displacer positioned inside the liquid container and configured to occupy a majority of the liquid container volume below the upper portion of the dry reagent composition. Thus, a method of using the subject system can include reconstituting a dry reagent composition. In some cases, the dry reagent composition is a dye composition, such as a polymer dye composition. The polymer dye composition can be a dry polymer dye composition. Thus, a method of using the system can include reconstituting a dye composition. In certain embodiments, the method includes introducing a volume of liquid into the liquid container and then introducing the solid volume displacer into the liquid container in a manner sufficient to displace the liquid onto the inner surface of the wall of the liquid container to reconstitute the dry reagent composition. Any convenient liquid handling device, including but not limited to a syringe, needle, pipette, aspirator, etc., can be used to add the volume of liquid to the container.

[0087] In practicing embodiments of the subject method, the method can include introducing a volume of liquid into a liquid container, where the liquid container includes an open end and a bottom separated by an intervening wall, and the inner surface of the wall includes the dry reagent composition, and introducing the solid volume displacer into the liquid container in a manner sufficient to displace the liquid onto the inner surface of the wall of the liquid container to reconstitute the dry reagent composition. In practicing embodiments of the subject method, the method can include mixing the liquid and the dry reagent composition by moving the solid volume displacer. In these cases, the movement can include rotating and / or reciprocating the solid volume displacer. In practicing some embodiments of the subject method, the movement can include rotating the solid volume displacer. In practicing some embodiments of the subject method, the movement can include reciprocating the solid volume displacer.

[0088] In certain embodiments, the liquid comprises a biological sample. In some cases, the biological sample can be derived from specific biological fluids including, but not limited to, blood, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, bronchoalveolar lavage, amniotic fluid, amniotic cord blood, urine, vaginal fluid, and semen. In some embodiments, the biological sample comprises whole blood or a fraction thereof. In some embodiments, the biological sample comprises plasma.

[0089] In certain embodiments, the liquid container is a sealed container (e.g., the liquid container is sealed), such as when the container includes a seal (e.g., a watertight and / or airtight seal). In these cases, the method can include removing the seal prior to positioning or introducing the volume of liquid within the liquid container. Removing the seal of the liquid container exposes the contents of the liquid container to the ambient environment and can enable utilization of the internal volume of the liquid container. Thus, a user who can utilize the internal volume of the liquid container can position the volume of liquid within the liquid container (e.g., by displacing the liquid onto the inner surface of the wall of the liquid container and introducing a solid volume displacer into the liquid container in a manner sufficient to reconstitute the dry reagent composition) to reconstitute the dry polymer dye composition within the liquid container. In practicing some embodiments of the subject method, the method can include sealing the liquid container after all of the dry reagent compositions have been reconstituted. In certain embodiments, the seal can be a removable cap. In these cases, the method can include removing the seal prior to positioning or introducing the volume of liquid within the liquid container and resealing the liquid container after all of the dry reagent compositions have been reconstituted.

[0090] In certain embodiments, a solid volume displacer can include a first end, a second end, and a body therebetween. In certain embodiments, the first end of the solid volume displacer is configured to be attached to a mechanical mixer. In practicing some embodiments of the subject methods, the method can include positioning the solid volume displacer within a liquid container (e.g., by attaching the first end of the solid volume displacer to a mechanical mixer) and then mixing the contents of the liquid container. Mixing can be performed using any convenient protocol. For example, mixing can be performed using a stirrer. The stirrer can be any convenient stirrer sufficient to mix the liquid within the liquid container, including, but not limited to, a vortexer, ultrasonic generator, shaker (e.g., manual, mechanical, or electric shaker), rocker, vibratory plate, magnetic stirrer, static mixer, rotor, blender, mixer, tumbler, orbital shaker, among other stirring protocols. In practicing some embodiments of the subject methods, reconstituting a dry reagent composition using a solid volume displacer involves introducing a smaller volume of liquid compared to using a stirrer (e.g., a vortexer) alone (i.e., without a solid volume displacer). For example, reconstituting a dry reagent composition using a solid volume displacer reduces the volume of liquid that needs to be introduced into the liquid container by 50% or more, e.g., 60% or more, or 70% or more, or 80% or more, or 90% or more, compared to using a vortexer alone without a solid volume displacer or the like. In practicing some embodiments of the subject methods, reconstituting a dry reagent composition using a vortexer together with a solid volume displacer involves introducing a smaller volume of liquid compared to using the solid volume displacer alone (i.e., without a vortexer).

[0091] In some cases, the method also includes assaying the reconstituted composition, such as a reconstituted dye composition. Assaying the reconstituted composition, such as a reconstituted dye composition, can be performed using any suitable assay device. For example, the assay device can be a flow cytometer. In these embodiments, the assay includes analyzing the reconstituted composition, such as a reconstituted dye composition, by flow cytometry. In some instances, the assay includes contacting the reconstituted composition, such as a reconstituted dye composition, with electromagnetic radiation (e.g., light), such as electromagnetic radiation having a wavelength corresponding to the excitation maximum of the reconstituted composition, such as a reconstituted dye composition. The assay can further include detecting the light emitted from the excited reagent composition, such as a dye composition. For example, the method can include detecting the light emitted from the excited reagent composition, such as a dye composition, at one or more wavelengths corresponding to the emission maximum of the reagent composition, such as a dye composition.

[0092] Suitable flow cytometry systems can include, but are not limited to, those described in Ormerod (ed.), Flow Cytometry: A Practical Approach, Oxford Univ. Press (1997), Jaroszeski et al. (eds.), Flow Cytometry Protocols, Methods in Molecular Biology No. 91, Humana Press (1997), Practical Flow Cytometry, 3rd ed., Wiley-Liss (1995), Virgo, et al. (2012) Ann Clin Biochem. Jan;49(pt1):17-28, Linden, et.al., Semin Throm Hemost. 2004 Oct;30(5):502-11, Alison, et al. J Pathol, 2010 Dec;222(4):335-344; and Herbig, et al. (2007) Crit Rev Ther Drug Carrier Syst. 24(3):203-255 (the disclosures of which are incorporated herein by reference).In certain instances, the flow cytometry systems of interest include the BD Biosciences FACSCanto(TM) II flow cytometer, BD Accuri(TM) flow cytometer, BD Biosciences FACSCelesta(TM) flow cytometer, BD Biosciences FACSLyric(TM) flow cytometer, BD Biosciences FACSVerse(TM) flow cytometer, BD Biosciences FACSymphony(TM) flow cytometer, BD Biosciences LSRFortessa(TM) flow cytometer, BD Biosciences LSRFortess(TM) X-20 flow cytometer, and BD Biosciences FACSCalibur(TM) flow cytometer, BD Biosciences FACSCount(TM) cell sorter, BD Biosciences FACSLyric(TM) cell sorter, and BD Biosciences Via(TM) cell sorter, BD Biosciences Influx(TM) cell sorter, BD Biosciences Jazz(TM) cell sorter, BD Biosciences Aria(TM) cell sorter, and BD Biosciences FACSMelody(TM) cell sorter, and the like.

[0093] In certain embodiments, the subject flow cytometry system is configured to sort one or more of the particles (e.g., cells) of a sample. The term "sort" as used herein is used in its conventional sense and refers to separating the components of a sample (e.g., non-cellular particles such as cells, biopolymers), and in some instances, delivering the separated components to one or more sample collection containers. For example, the subject system can be configured to sort a sample having two or more components, including, for example, three or more components, for example, four or more components, for example, five or more components, for example, ten or more components, for example, fifteen or more components, and twenty-five or more components. One or more of the sample components, for example, two or more sample components, for example, three or more sample components, for example, four or more sample components, for example, five or more sample components, for example, ten or more sample components can be separated from the sample and delivered to a sample collection container, and fifteen or more sample components can be separated from the sample and delivered to a sample collection container.

