Systems for reconstitution of dry reagent compositions and methods of use thereof

Abstract

A system for reconstitution of a dry reagent composition is presented. Aspects of the system include a liquid container having an inner wall with a dry reagent composition and a solids volume displacer placed inside the liquid container that occupies a majority of the volume of the liquid container below the top of the dry reagent composition. Aspects of the invention further include methods of using the system and kits including the system.

Description

[0001] Cross-references to related patent applications

[0002] Pursuant to Title 35 of the United States Patent Act, Section 119(e), this application claims the benefit of priority to the filing date of U.S. Provisional Patent Application No. 63 / 350,794, filed on June 9, 2022; the disclosure of U.S. Provisional Patent Application No. 63 / 350,794 is incorporated herein by reference in its entirety and becomes a part of the present invention. Background Art

[0003] Assays for determining the presence and concentration of an analyte in a biological sample fluid typically rely on specific binding of a detectable label to the target analyte. The detectable label can be a marker that can be observed by the naked eye or detected by spectroscopy (e.g., fluorescence or UV-visible spectroscopy). Typically, fluorescent dyes can be used as detectable labels, wherein the fluorescent dyes include specific fluorochromes. Fluorochromes can have certain properties, such as absorption spectra, extinction coefficients at wavelengths that are convenient for excitation, emission spectra, and quantum efficiency. Quantum efficiency is the number of photons emitted for each photon absorbed.

[0004] Multiplexing allows for the simultaneous detection of multiple analytes in a single test. For immunoassays, multiplexing involves a cocktail of antibodies, with each antibody labeled with a different dye. Each antibody binds to a specific analyte or antigen in the sample. This allows the different analytes or antigens to be distinguished and quantified based on the different dyes.

[0005] For convenience, multiplex detection reagents can be provided as premixed cocktails of individual binding molecules (e.g., antibodies). The cocktail may also contain other reaction components, such as buffers, salts, surfactants, etc. The most convenient cocktail is one that contains all the necessary components (single detection reagent). In this case, the sample is simply added to the reaction vessel containing the cocktail and the test is performed.

[0006] Stable assay reagents can be conveniently provided, especially those that are stable at room temperature. This allows for the transport and storage of reagents under non-refrigerated conditions. This is particularly important for testing in rural areas where resources, including electricity, may be limited. To this end, stable reagents can be provided by drying or lyophilizing aqueous solutions of reagents. However, some labeling reagents are problematic. If these reagents are in physical contact with each other, they will cross-link. After resuspending in a liquid solution, they remain cross-linked, so that the labels are no longer associated with a single antibody, resulting in erroneous results. This problem is particularly prominent in the field of flow cytometry, where multiple polymer dyes are used to create multiple (e.g., 10 or more) fluorescent channels using only one excitation wavelength. However, due to the chemical properties of these polymer dyes, multiple dyes cannot be stored in a mixed cocktail. It has been observed that polymer entities bind to each other in solution. This results in erroneous or abnormal cell populations that cannot exist in reality.

[0007] The 10th,545,137th United States Patent (USP) discloses a kind of reagent device, it has solid carrier and the first and second dry macromolecular dye compositions that are placed separately relative to the solid carrier surface, compared with providing two or more dye compositions but not being placed separately relative to each other, reduced the interaction between dyes. During use, the aqueous liquid is contacted with the dye composition, and the dye composition that is placed separately can be reconstructed. Reagent concentration must be kept, to obtain good detection performance. Therefore, the aqueous liquid that contacts with the dye composition can be a biological sample liquid, so that the analyte of reagent and sample is maintained at required concentration. Summary of the invention

[0008] However, in the device of U.S. Pat. No. 10,545,137, although the interaction between dyes is reduced, the inventors have recognized that a large amount of liquid is required to reconstitute the separately placed dye composition, which can lead to a decrease in dye concentration. This problem is amplified as the number of separately placed dye compositions increases, effectively limiting the number of dye compositions that can be used when using the device for detection.

[0009] Accordingly, the present inventors have recognised that it would be desirable to be able to reconstitute dye compositions using small amounts of liquid.

[0010] In view of this situation, the present disclosure is proposed, and its purpose is to provide a system for reconstructing a dry reagent composition, which 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 the dry reagent composition.

[0011] The present invention proposes a system for reconstructing a dry reagent composition. Various aspects of the system include a liquid container having an inner wall with a dry reagent composition and a solid volume displacer placed inside the liquid container to occupy most of the liquid container volume below the top of the dry reagent composition. Various aspects of the present invention further include a method of using the system and a kit including the system. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 A system according to an embodiment of the present invention is shown.

[0013] Figure 2A-2C Shows the use Figure 1 Methods for systematic reconstitution of dried reagent compositions.

[0014] Figure 3A-3B The volume of liquid required to reconstitute a dried reagent composition using a system according to an embodiment of the present invention and not using a system of the present invention were compared.

[0015] Figure 4A-4B A stepped solid volume displacer according to an embodiment of the present invention is described.

[0016] Figure 5A-5B A straight cylindrical solid volume displacer according to an embodiment of the present invention is described. DETAILED DESCRIPTION

[0017] The present invention proposes a system for reconstructing a dry reagent composition. Various aspects of the system include a liquid container having an inner wall with a dry reagent composition and a solid volume displacer placed inside the liquid container to occupy most of the liquid container volume below the top of the dry reagent composition. Various aspects of the present invention further include a method of using the system and a kit including the system.

[0018] Before describing the present invention in more detail, it should be understood that the present invention is not limited to the specific embodiments described, which may, of course, vary. It should also be understood that since the scope of the present invention will be limited only by the appended claims, the terms in the present invention are only used to describe specific embodiments and are not intended to be limiting.

[0019] If the present invention proposes a range of values, it should be understood that the present invention includes every intermediate value between the upper and lower limits of this range (unless the context clearly stipulates otherwise, it is accurate to one tenth of the unit of the lower limit), as well as any other specified values ​​or intermediate values ​​within this specified range. The upper and lower limits of these smaller ranges may be independently included in the smaller ranges and are also included in the scope of the present invention, but are subject to any explicitly excluded limits in the specified range. If the specified range includes one or two of the limits, the present invention also includes a range excluding any or both of these included limits.

[0020] Certain ranges in the present invention are expressed as numerical values ​​preceded by the term "about". The term "about" as used in the present invention is used to provide literal support for the exact number that follows it, as well as numbers that are close to or approximately the number that follows it. In determining whether a number is close to or approximately a number that is explicitly recited, the close or approximate unrecited number can be a number that is substantially equivalent to the explicitly recited number in the context in which it is presented.

[0021] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention belongs. Although any methods and materials similar or equivalent to the methods or materials described herein may also be used in the practice or testing of the present invention, representative illustrative methods and materials are described below.

[0022] All publications and patents cited in this specification are incorporated herein by reference as if each individual publication or patent was specifically and individually indicated to be incorporated herein by reference for the purpose of disclosing and describing the methods and / or materials in connection with the cited publication. Citation of any publication is for the purpose of disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Furthermore, the dates of publication provided may be different from the actual publication dates which need to be separately confirmed.

[0023] It is noted that, as used in the present disclosure and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that any optional elements may be excluded when drafting the claims. Therefore, this statement is intended to serve as a precondition for using exclusive terminology such as "only," "only" and the like in connection with the recitation of claim elements or for using a "negative" limitation.

[0024] After reading this disclosure, it will be apparent to those skilled in the art that each individual embodiment described and illustrated in the present invention has discrete components and features that are readily separable or combined with the features of any of the other multiple embodiments without departing from the scope or spirit of the present invention. Any method recited may be implemented in the order of events recited or in any other order that is logically possible.

[0025] Although methods and compositions have been or will be described for grammatical fluency and functional explanation, it should be clearly understood that, except as expressly provided in 35 U.S. Patent Act § 112, the claims should not be construed as necessarily being limited in any respect to the limitations of “means” or “steps” but rather be given the full meaning and scope of equivalents provided by the claims in accordance with the doctrine of equivalents, and in the event that a claim is expressly provided in 35 U.S. Patent Act § 112, it should be given full legal equivalents in accordance with 35 U.S. Patent Act § 112.

[0026] Based on the above summary, the present invention proposes a system for reconstructing a dry reagent composition. When further describing various embodiments of the present invention, the subject system is first described in more detail. Next, a method for using the subject system and a kit including the system are proposed.

[0027] System for reconstitution of dried reagent compositions

[0028] Embodiments of the present invention include a system for reconstructing a dry reagent composition, which is optimized for reconstructing or rehydrating the dry reagent composition and ultimately for detection, such as liquid sample detection. Various aspects of the system include a liquid container having an inner wall with a dry reagent composition and a solid volume displacer placed inside the liquid container to occupy most of the liquid container volume below the top of the dry reagent composition. The system allows a certain volume of liquid to be controlled and displaced in a manner sufficient to reconstruct the dry reagent composition, and this displacement method has many advantages, including but not limited to improvements in usability, cost and performance.

[0029] Liquid Container

[0030] According to the above summary, the system of the present invention includes a liquid container. In certain embodiments, the liquid container can be used for detection, such as detecting a liquid sample (such as a biological sample), such as detecting whether one or more analytes are present in the sample. According to certain embodiments of the present disclosure, the liquid container includes an open end and a bottom, which are separated by a wall, wherein the inner surface of the wall comprises a dry reagent composition.

[0031] The liquid container can be any convenient container that is compatible with the liquid sample and / or the reagents or analytes that may be in contact with the container. For example, the liquid container can be a liquid-compatible container that can hold the liquid sample. In some cases, the liquid sample can be an aqueous liquid sample, in which case the liquid container can be compatible with the aqueous sample. By "compatible" is meant that the liquid container (e.g., the material from which the container is made) is substantially inert (e.g., does not significantly react with) to the liquid and / or the reagents or analytes that may be in contact with the container.

[0032] The liquid container may include an open end and a bottom relative to the open end. In these examples, the open end of the liquid container (e.g., by an opening) exposes the inside of the liquid container to the surrounding environment, so that the contents of the liquid container are under the same atmospheric pressure as the surrounding environment. The longest cross-sectional dimension (e.g., diameter) of the liquid container may be different, and in some examples, the range is 0.5cm-5cm, such as 0.5cm-4.5cm, or 0.5cm-4cm, or 0.5cm-3.5cm, or 0.5cm-3cm, or 0.5cm-2.5cm, or 0.5cm-2cm, or 0.5cm-1.5cm, or 0.5cm-1.2cm, or 0.9cm-1.0cm. In certain embodiments, the open end of the liquid container may include a protrusion so that the open end can be installed with a snap cap. In other embodiments, the open end of the liquid container may include a thread, and a threaded cap may be screwed on the opening of the open end.

[0033] In some instances, the bottom of the liquid container opposite the open end may include a closed end, and the wall rises from the closed end. Therefore, the liquid container can accommodate a certain volume of fluid (such as gas or liquid). In some embodiments, the liquid container can accommodate a certain volume of liquid. The size of the liquid container may depend on the volume of liquid to be accommodated in the liquid container. For example, the volume (e.g., liquid volume) that the liquid container can contain ranges from 0.1nl to 1000ml, for example, 0.1ml to 900ml, or 0.1ml to 800ml, or 0.1ml to 700ml, or 0.1ml to 600ml, or 0.1ml to 500ml, or 0.1ml to 400ml, or 0.1ml to 300ml, or 0.1ml to 200ml, or 0.1ml to 100ml, or 0.1ml to 50ml, or 0.1ml to 25ml, or 0.1ml to 10ml, or 0.1ml to 5ml, or 0.1ml to 1.5ml, or 0.1ml to 1ml, or 0.1ml to 0.5ml. In some examples, the liquid container can contain a volume (eg, liquid volume) ranging from 0.1 ml to 200 ml, such as 0.5 ml to 100 ml, such as 1.0 ml to 50 ml.

[0034] The shape of the liquid container may vary, depending on the purpose of the container. For example, as described in the present invention, the liquid container may be used for detection, such as detection of liquid samples (such as biological samples). In these cases, the liquid container may have a shape compatible with the detection and / or the method or other device for performing the detection. For example, the liquid container may have a typical laboratory equipment shape for performing the detection, or a shape compatible with other devices for performing the detection. In some embodiments, the wall of the liquid container is in the shape of a cylinder, wherein the cylinder has an opening at a first end and a closed bottom connected to the cylinder at a second end opposite to the first end. In some cases, the liquid container has a circular cross-section. The shape of the liquid container may vary, depending on the purpose of the solid volume displacer. For example, the liquid container may have a shape compatible with the disclosed method. In some embodiments, the wall of the liquid container is conical. In some cases, the liquid container may be conical, or may include a conical portion. For example, the liquid container may include a substantially cylindrical wall portion adjacent to the first end opening and a conical wall portion adjacent to the second closed end opposite to the first end. In certain embodiments, the bottom of the liquid container is rounded. In certain embodiments, the liquid container can be a vial, a test tube or a centrifuge tube (such as a microcentrifuge tube). In some cases, the liquid container is a vial. In some cases, the liquid container is a test tube. As described above, the liquid container can hold a certain volume (such as a liquid volume). In the embodiment that the liquid container is a vial or a test tube, the volume (e.g., liquid volume) that the liquid container can accommodate ranges from 1*10^-7ml to 1000ml, such as 0.5ml-900ml, or 0.5ml-800ml, or 0.5ml-700ml, or 0.5ml-600ml, or 0.5ml-500ml, or 0.5ml-400ml, or 0.5ml-300ml, or 0.5ml-200ml, or 0.5ml-100ml, or 0.5ml-50ml, or 0.5ml-25ml, or 0.5ml-10ml, or 0.5ml-5ml, or 1ml-5ml. In some instances, the volume (e.g., liquid volume) that the vial or test tube can accommodate ranges from 0.5ml to 5ml.

[0035] In some embodiments, the width of the liquid container (also referred to as the diameter of a cylindrical liquid container) ranges from 0.5 cm to 5 cm, such as 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 instances, the width (or diameter) of the liquid container ranges from 0.5 cm to 2.5 cm. In some instances, the width (or diameter) of the liquid container ranges from 0.5 cm to 1.2 cm, such as 0.9 cm to 1.0 cm. In some embodiments, the length of the liquid container ranges from 1 cm to 20 cm, such as 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 instances, the length of the liquid container ranges from 1 cm to 10 cm. In some instances, the length of the liquid container ranges from 1 cm to 5 cm. In some instances, the length of the liquid container ranges from 1 cm to 2.5 cm.

[0036] As described above, embodiments of the liquid container may be compatible with the liquid sample and / or the reagent or analyte in contact with 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 made of glass, such as, but not limited to, silicate glass, borosilicate glass, sodium borosilicate glass (e.g., PYREX TM ), fused quartz glass, fused silica glass, etc. Other examples of suitable container materials include polymeric materials. The polymeric material can be a hydrophobic polymeric material. The hydrophobic polymeric material can be a plastic, such as but not limited to polystyrene, polypropylene, polymethylpentene, polytetrafluoroethylene (PTFE), perfluoroether (PFE), fluorinated ethylene propylene (FEP), perfluoroalkoxy resin (PFA), polyethylene terephthalate (PET), polyethylene (PE), low-density polyethylene (LDPE), polyetheretherketone (PEEK), etc.