[0094] In some embodiments, the particle sorting system of interest is configured to sort particles using a closed particle sorting module, such as that described in U.S. Patent Publication No. 2017 / 0299493, filed Mar. 28, 2017, the disclosure of which is incorporated herein by reference. In certain embodiments, the particles (e.g., cells) of a sample are sorted using a sorting determination module having a plurality of sorting determination units, such as that described in U.S. Patent Application No. 16 / 725,756, filed Dec. 23, 2019, the disclosure of which is incorporated herein by reference.

[0095] In some embodiments, the flow cytometer system is a flow cytometry system such as those described in U.S. Patent Nos. 10,006,852; 9,952,076; 9,933,341; 9,784,661; 9,726,527; 9,453,789; 9,200,334; 9,097,640; 9,095,494; 9,092,034; 8,975,595; 8,753,573; 8,233,146; 8,140,300; 7,544,326; 7,201,875; 7,129,505; 6,821,740; 6,813,017; 6,809,804; 6,372,506; 5,700,692; 5,643,796; 5,627,040; 5,620,842; 5,602,039, the disclosures of which are incorporated herein by reference in their entirety.

[0096] Other analytical methods may also be used, including but not limited to liquid chromatography - mass spectrometry or gas chromatography - mass spectrometry systems. For example, an assay may involve the use of an analytical separation device such as a liquid chromatograph (LC), including but not limited to a high performance liquid chromatograph (HPLC), a micro or nano liquid chromatograph, or an ultra - high pressure liquid chromatograph (UHPLC) device, capillary electrophoresis (CE), or a capillary electrophoresis chromatograph (CEC) device. A mass spectrometer (MS) system may also be used to assay a reagent composition, such as a dye composition. Examples of mass spectrometers may include, but are not limited to, electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI), electron impact (EI), atmospheric pressure photoionization (APPI), matrix - assisted laser desorption ionization (MALDI), or inductively coupled plasma (ICP) ionization, or any combination thereof. Similarly, any of a variety of different mass spectrometers, including but not limited to time - of - flight (TOF), Fourier transform ion cyclotron resonance (FTICR), ion trap, quadrupole or double - focusing magnetic sector mass analyzers, or any hybrid thereof, may be used.

[0097] In certain embodiments, the subject system is included in a fully automated apparatus. "Fully automated" means that the apparatus houses the subject system and prepares a reconstituted composition, such as a reconstituted dye composition, with little or no human intervention or manual input to the subject system. In certain embodiments, the subject system is configured to prepare and analyze a reconstituted composition, such as a reconstituted dye composition, without any human intervention.

[0098] In certain embodiments, the method also includes storing a reconstituted composition, such as a reconstituted dye composition, for a period of time. The reconstituted composition, such as a reconstituted dye composition, can be stored for a period of time before, during, and / or after assaying the reconstituted composition, such as a reconstituted dye composition. In some cases, the reconstituted composition, such as a reconstituted dye composition, is stored for a period of time of 24 hours or more, or 48 hours or more, or 72 hours or more, or 4 days or more, or 5 days or more, or 6 days or more, or 1 week or more, or 2 weeks or more, or 3 weeks or more, or 4 weeks or more, or 2 months or more, or 3 months or more, or 4 months or more, or 5 months or more, or 6 months or more, or 9 months or more, or 1 year or more. In certain cases, the reconstituted composition is stored for 24 hours or more. In certain cases, the reconstituted composition is stored for 48 hours or more. In certain cases, the reconstituted composition is stored for 72 hours or more. In certain cases, the reconstituted composition is stored for 1 week or more. In certain cases, the reconstituted composition is stored for 2 weeks or more. In certain cases, the reconstituted composition is stored for 3 weeks or more.

[0099] Embodiments of this method may further include transporting the reconstituted composition to a remote location. A "remote location" is a location other than the location where the dye composition is reconstituted. For example, the remote location can be another location within the same city (e.g., an office, a laboratory, etc.), another location in a different city, another location in a different state, another location in a different country, etc. Thus, when one article is shown to be "remote" from another article, it means that the two articles can be separated while being in the same room, or at least in different rooms or different buildings, and may be at least 1 mile, 10 miles, or 100 miles or more apart.

[0100] Figures 2A - 2C provide a depiction of a method of using a system for reconstituting a dry reagent composition according to an embodiment of the present invention. In Figure 2A, a volume of liquid 230 (e.g., a biological liquid sample) is introduced into liquid container 210. In Figure 2B, solid volume displacer 220 is introduced into liquid container 210 in a manner sufficient to displace the volume of liquid 230 onto the inner surface of the wall 213 of the liquid container to reconstitute dry reagent composition 214. In Figure 2C, after reconstitution, the assembly of liquid container 210, solid volume displacer 220, and the volume of liquid 230 is disassembled by removing solid volume displacer 220. The liquid 230 containing the reconstituted reagent can then be collected for use in an analytical application.

[0101] Kit Aspects of the present disclosure also include kits. The kits can include, for example, a system for reconstituting a dry reagent composition as described in any one of the embodiments described herein. Aspects of the system for reconstituting a dry reagent composition can include, for example, a liquid container and a solid volume displacer as described above. Aspects of the liquid container can include an open end and a bottom separated by an intervening wall, and the inner surface of the wall can include the dry reagent composition. Aspects of the solid volume displacer can include a first end, a second end, and a body therebetween, and the second end and the body are configured to be positioned inside the liquid container.

[0102] In certain embodiments, the kit includes the subject system for reconstituting the dry reagent composition and a package configured to hold the system. The package can be an enclosed package, such as a moisture-resistant container, optionally under an airtight and / or vacuum seal. In some embodiments, the package can protect its internal contents (e.g., the subject system) from light. In these cases, the package can protect against light at any range or spectrum of light wavelengths, including, for example, visible light, UV light, light within and / or near the excitation spectrum of any encapsulating dye. In certain cases, the package is a sterile package configured to maintain a device sealed within the package in a sterile environment. "Sterile" means substantially free of microorganisms (such as fungi, bacteria, viruses, spore forms, etc.). In some embodiments, the liquid container and the solid volume displacer are included in the same package. In other cases, the liquid container and the solid volume displacer are included in separate packages.

[0103] The kit can further include a buffer. For example, the kit can include buffers such as sample buffers, wash buffers, assay buffers, etc. The kit can further include additional reagents such as, but not limited to, detectable labels (e.g., fluorescent labels, colorimetric labels, chemiluminescent labels, multicolor reagents, avidin-streptavidin related detection reagents, radioactive labels, gold particles, magnetic labels, etc.). In certain embodiments, the kit can also include a calibration standard. For example, the kit can include a set of labeled beads, such as a set of standard fluorescently labeled beads.

[0104] In addition to the above components, the kit of the subject matter may further include instructions for practicing the method of the subject matter. These instructions may exist in various forms within the kit of the subject matter, and one or more of them may be present within the kit. One form in which these instructions may exist is as printed information on a suitable medium or substrate, such as one or more sheets of paper on which the information is printed, the package of the kit, an attached document, and the like. Another means is a computer-readable medium on which the information is recorded or stored, such as a CD, DVD, Blu-ray (registered trademark), computer-readable memory (e.g., flash memory), and the like. Yet another form that may exist is a website address that may be used via the Internet to access information at a deleted site. Instructions in any convenient form may be present within the kit.

[0105] Utility The systems and methods of the subject matter find use in applications where cell analysis from biological samples may be desired for research, laboratory testing, or therapeutic use. In some embodiments, the systems and methods of the subject matter facilitate the analysis of cells obtained from fluid or tissue samples, such as specimens for diseases including, but not limited to, cancer. The systems and methods of the present disclosure also enable the analysis of cells from biological samples (e.g., organs, tissues, tissue fragments, fluids) with improved efficiency and at low cost.

[0106] The subject systems and methods find use in applications where analysis of a sample using a dry reagent composition (e.g., a dye composition) is desired. For example, the subject systems and methods find use in applications where analysis of a sample using a dry polymer dye composition is desired. Embodiments of the subject systems and methods also find use in applications where analysis of a sample using a combination of a dry polymer dye composition and a dry non-polymer dye composition is desired. Thus, the subject systems and methods find use in applications where a sample is analyzed for a target analyte using a corresponding dry polymer dye composition. In some cases, where a non-polymer dye composition is also included in the liquid container of the system, the subject systems and methods find use in applications where a sample is analyzed for a target analyte using a corresponding dry polymer dye composition and a dry non-polymer dye composition.

[0107] The subject systems and methods find use in applications where it is desirable to minimize the volume of liquid required to reconstitute or rehydrate a dry reagent composition (e.g., a polymeric dye composition). As described herein, embodiments of the subject systems and methods provide a liquid container having an inner wall with a dry reagent composition, and a solid volume displacer positioned inside the liquid container and configured to occupy a majority of the liquid container volume below the upper portion of the dry reagent composition. Thus, by introducing the solid volume displacer into the liquid container in a manner sufficient to displace liquid onto the inner surface of the wall of the liquid container to reconstitute the dry reagent composition, it is facilitated to minimize the volume of liquid required to reconstitute or rehydrate a dry reagent composition (e.g., a polymeric dye composition). Minimizing the volume of liquid required to reconstitute or rehydrate a dry reagent composition can promote a higher reagent concentration after reconstitution or rehydration. A higher reagent concentration after reconstitution or rehydration can facilitate collection of more precise and / or accurate data regarding an assay performed using the subject system, and / or can reduce the cost of performing an assay using the subject system. For example, the subject systems and methods can facilitate a reduction in the volume of liquid required to reconstitute or rehydrate a dry reagent composition as compared to a system in which the dry reagent composition is provided, whereas other methods of moving liquid into the dry reagent composition (e.g., vortexing) are based on reconstituting the dry reagent composition.

[0108] The subject systems and methods find use in applications where analysis of a sample using two or more reagents (e.g., dye compositions) is desired. For example, the subject systems and methods find use in applications where analysis of a sample using two or more polymeric dye compositions is desired. Embodiments of the subject systems and methods may also find use in applications where analysis of a sample using two or more polymeric dye compositions in combination with one or more non-polymeric dye compositions is desired. Thus, the subject systems and methods find use in applications where a sample is analyzed for two or more analytes of interest using two or more corresponding polymeric dye compositions. In some cases, where a non-polymeric dye composition is also included in the liquid container, the subject systems and methods find use in applications where a sample is analyzed for two or more analytes of interest using two or more corresponding polymeric and non-polymeric dye compositions.

[0109] The subject systems and methods find use in applications where minimization of dye-dye interactions is desired. As described herein, embodiments of the subject systems and methods provide two or more dry polymer dye compositions that are precisely positioned on the inner surface of the wall of a liquid container. Thus, the precise positioning of the dye compositions relative to one another on the inner surface of the wall of the liquid container facilitates minimization of dye-dye interactions. Minimization of dye-dye interactions can facilitate collection of more precise and / or accurate data regarding an assay performed using the subject systems. For example, the subject systems and methods can facilitate reduction of dye-dye interactions as compared to containers where two or more dye compositions are provided but not precisely positioned relative to one another. Further, precise positioning of the dye compositions relative to one another on the inner surface of the wall of the liquid container allows the dye compositions to be positioned further from the bottom of the liquid container as additional dye compositions are added. Positioning the dye compositions further from the bottom of the liquid container can require a greater volume of liquid to reconstitute or rehydrate the dye compositions. Thus, when combined with introducing a solid volume displacer into the liquid container in a manner sufficient to displace the volume of liquid onto the inner surface of the wall of the liquid container to reconstitute the dye compositions, precise positioning of the dye compositions on the inner surface of the wall of the liquid container relative to one another can facilitate collection of more precise and / or accurate data regarding an assay performed using two or more dye compositions in the subject systems.

[0110] The following examples are presented by way of illustration and not by way of limitation.

Example

[0111] The following examples are presented to provide a complete disclosure and description of the methods of making and using the present invention to those skilled in the art, and are not intended to limit the scope that the inventors regard as the invention, nor are they intended to represent that the following experiments are all or the only experiments performed. Although efforts have been made to ensure the accuracy of the numbers used (e.g., amounts, temperatures, etc.), some experimental error and deviation should be considered. Unless otherwise indicated, parts are by weight, molecular weight is weight-average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure. Additionally, abbreviations for common laboratory protocols can be used (e.g., hr = hour, min = minute, ml = milliliter, ul = microliter, rpm = revolutions per minute, g (in the context of centrifugation) = multiples of gravity, etc.).

[0112] The following experiments compare a system for reconstituting the dry reagent composition of the present invention with prior art systems and methods for reconstituting dry reagent compositions.

[0113] Materials and Methods The dye composition is dispensed onto the inner surface of the wall of a 12×75 mm tube, for example using a liquid handler device, to create a liquid container according to one embodiment of the present invention. The dye composition is dispensed to a distance of 20 mm from the bottom of the tube and then dried.

[0114] Three embodiments of the solid volume displacer according to the embodiments of the present invention are machined. The solid volume displacer 1 is machined such that the diameter of the outer surface of the body of the displacer is 1 mm smaller than the diameter of the inner surface of the wall of the liquid container. The first end of the solid volume displacer 1 is configured as a flange that prevents the second end of the displacer from contacting the bottom of the liquid container. Both the solid volume displacers 2 and 3 are machined such that the diameter of the outer surface of the body of the displacer is 0.5 mm smaller than the diameter of the inner surface of the wall of the liquid container. The first end of the solid volume displacer 2 is configured as a flange that prevents the second end of the displacer from contacting the bottom of the liquid container. The first end of the solid volume displacer 3 is configured as a flange that allows the second end of the displacer to contact the bottom of the liquid container.

[0115] The control method is tested by filling the liquid container with liquid until it is observed that all of the dry dye composition in the liquid container has been reconstituted.

[0116] Figure 3A provides a depiction of a method for testing a vortex treatment for reconstituting the dry dye composition in a liquid container. From left to right, first, a volume of liquid is introduced into the liquid container. Next, the liquid container is vortexed to move the introduced volume of liquid up the wall of the liquid container. After the vortex treatment, the liquid container is observed to determine whether the introduced volume of liquid was sufficient to reconstitute all of the dry dye composition in the liquid container.