[0037] In some embodiments, as described above, the liquid container can hold a certain volume of fluid (such as gas or liquid). In some instances, the liquid container can hold a certain volume of liquid. In some embodiments, the liquid container can be sealed. That is, the liquid container can include a seal that can substantially prevent the contents of the liquid container (such as the liquid inside the liquid container) from flowing out of the liquid container. The seal of the liquid container can also substantially prevent other substances from entering the liquid container. For example, the seal can be a watertight seal that can substantially prevent liquid from entering and exiting the container, or it can be an airtight seal that can substantially prevent gas from entering and exiting the container. In some instances, the seal is a removable or frangible seal, and the contents of the liquid container can be exposed to the surrounding environment when needed (such as if a portion of the contents of the liquid container needs to be removed). In some instances, the seal is made of an elastic material, which can provide a barrier (such as a watertight and / or airtight seal) to retain the sample in the container. Depending on the type of container, specific types of seals include but are not limited to films, such as polymer films, lids, etc. Suitable seal materials include rubber or polymer seals, such as but not limited to silicone rubber, natural rubber, styrene-butadiene rubber, ethylene-propylene copolymer, polychloroprene, polyacrylate, polybutadiene, polyurethane, styrene-butadiene, etc. and combinations thereof. For example, in certain embodiments, the seal is a diaphragm that can be pierced with a needle, a syringe, or a cannula. The seal can also facilitate access to the sample in the liquid container and provide a protective barrier covering the container opening. In certain instances, the seal is a removable seal, such as a threaded cap, a snap cap, or other suitable sealing member that can be used for the opening of the liquid container. For example, before or after the sample has been added to the liquid container, a threaded cap can be screwed on top of the opening.

[0038] As mentioned above, the liquid container can hold a certain volume of fluid (such as gas or liquid). In some instances, the bottom of the liquid container has an inner surface and an outer surface. In some instances, the wall of the liquid container has an inner surface and an outer surface. In these embodiments, the inner surface of the liquid container is the surface of the liquid container facing the interior of the container. The inner surface can contact the liquid container contents. The outer surface of the liquid container is the surface of the liquid container facing away from the interior of the container. The outer surface does not contact the liquid container contents.

[0039] As mentioned above, the liquid container may include a dry reagent composition. In certain embodiments, the dry reagent composition is placed on a surface (such as an inner surface) of a liquid container. So-called "reagent" refers to a substance or compound that interacts with other substances or compounds in order to promote a chemical reaction or test whether a reaction occurs. For example, the substance or compound may include but is not limited to microspheres, preservatives, surfactants, defoamers, buffers (such as pH buffers), reactants, specific binding members (such as antibodies), dyes, etc. So-called "drying" refers to that the reagent composition contains a small amount of solvent (or, for example, substantially free of solvent), or the reagent composition is combined with a certain amount of solvent to reduce its own viscosity. For example, in certain embodiments, the dry reagent composition can be a relatively high viscosity liquid (such as syrup), and under standard conditions, adding a certain volume of solvent (such as water) can reduce the viscosity of the reagent composition 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 includes 25 wt% or less of solvent, such as 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 non-fluid. In some cases, the dry reagent composition is substantially solid. For example, under standard conditions, the dry reagent composition can have high viscosity, such as 10,000cP or more, or 25,000cP or more, or 50,000cP or more, or 75,000cP or more, or 100,000cP or more, or 150,000cP or more, or 200,000cP or more, or 250,000cP or more. In certain embodiments, the dry reagent can be partially or completely dissolved in a certain volume of solvent. For example, the dry reagent composition can be rehydrated or reconstructed in water. In certain embodiments, the dry reagent composition can maintain its shape and position (for example, on the inner surface of the liquid container adhered with the composition, see below) before contacting with the solvent.

[0040] In some examples, the dry reagent composition is placed on the inner surface of the liquid container wall. The liquid container may include one or more dry reagent compositions (e.g., dye compositions) on the container surface (e.g., the inner surface of the liquid container wall), such as 2 or more, or 3 or more, or 4 or more, or 5 or more, or 6 or more, or 7 or more, or 8 or more, or 9 or more, or 10 or more, or 11 or more, or 12 or more on the inner surface of the liquid container wall. In some embodiments, the container comprises 2-50 kinds of dry reagent compositions on the inner surface, such as 2-40 kinds, 2-30 kinds, 2-20 kinds, 2-15 kinds, 2-10 kinds, 2-7 kinds, or 2-5 kinds of dry reagent compositions on the inner surface of the wall. For example, the container may include 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 dry reagent compositions on the inner surface of the liquid container wall. In some cases, the liquid container includes 2 dry reagent compositions on the inner surface of the liquid container wall. In some cases, the liquid container includes 5 dry reagent compositions on the inner surface of the liquid container wall. In some cases, the liquid container includes 7 dry reagent compositions on the inner surface of the liquid container wall. In some cases, the liquid container includes 10 dry reagent compositions on the inner surface of the liquid container wall. In some cases, the liquid container includes 15 dry reagent compositions on the inner surface of the liquid container wall.

[0041] As described above, the container may include two or more dry reagent compositions placed relative to the container surface (such as the inner surface of the liquid container wall). In some instances, the liquid container wall may include a second dry reagent composition placed on its inner surface. In some instances, the first and second (e.g., two or more) dry reagent compositions adhere to the inner surface of the liquid container wall. The so-called "adhesion" refers to the dry reagent composition being stably associated with the inner surface position where it is located, so that it will not deviate from the inner surface position where it is located when it is in a dry state. During reconstruction, the reagent (e.g., dye) in the dry reagent composition is separated from the surface position where it is located, so that it is present in the liquid used to reconstruct the dry reagent composition. The dry reagent composition may be located at different positions on the inner surface of the liquid container wall. For example, the first and second dry reagent compositions may be placed separately on the inner surface of the liquid container wall. The so-called "different positions" or "separate placement" refers to a dry reagent composition being arranged at a position different from the position where another dry reagent composition is located. The position of the dry reagent composition may refer to the position of the dry reagent composition on the surface of the liquid container, and / or may refer 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 space of a limited volume. For example, the dry reagent composition may occupy a certain volume of space on the surface of the liquid container. The space of a certain volume occupied by a separately placed dry reagent composition does not significantly overlap or overlap with the space of a certain volume occupied by another dry reagent composition. In some cases, the space of a certain volume occupied by the separately placed dry reagent composition and another dry reagent composition does not overlap or overlap at all. Embodiments in which the dry reagent compositions are placed separately can minimize the interaction between each dry reagent composition (such as interaction between dyes).

[0042] In other words, the separately placed dry reagent composition is not significantly mixed with another dry reagent composition (such as a polymeric dye composition), for example, any part of the separately placed dry reagent composition is not substantially mixed with a part of another dry reagent composition (such as a polymeric dye composition). In some instances, the separately placed dry reagent composition is not mixed with another dry reagent composition (such as a polymeric dye composition), for example, any part of the separately placed dry reagent composition is not mixed with a part of another dry reagent composition (such as a polymeric dye composition). In certain embodiments, the separately placed dry reagent composition includes a single reagent. For example, the separately placed dry reagent composition can be substantially composed of a single reagent, and does not include another reagent with a large amount. Relative to any other reagent that may be present in the dry reagent composition, the separately placed dry reagent composition can include a large amount of excess reagent, for example, relative to any other reagent that may be present in the dry reagent composition, 75wt% or more (such as 80wt% or more, or 85wt% or more, or 90wt% or more, or 95wt% or more, or 97wt% or more, or 99wt% or more, or 100wt%) of reagent. In certain embodiments, the dry reagent composition placed separately includes a single dye.For example, the dry reagent composition placed separately can be basically composed of a single dye, and does not include another very large dye in consumption.Relative to any other dye that may exist in the dry reagent composition, the dry reagent composition placed separately can include a large amount of excessive dyes, such as relative to any other dye that may exist in the dry reagent composition, 75wt% or more (such as 80wt% or more, or 85wt% or more, or 90wt% or more, or 95wt% or more, or 97wt% or more, or 99wt% or more, or 100wt%) dye.In some instances, the dry reagent composition includes two or more reagents.In some instances, the first and / or second dry reagent composition includes two or more reagents, such as including 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 instances, the dry reagent composition includes two or more dyes. In some examples, the first and / or second dry reagent composition includes 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.

[0043] In some instances, the separately placed dry reagent composition is spaced apart from the liquid container bottom. The dry reagent composition spaced apart from the liquid container bottom can be physically separated from the liquid container bottom. For example, the separately placed dry reagent composition can be placed on the inner surface of the liquid container wall, and the bottom of the liquid container is spaced apart by a certain distance, so that there is a certain distance between the dry reagent composition edge closest to the liquid container bottom and the liquid container bottom edge closest to the dry reagent composition. The liquid container bottom edge can be any point that the liquid container bottom contacts the liquid container wall. In certain embodiments, when the liquid container wall no longer has a specific shape or no longer continues at a specific angle, the liquid container bottom can be considered as contacting with the liquid container wall. In the embodiment in which the liquid container wall is cylindrical (such as a tapered cylinder) and the liquid container bottom is circular, the liquid container bottom can be considered as no longer parallel to each other at the opposite horizontal end of the liquid container wall, or in the embodiment in which the cylinder is tapered, no longer substantially parallel to each other (that is, the liquid container is no longer a cylinder, but is similar to a hemisphere) position contacts with the liquid container wall. In certain embodiments, the distance between the dry reagent composition on the inner surface of the liquid container wall 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. m or more, or 30mm or more, or 35mm or more, or 40mm or more, or 50mm or more, or 60mm or more, or 70mm or more, or 80mm or more, or 90mm or more, or 100mm or more, or 110mm or more, or 120mm or more, or 130mm or more, or 140mm or more, or 150mm or more, or 160mm or more, or 170mm, or more, or 180mm or more, or 190mm or more, or 200mm or more.For example, the distance between the dry reagent composition on the inner surface of the liquid container wall and the bottom of the liquid container can range from 0.1 mm to 200 mm, such as 0.1 mm to 190 mm, or 0.1 mm to 180 mm, or 0.1 mm to 170 mm, or 0.1 mm to 160 mm, or 0.1 mm to 150 mm, or 0.1 mm to 140 mm, or 0.1 mm to 130 mm, or 0.1 mm to 120 mm, or 0.1 mm to 110 mm, or 0.1 mm to 100 mm, or 0.1 mm to 90 mm, or 0.1 mm to 80 mm. In some embodiments, the dry reagent composition on the inner surface of the liquid container wall and the distance between the bottom of the liquid container ranges from 0.1mm to 200mm. In some cases, the dry reagent composition on the inner surface of the liquid container wall and the distance between the bottom of the liquid container ranges from 0.1mm to 10mm.

[0044] In some instances, the separately placed dry reagent compositions are spaced apart from each other at separate positions on the surface of the liquid container. The dry reagent compositions spaced apart from another dry reagent composition can be physically separated from adjacent dry reagent compositions. For example, the separately placed dry reagent compositions can be placed at separate positions on the surface of the liquid container so that there is a certain distance between the edge of the dry reagent composition and the edge of the adjacent dry reagent composition. In certain embodiments, the distance between each separate position of the dry reagent composition on the inner surface of the liquid container is 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. m or more, or 30mm or more, or 35mm or more, or 40mm or more, or 50mm or more, or 60mm or more, or 70mm or more, or 80mm or more, or 90mm or more, or 100mm or more, or 110mm or more, or 120mm or more, or 130mm or more, or 140mm or more, or 150mm or more, or 160mm or more, or 170mm, or more, or 180mm or more, or 190mm or more, or 200mm or more. For example, the distance between individual locations of the dried reagent composition on the surface of the liquid container can range from 0.1 mm to 200 mm, such as 0.1 mm to 190 mm, or 0.1 mm to 180 mm, or 0.1 mm to 170 mm, or 0.1 mm to 160 mm, or 0.1 mm to 150 mm, or 0.1 mm to 140 mm, or 0.1 mm to 130 mm, or 0.1 mm to 120 mm, or 0.1 mm to 110 mm, or 0.1 mm to 100 mm, or 0.1 mm to 90 mm, or 0.1 mm to 80 mm. In some embodiments, the distance between each individual position 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 individual locations of the dried reagent composition on the surface of the liquid container ranges from 0.1 mm to 10 mm.

[0045] In certain embodiments, the separately placed dry reagent compositions are placed adjacent to each other on the inner surface of the liquid container wall, but are not spaced apart from each other. In these examples, the edge of a dry reagent composition can contact with the edge of an adjacent dry reagent composition. For example, the space volume occupied by a dry reagent composition can contact with the space volume occupied by another (adjacent) dry reagent composition, but there is no significant overlap. In these embodiments, adjacent dry reagent compositions can contact each other, but are not significantly mixed together, for example, any part of the separately placed dry reagent compositions is not mixed with a part of another (adjacent) dry reagent composition substantially.

[0046] The dry reagent composition placed separately can be present on the same surface of the liquid container (such as the inner surface of the liquid container wall), but can also be arranged on the liquid container surface (such as the inner surface of the liquid container wall) or in different positions relative to the liquid container surface. For example, as mentioned above, the liquid container can include an inner surface and an outer surface. In some instances, the dry reagent composition is placed on the surface of the liquid container (such as the inner surface of the liquid container wall). In some cases, the dry reagent composition is placed separately on the inner surface of the liquid container (such as the inner surface of the liquid container wall).

[0047] The example of the separately placed dry reagent composition includes the following embodiments: a dry reagent composition is arranged at a certain position on the surface of the liquid container (e.g., the inner surface of the liquid container wall), and another dry reagent composition is also arranged at different positions on the container surface. Therefore, the separately placed dry reagent composition can be placed at a separate position on the surface of the liquid container. For example, an embodiment of a container may include the first and second dry reagent compositions, wherein the first dry reagent composition is placed at a certain position on the inner surface of the liquid container wall, and the second dry reagent composition is placed at a position different from the first dry reagent composition on the inner surface of the liquid container wall. As described above, the first and second dry reagent compositions can be spaced apart from each other so that there is a certain distance between the separate positions of the first and second dry reagent compositions on the surface of the liquid container (e.g., the inner surface of the liquid container wall). The distance between the first and second dry reagent compositions can be determined according to the above range and numerical value.

[0048] The dry reagent composition can have any suitable size. In some instances, the first and second dry reagent compositions have the same size. In some embodiments, the first and second dry reagent compositions include different sizes. The dry reagent composition can have any suitable cross-sectional shape, for example, including a straight 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 parabola). In some instances, the diameter range of the first and second dry reagent compositions is 0.01mm-5mm, for example, including 0.01mm-4mm, 0.01mm-3mm, 0.01-2mm, 0.01-1mm, 0.1mm-5mm, 0.1mm-4mm, 0.1mm-3mm, 0.1-2mm, 0.1-1mm, 0.5mm-5mm, 0.5mm-4mm, 0.5mm-3mm, 0.5-2mm, 0.5-1mm. In some embodiments, the first and second dry reagent compositions include a range of 5×10 -5 mm 2 -20 mm 2 (For example, including 5×10 -5 mm 2 -20 mm 2 , 5×10 -3 mm 2 -20 mm 2 , 0.1mm 2 -20 mm 2 , 5×10 -5 mm 2 -0.5mm 2 , 5×10 -3 mm 2 -0.5mm 2 , 0.1mm 2 -0.5mm 2 )’s surface area.

[0049] The inner surface of the liquid container (for example the inner surface of the liquid container wall) can provide an additional dry reagent composition. For example, the container may include the 3rd dry reagent composition (such as dye composition) placed separately on the inner surface of the liquid container wall. The 3rd dry reagent composition can be placed separately relative to the first dry reagent composition, and can also be placed separately relative to the second dry reagent composition. Therefore, as described in the present invention, each dry reagent composition (for example the first, second and third dry reagent composition) can be placed separately relative to each other on the inner surface of the liquid container wall, as described above. The dry reagent composition placed separately can be additionally arranged on the surface of the liquid container, such as 4 or more dry reagent compositions placed separately, or 5 or more, 7 or more, 10 or more etc. In some instances, the dry reagent composition adheres to the inner surface of the liquid container wall.