[0117] Figures 2A - 2C provide a depiction of a method of testing a solid volume displacer (e.g., according to one embodiment of the present invention). In Figure 2A, a volume of liquid 230 is introduced into liquid container 210. In Figure 2B, solid volume displacer 220 is introduced into liquid container 210 in a manner sufficient to displace the volume of liquid 230 onto the inner surface of the wall 213 of the liquid container. In Figure 2C, solid volume displacer 220 has been removed from liquid container 210, and the liquid container is observed to determine whether the introduced volume of liquid was sufficient to reconstitute all of the dry dye composition in the liquid container.

[0118] [Table 1]

[0119] The previous calculations were performed for the volume of the sphere (twice the volume of the bottom end or the second end) and the volume of the cylinder using the following equations, respectively.

[0120] [Number]

[0121] Results The control method requires 1463.2 ul of liquid to reconstitute all of the dry dye composition. The vortexing process for reconstituting the dry dye composition in the liquid container requires significantly less liquid than the control. To reconstitute all of the dry dye composition by vortexing, 827 ul of liquid needs to be introduced into the liquid container before the liquid container is vortexed.

[0122] Solid volume displacer 1 requires 323.8 ul of liquid to reconstitute all of the dry dye composition in the liquid container. Thus, solid volume displacer 1 requires 77.9% less liquid volume than the control and 60.8% less liquid volume than the vortexing process to reconstitute all of the dry dye composition.

[0123] The solid volume displacer 2 requires 167.5 ul of liquid to reconstitute all of the dry dye composition in the liquid container. Thus, the solid volume displacer 2 requires 88.6% less liquid volume than the control and 79.7% less liquid volume than vortexing to reconstitute all of the dry dye composition. FIGS. 3A and 3B depict a comparison of vortexing to reconstitute the dry dye composition as compared to using the solid volume displacer 2. FIG. 3A depicts, from left to right, the introduced liquid volume that fills the liquid container up to a height of 10 mm on the wall of the liquid container, the liquid container having been vortexed, and the resulting liquid volume containing a relatively diluted reconstituted dye composition. FIG. 3B depicts, from left to right, the introduced liquid volume that only partially fills the rounded bottom of the liquid container, the solid volume displacer 2 being introduced into the liquid container in a manner sufficient to displace the volume of liquid up onto the inner surface of the wall of the liquid container, and the resulting liquid volume containing a reconstituted dye composition at a relatively high concentration after the solid volume displacer 2 has been removed.

[0124] The solid volume displacer 3 requires 139.1 ul of liquid to reconstitute all of the dry dye composition in the liquid container. Thus, the solid volume displacer 3 requires 90.5% less liquid volume than the control and 83.2% less liquid volume than vortexing to reconstitute all of the dry dye composition.

[0125] Notwithstanding the appended claims, the disclosure is also defined by the following appendices. 1. A system for reconstituting a dry reagent composition, comprising a liquid container having an open end and a bottom separated by an intervening wall, the inner surface of the wall containing the dry reagent composition, and a solid volume displacer positioned inside the liquid container and configured to occupy a majority of the liquid container volume below the upper portion of the dry reagent composition. A system comprising. 2. The system according to appended claim 1, wherein the solid volume displacer is configured to occupy 60% or more of the volume of the liquid container below the upper part of the dry reagent composition. 3. The system according to appended claim 2, wherein the solid volume displacer is configured to occupy 75% or more of the volume of the liquid container below the upper part of the dry reagent composition. 4. The system according to appended claim 3, wherein the solid volume displacer is configured to occupy 85% or more of the volume of the liquid container below the upper part of the dry reagent composition. 5. The system according to appended claim 4, wherein the solid volume displacer is configured to occupy 90% or more of the volume of the liquid container below the upper part of the dry reagent composition.

[0126] 6. The system according to any one of appended claims 1 to 5, wherein the dry reagent composition contains a dye and / or beads. 7. The system according to appended claim 6, wherein the dry reagent composition contains a dye. 8. The system according to appended claim 6 or 7, wherein the dye is a polymer dye. 9. The system according to appended claim 8, wherein the polymer dye contains a conjugated polymer. 10. The system according to any one of appended claims 1 to 9, wherein the liquid container contains a volume of liquid greater than or equal to the volume of the liquid container below the upper part of the dry reagent composition, and is not configured to be occupied by the solid volume displacer.

[0127] 11. The system according to any one of appended claims 1 to 10, wherein the dry reagent composition is positioned 3 mm or more from the bottom of the liquid container. 12. The system according to appended claim 11, wherein the dry reagent composition is positioned 10 mm or more from the bottom of the liquid container. 13. The system according to appended claim 12, wherein the dry reagent composition is positioned 20 mm or more from the bottom of the liquid container. 14. The system according to any one of appended claims 1 to 13, wherein the inner surface of the wall of the liquid container contains two or more clearly positioned dry reagent compositions. 15. The system according to appended claim 14, wherein two or more dry reagent compositions are positioned at separate locations on the inner surface of the wall of the liquid container.

[0128] 16. The inner surface of the wall of the liquid container contains six or more clearly positioned dry reagent compositions, and the system according to any one of appendices 1 to 15. 17. Two or more of the dry reagent compositions contain a dye, and the system according to any one of appendices 14 to 16. 18. The dye is a polymer dye, and the system according to appendix 17. 19. The polymer dye contains a conjugated polymer, and the system according to appendix 18. 20. The liquid container includes a tube or a vial, and the system according to any one of appendices 1 to 19.

[0129] 21. The solid volume displacer is configured as a pestle, and the system according to any one of appendices 1 to 20. 22. The volume of the liquid container below the upper part of the dry reagent composition, which is not configured to be occupied by the solid volume displacer, is 350 μl or less, and the system according to any one of appendices 1 to 21. 23. The volume of the liquid container below the upper part of the dry reagent composition, which is not configured to be occupied by the solid volume displacer, is 150 μl or less, and the system according to appendix 22. 24. The solid volume displacer includes a first end, a second end, and a body therebetween, and the second end and the body are configured to be positioned inside the liquid container, and the system according to any one of appendices 1 to 23. 25. The body of the solid volume displacer is configured as a cylinder, and the system according to appendix 24.

[0130] 26. The body of the solid volume displacer includes an outer surface concentric with the inner surface of the wall of the liquid container, and the system according to appendix 25. 27. The difference between the diameter of the outer surface of the body of the solid volume displacer and the diameter of the inner surface of the wall of the liquid container is 1 mm or less, and the system according to appendix 26. 28. The system according to appended claim 26, wherein the difference between the outer diameter of the main body of the solid volume displacer and the inner diameter of the wall of the liquid container is 0.5 mm or less. 29. The system according to appended claim 26, wherein the difference between the outer diameter of the main body of the solid volume displacer and the inner diameter of the wall of the liquid container is 0.05 mm or less. 30. The system according to any one of appended claims 25 to 29, wherein the outer surface of the main body of the solid volume displacer and the wall of the liquid container are tapered.

[0131] 31. The system according to any one of appended claims 25 to 30, wherein when the first end of the solid volume displacer is positioned outside the liquid container and the main body and the second end of the solid volume displacer are positioned inside the liquid container. 32. The system according to appended claim 31, wherein the first end of the solid volume displacer includes a flange. 33. The system according to appended claim 32, wherein the flange is configured such that the second end of the solid volume displacer does not contact the bottom of the liquid container. 34. The system according to appended claim 33, wherein the flange is configured to allow the second end of the solid volume displacer to contact the bottom of the liquid container. 35. The system according to any one of appended claims 25 to 34, wherein the solid volume displacer further comprises a ring of filter medium positioned around the outside of the main body near the first end.

[0132] 36. The system according to appended claim 35, wherein the ring of filter medium is configured to prevent liquid from splashing out of the liquid container when the main body of the solid volume displacer is positioned inside the liquid container. 37. The system according to any one of appended claims 25 to 36, wherein the first end of the solid volume displacer includes a gripping portion. 38. The system according to any one of appended claims 1 to 37, wherein the solid volume displacer is hollow. 39. The system according to any one of appended claims 25 to 37, wherein the bottom of the liquid container is round. 40. The system according to appended claim 39, wherein the second end of the solid volume displacer is round and concentric with the bottom of the liquid container.

[0133] 41. The system according to any one of appended claims 1 to 40, wherein the solid volume displacer is configured to be attached to a mechanical mixer. 42. The system according to any one of appended claims 1 to 41, wherein the solid volume displacer comprises a first polymer material. 43. The system according to appended claim 42, wherein the first polymer material comprises a first hydrophobic polymer material. 44. The system according to appended claim 43, wherein the first hydrophobic polymer material comprises a first plastic, a first resin, a first metal, or a first ceramic. 45. The system according to appended claim 44, wherein the first hydrophobic polymer material comprises a first plastic.

[0134] 46. The system according to appended claim 45, wherein the first plastic comprises LDPE or PTFE. 47. The system according to any one of appended claims 1 to 46, wherein the liquid container comprises a second polymer material. 48. The system according to appended claim 47, wherein the second polymer material comprises a second hydrophobic polymer material. 49. The system according to appended claim 48, wherein the second hydrophobic polymer material comprises a second plastic. 50. The system according to appended claim 49, wherein the second plastic comprises LDPE or PTFE.

[0135] 51. The system according to any one of appended claims 1 to 50, wherein the liquid container comprises a seal. 52. The system according to appended claim 51, wherein the seal comprises a removable cap.

[0136] 53. A method for reconstituting a dry reagent composition, introducing a volume of liquid into a liquid container that includes an open end and a bottom separated by an intervening wall, the inner surface of the wall containing the dry reagent composition; Introducing a solid volume displacer into the liquid container in a manner sufficient to displace the liquid onto the inner surface of the wall of the liquid container to reconstitute the dry reagent composition A method comprising. 54. The method according to appendix 53, wherein the solid volume displacer occupies a majority of the liquid container volume below the upper part of the dry reagent composition. 55. The method according to appendix 54, wherein the solid volume displacer occupies 60% or more of the liquid container volume below the upper part of the dry reagent composition. 56. The method according to appendix 55, wherein the solid volume displacer occupies 75% or more of the liquid container volume below the upper part of the dry reagent composition. 57. The method according to appendix 56, wherein the solid volume displacer occupies 85% or more of the liquid container volume below the upper part of the dry reagent composition.

[0137] 58. The method according to appendix 57, wherein the solid volume displacer occupies 90% or more of the liquid container volume below the upper part of the dry reagent composition. 59. The method according to any one of appendices 53 to 58, wherein the dry reagent composition contains a dye and / or beads. 60. The method according to appendix 59, wherein the dry reagent composition contains a dye. 61. The method according to appendix 60, wherein the dye is a polymer dye. 62. The method according to appendix 61, wherein the polymer dye contains a conjugated polymer.

[0138] 63. The method according to any one of appendices 53 to 62, wherein the dry reagent composition is positioned 3 mm or more from the bottom of the liquid container. 64. The method according to appendix 63, wherein the dry reagent composition is positioned 10 mm or more from the bottom of the liquid container. 65. The method according to appendix 64, wherein the dry reagent composition is positioned 20 mm or more from the bottom of the liquid container. 66. The method according to any one of appendices 53 to 65, wherein the inner surface of the wall of the liquid container contains two or more clearly positioned dry reagent compositions. The method according to appended claim 66, wherein two or more than two drying reagent compositions are positioned at separate locations on the inner surface of the wall of the liquid container.

[0139] The method according to appended claim 67, wherein the inner surface of the wall of the liquid container contains six or more clearly positioned drying reagent compositions. The method according to any one of appended claims 66 to 68, wherein two or more of the drying reagent compositions contain a dye. The method according to appended claim 69, wherein the dye is a polymeric dye. The method according to appended claim 70, wherein the polymeric dye contains a conjugated polymer. The method according to any one of appended claims 53 to 71, wherein the liquid container includes a tube or a vial.

[0140] The method according to any one of appended claims 53 to 72, wherein the solid volume displacer is configured as a pestle. The method according to any one of appended claims 53 to 73, wherein the volume of the liquid introduced into the liquid container is 5000 μl or less. The method according to appended claim 74, wherein the volume of the liquid introduced into the liquid container is 350 μl or less. The method according to appended claim 75, wherein the volume of the liquid introduced into the liquid container is 150 μl or less. The method according to any one of appended claims 53 to 76, wherein the solid volume displacer includes a first end, a second end, and a body therebetween, and the second end and the body are configured to be positioned inside the liquid container.

[0141] The method according to appended claim 77, wherein the body of the solid volume displacer is configured as a cylinder. The method according to appended claim 78, wherein the body of the solid volume displacer includes an outer surface that is concentric with the inner surface of the wall of the liquid container. The method according to appended claim 79, wherein the difference between the diameter of the outer surface of the body of the solid volume displacer and the diameter of the inner surface of the wall of the liquid container is 1 mm or less. 81. The method according to appended note 80, wherein the difference between the outer diameter of the main body of the solid volume displacer and the inner diameter of the wall of the liquid container is 0.5 mm or less. 82. The method according to appended note 81, wherein the difference between the outer diameter of the main body of the solid volume displacer and the inner diameter of the wall of the liquid container is 0.05 mm or less.

[0142] 83. The method according to any one of appended notes 77 to 82, wherein the outer surface of the main body of the solid volume displacer and the wall of the liquid container are tapered. 84. The method according to any one of appended notes 77 to 83, wherein when the first end of the solid volume displacer is positioned inside the main body of the solid volume displacer and the second end is positioned inside the liquid container, it is configured to be positioned outside the liquid container. 85. The method according to appended note 84, wherein the first end of the solid volume displacer includes a flange. 86. The method according to appended note 85, wherein the flange is configured such that the second end of the solid volume displacer does not contact the bottom of the liquid container. 87. The method according to appended note 86, wherein the flange is configured to allow the second end of the solid volume displacer to contact the bottom of the liquid container.

[0143] 88. The method according to any one of appended notes 84 to 87, wherein the flange includes a ring of filter medium positioned around the outside of the main body near the first end. 89. The method according to appended note 88, wherein the ring of filter medium is configured to prevent liquid from splashing out of the liquid container when the main body of the solid volume displacer is positioned inside the liquid container. 90. The method according to any one of appended notes 77 to 89, wherein the first end of the solid volume displacer includes a gripping portion. 91. The method according to any one of appended notes 53 to 90, wherein the solid volume displacer is hollow. 92. The method according to any one of appended notes 77 to 90, wherein the bottom of the liquid container is round.

[0144] 93. The method according to appendix 92, wherein the second end of the solid volume displacer is round and concentric with the bottom of the liquid container. 94. The method according to any one of appendices 53 to 93, further comprising mixing the liquid and the dry reagent composition by moving the solid volume displacer. 95. The method according to appendix 94, wherein the movement includes rotating the solid volume displacer. 96. The method according to appendix 94 or 95, wherein the movement includes reciprocating the solid volume displacer. 97. The method according to any one of appendices 53 to 96, wherein the solid volume displacer is configured to be attached to a mechanical mixer.