[0050] Other examples of separately placed dry reagent compositions include the following embodiments: a dry reagent composition is arranged at a certain position on the surface of a liquid container (for example, the inner surface of a liquid container wall), and another dry reagent composition is placed at the same position. Therefore, the separately placed dry reagent composition can be co-located at the same position of the surface of a liquid container (for example, the inner surface of a liquid container wall). The dry reagent composition can be co-located at the same position, but can still be separately placed. For example, the dry reagent composition can be separated from each other with a non-dye material. In some cases, the non-dye material is sandwiched between the separately placed dry reagent compositions. The non-dye material can substantially cover the surface of the dry reagent composition so that adjacent dry reagent compositions are separated from the dry reagent composition. For example, a non-dye material can be placed above the dry reagent composition surface, and another dry reagent composition can be placed on the non-dye material surface. In these examples, the dry reagent composition can be physically separated from other dry reagent compositions with a non-staining material. In some cases, the separately placed dry reagent composition can be placed on the container surface as an alternating layer of a dry reagent composition and a non-dye material. Therefore, in certain embodiments, two or more dry reagent compositions are separated relative to each other, and are also co-located at the same position on the surface of a liquid container. In certain embodiments, a non-dye material (eg, a non-reagent material) can be placed in a liquid container, spaced apart from separately placed dry reagent compositions, such that a distance exists between the non-dye material and one or more dry reagent compositions.

[0051] In certain embodiments, the non-dye material is a material compatible with other detection components (e.g., reagents, buffers, analytes, etc.) that may be present in the liquid container during use. For other detection components (e.g., reagents, buffers, analytes, etc.) that may be present in the container during use, the non-dye material may be substantially inert, so that there is no significant reaction between the non-dye material and other detection components. Examples of non-dye materials include, but are not limited to, any reagent, non-reagent material, stabilizer, buffer, soluble inert material (e.g., water-soluble inert material), microspheres, etc. described above. Target stabilizers include, but are not limited to, sugars and polyols. Sugars and polyols suitable for freeze-dried dye compositions include sugars compatible with other reagents, buffers, dyes, and sample components used. Suitable examples of 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, a disaccharide can be sucrose. Examples of suitable polyols include, but are not limited to, mannitol, glycerol, erythritol, threitol, xylitol, sorbitol, and the like, and combinations thereof. Non-dye materials may include bovine serum albumin (BSA), sodium azide, glycerol, phenylmethylsulfonyl fluoride (PMSF), ethylenediaminetetraacetic acid (EDTA), buffered citrate, phosphate buffered saline (PBS), sodium chloride, paraformaldehyde, and the like, and combinations thereof.

[0052] For example, an embodiment of the liquid container may include a first and a second dry reagent composition, wherein the first dry reagent composition is arranged at a certain position on the surface of the liquid container (e.g., the inner surface of the liquid container wall), and the second dry reagent composition is co-located with the first dry reagent composition at the same position. As described above, the first and second dry reagent compositions may be spaced apart from each other so that there is a certain distance between the first and second dry reagent compositions. For example, as described above, the first and second dry reagent compositions may be separated from each other by non-dye materials. The distance between the first and second dry reagent compositions may be determined according to the above range and numerical value. For example, a non-dye material may be sandwiched between the first and second dry reagent compositions placed separately. In these embodiments, the first dry reagent composition may be placed on the surface of the liquid container, a layer of non-dye material may be arranged on the surface of the first dry reagent composition, and the second dry reagent composition may be arranged on the surface of the non-dye composition. In these examples, the first dry reagent composition may be physically separated from the second dry reagent composition by a non-dye material. Therefore, in certain embodiments, the first and second dry reagent compositions are placed separately relative to each other and are also co-located at the same position on the surface of the liquid container. For example, the non-dye material layer on the surface of the first dry reagent composition may substantially cover the entire surface of the first dry reagent composition. In these examples, the second dry reagent composition arranged on the non-dye composition surface may not significantly contact with the first dry reagent composition. In some cases, the non-dye material is a substantially continuous non-dye material of a layer on the first dry reagent composition surface. For example, the non-dye material can cover a considerable portion on the first dry reagent composition surface, for example 75% or more on the first dry reagent composition surface, or 80% or more on the first dry reagent composition surface, or 85% or more, or 90% or more, or 95% or more, or 97% or more, or 99% or more. The embodiment that the surface of the first dry reagent composition is substantially covered by the non-dye material can minimize the interaction between the dyestuffs of the first and second dry reagent compositions.

[0053] In certain embodiments, the thickness range of the non-dye material is 0.01mm-5mm, for example 0.05mm-5mm, or 0.1mm-5mm, or 0.1mm-4mm, or 0.1mm-3mm, or 0.1mm-2mm, or 0.1mm-1mm, or 0.1mm-0.9mm, or 0.1mm-0.8mm, or 0.1mm-0.7mm, or 0.1mm-0.6mm, or 0.1mm-0.5mm. In certain instances, the thickness of the non-dye material is 0.1mm-1mm. In some cases, the thickness of the non-dye material is 0.1mm-0.05mm.

[0054] Other dry reagent compositions may also be provided. For example, the liquid container may include a third dry reagent composition that is placed separately relative to the first and second dry reagent compositions. Therefore, the third dry reagent composition may be placed separately relative to the first dry reagent composition, or may be placed separately relative to the second dry reagent composition. Therefore, as described in the present invention, each dry reagent composition (e.g., the first, second, and third dry reagent compositions) may be placed separately relative to each other. In some cases, each dry reagent composition may be separated from each other by a non-dye material. For example, each dry reagent composition may be separated from each other by a non-dye material. In some cases, the non-dye material is sandwiched between each separately placed dry reagent composition. In some instances, each separately placed dry reagent composition is a layer, and there is a layer of non-dye material between each separately placed dry reagent composition. As described above, several layers of separately placed dry reagent compositions may be additionally provided, such as 4 or more separately placed dry reagent compositions, or 5 or more, 7 or more, 10 or more, etc. Therefore, a variety of dry reagent compositions may be placed separately relative to each other, or may be co-located at the same position on the surface of the liquid container (e.g., the inner surface of the liquid container wall).

[0055] In certain embodiments, the dry reagent composition on the liquid container surface is a dry dye composition comprising dye.Dry dye composition is a dye composition comprising a small amount of solvent.For example, the dry dye composition can comprise a small amount of liquid, such as water.In some cases, the dry dye composition does not comprise solvent substantially.For example, the dry dye composition can not comprise liquid substantially, such as water.In certain embodiments, the dry dye composition comprises 25wt% or following solvent, for example 20wt% or following, or 15wt% or following, or 10wt% or following, or 5wt% or following, or 3wt% or following, or 1wt% or following, or 0.5wt% or following solvent.In some cases, the dry dye composition is non-fluid.In some cases, the dry reagent composition is solid substantially. For example, under standard conditions, the dried dye 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.

[0056] In some instances, the dye composition is a freeze-dried dye composition. In some cases, the freeze-dried dye composition is a dye composition obtained by removing the water in the dye composition by sublimation, wherein the water in the dye composition has undergone a phase change from solid to gaseous. For example, the freeze-dried dye composition can be a dye composition obtained by freezing the dye composition (for example freezing the water in the dye composition), then reducing the pressure around the dye composition, sublimating the water in the dye composition, thereby removing the water in the composition. In some instances, according to the measurement result of Karl Fischer (KF) titration, the freeze-dried dye composition comprises a small amount of water, for example 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. In some cases, according to the measurement results of Karl Fischer titration, the freeze-dried dye composition contains 3% or less water. In some cases, according to the measurement results of Karl Fischer titration, the freeze-dried dye composition contains 1% or less water. In some cases, according to the measurement results of Karl Fischer titration, the freeze-dried dye composition contains 0.5% or less water. The freeze-dried dye composition may include additives and / or adjuvants, such as stabilizers. In some cases, the freeze-dried dye composition includes stabilizers, such as sugars or polyols. Sugars and polyols suitable for freeze-dried dye compositions include sugars compatible with other reagents, buffers, dyes and sample components used. Suitable examples of 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 some instances, the sugar is a disaccharide. For example, a disaccharide can be sucrose. Suitable examples of polyols include, but are not limited to, mannitol, glycerol, erythritol, threitol, xylitol, sorbitol, etc. and combinations thereof.

[0057] The dye in the dye composition can be used as a detectable label. In some cases, the dye includes a detectable portion or marker that can be detected based on fluorescence emission maximum, fluorescence polarization, fluorescence lifetime, light scattering, mass, molecular mass or a combination thereof. In certain embodiments, the detectable label is a fluorescent group (i.e., a fluorescent marker, a fluorescent dye, etc.). The target fluorescent group may include, but is not limited to, a dye suitable for analytical applications (such as flow cytometry, imaging, etc.).

[0058] In some instances, the fluorescent group is a polymeric dye. In some instances of the method, the polymer dye includes a coupled polymer. The coupled polymer (CP) is characterized by a delocalized electronic structure, which includes a main chain in which unsaturated bonds (such as double bonds and / or triple bonds) and saturated bonds (such as single bonds) are alternately arranged, wherein π electrons can move from one bond to another. Therefore, the coupled main chain can give the polymer dye an extended linear structure with limited bond angles between polymer repeating units. For example, although proteins and nucleic acids are also polymerized, in some cases they do not form extended rod-like structures, but fold into higher-order three-dimensional shapes. In addition, CP may form a "rigid rod" polymer main chain and experience limited twisting (such as torsion) angles between monomer repeating units along the polymer main chain. In some instances, the polymer dye includes a CP with a rigid rod structure. The structural characteristics of the polymer dye will affect the fluorescence properties of the molecule.

[0059] Targeted polydyes include, but are not limited to, those disclosed in Gaylord et al., No. 20040142344, No. 20080293164, No. 20080064042, No. 20100136702, No. 20110256549, No. 20110257374, No. 20120028828, No. 20120252986, No. 20130190193, No. 20160264737, No. 20160266131, No. 20180231530, No. 20180009990, No. 20180009989, and No. The polymeric dyes described in U.S. Publication No. 20180163054, the disclosure of which is incorporated herein by reference as a whole, become a part of the present invention; Gaylord et al., Journal of the American Chemical Society, 2001, 123(26), pp. 6417-6418; Feng et al., Reviews of the Chemical Society, 2010, 39, 2411-2419; Traina et al., Journal of the American Chemical Society, 2011, 133(32), the polymeric dyes described in which are incorporated herein by reference as a whole, become a part of the present invention.

[0060] The polymeric dye may have one or more desirable spectral properties, such as a specific maximum absorption wavelength, a specific maximum emission wavelength, an extinction coefficient, a quantum yield, etc. (see Chattopadhyay et al., "Brilliant violet fluorophores: novel ultra-bright fluorescent compounds for immunofluorescence experiments." Flow Cytometry Part A, 81A(6), 456-466, 2012). In some embodiments, the polymeric dye has an absorption curve between 280 nm and 475 nm. In some embodiments, the polymeric dye has a maximum absorption value (maximum excitation value) within the range of 280 nm and 475 nm. In some embodiments, the polymeric dye absorbs incident light with a wavelength range of between 280 nm and 475 nm. In some embodiments, the polymeric dye has a maximum emission wavelength in the range of 400 nm to 850 nm (e.g., 415 nm to 800 nm), wherein specific examples of target maximum emission values ​​include, but are not limited to, 421 nm, 510 nm, 570 nm, 602 nm, 650 nm, 711 nm, and 786 nm. In some instances, the polydye has a maximum emission wavelength in a range selected from the group consisting of 410nm-430nm, 500nm-520nm, 560nm-580nm, 590nm-610nm, 640nm-660nm, 700nm-720nm, and 775nm-795nm. In some embodiments, the polydye has a maximum emission wavelength of 421nm. In some instances, the polydye has a maximum emission wavelength of 510nm. In some cases, the polydye has a maximum emission wavelength of 570nm. In some embodiments, the polydye has a maximum emission wavelength of 602nm. In some instances, the polydye has a maximum emission wavelength of 650nm. In some cases, the polydye has a maximum emission wavelength of 711nm. In some embodiments, the polydye has a maximum emission wavelength of 786nm. In some instances, the polydye has a maximum emission wavelength of 421nm±5nm. In some embodiments, the polydye has a maximum emission wavelength of 510nm±5nm. In some instances, the polydye has a maximum emission wavelength of 570nm ± 5nm. In some instances, the polydye has a maximum emission wavelength of 602nm ± 5nm. In some embodiments, the polydye has a maximum emission wavelength of 650nm ± 5nm. In some instances, the polydye has a maximum emission wavelength of 711nm ± 5nm. In some cases, the polydye has a maximum emission wavelength of 786nm ± 5nm. In some embodiments, the polydye has a maximum emission value selected from the group of 421nm, 510nm, 570nm, 602nm, 650nm, 711nm and 786nm.

[0061] Specific polydyes that may be used include, but are not limited to, BD Horizon Brilliant TM Dyes, such as BD Horizon Brilliant TM Violet dyes (e.g., BV421, BV480, BV510, BV570, BV605, BV650, BV711, BV750, BV786); BD Horizon Brilliant TM Ultraviolet dyes (e.g., BUV395, BUV496, BUV563, BUV615, BUV661, BUV737, BUV805); BD Horizon Brilliant TM Blue dyes (e.g., BB515, BB630-P2, BB660-P2, BB700, BB755-P, BB-790P) (BD Biosciences, San Jose, CA).

[0062] In certain embodiments, as described above, the liquid container (e.g., the inner surface of the liquid container wall) comprises more than one dye composition, such as two dye compositions (e.g., the first and second dye compositions). In these embodiments, as described above, the dye composition may be a polymeric dye composition. For example, the liquid container may include the first and second polymeric dye compositions. As described above, the first and second polymeric dyes may be coupled polymers (CP). In some cases, as described above, the first and second polymeric dyes are water-soluble coupled polymers. In certain examples, the dye compositions in the liquid container may be different dye compositions, such as different polymeric dye compositions. Different dye compositions may differ from each other in terms of one or more characteristics of chemical components and / or dyes. For example, at least one value of the excitation maximum and emission maximum of different dye compositions differs from each other. In some cases, the excitation maximum of different dye compositions differs from each other. In some cases, the emission maximum of different dye compositions differs from each other. In some cases, the excitation maximum and emission maximum of different dye compositions differ from each other. Therefore, in the embodiment comprising the first and second dyes, at least one value of the excitation maximum and emission maximum of the first and second dyes differs from each other. For example, the excitation maximum, emission maximum, or excitation maximum and emission maximum of the first and second dyes differ from each other. The liquid container may also include other dye compositions, and as described above, each dye composition in the container is different from another.

[0063] In certain embodiments, the liquid container (such as the inner surface of the liquid container wall) also includes other types of dye compositions, such as one or more non-polymeric dye compositions. As mentioned above, dye can include detectable parts or markers that can be detected based on fluorescence emission maximum, fluorescence polarization, fluorescence lifetime, light scattering, quality, molecular mass or its combination. In certain embodiments, the non-polymeric dye includes fluorescent group (i.e. fluorescent marker, fluorescent dye, etc.). The target fluorescent group can include but is not limited to the dye that is suitable for analytical applications (such as flow cytometry, imaging, etc.). Many non-polymeric dyes can be purchased from various sources (for example, Molecular Probes (Eugene, Oregon, USA) and Exciton (Dayton, Ohio, USA)). For example, the fluorescent group of the non-polymeric dye may include 4-acetamido-4'-isothiocyanatostilbene-2,2'disulfonic acid; acridine and its derivatives, such as acridine, acridine orange, acridine yellow, acridine red and acridine isothiocyanate; 5-(2'-aminoethyl)naphthylamine-1-sulfonic acid (EDANS); 4-amino-N-[3-vinylsulfone]phenyl]naphthalimide-3,5-disulfonate (fluorescein yellow VS); N-(4-anilino-1-naphthyl)maleimide; aminobenzamide; brilliant yellow; coumarin and its derivatives, Such as coumarin, 7-amino-4-methylcoumarin (AMC, coumarin 120), 7-amino-4-trifluoromethylcoumarin (coumarin 151); cyanide and its derivatives, such as tetrachlorotetrabromofluorescein, Cy3, Cy3.5, Cy5, Cy5.5 and Cy7; 4',6-diamidino-2-phenylindole (DAPI); 5',5"-dibromopyrogallolsulfonphthalein (bromopyrogallol red); 7-diethylamino-3-(4'-isothiocyanatophenyl)-4-methylcoumarin; diethylaminocoumarin; di Ethylenetriamine pentaacetate; 4,4'-diisothiocyanatodiphenylethane-2,2'-disulfonic acid; 4,4'-diisothiocyanato-2,2'-disulfonic acid; 5-[dimethylamino]naphthalene-1-sulfonyl chloride (DNS, dansyl chloride); 4-(4'-dimethylaminophenylazo)benzoic acid (DABCYL); 4-dimethylaminophenylazo-4'-isothiocyanate (DABITC); eosin and derivatives, such as eosin and eosin isothiocyanate; erythrosin and derivatives, such as erythrosin B and erythrosin isothiocyanate Esters; Ethidium; Fluorescein and its 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), chlorotriazinylfluorescein, naphthylfluorescein and QFITC ​​(XRITC); Fluorescein amine; IR144; IR1446; Green fluorescent protein (GFP); Coral reef fluorescent protein (RCFP); Lissamine TM; Lissamine rhodamine, fluorescent yellow; malachite green isothiocyanate; 4-methylumbelliferone; o-cresolphthalein; nitrotyrosine; p-aminobenzenesulfonic acid; Nile red; Oregon green; phenol red; B-phycoerythrin (PE); o-phthalaldehyde; pyrene and its derivatives, such as pyrene, pyrene butyrate and 1-pyrene butyric acid succinimidyl ester; Reactive Red 4 (Cibacron TM 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, sulfonyl chloride derivative of sulforhodamine 101 (Texas Red), N,N,N',N'-tetramethyl-6-carboxyrhodamine (TAMRA), tetramethylrhodamine and tetramethylrhodamine isothiocyanate (TRITC); riboflavin; rhodamine and terbium chelate derivatives; xanthene; carotenoid-protein complexes, such as peridinin-chlorophyll-protein (PerCP); allophycocyanin (APC); or combinations thereof.