[0145] 98. The method according to any one of appendices 53 to 97, wherein the solid volume displacer includes a first polymer material. 99. The method according to appendix 98, wherein the first polymer material includes a first hydrophobic polymer material. 100. The method according to appendix 99, wherein the first hydrophobic polymer material includes a first plastic, a first resin, a first metal, or a first ceramic. 101. The method according to appendix 100, wherein the first hydrophobic polymer material includes a first plastic. 102. The method according to appendix 100, wherein the first plastic includes LDPE or PTFE.

[0146] 103. The method according to any one of appendices 53 to 102, wherein the liquid container includes a second polymer material. 104. The method according to appendix 103, wherein the second polymer material includes a second hydrophobic polymer material. 105. The method according to appendix 104, wherein the second hydrophobic polymer material includes a second plastic. 106. The method according to appendix 105, wherein the second plastic includes LDPE or PTFE. 107. The method according to any one of appendices 53 to 106, wherein the liquid container is sealed and the method includes removing the seal before positioning the volume of liquid inside the liquid container.

[0147] 108. The method according to any one of appendices 53 to 107, further comprising sealing the liquid container after all the dry reagent compositions have been reconstituted. 109. The method according to appendix 107 or 108, wherein the seal comprises a removable cap.

[0148] 110. An assembly comprising an open end and a bottom separated by an intermediate wall, wherein the inner surface of the wall comprises a liquid container containing a dry reagent composition, the volume of the liquid introduced into the liquid container, and a solid volume displacer positioned inside the liquid container so as to displace the volume of the liquid onto the inner surface of the wall of the liquid container to contact the dry reagent composition. 111. The assembly according to appendix 110, wherein the solid volume displacer occupies most of the volume of the liquid container below the upper part of the dry reagent composition. 112. The assembly according to appendix 111, wherein the solid volume displacer occupies 60% or more of the volume of the liquid container below the upper part of the dry reagent composition. 113. The assembly according to appendix 112, wherein the solid volume displacer occupies 75% or more of the volume of the liquid container below the upper part of the dry reagent composition. 114. The assembly according to appendix 113, wherein the solid volume displacer occupies 85% or more of the volume of the liquid container below the upper part of the dry reagent composition.

[0149] 115. The assembly according to appendix 114, wherein the solid volume displacer occupies 90% or more of the volume of the liquid container below the upper part of the dry reagent composition. 116. The assembly according to any one of appendices 110 to 115, wherein the dry reagent composition comprises a dye and / or beads. 117. The assembly according to appendix 116, wherein the dry reagent composition comprises a dye. 118. The assembly according to appendix 117, wherein the dye is a polymeric dye. 119. The assembly according to appendix 118, wherein the polymeric dye comprises a conjugated polymer.

[0150] 120. The assembly according to any one of appendices 110 to 119, wherein the dry reagent composition is positioned 3 mm or more from the bottom of the liquid container. 121. The assembly according to appendix 120, wherein the dry reagent composition is positioned 10 mm or more from the bottom of the liquid container. 122. The assembly according to appendix 121, wherein the dry reagent composition is positioned 20 mm or more from the bottom of the liquid container. 123. The assembly according to any one of appendices 110 to 122, wherein the inner surface of the wall of the liquid container contains two or more clearly positioned dry reagent compositions. 124. The assembly according to appendix 123, wherein two or more dry reagent compositions are positioned at separate locations on the inner surface of the wall of the liquid container.

[0151] 125. The assembly according to appendix 123 or 124, wherein the inner surface of the wall of the liquid container contains six or more clearly positioned dry reagent compositions. 126. The assembly according to any one of appendices 123 to 125, wherein two or more dry reagent compositions contain a dye. 127. The assembly according to appendix 126, wherein the dye is a polymer dye. 128. The assembly according to appendix 127, wherein the polymer dye contains a conjugated polymer. 129. The assembly according to any one of appendices 110 to 128, wherein the liquid container includes a tube or a vial.

[0152] 130. The assembly according to any one of appendices 110 to 129, wherein the solid volume displacer is configured as a pestle. 131. The assembly according to any one of appendices 110 to 130, wherein the volume of the liquid introduced into the liquid container is 5000 μl or less. 132. The assembly according to appendix 131, wherein the volume of the liquid introduced into the liquid container is 350 μl or less. 133. The assembly according to appendix 132, wherein the volume of the liquid introduced into the liquid container is 150 μl or less. 134. The solid volume displacer includes a first end, a second end, and a body therebetween, and the second end and the body are positioned inside the liquid container, the assembly according to any one of appendices 110 to 133.

[0153] 135. The body of the solid volume displacer is configured as a cylinder, the assembly according to appendix 134. 136. The body of the solid volume displacer includes an outer surface concentric with the inner surface of the wall of the liquid container, the assembly according to appendix 135. 137. The difference between the diameter of the outer surface of the body of the solid volume displacer and the diameter of the inner surface of the wall of the liquid container is 1 mm or less, the assembly according to appendix 136. 138. The difference between the diameter of the outer surface of the body of the solid volume displacer and the diameter of the inner surface of the wall of the liquid container is 0.5 mm or less, the assembly according to appendix 137. 139. The difference between the diameter of the outer surface of the body of the solid volume displacer and the diameter of the inner surface of the wall of the liquid container is 0.05 mm or less, the assembly according to appendix 138.

[0154] 140. The outer surface of the body of the solid volume displacer and the wall of the liquid container are tapered, the assembly according to any one of appendices 134 to 139. 141. The first end of the solid volume displacer is positioned outside the liquid container, the assembly according to any one of appendices 134 to 140. 142. The first end of the solid volume displacer includes a flange, the assembly according to appendix 141. 143. The flange is in contact with the open end of the liquid container, and the second end of the solid volume displacer is not in contact with the bottom of the liquid container, the assembly according to appendix 142. 144. The flange is not in contact with the open end of the liquid container, and the second end of the solid volume displacer is in contact with the bottom of the liquid container, the assembly according to appendix 142.

[0155] 145. The assembly according to any one of appendices 136 to 144, wherein the solid volume displacer further comprises a ring of filter medium positioned around the outer surface of the body near the first end. 146. The assembly according to appendix 145, wherein the ring of filter medium is positioned between the outer surface of the body of the solid volume displacer and the inner surface of the wall of the liquid container in a manner sufficient to prevent liquid from spilling out of the liquid container. 147. The assembly according to any one of appendices 134 to 146, wherein the first end of the solid volume displacer includes a gripping portion. 148. The assembly according to any one of appendices 110 to 147, wherein the solid volume displacer is hollow. 149. The assembly according to any one of appendices 134 to 147, wherein the bottom of the liquid container is round.

[0156] 150. The assembly according to appendix 149, wherein the second end of the solid volume displacer is round and concentric with the bottom of the liquid container. 151. The assembly according to any one of appendices 110 to 150, wherein the solid volume displacer is configured to be attached to a mechanical mixer. 152. The assembly according to any one of appendices 110 to 151, wherein the solid volume displacer includes a first polymer material. 153. The assembly according to appendix 152, wherein the first polymer material includes a first hydrophobic polymer material. 154. The assembly according to appendix 153, wherein the first hydrophobic polymer material includes a first plastic, a first resin, a first metal, or a first ceramic.