[0064] In certain embodiments, the dye composition included in the liquid container (such as the inner surface of the liquid container wall) comprises a polymeric dye composition, as described above. In some cases, the dye composition included in the liquid container comprises a non-polymeric dye composition, as described above. In some instances, the dye composition included in the liquid container comprises a polymeric dye composition and a non-polymeric dye composition simultaneously. As described above, each dye composition (such as polymeric and non-polymeric dye composition) can be placed separately on the liquid container surface. In some cases, the liquid container comprises a plurality of dye compositions, as described above. For example, the liquid container may 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.

[0065] As mentioned above, the liquid container can include polymeric dye composition and non-polymeric dye composition simultaneously.In some instances, polymeric dye composition is mixed with non-polymeric dye composition.In certain embodiments, the mixture of polymeric dye composition and non-polymeric dye composition will not have significant dye interaction between polymeric dye composition and non-polymeric dye composition.For example, the fluorescence emission energy of polymeric dye composition will not be significantly quenched due to interaction with non-polymeric dye composition.In some cases, the fluorescence emission energy of polymeric dye composition will not be significantly dissipated due to non-radiative transition.In these embodiments, compared with the fluorescence of polymeric dye composition when there is no non-polymeric dye composition, the detectable fluorescence of polymeric dye composition will not be significantly lower than expectation.Equally, in certain embodiments, the fluorescence emission energy of non-polymeric dye composition will not be significantly quenched due to interaction with polymeric dye composition.For example, the fluorescence emission energy of non-polymeric dye composition will not be significantly dissipated due to non-radiative transition.In these embodiments, compared with the fluorescence of non-polymeric dye composition when there is no polymeric dye composition, the detectable fluorescence of non-polymeric dye composition will not be significantly lower than expectation.

[0066] In certain embodiments, the dye composition includes dyestuff, such as polymerization and / or non-polymeric dyestuff, as described above. The dye composition may also include other components, such as but not limited to solvent, buffer, stabilizer etc. For example, the dye composition may include stabilizer, for reducing and / or substantially preventing the dyestuff in the dye composition from degrading. In some cases, the stabilizer present in the dye composition is enough to reduce and / or substantially prevent the dyestuff in the dye composition from degrading in a certain time (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). The example of stabilizer includes but is not limited to bovine serum albumin (BSA), sodium azide, glycerol, phenylmethylsulfonyl fluoride (PMSF) etc. In addition, other additives may be present in the composition, such as additives for preserving whole blood cells, such as platelet stabilizing factors, etc. Examples of additives that may be included in the composition include anticoagulants, such as ethylenediaminetetraacetic acid (EDTA), buffer-treated citrate, heparin, etc. The composition may include these liquid or dry additives.

[0067] In certain examples, 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 coupled to each other (eg, covalently linked) via an optional linker at any suitable position of the two molecules.

[0068] As used herein, the term "specific binding member" refers to one member of a pair of molecules that have binding specificity to each other. The surface of one member of the pair may have an area or cavity that specifically binds to a surface area or cavity of the other member of the pair. Thus, the members of the pair have the property of specifically binding to each other, thereby producing a binding complex. In certain embodiments, the affinity between the specific binding members in the binding complex is characterized by a K. d (dissociation constant) is 10 -6 M or less, e.g. 10 -7 M or below, including 10 -8 M or less, e.g. 10 -9 M or below, 10 -10 M or below, 10 -11 M or below, 10 -12 M or below, 10 -13 M or below, 10 -14 M or below, including 10 -15 M or less. In certain embodiments, the specific binding member specifically binds with high affinity. The so-called "high affinity" refers to a binding member with an apparent K d is 10×10 -9 M or less (e.g. 1×10 -9 M or below, 3×10 -10 M or less, 1×10 -10 M or below, 3×10 -11 M or less, 1×10 -11 M or below, 3×10 -12 M or less, or 1×10 -12 M or less).

[0069] The specific binding member can be a protein member. The term "protein" used in the present invention refers to a portion consisting of amino acid residues. The protein portion can be a polypeptide. In some cases, the protein-specific binding member is an antibody. In certain embodiments, the protein-specific binding member is an antibody fragment, such as an antibody binding fragment that specifically binds to a polymer dye. The terms "antibody" and "antibody molecule" used in the present invention are used interchangeably and refer to a protein consisting of one or more polypeptides that are substantially encoded by all or part of an identified immunoglobulin gene. Taking human immunoglobulin genes as an example, the identified immunoglobulin genes include kappa (k), lambda (l) and heavy chain loci (which together constitute a large number of variable region genes), as well as constant region genes mu (u), delta (d), gamma (g), sigma (e) and alpha (a) (encoding IgM, IgD, IgG, IgE and IgA isotypes, respectively). The variable region of an immunoglobulin light chain or heavy chain consists of a "framework" region (FR) interrupted by three hypervariable regions (also known as "complementarity determining regions" or "CDRs"). The scope of the framework region and CDR has been precisely defined (see "Protein Sequences of Immunological Importance", E. Kabat et al., U.S. Department of Health and Human Services, (1991)). The numbering of all antibody amino acid sequences described in the present invention conforms to the Kabat system. The framework region sequences of different light chains or heavy chains within a species are relatively conserved. The framework region of an antibody (i.e., the combined framework region of the component light chain and heavy chain) is used to position and align the CDR. The CDR is primarily responsible for binding to the epitope of the antigen. "Antibody" includes full-length antibodies and may refer to natural antibodies from any organism, engineered antibodies, or antibodies reconstructed for experiments, treatments, or other purposes further defined below.

[0070] Target antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv, scFv or other antibody antigen binding subsequences, which can be produced by modifying the entire antibody or synthesized de novo using DNA reconstruction technology. Antibodies can be monoclonal or polyclonal antibodies and can have other specific activities on cells (such as antagonists, agonists, neutralizing antibodies, inhibitory antibodies or stimulatory antibodies). It should be understood that antibodies can have other conservative amino acid substitutions that have little effect on antigen binding or other antibody functions.

[0071] In certain embodiments, the specific binding member is a Fab fragment, a F(ab')2 fragment, a 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 murine antibody or a binding fragment thereof. In some instances, the specific binding member is a reconstituted antibody or a binding fragment thereof.

[0072] In certain embodiments, the liquid container further comprises a calibration standard. The calibration standard can be used to determine the accuracy of the test and ensure the consistency of subsequent tests. In some cases, the calibration standard comprises a labeled microsphere, such as a fluorescently labeled microsphere. The fluorescently labeled microsphere can be a standard fluorescently labeled microsphere, which is typically used as a calibration standard. Examples of standard fluorescently labeled microspheres include, but are not limited to, fluorescently labeled microparticles or nanoparticles. In some cases, the fluorescently labeled microspheres are configured to be suspended in the detection mixture without significant precipitation or aggregation. In certain embodiments, the fluorescently labeled microspheres include, but are not limited to, fluorescently labeled polystyrene microspheres, fluorescein microspheres, rhodamine microspheres, and other microspheres labeled with fluorescent dyes. Other examples of fluorescently labeled microspheres are described in U.S. Patents Nos. 6,350,619; 7,738,094; and 8,248,597, the disclosure of each of which is incorporated herein by reference as a whole.

[0073] In some cases, the liquid container is convenient for storing the dye composition over a long period of time. 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, wherein the dye composition is substantially stable over a long period of time. So-called "stable" or "storage-stable" or "substantially stable" refers to that the dye composition will not significantly degrade and / or lose activity over a long period of time. For example, the storage-stable dye composition will not significantly lose fluorescence activity due to the degradation of the dye composition over a long period of time, such as losing 10% or less fluorescence activity over a long period of time, 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 fluorescence activity. In some instances, the loss of fluorescence activity of the storage-stable dye composition over a long period of time is 5% or less. In some cases, the storage-stable dye composition maintains its fluorescent activity substantially over a long period of time, such as maintaining 100% activity over a long period of time, or 99% or more, 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 activity. For example, the storage-stable dye composition can maintain 90% or more fluorescent activity over a long period of time. In some cases, the storage-stable composition can maintain 95% or more fluorescent activity over a long period of time. "A longer period of time" refers to a period of time, such as 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. For example, a longer period of time can be 6 months or more. In some cases, a longer period of time is 9 months or more. In some cases, a longer period of time is 1 year (e.g., 12 months) or more. In some cases, a longer period of time is 1.5 years (e.g., 18 months) or more. In some cases, a longer period of time is 2 years (e.g., 24 months) or more. In certain instances, the longer period of time is 10 years or less, such as 7.5 years or less, including 5 years or less, such as 2 years or less.

[0074] Various aspects of liquid containers (including dry compositions, such as dry polymeric coupled dye compositions) that can be part of the system of embodiments of the present invention are further provided in U.S. Patents Nos. 10,545,137 and 11,320,437, U.S. pending application No. 2020 / 0147615A1, and U.S. pending applications Nos. 17 / 580,130 and 17 / 830,070; the disclosures of the above-mentioned documents are incorporated herein by reference and become a part of the present invention.

[0075] Solid volume displacer

[0076] According to the above summary, the system of the present invention also includes a solid volume displacer. The solid volume displacer according to certain embodiments of the present disclosure can be placed inside the liquid container, for example, as described above, the solid volume displacer occupies most of the liquid container volume below the top of the dry reagent composition placed on the inner surface of the liquid container wall.

[0077] The solid volume displacer can be compatible with a liquid sample and / or a reagent or analyte that can be in contact with (e.g., contained within) a liquid container. In some cases, the liquid sample can be an aqueous liquid sample, in which case the solid volume displacer can be compatible with the aqueous sample. By "compatible" is meant that the solid volume displacer (e.g., the material from which the solid volume displacer is made) is substantially inert to (e.g., does not significantly react with) the liquid and / or reagent or analyte that is in contact with the liquid container.

[0078] The solid volume displacer may include a first end and a second end and a body therebetween, wherein the second end and the body may be placed inside a liquid container, as described above, so that the second end and the body displace a certain volume of liquid upwardly along the inner surface of the liquid container wall, thereby reconstructing the dry reagent composition. For example, the second end and the body may be placed inside a liquid container so that the solid volume displacer occupies most of the liquid container volume below the top of the dry reagent composition. For example, in certain embodiments, the second end and the body may be placed inside a liquid container so that the solid volume displacer occupies more than 50% of the liquid container volume below the top of the dry reagent composition. In some cases, the second end and the body may be placed inside a liquid container so that the solid volume displacer occupies a portion of the liquid container volume below the top of the dry reagent composition, such as 55% or more of the volume, 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. In the embodiment of placing a variety of dry reagent compositions on the inner surface of the liquid container wall, the solid volume displacer can be placed inside the liquid container so that the solid volume displacer occupies most of the liquid container volume below the top of the dry reagent composition farthest from the bottom of the liquid container. In these cases, the solid volume displacer can displace a certain volume of liquid upward along the inner surface of the liquid container wall, thereby reconstructing any and all dry reagent compositions present on the inner surface of the wall. In some instances, the solid volume displacer can be removed from the liquid container so that the liquid in the liquid container is displaced in a manner sufficient to reconstruct the dry reagent composition, and the solid volume displacer can be removed from the liquid container. In these instances, the removable solid volume displacer allows access to the reconstructed reagent composition. The reconstructed reagent composition can be collected for analysis applications (e.g., flow cytometry, imaging, etc.).

[0079] As mentioned above, the liquid container can hold a certain volume of fluid (such as gas or liquid). In certain embodiments, the liquid container can hold a certain volume of liquid below the top of the dry reagent composition. In these examples, there may be a certain liquid container volume below the top of the dry reagent composition, and the solid volume displacer does not take the certain liquid container volume, for example, when the solid volume displacer is placed on the liquid container inside, the liquid of this volume is introduced into the liquid container and the liquid container is filled to the top of the dry reagent composition. For example, in some instances, the volume of the liquid container below the top of the dry reagent composition that is not occupied by the solid volume displacer (e.g., the volume of liquid required to fill the liquid container to the top of the dry reagent composition when the solid volume displacer is placed inside the liquid container) can be 5000 μl or less, 1000 μl or less, such as 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 volume (e.g., biological liquid sample) contained in the liquid container is greater than or equal to the volume of the liquid container below the top of the dry reagent composition that is not occupied by the solid volume displacer.

[0080] As mentioned above, when the solid volume displacer is placed on the liquid container inside, the liquid container volume below the dry reagent composition top may not be occupied by the solid volume displacer. In other words, when the solid volume displacer is placed on the liquid container inside, a certain volume of liquid may be required to fill the liquid container to the top of the dry reagent composition. Therefore, the solid volume displacer can reduce the liquid container to be filled to the required liquid volume of the dry reagent composition top, for example, contacting with the dry reagent composition. For example, the solid volume displacer can reduce the liquid container to be filled to the required liquid volume of the dry reagent composition top 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.

[0081] In certain embodiments, compared with other systems that move liquid upward along the liquid container wall, the solid volume displacer can reduce the liquid volume required for contact dry reagent composition in the liquid container. In certain embodiments, compared with using an agitator (such as a vortex oscillator) alone (i.e., without using a solid volume displacer), the solid volume displacer can reduce the liquid volume required for contact dry reagent composition. For example, compared with using a vortex oscillator, the solid volume displacer can reduce the liquid volume required for contact dry reagent composition by 50% or more, such as 60% or more, or 70% or more, or 80% or more, or 90% or more. In certain embodiments, the solid volume displacer can be fixed to a mechanical mixer. For example, the first end of the solid volume displacer can be fixed to a mechanical mixer. In certain embodiments, the first end of the solid volume displacer can be fixed to an agitator (such as a vortex oscillator).

[0082] The shape of the solid volume displacer can be different, depending on the purpose of the solid volume displacer. For example, as described in the present invention, the solid volume displacer can be introduced into the above-mentioned liquid container in a manner sufficient to displace a certain volume of liquid upward along the inner surface of the liquid container wall, thereby reconstructing the above-mentioned dry reagent composition. In these cases, the solid volume displacer can be configured to be placed in a shape compatible with the solid volume displacer inside the liquid container. For example, in an embodiment in which the liquid container wall is cylindrical, the main body of the solid volume displacer can be configured to be cylindrical. In these embodiments, the solid volume displacer may include an outer surface concentric with the inner surface of the liquid container wall. In these examples, the outer surface of the solid volume displacer may have a diameter different from the diameter of the inner surface of the liquid container wall (for example, a diameter smaller than the diameter of the inner surface of the liquid container wall). For example, the difference (i.e., the gap) between the outer diameter of the solid volume displacer body and the inner diameter of the liquid container wall 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 multiple parts with different diameters, or the outer surface may be stepped. In embodiments where the liquid container wall is conical, the outer surface of the solid volume displacer body may be conical with the same or similar angles. In embodiments where the bottom of the liquid container is circular, the second end of the solid volume displacer may be circular and concentric with the bottom of the liquid container. In some embodiments, the solid volume displacer may be configured as a pestle. In some cases, the solid volume displacer is a hollow solid volume displacer.