[0157] 155. The assembly according to appendix 154, wherein the first hydrophobic polymer material includes a first plastic. 156. The assembly according to appendix 155, wherein the first plastic includes LDPE or PTFE. 157. The assembly according to any one of appendices 110 to 156, wherein the liquid container includes a second polymer material. 158. The assembly according to appendix 157, wherein the second polymer material comprises a second hydrophobic polymer material. 159. The assembly according to appendix 158, wherein the second hydrophobic polymer material comprises a second plastic.

[0158] 160. The assembly according to appendix 159, wherein the second plastic comprises LDPE or PTFE. 161. The assembly according to any one of appendices 110 to 160, wherein the liquid container comprises a removable seal attached to the liquid container. 162. The assembly according to appendix 161, wherein the removable seal comprises a removable cap.

[0159] 163. A kit comprising: a liquid container including an open end and a bottom separated by an intermediate wall, the inner surface of the wall containing a dry reagent composition; a solid volume displacer positioned inside the liquid container and configured to occupy a majority of the liquid container volume below the upper portion of the dry reagent composition; and a package configured to hold the liquid container and the solid volume displacer. 164. The kit according to appendix 163, wherein the solid volume displacer is configured to occupy 75% or more of the liquid container volume below the upper portion of the dry reagent composition. 165. The kit according to appendix 164, wherein the solid volume displacer is configured to occupy 85% or more of the liquid container volume below the upper portion of the dry reagent composition. 166. The kit according to appendix 165, wherein the solid volume displacer is configured to occupy 90% or more of the liquid container volume below the upper portion of the dry reagent composition. 167. The kit according to any one of appendices 163 to 166, wherein the dry reagent composition comprises a dye and / or beads.

[0160] 168. The kit according to appendix 167, wherein the dry reagent composition comprises a dye. 169. The kit according to appendix 168, wherein the dye is a polymer dye. 170. The kit according to appended item 169, wherein the polymeric dye contains a conjugated polymer. 171. The kit according to any one of appended items 163 to 170, wherein the dry reagent composition is positioned 3 mm or more from the bottom of the liquid container. 172. The kit according to appended item 171, wherein the dry reagent composition is positioned 10 mm or more from the bottom of the liquid container.

[0161] 173. The kit according to appended item 172, wherein the dry reagent composition is positioned 20 mm or more from the bottom of the liquid container. 174. The kit according to any one of appended items 163 to 173, wherein the inner surface of the wall of the liquid container contains two or more clearly positioned dry reagent compositions. 175. The kit according to appended item 174, wherein two or more dry reagent compositions are positioned at separate locations on the inner surface of the wall of the liquid container. 176. The kit according to appended item 174 or 175, wherein the inner surface of the wall of the liquid container contains six or more clearly positioned dry reagent compositions. 177. The kit according to any one of appended items 174 to 176, wherein two or more dry reagent compositions contain a dye.

[0162] 178. The kit according to appended item 177, wherein the dye is a polymeric dye. 179. The kit according to appended item 178, wherein the polymeric dye contains a conjugated polymer. 180. The kit according to any one of appended items 163 to 179, wherein the liquid container includes a tube or a vial. 181. The kit according to any one of appended items 163 to 180, wherein the solid volume displacer is configured as a pestle. 182. The kit according to any one of appended items 163 to 181, wherein the volume of the liquid container below the upper part of the dry reagent composition, which is not configured to be occupied by the solid volume displacer, is 350 μl or less.

[0163] The kit according to appended note 182, wherein the volume of the liquid container below the upper part of the dry reagent composition, which is not configured to be occupied by the solid volume displacer, is 150 μl or less. 184. The kit according to any one of appended notes 163 to 183, wherein the solid volume displacer includes a first end, a second end, and a body therebetween, and the second end and the body are configured to be positioned inside the liquid container. 185. The kit according to appended note 184, wherein the body of the solid volume displacer is configured as a cylinder. 186. The kit according to appended note 185, wherein the body of the solid volume displacer includes an outer surface concentric with the inner surface of the wall of the liquid container. 187. The kit according to appended note 186, wherein the difference between the diameter of the outer surface of the body of the solid volume displacer and the diameter of the inner surface of the wall of the liquid container is 1 mm or less.

[0164] 188. The kit according to appended note 187, wherein the difference between the diameter of the outer surface of the body of the solid volume displacer and the diameter of the inner surface of the wall of the liquid container is 0.5 mm or less. 189. The kit according to appended note 188, wherein the difference between the diameter of the outer surface of the body of the solid volume displacer and the diameter of the inner surface of the wall of the liquid container is 0.05 mm or less. 190. The kit according to any one of appended notes 184 to 189, wherein the outer surface of the body of the solid volume displacer and the wall of the liquid container are tapered. 191. The kit according to any one of appended notes 184 to 190, wherein the first end of the solid volume displacer is configured to be positioned outside the liquid container when the body of the solid volume displacer and the second end are positioned inside the liquid container. 192. The kit according to appended note 191, wherein the first end of the solid volume displacer includes a flange.

[0165] 193. The kit according to appended note 192, wherein the flange is configured such that the second end of the solid volume displacer does not contact the bottom of the liquid container. 194. The kit according to appendix 192, wherein the flange is configured to enable the second end of the solid volume displacer to contact the bottom of the liquid container. 195. The kit according to any one of appendices 184 to 194, wherein the solid volume displacer further comprises a ring of filter medium positioned around the outside of the body near the first end. 196. The kit according to appendix 195, wherein the ring of filter medium is configured to prevent liquid from splashing out of the liquid container when the body of the solid volume displacer is positioned inside the liquid container. 197. The kit according to any one of appendices 184 to 196, wherein the first end of the solid volume displacer includes a gripping portion.

[0166] 198. The kit according to any one of appendices 163 to 197, wherein the solid volume displacer is hollow. 199. The kit according to any one of appendices 184 to 197, wherein the bottom of the liquid container is round. 200. The kit according to appendix 199, wherein the second end of the solid volume displacer is round and concentric with the bottom of the liquid container. 201. The kit according to any one of appendices 163 to 200, wherein the solid volume displacer is configured to be attached to a mechanical mixer. 202. The kit according to any one of appendices 163 to 201, wherein the solid volume displacer includes a first polymer material.

[0167] 203. The kit according to appendix 202, wherein the first polymer material includes a first hydrophobic polymer material. 204. The kit according to appendix 203, wherein the first hydrophobic polymer material includes a first plastic, a first resin, a first metal, or a first ceramic. 205. The kit according to appendix 204, wherein the first hydrophobic polymer material includes a first plastic. 206. The kit according to appendix 205, wherein the first plastic includes LDPE or PTFE. Kit according to any one of appendices 163 to 206, wherein the liquid container contains a second polymer material.

[0168] Kit according to appendix 207, wherein the second polymer material contains a second hydrophobic polymer material. Kit according to appendix 208, wherein the second hydrophobic polymer material contains a second plastic. Kit according to appendix 209, wherein the second plastic contains LDPE or PTFE. Kit according to any one of appendices 163 to 210, wherein the liquid container contains a seal. Kit according to appendix 211, wherein the seal contains a removable cap.

[0169] In at least some of the above embodiments, one or more elements used in one embodiment can be used interchangeably in another embodiment as long as the substitution is technically feasible. Those skilled in the art should understand that various other omissions, additions, and modifications can be made to the above methods and structures without departing from the scope of the claimed subject matter. It is intended that all such modifications and changes fall within the scope of the subject matter defined by the appended claims.