[0083] As described above, the distance between the dry reagent composition on the inner surface of the liquid container wall and the bottom of the liquid container can be different. In some cases, the solid volume displacer can displace a certain volume of liquid along the inner surface upward by a certain distance, and this distance is greater than or equal to the distance between the dry reagent composition on the inner surface of the liquid container wall and the bottom of the liquid container. For example, the solid volume displacer can displace a certain volume of liquid along the inner surface of the liquid container wall upward by 0.1mm or more, such as 0.5mm or more, or 1mm or more, or 2mm or more, or 3mm or more, or 4mm or more, or 5mm or more, or 6mm or more, or 7mm or more, or 8mm or more, or 9mm or more, or 10mm or more, or 12mm or more, or 14mm or more, or 16mm or more, or 18mm or more, or 20mm or more, or 25mm or more, or 30mm or more, or 35mm or more, or 40mm or more, or 50mm or more, or 60mm or more, or 70mm or more, or 80mm or more, or 90mm or more, or 100mm or more, or 110mm or more, or 120mm or more, or 130mm or more, or 140mm or more, or 150mm or more, or 160mm or more, or 170mm, or more, or 180mm or more, or 190mm or more, or 200mm or more.

[0084] As described above, embodiments of the solid volume displacer can be compatible with liquid samples and / or reagents or analytes that are in contact with (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 made of glass, such as, but not limited to, silicate glass, borosilicate glass, sodium borosilicate glass (e.g., PYREX TM ), fused quartz glass, fused silica glass, etc. 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 but not limited to polyurethane acrylate (UMA), 9120, WaterShed XC 11122 and In some cases, the hydrophobic polymer material can be a plastic, such as but not limited to polystyrene, polypropylene, polymethylpentene, polytetrafluoroethylene (PTFE), perfluoroether (PFE), fluorinated ethylene propylene (FEP), perfluoroalkoxy resin (PFA), polyethylene terephthalate (PET), polyethylene (PE), low-density polyethylene (LDPE), polyetheretherketone (PEEK), etc. The material of the solid volume displacer can be transparent, translucent and / or any color, such as gray, black, white, yellow, etc.

[0085] In some embodiments, as described above, the solid volume displacer includes a first end and a second end and a body therebetween, wherein the second end and the body can be placed inside a liquid container. In some embodiments, when the body and the second end of the solid volume displacer are placed inside a liquid container, the first end of the solid volume displacer can be placed outside the liquid container. In some instances, the first end of the solid volume displacer can include a flange. In some embodiments, the flange can prevent the second end of the solid volume displacer from contacting the bottom of the liquid container, for example, by contacting the open end on the liquid container. In other embodiments, the flange can 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, which is placed around the outside of the body near the first end. In these instances, when the body of the solid volume displacer is placed inside a liquid container, the ring of filter media can prevent liquid from splashing out of the liquid container. For example, when the body of the solid volume displacer is placed inside a liquid container, the ring of filter media can prevent liquid from splashing out of the container by eliminating the gap between the outer surface of the solid volume displacer body and the inner surface of the liquid container wall near the open end of the liquid container. In other examples, when the body of the solid volume displacer is placed inside a liquid container, the ring of filter media can prevent liquid from splashing out of the container by connecting with the open end of the liquid container or sliding over and outside the open end of the liquid container (e.g., while maintaining contact with the open end).

[0086] In certain embodiments, the solid volume displacer can prevent the generation of aerosols or liquid sprays during use (e.g., when the body of the solid volume displacer is inserted into and subsequently removed from the interior of a liquid container). In certain cases, when the body of the solid volume displacer is placed inside a liquid container, a ring of filter media of the displacer can equalize air pressure between the inside and outside of the container. For example, the ring of filter media can be made of a material that allows air to pass but hinders or prevents liquid from passing, such as a fibrous material or a porous material. In certain cases, the fibrous material or porous material can selectively allow certain components of the liquid to pass, but prevent other components of the liquid from passing. For example, the fibrous material or porous material can allow liquid and various substances dissolved therein to pass, while preventing cells (such as mammalian and / or bacterial cells) and virus particles from passing. In certain embodiments, the flange of the displacer includes surface features that can equalize air pressure between the inside and outside of the container, such as protrusions or recesses on the surface of the flange, wherein the flange can contact the liquid container when the body of the solid volume displacer is placed inside the liquid container. In certain embodiments, the second end of the solid volume displacer may include protrusions (e.g., a series of ribs) and / or recesses (e.g., concentric portions of reduced diameter) that prevent liquid from being ejected from an opening of a liquid container when the displacer is used to reconstitute a dry reagent composition, as described below.

[0087] In some embodiments, the first end of the solid volume displacer may include a handle. In some cases, the handle may include a protrusion (e.g., a rib or a bump), or may include a non-slip material to enhance the user's grip and / or control. In some embodiments, the solid volume displacer may include a handle and a flange (e.g., as described above). In other cases, the solid volume displacer may include only one of the handle or the flange. In some embodiments, when the second end and the main body of the solid volume displacer are placed inside the liquid container, the first end of the solid volume displacer may seal the open end of the liquid container. That is, the first end of the solid volume displacer can basically prevent the contents of the liquid container (e.g., the liquid in the liquid container) from flowing out of the liquid container. The first end of the solid volume displacer can also basically prevent other substances from entering the liquid container. For example, the first end of the solid volume displacer can form a watertight seal that basically prevents liquid from entering and exiting the container, or can form an airtight seal that basically prevents gas from entering and exiting the container. In some embodiments, the first end of the solid volume displacer may form a watertight seal that substantially prevents liquid from entering or exiting the liquid container, but allows air to flow out of and into the container to balance air pressure and prevent the generation of aerosols, as described above. In some instances, 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 cap or a snap cap or other suitable sealing member that can be used for 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 on over the opening of the liquid container when the second end and body of the solid volume displacer are placed inside the liquid container.

[0088] In some embodiments, one or more components of the solid volume displacer may be adjustable or removable components. In some cases, the flange may be an adjustable or removable flange. For example, the flange may include a latch or clamp, and the user may fix or remove the flange depending on whether contact is required between the second end of the solid volume displacer and the bottom of the liquid container. In some cases, the latch or clamp may allow the user to adjust the position of the flange on the solid volume displacer according to the required gap between the second end of the solid volume displacer and the bottom of the liquid container. In some cases, the circle of filter media can be removable. In some embodiments, the solid volume displacer can be cleaned and / or sterilized for subsequent use (e.g., using an autoclave), or can be discarded. Therefore, in some embodiments, the solid volume displacer described in the present invention is a disposable product, such as after one use.

[0089] As mentioned above, the solid volume displacer can be placed inside the above-mentioned liquid container, occupying most of the volume of the liquid container 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, when the body of the solid volume displacer is placed inside the liquid container, the outer surface of the solid volume displacer body is towards the inner surface of the liquid container wall. The outer surface of the solid volume displacer body can contact the content of the liquid container.

[0090] Figure 1 A system 100 is shown according to an embodiment of the present invention. Figure 1 The system 100 described in the invention consists 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, which are separated by a wall 113. The inner surface of the wall 113 includes a dry reagent composition 114, which can be a polymeric dye composition (such as a coupled polymeric dye composition) or other dye or non-dye reagent composition. The solid volume displacer 120 includes a first end 121 and a second end 122 and a body 123 therebetween.

[0091] The liquid container 110 is configured as a test tube or a centrifuge tube, having a cylindrical wall 113 and a circular bottom 112. The dry reagent composition 114 that is farthest from the bottom of the liquid container 112 is approximately 20 mm from the bottom of the liquid container 112. The first end of the solid volume displacer 121 includes a flange for preventing the second end of the solid volume displacer 122 from contacting the bottom of the liquid container 112, and a handle 124 for improving usability / enhancing the user's grip. The body of the solid volume displacer 123 is configured as a cylinder, and the outer surface of the cylinder is concentric with the inner surface of the wall of the liquid container 113. The second end of the solid volume displacer 122 is circular and concentric with the bottom of the liquid container 112. Component 130 shows the system 100, wherein the solid volume displacer is placed inside the liquid container.

[0092] Figure 4A-4B The present invention describes a stepped solid volume displacer according to an embodiment of the present invention. Figure 4A, a side view of a solid volume displacer 400 is provided, the solid volume displacer 400 having cylindrical portions or steps 410, 420 and 430 of different diameters. Step 410 is adjacent to the first end of the solid volume displacer 400 and is 60 mm long and 9.9 mm in diameter. Step 410 can act as a handle when the displacer 400 is placed in a liquid container. Step 420 is 20 mm long and 9.6 mm in diameter. Step 430 is adjacent to the second end of the solid volume displacer 400 and is 40 mm long and 8.9 mm in diameter. The second end of the solid volume displacer 400 is a spherical cap having a hemispherical shape with a radius of 4.6 mm, wherein the hemispherical portion protruding outside the step 430 is configured as a continuation of the step 430. In Figure 4B , a three-dimensional rendering of a solid volume displacer 400 is provided, alongside a slice view of a liquid container 450 that is compatible with the displacer. In some cases, the liquid container 450 is approximately 75.5 mm long, has a tapered cylindrical portion, and a hemispherical bottom that is 1.0 mm thick. Therefore, 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 displacer spherical cap (i.e., the endmost end of the second end of the displacer). In some cases, when the second end of the displacer contacts the bottom of the container, the gap between the outer surface diameter of the solid volume displacer 400 and the inner surface diameter of the liquid container 450 near the bottom of the container 450 may be approximately 0.2 mm. The portion of the solid volume displacer 400 that extends outside the container 450 when the second end of the solid volume displacer 400 contacts the bottom of the container is configured as a handle.

[0093] Figure 5A-5B A straight cylindrical (i.e. non-stepped) solid volume displacer according to an embodiment of the present invention is described. Figure 5A In FIG. 5 , a side view of a solid volume displacer 500 is provided, which has a first end, a single cylindrical portion of 120 mm in length and 9.0 mm in diameter, and a spherical cap. The spherical cap is hemispherical with a radius of 4.6 mm, wherein the portion of the hemispherical shape protruding from the cylindrical portion (i.e., the diameter of 9.0 mm) is configured as a continuation of the cylindrical portion. Figure 5B, a three-dimensional rendering of a solid volume displacer 500 is provided, alongside a slice view of a liquid container 530 that is compatible with the displacer. In some cases, the liquid container 530 is approximately 75.5 mm long, has a tapered cylindrical portion, and a hemispherical bottom that is 1.0 mm thick. Therefore, when the second end of the solid volume displacer 500 contacts the bottom of the container 530, the open end of the container is approximately 69.9 mm from the center point of the displacer spherical cap (i.e., the endmost end of the second end of the displacer). In some cases, when the second end of the displacer contacts the bottom of the container, the gap between the outer surface diameter of the solid volume displacer 500 and the inner surface diameter of the liquid container 530 near the bottom of the container 530 may be approximately 0.1 mm. The portion of the solid volume displacer 400 that extends to the outside of the container 530 when the second end of the solid volume displacer 500 contacts the bottom of the container is configured as a handle.

[0094] How to use

[0095] According to the above summary, various aspects of the present disclosure also include the method of reconstructing dry reagent composition using the subject system.As mentioned above, the system for reconstructing dry reagent composition may include one or more dry reagent compositions (such as the first and second polymerized dye compositions) placed separately on the liquid container surface (such as the inner surface of the liquid container wall), and may be placed inside the liquid container to occupy the solid volume displacer of most of the liquid container volume below the top of the dry reagent composition.Therefore, the use method of the subject system may include reconstructing dry reagent composition.In some instances, the dry reagent composition is a dye composition, such as a polymerized dye composition. The polymerized dye composition may be a dry polymerized dye composition.Therefore, the use method of the system may include reconstructing dye composition.In certain embodiments, the method includes introducing a certain volume of liquid into the liquid container, then in a manner that is enough to displace the liquid upwards along the inner surface of the liquid container wall, the solid volume displacer is introduced into the liquid container, thereby reconstructing the dry reagent composition.Any convenient liquid handling device (such as but not limited to syringe, needle, pipette, aspirator and other liquid handling devices) may be used to add the certain volume of liquid into the container.

[0096] In the implementation of embodiments of the subject method, the method may include introducing a volume of liquid into a liquid container, the liquid container comprising: an open end and a bottom, separated by a wall, wherein the inner surface of the wall comprises a dry reagent composition; introducing a solid volume displacer into the liquid container in a manner sufficient to displace the liquid upward along the inner surface of the liquid container wall, thereby reconstructing the dry reagent composition. In the implementation of embodiments of the subject method, the method may include mixing the liquid and the dry reagent composition by moving the solid volume displacer. In these examples, the movement may include rotating and / or reciprocating the solid volume displacer. In the implementation of certain embodiments of the subject method, the movement may include rotating the solid volume displacer. In the implementation of certain embodiments of the subject method, the movement may include reciprocating the solid volume displacer.

[0097] In some embodiments, the liquid comprises a biological sample. In some cases, the biological sample may be derived from a specific biological fluid, such as, but not limited to, blood, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, bronchoalveolar lavage fluid, 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.

[0098] In certain embodiments, the liquid container is a sealed container (for example, the liquid container is through a sealing process), for example, the container includes a seal (for example, impermeable and / or airtight seal). In these examples, the method may include: before a certain volume of liquid is placed in or introduced into the liquid container, the seal is disassembled. The seal on the liquid container is removed and the contents of the liquid container can be exposed to the surrounding environment, and the internal volume of the liquid container is allowed to be touched. Therefore, the user who can touch the internal volume of the liquid container can place a certain volume of liquid in the liquid container inside, so that the liquid container is reconstructed to dry polymeric dye composition (for example, in a manner that is enough to displace the inner surface of the liquid container wall upwards, the solid volume displacer is introduced into the liquid container, thereby reconstructing the dry reagent composition). When implementing some embodiments of the subject method, the method may include in that after all dry reagent compositions have been reconstructed, the liquid container is sealed. In certain embodiments, the seal may be a removable cover. In these examples, the method may include: before a certain volume of liquid is placed in or introduced into the liquid container inside, the seal is disassembled, and after all dry reagent compositions have been reconstructed, the liquid container is resealed.

[0099] In certain embodiments, the solid volume displacer may include a first end and a second end and a main body therebetween. In certain embodiments, the first end of the solid volume displacer may be fixed to a mechanical mixer. When implementing certain embodiments of the subject method, the method may include: after the solid volume displacer is placed inside the liquid container, the contents of the mixed liquid container (e.g., by fixing the first end of the solid volume displacer to a mechanical mixer). Any convenient scheme may be used to mix. For example, a stirrer may be used to mix. The stirrer may be any convenient stirrer sufficient to mix the liquid inside the liquid container, including but not limited to a vortex oscillator, a remote sonic locator, a shaking table (e.g., a manual, mechanical or electric shaking table), a rocker, an oscillating plate, a magnetic stirrer, a static mixer, a rotator, an admixture, a mixer, a drum, an orbital shaking table and other stirring schemes. When implementing certain embodiments of the subject method, compared with using a stirrer (e.g., a vortex oscillator) alone (i.e., without using a solid volume displacer), using a solid volume displacer to reconstruct the dry reagent composition includes introducing a smaller volume of liquid. For example, compared to using a vortex oscillator alone (e.g., without using a solid volume displacer), reconstructing a dry reagent composition using a solid volume displacer can reduce the volume of liquid that needs to be introduced into a liquid container by 50% or more, such as 60% or more, or 70% or more, 80% or more, or 90% or more. In certain embodiments of the subject method, reconstructing a dry reagent composition using a vortex oscillator with a solid volume displacer includes introducing a smaller volume of liquid compared to using a solid volume displacer alone (i.e., without using a vortex oscillator).

[0100] In some cases, the method further includes detecting a reconstructed composition, such as a reconstructed dye composition. Any suitable detection device can be used to detect the reconstructed composition (such as a reconstructed dye composition). For example, the detection device can be a flow cytometer. In these embodiments, the detection includes performing flow cytometry analysis on the reconstructed composition (such as a reconstructed dye composition). In some instances, the detection includes contacting the reconstructed composition (such as a reconstructed dye composition) with electromagnetic radiation (such as light) (such as electromagnetic radiation with a wavelength corresponding to the excitation maximum of the reconstructed composition (such as a reconstructed dye composition)). The detection may further include detecting the light emitted by the excitation reagent composition (such as a dye composition). For example, the method may include: detecting the emitted light of the excitation 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).

[0101] Suitable flow cytometer systems may include, but are not limited to, those described in Ormerod (ed.), Flow Cytometry: A Practical Approach, Oxford University Press (1997); Jaroszeski et al. (eds.), Flow Cytometry Protocols, Methods in Molecular Biology 91, Humana Press (1997); Practical Flow Cytometry, 3rd ed., Wiley-Liss (1995); Virgo et al. (2012), Annals of Clinical Biochemistry Jan;49(pt 1):17-28; Linden et al., Seminars in Thrombosis and Hemostasis, Oct 2004;30(5):502-11; Alison et al., J Pathol, Dec 2010;222(4):335-344; and Herbig et al. (2007), Critical Reviews of Therapeutic Drug Delivery Systems 24(3):203-255; the disclosures of the above documents are incorporated herein by reference. In some examples, the flow cytometry system of interest includes a 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 LSR Fortess TM X-20 Flow Cytometer and BD Biosciences FACSCalibur TM Cell sorter, 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 TMCell sorter and BD Biosciences FACSMelody TM Cell sorter, etc.

[0102] In certain embodiments, the subject flow cytometry system can sort one or more particles (e.g., cells) in a sample. In the present invention, the term "sorting" in its conventional sense refers to separating components of a sample (e.g., cells and non-cellular particles such as biomacromolecules), and in some cases transferring the separated components to one or more sample collection containers. For example, the subject system can be used to sort samples having 2 or more components (e.g., 3 or more components, 4 or more components, 5 or more components, 10 or more components, 15 or more components), including sorting samples having 25 or more components. One or more sample components (e.g., 2 or more sample components, 3 or more sample components, 4 or more sample components, 5 or more sample components, 10 or more sample components) can be separated from a sample and transferred to a sample collection container, including 15 or more sample components can be separated from a sample and transferred to a sample collection container.

[0103] In some embodiments, the target particle sorting system sorts particles by a closed particle sorting module, such as the particle sorting module described in U.S. Patent Publication No. 2017 / 0299493 filed on March 28, 2017, the contents of which are incorporated herein by reference. In certain embodiments, a sorting decision module having a plurality of sorting decision units is used to sort particles (e.g., cells) of a sample, such as the sorting decision module described in U.S. Patent Application No. 16 / 725,756 filed on December 23, 2019, the disclosure of which is incorporated herein by reference.

[0104] In certain embodiments, the flow cytometer system is a flow cytometer system as described in the following U.S. Patents: 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; The disclosures of the above-mentioned U.S. patents are incorporated herein by reference in their entirety and become a part of the present invention.

[0105] Other analytical methods may also be used, such as but not limited to liquid chromatography-mass spectrometry or gas chromatography-mass spectrometry systems. For example, detection may include the use of analytical separation devices such as liquid chromatographs (LC), including high performance liquid chromatographs (HPLC), micro or nano liquid chromatographs or ultra-high pressure liquid chromatographs (UHPLC) devices, capillary electrophoresis (CE) or capillary electrophoresis chromatographs (CEC) devices. A mass spectrometer (MS) system may also be used to detect reagent compositions, such as dye compositions. 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, etc. or any combination thereof. Likewise, any of a variety of mass analyzers may be employed, including a time-of-flight (TOF) analyzer, a Fourier transform ion cyclotron resonance (FTICR) analyzer, an ion trap analyzer, a quadrupole or double focusing magnetic electric sector mass analyzer, or any hybrid thereof.

[0106] In certain embodiments, the subject system is included in a fully automated device. By "fully automated" is meant that the device receives the subject system and prepares a reconstructed composition, such as a reconstructed dye composition, but requires little human intervention or human input to the subject system. In certain embodiments, the subject system can prepare and analyze a reconstructed composition, such as a reconstructed dye composition, without requiring any human intervention.

[0107] In certain embodiments, the method also includes storing the reconstructed composition (e.g., reconstructed dye composition) for a period of time. Before, during, and / or after detecting the reconstructed composition (e.g., reconstructed dye composition), the reconstructed composition (e.g., reconstructed dye composition) can be stored for a period of time. In some examples, the reconstructed composition (e.g., reconstructed dye composition) is stored for a period of time, 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. In some cases, the reconstructed composition is stored for 24 hours or more. In some cases, the reconstructed composition is stored for 48 hours or more. In some cases, the reconstructed composition is stored for 72 hours or more. In some cases, the reconstructed composition is stored for 1 week or more. In some cases, the reconstructed composition is stored for 2 weeks or more. In some cases, the reconstituted composition is stored for 3 weeks or more.

[0108] Embodiments of the method may further include transporting the reconstructed composition to a remote location. "Remote location" refers to a location outside the dye composition reconstruction location. For example, the remote location can be another location (such as an office, laboratory, etc.) in the same city, another location in a different city, another location in different states, another location in different countries, etc. Therefore, when an article is noted as "far away" from another article, it means that the two articles can be in the same room but separated from each other, or at least in different rooms or different buildings, and can be at least one mile, ten miles, or one hundred miles or more apart.

[0109] Figure 2A-2C A method for reconstructing a dry reagent composition using a system according to an embodiment of the present invention is described. Figure 2A In the embodiment of the present invention, a volume of liquid 230 (e.g., a biological liquid sample) is introduced into the liquid container 210. Figure 2B In the embodiment of the present invention, the solid volume displacer 220 is introduced into the liquid container 210 in a manner sufficient to displace the volume of liquid 230 upward along the inner surface of the wall of the liquid container 213, thereby reconstructing the dry reagent composition 214. Figure 2C In the embodiment of the present invention, after reconstitution, the solid volume displacer 220 is removed, thereby disassembling the assembly consisting of the liquid container 210, the solid volume displacer 220 and a certain volume of liquid 230. The liquid 230 containing the reconstitution reagent can then be collected for analysis.

[0110] Kits

[0111] Various aspects of the present disclosure also include a kit. The kit may include a system for reconstructing a dry reagent composition according to any embodiment of the present invention. Various aspects of the system for reconstructing a dry reagent composition may include a liquid container and a solid volume displacer, as described above. Various aspects of the liquid container may include an open end and a bottom, separated by a wall, wherein the inner surface of the wall comprises a dry reagent composition. Various aspects of the solid volume displacer may include a first end and a second end and a body therebetween, wherein the second end and the body may be placed inside the liquid container.

[0112] In certain embodiments, the kit includes a subject system for reconstructing a dry reagent composition and a package that can accommodate the system. The package can be a sealed package, such as a waterproof steam container, or can be airtight and / or vacuum sealed. In certain embodiments, the package can protect its internal contents (such as the subject system) from light exposure. In these examples, the package can resist light of any wavelength range or spectrum, including visible light, ultraviolet light, light within and / or near the excitation spectrum of any encapsulated dye, etc. In certain instances, the package is a sterile package, and the device encapsulated in the package can be maintained in a sterile environment. "Sterile" refers to substantially no microorganisms, such as fungi, bacteria, viruses, spore forms, etc. In certain embodiments, the liquid container and the solid volume displacer are contained in the same package. In other examples, the liquid container and the solid volume displacer are contained in separate packages.

[0113] The kit may further include a buffer. For example, the kit may include a buffer, such as a sample buffer, a wash buffer, a detection buffer, etc. The kit may further include other reagents, such as but not limited to detectable labels (such as 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 also includes calibration standards. For example, the kit may include a set of labeled microspheres, such as a set of standard fluorescent labeled microspheres.

[0114] In addition to the above-mentioned components, the theme kit may further include instructions for implementing the theme method. The instructions may be present in the theme kit in various forms, wherein one or more instruction forms may be present in the kit. A form of existence of these instructions is the information printed on a suitable medium or substrate, for example, a sheet or multiple sheets of paper with information printed in the packaging, package instructions, etc. of the kit. Another way is a computer-readable medium, such as a CD, DVD, blue disc, computer-readable memory (such as a flash memory), etc., and information has been recorded or stored on these media. Another form that can exist is a website address, which can be used to access information on a deleted website via the Internet. Any convenient instruction form may be present in the kit.

[0115] Useful functions

[0116] The subject systems and methods are applicable to research, laboratory tests or treatments requiring cell analysis of biological samples. In certain embodiments, the subject systems and methods facilitate analysis of cells obtained from fluid or tissue samples (such as disease specimens, including but not limited to cancer). The systems and methods of the present disclosure also allow for analysis of cells from biological samples (e.g., organs, tissues, tissue fragments, fluids) at high efficiency and low cost.

[0117] The subject system and method can be used for applications where a dry reagent composition (such as a dye composition) is required for sample analysis. For example, the subject system and method can be used for applications where a dry polymeric dye composition is required for sample analysis. Embodiments of the subject system and method can also be used for applications where a dry polymeric dye composition is required to be combined with a dry non-polymeric dye composition for sample analysis. Therefore, the subject system and method can be used for applications where a corresponding dry polymeric dye composition is used to analyze target analytes in a sample. In some cases, if a non-polymeric dye composition is also included in the liquid container of the system, the subject system and method can be used for applications where a corresponding dry polymeric dye composition and a dry non-polymeric dye composition are used to analyze target analytes in a sample.

[0118] The subject system and method can be used for the application occasions that need to reduce the required liquid volume of dry reagent composition (such as polymeric dye composition) reconstruction or rehydration to the greatest extent.As described in the present invention, the embodiment of the subject system and method provides the liquid container with dry reagent composition on inner wall and is placed on the solid volume displacer of the liquid container volume of most of liquid container below the top of the liquid container internal occupation dry reagent composition.Therefore, in the mode that is enough to make liquid displace upward along the inner surface of liquid container wall, solid volume displacer is introduced into liquid container, thereby reconstructs dry reagent composition, helps to reduce the required liquid volume of dry reagent composition (such as polymeric dye composition) reconstruction or rehydration to the greatest extent.Reduce the required liquid volume of dry reagent composition reconstruction or rehydration to the greatest extent, help to improve the reagent concentration after reconstruction or rehydration.Improve the reagent concentration after reconstruction or rehydration and help to collect more accurate and / or accurate data related to detecting using the subject system, and / or reduce the cost of detecting using the subject system.For example, compared with providing dry reagent composition but relying on other methods to move liquid into dry reagent composition (such as vortex method) thereby reconstructing the system of dry reagent composition, the subject system and method help to reduce the required liquid volume of dry reagent composition reconstruction or rehydration.

[0119] The subject system and method can be used for applications where two or more reagents (such as dye compositions) are required for sample analysis. For example, the subject system and method can be used for applications where two or more polymeric dye compositions are required for sample analysis. Embodiments of the subject system and method can also be used for applications where two or more polymeric dye compositions are combined with one or more non-polymeric dye compositions for sample analysis. Therefore, the subject system and method can be used for applications where two or more corresponding polymeric dye compositions are used to analyze two or more target analytes in a sample. In some cases, if the liquid container also contains a non-polymeric dye composition, the subject system and method can be used for applications where two or more corresponding polymeric dye compositions and non-polymeric dye compositions are used to analyze two or more target analytes in a sample.

[0120] The subject system and method can be used for applications where it is necessary to minimize the interaction between dyes. As described in the present invention, the embodiment of the subject system and method provides two or more dry polymeric dye compositions placed separately on the inner surface of the liquid container wall. Therefore, the dye composition is placed separately relative to each other on the inner surface of the liquid container wall to help minimize the interaction between dyes. Minimizing the interaction between dyes can help collect more accurate and / or accurate data related to detection using the subject system. For example, compared with providing two or more dye compositions but not placed separately relative to each other, the subject system and method can help reduce the interaction between dyes. In addition, the dye composition is placed separately relative to each other on the inner surface of the liquid container wall to make the dye composition farther and farther away from the bottom of the liquid container as the dye composition is added additionally. The dye composition is farther and farther away from the bottom of the liquid container to make the liquid volume required for the reconstruction or rehydration of the dye composition increase. Therefore, when a solid volume displacer is introduced into a liquid container to cooperate with a certain volume of liquid along the inner surface of the liquid container wall to displace upwards thereby reconstructing the dye composition, the dye composition is placed separately relative to each other on the inner surface of the liquid container wall to help collect more accurate and / or accurate data related to detection using two or more dye compositions in the subject system.

[0121] The following examples are offered by way of illustration and not by way of limitation.

[0122] Example

[0123] The following examples are presented to provide a person of ordinary skill in the art with a complete disclosure and description of how to make and use the invention, but are not intended to limit the scope of the invention as recognized by the inventors, nor are they intended to represent that the following experiments are all or the only experiments performed. We have tried to ensure the accuracy of the numbers used (e.g., quantities, temperatures, etc.), but some experimental errors and deviations should be taken into account. Unless otherwise stated, parts are parts by weight, molecular weights are weight average molecular weights, temperatures are in degrees Celsius, and pressures are atmospheric pressure or near atmospheric pressure. In addition, common laboratory protocol abbreviations may also be used (e.g., hr = hours, min = minutes, ml = milliliters, μl = microliters, rpm = revolutions per minute, g (in the case of centrifugation) = multiples of gravity, etc.).

[0124] The following experiments compared the system of the present invention for reconstituting a dry reagent composition with prior art systems and methods for reconstituting a dry reagent composition.

[0125] Materials and methods

[0126] The dye composition was dispensed onto the inner surface of a 12×75 mm tube wall using a liquid handling device to form a liquid container according to an embodiment of the present invention. The dye composition was dispensed within a range of 20 mm from the bottom of the tube and then dried.

[0127] Three embodiments of the solid volume displacer according to the embodiment of the present invention are machined. Solid volume displacer 1 is machined so that the diameter of the outer surface of the displacer body is 1 mm smaller than the diameter of the inner surface of the liquid container wall. The first end of solid volume displacer 1 is configured as a flange to prevent the second end of the displacer from contacting the bottom of the liquid container. At the same time, solid volume displacers 2 and 3 are machined so that the diameter of the outer surface of the displacer body is 0.5 mm smaller than the diameter of the inner surface of the liquid container wall. The first end of solid volume displacer 2 is configured as a flange to prevent the second end of the displacer from contacting the bottom of the liquid container. The first end of solid volume displacer 3 is configured as a flange to enable the second end of the displacer to contact the bottom of the liquid container.

[0128] A control method was tested by filling a liquid container with liquid until all of the dry dye composition of the liquid container was observed to be reconstituted.

[0129] Figure 3A A method for testing vortexing for the purpose of reconstructing a dry dye composition in a liquid container is described. From left to right, a certain volume of liquid is first introduced into the liquid container. The liquid in the liquid container is then vortexed to move the introduced certain volume of liquid upward along the wall of the liquid container. After the vortexing, the liquid container is observed to determine whether the volume of liquid introduced is sufficient to reconstruct all the dry dye composition in the liquid container.