[0170] Generally, the terms used herein, and in particular the terms used in the appended claims (e.g., the body of the appended claims), are generally intended to be open terms (e.g., the term "including" should be construed as "including but not limited to", the term "has" should be construed as "has at least", the term "includes" should be construed as "including but not limited to", etc.), which is understood by those skilled in the art. When the introduction of a specific number of claim recitations is intended, such intention is explicitly recited in the claims, and when there is no such enumeration, it is further understood by those skilled in the art that such intention does not exist. For example, for purposes of illustration, the following appended claims may include the use of the introductory phrases "at least one" and "one or more" to introduce claim recitations. However, the use of such phrases should not be construed to mean that the introduction of a claim recitation by the indefinite article "a" or "an" limits a particular claim that includes such introduced claim recitation to an embodiment that includes only one such recitation. The same applies to the use of the definite article in introducing claim recitations (e.g., "a" and / or "an" is "at least one" or "one or more"). Further, even if the introduction of a specific number of claim recitations is explicitly enumerated, those skilled in the art will recognize that such enumeration should be construed to mean at least the number enumerated (e.g., the bare enumeration of "two recitations" without other modifiers means at least two recitations, or two or more recitations).Furthermore, when a convention similar to “at least one of A, B, and C, etc.” is used, generally, such a construction is intended in the sense that a person skilled in the art would understand the convention (e.g., “a system having at least one of A, B, and C” includes, but is not limited to, a system having only A, only B, only C, both A and B, both A and C, both B and C, and / or all of A, B, and C, etc.). When a convention similar to “at least one of A, B, or C, etc.” is used, generally, such a construction is intended in the sense that a person skilled in the art would understand the convention (e.g., “a system having at least one of A, B, or C” includes, but is not limited to, a system having only A, only B, only C, both A and B, both A and C, both B and C, and / or all of A, B, and C, etc.). It is further understood by a person skilled in the art that, regardless of the specification, claims, or drawings, substantially any separate words and / or phrases presenting two or more alternative terms should be understood to contemplate the possibility of including one of the terms, any of the terms, or both terms. For example, the phrase “A or B” is understood to include the possibilities of “A” or “B” or “A and B”.

[0171] In addition, when a feature or aspect of the present disclosure is described from the perspective of a Markush group, thus, it is recognized by a person skilled in the art that the present disclosure is also described from the perspective of any individual member of the Markush group or any subgroup of the members.

[0172] As will be understood by those skilled in the art, for any purpose, for example, from the perspective of providing a written description, all ranges disclosed in this specification also include any possible sub-ranges and combinations of those sub-ranges. Any listed range sufficiently indicates that the same range can be decomposed into at least equal halves, thirds, fourths, fifths, tenths, etc., and can be easily recognized as enabling such decomposition. As a non-limiting example, each range considered in this specification can be easily decomposed into, for example, the lower third, the middle third, and the upper third. As will also be understood by those skilled in the art, all terms such as "maximum", "at least", "greater than", "less than", etc. include the recited number and refer to ranges that can be later decomposed into sub-ranges as discussed above. Finally, as will be understood by those skilled in the art, a range includes each individual member. Thus, for example, a group having 1 to 3 articles refers to a group having 1, 2, or 3 articles. Similarly, a group having 1 to 5 articles refers to a group having 1, 2, 3, 4, or 5 articles, etc.

[0173] The above invention has been described in some detail by way of illustration and example for the purpose of clear understanding. However, it will be readily apparent to those skilled in the art that certain changes and modifications can be made to those inventions without departing from the spirit or scope of the appended claims in light of the teachings of the present invention.

[0174] Accordingly, the foregoing merely illustrates the principles of the invention. It will be appreciated by those skilled in the art that, although not explicitly described or shown herein, various arrangements embodying the principles of the invention can be devised that are within its spirit and scope. Further, all of the 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 further the art, and are to be construed as not being limited to such specifically recited examples and conditions. Additionally, all descriptions herein of the principles, aspects, and embodiments of the invention, as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Moreover, such equivalents are intended to include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function regardless of structure. Further, nothing disclosed herein is intended to be dedicated to the public, whether or not such disclosure is explicitly recited in the claims.

[0175] Accordingly, the scope of the invention is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of the invention are embodied by the appended claims. In the claims, 35 U.S.C. § 112(f) or 35 U.S.C. § 112(6) is expressly defined to be used to limit a claim only when the exact phrase "means for" or the exact phrase "step for" begins the limitation in the claim, and such exact phrases are not used in the limitation of a claim, 35 U.S.C. § 112(f) or 35 U.S.C. § 112(6) is not applied.

[0176] Cross - Reference to Related Applications This application claims priority under 35 U.S.C. § 119(e) to the filing date of U.S. Provisional Patent Application No. 63 / 350,794, filed on June 9, 2022, the disclosure of which is hereby incorporated by reference in its entirety.

Claims

1. A system for reconstituting a dry reagent composition, comprising: a liquid container having an open end and a bottom separated by a wall therebetween, the inner surface of the wall containing the dry reagent composition; and a solid volume displacer positioned inside the liquid container and configured to occupy a majority of the volume of the liquid container below the upper portion of the dry reagent composition. The system.

2. The system according to any one of the preceding claims, wherein the dry reagent composition comprises a dye and / or beads.

3. The system according to claim 2, wherein the dry reagent composition comprises a dye.

4. The system according to claim 3, wherein the dye is a polymeric dye.

5. The system according to claim 4, wherein the polymeric dye comprises a conjugated polymer.

6. The system according to any one of the preceding claims, wherein the liquid container contains a volume of liquid greater than the volume of the liquid container below the upper portion of the dry reagent composition, which is not configured to be occupied by the solid volume displacer.

7. The system according to any one of the preceding claims, wherein the inner surface of the wall of the liquid container contains two or more clearly positioned dry reagent compositions.

8. The system according to any one of the preceding claims, wherein the liquid container comprises a tube or a vial.

9. The system according to any one of the preceding claims, wherein the solid volume displacer is configured as a pestle.

10. The system according to any one of the preceding claims, wherein the solid volume displacer comprises a first end, a second end, and a body therebetween, and the second end and the body are configured to be positioned inside the liquid container.

11. The system according to any one of the preceding claims, wherein the solid volume displacer is hollow.

12. The system according to any one of the preceding claims, wherein the liquid container comprises a seal.

13. A method for reconstituting a dry reagent composition, comprising: introducing a volume of liquid into a liquid container having an open end and a bottom separated by a wall therebetween, the inner surface of the wall containing the dry reagent composition; and introducing a solid volume displacer into the liquid container in a manner sufficient to displace the liquid onto the inner surface of the wall of the liquid container to reconstitute the dry reagent composition. The method.

14. A liquid container including an open end and a bottom separated by an intervening wall, the inner surface of said wall containing a dry reagent composition, and the volume of liquid introduced into said liquid container, and a solid volume displacer positioned inside said liquid container to displace said volume of liquid onto said inner surface of said wall of said liquid container to contact said dry reagent composition An assembly comprising.

15. A liquid container including an open end and a bottom separated by an intervening wall, the inner surface of said wall containing a dry reagent composition, and a solid volume displacer positioned inside said liquid container and configured to occupy a majority of the liquid container volume below the upper portion of said dry reagent composition, and a package configured to hold said liquid container and said solid volume displacer A kit comprising.

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