[0130] Figure 2A-2C A method for testing a solid volume displacer (e.g., according to an embodiment of the present invention) is described. Figure 2A In the embodiment, a certain volume of liquid 230 is introduced into the liquid container 210. Figure 2B In the embodiment, the solid volume displacer 220 is introduced into the liquid container 210 in a manner sufficient to displace the volume of liquid 230 upward along the inner surface of the wall of the liquid container 213. Figure 2C In the embodiment of the present invention, the solid volume displacer 220 is removed from the liquid container 210 and the liquid container is observed to determine whether the volume of liquid introduced is sufficient to reconstitute all of the dry dye composition in the liquid container.

[0131] Table 1: Solid volume displacer dimensions and experimental results.

[0132]

[0133] For the volume of a sphere (twice the volume of the base or second end) and the volume of a cylinder, use the following formulas for the above calculations:

[0134] Sphere: Cylinder: V = πr 2 h

[0135] result

[0136] The control method requires the introduction of 1463.2 μl of liquid to reconstruct all dry dye compositions. The liquid required for reconstructing the liquid container dry dye compositions by vortexing is significantly less than that of the control group. In order to reconstruct all dry dye compositions by vortexing, 827 μl of liquid needs to be introduced into the liquid container before the liquid container is vortexed.

[0137] Solid volume displacer 1 required 323.8 μl of liquid to reconstitute all the dry dye composition in the liquid container. Accordingly, the liquid volume required to reconstitute all the dry dye composition by solid volume displacer 1 was 77.9% less than the control group and 60.8% less than the vortex method.

[0138] Solid volume displacer 2 required 167.5 μl of liquid to reconstitute all the dried dye composition in the liquid container. Accordingly, the liquid volume required to reconstitute all the dried dye composition by solid volume displacer 2 was 88.6% less than the control group and 79.7% less than the vortex method. Figure 3A-3B A comparison of the vortexing method for reconstituting a dried dye composition compared to the use of a solid volume displacer 2 is described. Figure 3A Depicted from left to right is the introduction of a volume of liquid, the filling of the liquid container up to a height of 10 mm along the liquid container wall, the vortexing of the liquid container, and the resulting liquid volume containing a relatively diluted reconstituted dye composition. Figure 3B From left to right, it is described that the introduced liquid volume only partially fills the circular bottom of the liquid container, and the solid volume displacer 2 is introduced into the liquid container in a manner sufficient to displace the certain volume of liquid upward along the inner surface of the liquid container wall. After removing the solid volume displacer 2, the resulting liquid volume contains a relatively high concentration of the reconstructed dye composition.

[0139] Solid volume displacer 3 required 139.1 μl of liquid to reconstitute all the dry dye composition in the liquid container. Accordingly, the solid volume displacer 3 required 90.5% less liquid volume to reconstitute all the dry dye composition than the control group and 83.2% less than the vortex method.

[0140] Notwithstanding the appended claims, the present disclosure is defined by the following terms:

[0141] 1. A system for reconstitution of a dry reagent composition, the system comprising:

[0142] a liquid container comprising an open end and a bottom separated by a wall, wherein an inner surface of the wall comprises a dry reagent composition;

[0143] A solid volume displacer is placed inside the liquid container to occupy a majority of the liquid container volume below the top of the dry reagent composition.

[0144] 2. A system according to claim 1, wherein the solid volume displacer can occupy 60% or more of the volume of the liquid container below the top of the dry reagent composition.

[0145] 3. A system according to claim 2, wherein the solid volume displacer can occupy 75% or more of the volume of the liquid container below the top of the dry reagent composition.

[0146] 4. A system according to claim 3, wherein the solid volume displacer can occupy 85% or more of the volume of the liquid container below the top of the dry reagent composition.

[0147] 5. A system according to claim 4, wherein the solid volume displacer can occupy 90% or more of the volume of the liquid container below the top of the dry reagent composition.

[0148] 6. A system according to any of the preceding clauses, wherein the dry reagent composition comprises a dye and / or microspheres.

[0149] 7. A system according to clause 6, wherein the dry reagent composition includes a dye.

[0150] 8. A system according to clause 6 or 7, wherein the dye is a polymeric dye.

[0151] 9. The system of clause 8, wherein the polymeric dye comprises a coupled polymer.

[0152] 10. A system according to any of the preceding clauses, wherein the liquid container contains a liquid volume that is greater than or equal to the liquid container volume below the top of the dry reagent composition that is not occupied by the solid volume displacer.

[0153] 11. A system according to any of the preceding clauses, wherein the dry reagent composition is located 3 mm or more from the bottom of the liquid container.

[0154] 12. A system according to clause 11, wherein the position of the dry reagent composition is 10 mm or more from the bottom of the liquid container.

[0155] 13. A system according to clause 12, wherein the position of the dry reagent composition is 20 mm or more from the bottom of the liquid container.

[0156] 14. A system according to any of the preceding clauses, wherein the inner surface of the liquid container wall comprises two or more separately placed dry reagent compositions.

[0157] 15. The system of clause 14, wherein the two or more dry reagent compositions are placed at separate locations on the inner surface of the liquid container wall.

[0158] 16. A system according to any of the preceding clauses, wherein the inner surface of the liquid container wall comprises six or more separately placed dry reagent compositions.

[0159] 17. The system of any of clauses 14-16, wherein the two or more dry reagent compositions comprise a dye.

[0160] 18. The system of clause 17, wherein the dye is a polymeric dye.

[0161] 19. The system of clause 18, wherein the polymeric dye comprises a coupled polymer.

[0162] 20. A system according to any of the preceding clauses, wherein the liquid container comprises a tube or a vial.

[0163] 21. A system according to any of the preceding clauses, wherein the solid volume displacer is configured as a pestle.

[0164] 22. A system according to any of the preceding clauses, wherein the volume of the liquid container below the top of the dry reagent composition not occupied by the solid volume displacer is 350 μl or less.

[0165] 23. A system according to clause 22, wherein the volume of the liquid container below the top of the dry reagent composition not occupied by the solid volume displacer is 150 μl or less.

[0166] 24. The system of any of the preceding clauses, wherein the solid volume displacer comprises a first end and a second end and a body therebetween, the second end and the body being positionable inside a liquid container.

[0167] 25. The system of clause 24, wherein the body of the solid volume displacer is configured as a cylinder.

[0168] 26. The system of clause 25, wherein the two or more dry reagent compositions are placed at separate locations on the inner surface of the liquid container wall.

[0169] 27. The system of clause 26, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 1 mm or less.

[0170] 28. The system of clause 26, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 0.5 mm or less.

[0171] 29. The system of clause 26, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 0.05 mm or less.

[0172] 30. The system according to any of clauses 25-29, wherein the outer surface of the body of the solid volume displacer and the wall of the liquid container are tapered.

[0173] 31. The system of any of clauses 25-30, wherein the first end of the solid volume displacer is positionable outside of a liquid container while the body and second end of the solid volume displacer are positionable inside of a liquid container.

[0174] 32. The system of clause 31, wherein the first end of the solid volume displacer comprises a flange.

[0175] 33. The system of clause 32, wherein the flange prevents the second end of the solid volume displacer from contacting the bottom of the liquid container.

[0176] 34. The system of clause 33, wherein the flange enables the second end of the solid volume displacer to contact the bottom of the liquid container.

[0177] 35. The system of any of clauses 25-34, wherein the solid volume displacer further comprises a ring of filter media disposed around the exterior of the body proximate the first end.

[0178] 36. The system of clause 35, wherein the ring of filter media prevents liquid from splashing out of the liquid container when the body of the solid volume displacer is placed inside the liquid container.

[0179] 37. The system of any of clauses 25-36, wherein the first end of the solid volume displacer comprises a handle.

[0180] 38. The system of any of the preceding clauses, wherein the solid volume displacer is a hollow solid volume displacer.

[0181] 39. The system according to any one of clauses 25-37, wherein the liquid container has a rounded bottom.

[0182] 40. The system of clause 39, wherein the second end of the solid volume displacer is rounded and concentric with the bottom of the liquid container.

[0183] 41. A system according to any of the preceding clauses, wherein the solid volume displacer is fixable to a mechanical mixer.

[0184] 42. The system of any of the preceding clauses, wherein the solid volume displacer comprises a first polymeric material.

[0185] 43. The system of clause 42, wherein the first polymeric material comprises a first hydrophobic polymeric material.

[0186] 44. The system of any of clause 43, wherein the first hydrophobic polymeric material comprises a first plastic, a first resin, a first metal, or a first ceramic.

[0187] 45. The system of any of clause 44, wherein the first hydrophobic polymeric material comprises a first plastic.

[0188] 46. ​​The system of clause 45, wherein the first plastic comprises LDPE or PTFE.

[0189] 47. A system according to any of the preceding clauses, wherein the liquid container comprises a second polymeric material.

[0190] 48. The system of clause 47, wherein the second polymeric material comprises a second hydrophobic polymeric material.

[0191] 49. The system of clause 48, wherein the second hydrophobic polymeric material comprises a second plastic.

[0192] 50. The system of clause 49, wherein the second plastic comprises LDPE or PTFE.

[0193] 51. A system according to any of the preceding clauses, wherein the liquid container includes a seal.

[0194] 52. The system of clause 51 , wherein the seal comprises a removable cover.

[0195] 53. A method for reconstituted a dry reagent composition, the method comprising:

[0196] introducing a volume of liquid into a liquid container, the liquid container comprising an open end and a bottom separated by a wall, wherein an inner surface of the wall comprises a dry reagent composition;

[0197] The solid volume displacer is introduced into the liquid container in a manner sufficient to displace liquid upwardly along the interior surface of the liquid container wall, thereby reconstituting the dry reagent composition.

[0198] 54. A method according to clause 53, wherein the solid volume displacer occupies a majority of the liquid container volume below the top of the dry reagent composition.

[0199] 55. A method according to clause 54, wherein the solid volume displacer occupies 60% or more of the volume of the liquid container below the top of the dry reagent composition.

[0200] 56. A method according to clause 55, wherein the solid volume displacer occupies 75% or more of the volume of the liquid container below the top of the dry reagent composition.

[0201] 57. A method according to clause 56, wherein the solid volume displacer occupies 85% or more of the volume of the liquid container below the top of the dry reagent composition.

[0202] 58. A method according to clause 57, wherein the solid volume displacer occupies 90% or more of the volume of the liquid container below the top of the dry reagent composition.

[0203] 59. A method according to any one of clauses 53 to 58, wherein the dry reagent composition comprises a dye and / or microspheres.

[0204] 60. The method of clause 59, wherein the dry reagent composition comprises a dye.

[0205] 61. The method of clause 60, wherein the dye is a polymeric dye.

[0206] 62. The method of clause 61, wherein the polymeric dye comprises a coupled polymer.

[0207] 63. A method according to any one of clauses 53 to 62, wherein the dried reagent composition is located 3 mm or more from the bottom of the liquid container.

[0208] 64. A method according to clause 63, wherein the position of the dry reagent composition is 10 mm or more from the bottom of the liquid container.

[0209] 65. A method according to clause 64, wherein the position of the dry reagent composition is 20 mm or more from the bottom of the liquid container.

[0210] 66. A method according to any of clauses 53-65, wherein the inner surface of the liquid container wall comprises two or more separately placed dry reagent compositions.

[0211] 67. A method according to clause 66, wherein the two or more dry reagent compositions are placed at separate locations on the inner surface of the liquid container wall.

[0212] 68. The method of clause 67, wherein the inner surface of the liquid container wall comprises six or more separately placed dry reagent compositions.

[0213] 69. The method of any one of clauses 66-68, wherein the two or more dry reagent compositions comprise a dye.

[0214] 70. The method of clause 69, wherein the dye is a polymeric dye.

[0215] 71. The method of clause 70, wherein the polymeric dye comprises a coupled polymer.

[0216] 72. A method according to any of clauses 53-71, wherein the liquid container comprises a tube or a vial.

[0217] 73. A method according to any of clauses 53-72, wherein the solid volume displacer is configured as a pestle.

[0218] 74. A method according to any one of clauses 53 to 73, wherein the volume of liquid introduced into the liquid container is 5000 μl or less.

[0219] 75. A method according to clause 74, wherein the volume of liquid introduced into the liquid container is 350 μl or less.

[0220] 76. A method according to clause 75, wherein the volume of liquid introduced into the liquid container is 150 μl or less.

[0221] 77. The method of any of clauses 53-76, wherein the solid volume displacer comprises a first end and a second end and a body therebetween, the second end and the body being positionable within a liquid container.

[0222] 78. The method of clause 77, wherein the body of the solid volume displacer is configured as a cylinder.

[0223] 79. The method of clause 78, wherein the two or more dry reagent compositions are placed at separate locations on the inner surface of the liquid container wall.

[0224] 80. The method of clause 79, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 1 mm or less.

[0225] 81. A method according to clause 80, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 0.5 mm or less.

[0226] 82. A method according to clause 81, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 0.05 mm or less.

[0227] 83. A method according to any of clauses 77-82, wherein the outer surface of the body of the solid volume displacer and the wall of the liquid container are tapered.

[0228] 84. A method according to any of clauses 77-83, wherein the first end of the solid volume displacer is positionable outside of a liquid container while the body and second end of the solid volume displacer are positionable inside of a liquid container.

[0229] 85. The method of clause 84, wherein the first end of the solid volume displacer comprises a flange.

[0230] 86. The method of clause 85, wherein the flange prevents the second end of the solid volume displacer from contacting the bottom of the liquid container.

[0231] 87. The method of clause 86, wherein the flange enables the second end of the solid volume displacer to contact the bottom of the liquid container.

[0232] 88. A method according to any of clauses 84-87, wherein the flange comprises a ring of filter media disposed around the exterior of the body proximate the first end.

[0233] 89. The method of clause 88, wherein the ring of filter media prevents liquid from splashing out of the liquid container when the body of the solid volume displacer is placed inside the liquid container.

[0234] 90. The method of any of clauses 77-89, wherein the first end of the solid volume displacer comprises a handle.

[0235] 91. The method of any one of clauses 53-90, wherein the solid volume displacer is a hollow solid volume displacer.

[0236] 92. A method according to any one of clauses 77-90, wherein the bottom of the liquid container is circular.

[0237] 93. A method according to clause 92, wherein the second end of the solid volume displacer is circular and concentric with the bottom of the liquid container.

[0238] 94. A method according to any one of clauses 53-93, wherein the method further comprises mixing the liquid and dry reagent compositions by moving a solid volume displacer.

[0239] 95. The method of clause 94, wherein the moving comprises rotating a solid volume displacer.

[0240] 96. A method according to any of clauses 94-95, wherein the moving comprises reciprocating the solid volume displacer.

[0241] 97. A method according to any of clauses 53 to 96, wherein the solid volume displacer is fixable to a mechanical mixer.

[0242] 98. The method of any of clauses 53-97, wherein the solid volume displacer comprises a first polymeric material.

[0243] 99. The method of clause 98, wherein the first polymeric material comprises a first hydrophobic polymeric material.

[0244] 100. The method of clause 99, wherein the first hydrophobic polymeric material comprises a first plastic, a first resin, a first metal, or a first ceramic.

[0245] 101. The method of clause 100, wherein the first hydrophobic polymeric material comprises a first plastic.

[0246] 102. The method of clause 100, wherein the first plastic comprises LDPE or PTFE.

[0247] 103. A method according to any of clauses 53-102, wherein the liquid container comprises a second polymeric material.

[0248] 104. The method of clause 103, wherein the second polymeric material comprises a second hydrophobic polymeric material.

[0249] 105. The method of clause 104, wherein the second hydrophobic polymeric material comprises a second plastic.

[0250] 106. The method of clause 105, wherein the second plastic comprises LDPE or PTFE.

[0251] 107. A method according to any of clauses 53-106, wherein the liquid container is a sealed container, and the method includes removing a seal before placing the certain volume of liquid into the interior of the liquid container.

[0252] 108. The method of any one of clauses 53-107, further comprising sealing the liquid container after all of the dry reagent composition has been reconstituted.

[0253] 109. A method according to any of clauses 107-108, wherein the seal comprises a removable cover.

[0254] 110. An assembly comprising:

[0255] a liquid container comprising an open end and a bottom separated by a wall, wherein an inner surface of the wall comprises a dry reagent composition;

[0256] A certain volume of liquid is introduced into the liquid container;

[0257] The solid volume displacer is placed inside the liquid container to displace the certain volume of liquid upward along the inner surface of the liquid container wall, thereby contacting with the dry reagent composition.

[0258] 111. An assembly according to clause 110, wherein the solid volume displacer occupies a majority of the liquid container volume below the top of the dry reagent composition.

[0259] 112. An assembly according to clause 111, wherein the solid volume displacer occupies 60% or more of the volume of the liquid container below the top of the dry reagent composition.

[0260] 113. An assembly according to clause 112, wherein the solid volume displacer occupies 75% or more of the volume of the liquid container below the top of the dry reagent composition.

[0261] 114. An assembly according to clause 113, wherein the solid volume displacer occupies 85% or more of the volume of the liquid container below the top of the dry reagent composition.

[0262] 115. An assembly according to clause 114, wherein the solid volume displacer occupies 90% or more of the volume of the liquid container below the top of the dry reagent composition.

[0263] 116. An assembly according to any of clauses 110-115, wherein the dry reagent composition comprises a dye and / or microspheres.

[0264] 117. A component according to clause 116, wherein the dry reagent composition includes a dye.

[0265] 118. The assembly of clause 117, wherein the dye is a polymeric dye.

[0266] 119. The assembly of clause 118, wherein the polymeric dye comprises a coupled polymer.

[0267] 120. An assembly according to any of clauses 110-119, wherein the dry reagent composition is located 3 mm or more from the bottom of the liquid container.

[0268] 121. An assembly according to clause 120, wherein the dry reagent composition is located 10 mm or more from the bottom of the liquid container.

[0269] 122. A component according to claim 121, wherein the dry reagent composition is located 20 mm or more from the bottom of the liquid container.

[0270] 123. An assembly according to any of clauses 110-122, wherein the inner surface of the liquid container wall comprises two or more separately placed dry reagent compositions.

[0271] 124. An assembly according to clause 123, wherein the two or more dry reagent compositions are placed at separate locations on the inner surface of the liquid container wall.

[0272] 125. An assembly according to clause 123 or 124, wherein the inner surface of the liquid container wall comprises six or more separately placed dry reagent compositions.

[0273] 126. An assembly according to any of clauses 123 or 125, wherein the two or more dry reagent compositions include a dye.

[0274] 127. The assembly of clause 126, wherein the dye is a polymeric dye.

[0275] 128. The assembly of clause 127, wherein the polymeric dye comprises a coupled polymer.

[0276] 129. An assembly according to any of clauses 110-128, wherein the liquid container comprises a tube or a vial.

[0277] 130. An assembly according to any of clauses 110-129, wherein the solid volume displacer is configured as a pestle.

[0278] 131. An assembly according to any of clauses 110-130, wherein the volume of liquid introduced into the liquid container is 5000 μl or less.

[0279] 132. A component according to clause 131, wherein the volume of liquid introduced into the liquid container is 350 μl or less.

[0280] 133. A component according to clause 132, wherein the volume of liquid introduced into the liquid container is 150 μl or less.

[0281] 134. An assembly according to any of clauses 110-133, wherein the solid volume displacer includes a first end and a second end and a body therebetween, the second end and the body being positioned inside the liquid container.

[0282] 135. An assembly according to clause 134, wherein the body of the solid volume displacer is configured as a cylinder.

[0283] 136. An assembly according to clause 135, wherein the two or more dry reagent compositions are placed at separate locations on the inner surface of the liquid container wall.

[0284] 137. A component according to clause 136, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 1 mm or less.

[0285] 138. A component according to clause 137, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 0.5 mm or less.

[0286] 139. A component according to clause 138, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 0.05 mm or less.

[0287] 140. An assembly according to any of clauses 134-139, wherein the outer surface of the body of the solid volume displacer and the wall of the liquid container are tapered.

[0288] 141. An assembly according to any of clauses 134-140, wherein the first end of the solid volume displacer is placed outside the liquid container.

[0289] 142. An assembly according to clause 141, wherein the first end of the solid volume displacer includes a flange.

[0290] 143. An assembly according to clause 142, wherein the flange contacts the open end of the liquid container and the second end of the solid volume displacer does not contact the bottom of the liquid container.

[0291] 144. An assembly according to clause 142, wherein the flange does not contact the open end of the liquid container and the second end of the solid volume displacer contacts the bottom of the liquid container.

[0292] 145. An assembly according to any of clauses 136-144, wherein the solid volume displacer further comprises a ring of filter media disposed around the outer surface of the body proximate the first end.

[0293] 146. An assembly according to clause 145, wherein the ring of filter media is positioned between the outer surface of the solid volume displacer body and the inner surface of the liquid container wall in a manner sufficient to prevent liquid from splashing out of the liquid container.

[0294] 147. An assembly according to any of clauses 134-146, wherein the first end of the solid volume displacer includes a handle.

[0295] 148. An assembly according to any of clauses 110-147, wherein the solid volume displacer is a hollow solid volume displacer.

[0296] 149. An assembly according to any of clauses 134-147, wherein the bottom of the liquid container is circular.

[0297] 150. An assembly according to clause 149, wherein the second end of the solid volume displacer is circular and concentric with the bottom of the liquid container.

[0298] 151. An assembly according to any of clauses 110-150, wherein the solid volume displacer is securable to a mechanical mixer.

[0299] 152. An assembly as described in any of clauses 110-151, wherein the solid volume displacer comprises a first polymeric material.

[0300] 153. An assembly as described in claim 152, wherein the first polymeric material comprises a first hydrophobic polymeric material.

[0301] 154. The assembly of clause 153, wherein the first hydrophobic polymeric material comprises a first plastic, a first resin, a first metal, or a first ceramic.

[0302] 155. An assembly as described in clause 154, wherein the first hydrophobic polymeric material comprises a first plastic.

[0303] 156. An assembly as described in clause 155, wherein the first plastic comprises LDPE or PTFE.

[0304] 157. An assembly as described in any of clauses 110-156, wherein the liquid container comprises a second polymeric material.

[0305] 158. An assembly as described in clause 157, wherein the second polymeric material comprises a second hydrophobic polymeric material.

[0306] 159. An assembly as described in clause 158, wherein the second hydrophobic polymeric material comprises a second plastic.

[0307] 160. An assembly as described in clause 159, wherein the second plastic comprises LDPE or PTFE.

[0308] 161. An assembly according to any of clauses 110-160, wherein the liquid container includes a removable seal secured to the liquid container.

[0309] 162. An assembly as described in clause 161, wherein the removable seal includes a removable cover.

[0310] 163. A kit comprising:

[0311] a liquid container comprising an open end and a bottom separated by a wall, wherein an inner surface of the wall comprises a dry reagent composition;

[0312] a solid volume displacer, positioned within the liquid container to occupy a majority of the liquid container volume below the top of the dry reagent composition;

[0313] A package can accommodate the liquid container and the solid volume displacer.

[0314] 164. A kit according to claim 163, wherein the solid volume displacer can occupy 75% or more of the volume of the liquid container below the top of the dry reagent composition.

[0315] 165. A kit according to claim 164, wherein the solid volume displacer can occupy 85% or more of the volume of the liquid container below the top of the dry reagent composition.

[0316] 166. A kit according to claim 165, wherein the solid volume displacer can occupy 90% or more of the volume of the liquid container below the top of the dry reagent composition.

[0317] 167. A kit according to any of clauses 163-166, wherein the dry reagent composition comprises a dye and / or microspheres.

[0318] 168. A kit according to clause 167, wherein the dry reagent composition includes a dye.

[0319] 169. A kit according to clause 168, wherein the dye is a polymeric dye.

[0320] 170. The kit of clause 169, wherein the polymeric dye comprises a coupled polymer.

[0321] 171. A kit according to any of clauses 163-170, wherein the dry reagent composition is positioned 3 mm or more from the bottom of the liquid container.

[0322] 172. A kit according to claim 171, wherein the dry reagent composition is located 10 mm or more from the bottom of the liquid container.

[0323] 173. A kit according to claim 172, wherein the dry reagent composition is located 20 mm or more from the bottom of the liquid container.

[0324] 174. A kit according to any of clauses 163-173, wherein the inner surface of the liquid container wall comprises two or more separately placed dry reagent compositions.

[0325] 175. A kit according to clause 174, wherein the two or more dry reagent compositions are placed at separate locations on the inner surface of the liquid container wall.

[0326] 176. A kit according to clause 174 or 175, wherein the inner surface of the liquid container wall comprises six or more separately placed dry reagent compositions.

[0327] 177. A kit according to any of clauses 174-176, wherein the two or more dry reagent compositions include a dye.

[0328] 178. A kit according to clause 177, wherein the dye is a polymeric dye.

[0329] 179. The kit of clause 178, wherein the polymeric dye comprises a coupled polymer.

[0330] 180. A kit according to any of clauses 163-179, wherein the liquid container comprises a tube or a vial.

[0331] 181. A kit according to any of clauses 163-180, wherein the solid volume displacer is configured as a pestle.

[0332] 182. A kit according to any of clauses 163-181, wherein the volume of the liquid container below the top of the dry reagent composition not occupied by the solid volume displacer is 350 μl or less.

[0333] 183. A kit according to clause 182, wherein the volume of the liquid container below the top of the dry reagent composition not occupied by the solid volume displacer is 150 μl or less.

[0334] 184. A kit according to any of clauses 163-183, wherein the solid volume displacer includes a first end and a second end and a body therebetween, the second end and the body being positionable within a liquid container.

[0335] 185. A kit according to clause 184, wherein the body of the solid volume displacer is configured as a cylinder.

[0336] 186. A kit according to clause 185, wherein the two or more dry reagent compositions are placed at separate locations on the inner surface of the liquid container wall.

[0337] 187. A kit according to clause 186, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 1 mm or less.

[0338] 188. A kit according to clause 187, wherein the difference between the diameter of the outer surface of the solid volume displacer body and the diameter of the inner surface of the liquid container wall is 0.5 mm or less.

[0339] 189. A kit according to clause 188, wherein the difference between the outer surface diameter of the solid volume displacer body and the inner surface diameter of the liquid container wall is 0.05 mm or less.

[0340] 190. A kit according to any of clauses 184-189, wherein the outer surface of the body of the solid volume displacer and the wall of the liquid container are tapered.

[0341] 191. A kit according to any of clauses 184-190, wherein the first end of the solid volume displacer can be placed outside the liquid container when the body and second end of the solid volume displacer are placed inside the liquid container.

[0342] 192. A kit according to clause 191, wherein the first end of the solid volume displacer includes a flange.

[0343] 193. A kit according to clause 192, wherein the flange prevents the second end of the solid volume displacer from contacting the bottom of the liquid container.

[0344] 194. A kit according to clause 192, wherein the flange enables the second end of the solid volume displacer to contact the bottom of the liquid container.

[0345] 195. A kit according to any of clauses 184-194, wherein the solid volume displacer further comprises a ring of filter media disposed around the exterior of the body proximate the first end.

[0346] 196. A kit according to clause 195, wherein the ring of filter media prevents liquid from splashing out of the liquid container when the body of the solid volume displacer is placed inside the liquid container.

[0347] 197. A kit according to any of clauses 184-196, wherein the first end of the solid volume displacer includes a handle.

[0348] 198. A kit according to any of clauses 163-197, wherein the solid volume displacer is a hollow solid volume displacer.

[0349] 199. A kit according to any of clauses 184-197, wherein the bottom of the liquid container is circular.

[0350] 200. A kit according to clause 199, wherein the second end of the solid volume displacer is circular and concentric with the bottom of the liquid container.

[0351] 201. A kit according to any of clauses 163-200, wherein the solid volume displacer is securable to a mechanical mixer.

[0352] 202. A kit according to any of clauses 163-201, wherein the solid volume displacer comprises a first polymeric material.

[0353] 203. A kit according to clause 202, wherein the first polymeric material comprises a first hydrophobic polymeric material.

[0354] 204. A kit according to clause 203, wherein the first hydrophobic polymeric material comprises a first plastic, a first resin, a first metal, or a first ceramic.

[0355] 205. A kit according to clause 204, wherein the first hydrophobic polymeric material comprises a first plastic.

[0356] 206. A kit according to clause 205, wherein the first plastic comprises LDPE or PTFE.

[0357] 207. A kit according to any of clauses 163-206, wherein the liquid container comprises a second polymeric material.

[0358] 208. A kit according to clause 207, wherein the second polymeric material comprises a second hydrophobic polymeric material.

[0359] 209. A kit according to clause 208, wherein the second hydrophobic polymeric material comprises a second plastic.

[0360] 210. A kit according to clause 209, wherein the second plastic comprises LDPE or PTFE.

[0361] 211. A kit according to any of clauses 163-210, wherein the liquid container includes a seal.

[0362] 212. A kit according to clause 211, wherein the seal comprises a removable cover.

[0363] Unless replacement is technically infeasible, in at least some of the foregoing embodiments, one or more components used in one embodiment may be used interchangeably in another embodiment. It will be appreciated by those skilled in the art that various other omissions, additions, and modifications may be made to the above methods and structures without departing from the scope of the claimed subject matter. All such modifications and variations fall within the scope of the subject matter defined in the appended claims.

[0364] Those skilled in the art will understand that, in general, the terms used in the present invention, and in particular the terms used in the appended claims (e.g., the body of the appended claims), are generally "open" terms (e.g., the term "including" should be interpreted as "including but not limited to", the term "having" should be interpreted as "having at least", the term "includes" should be interpreted as "including but not limited to", etc.). Those skilled in the art will further understand that if a specific number in a claim recitation is intended to be introduced, such intention will be explicitly stated in the claim, and if there is no such statement, such intention does not exist. For example, to aid understanding, the following appended claims may include the use of the introductory phrases "at least one" and "one or more" to introduce claim recitations. However, even if the same claim contains introductory phrases such as "one or more" or "at least one" and indefinite articles such as "a" or "an" (for example, "a" and / or "an" should be interpreted as meaning "at least one" or "one or more"), the use of such phrases should not be interpreted as implying that the indefinite articles "a" or "an" are used to introduce claim recitations, limiting any particular claim containing such introduced claim recitations to embodiments containing only one such recitation; the same is true for the use of definite articles to introduce claim recitations. In addition, even if a specific number of introduced claim recitations is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted as indicating at least the recited number (for example, the simple recitation "two recitations" without other modifiers means at least two recitations, or two or more recitations). In addition, in instances where a convention similar to "at least one of A, B, and C, etc." is used, the intended meaning of such a construction is generally the understanding of the convention by those skilled in the art (e.g., "a system having at least one of A, B, and C" includes, but is not limited to, a system having A alone, B alone, C alone, A and B at the same time, A and C at the same time, B and C at the same time, and / or A, B, and C at the same time, etc.). In instances where a convention similar to "at least one of A, B, and C, etc." is used, the intended meaning of such a construction is generally the understanding of the convention by those skilled in the art (e.g., "a system having at least one of A, B, or C" includes, but is not limited to, a system having A alone, B alone, C alone, A and B at the same time, A and C at the same time, B and C at the same time, and / or A, B, and C at the same time, etc.). Those skilled in the art will further understand that almost any antonym and / or phrase that proposes two or more alternative terms, whether in the specification, claims, or drawings, should be understood to include the three possibilities of one of the terms, either of the terms, or both of the terms. For example, the phrase "A or B" should be understood to include the three possibilities of "A" or "B" or "A and B."

[0365] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also capable of being described in terms of any individual member or subgroup of members of the Markush group.

[0366] Those skilled in the art will understand that for any and all purposes, such as from the perspective of providing a written description, all ranges disclosed in the present invention also include any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily identified as fully describing and achieving the same range, which can be decomposed into at least equal halves, thirds, quarters, fifths, ten-tenths, etc. As a non-limiting example, each range described in the present invention can be easily decomposed into a lower third, a middle third, and an upper third, etc. Those skilled in the art will also understand that "at most", "at least", "greater than", "less than", etc., all include the stated quantity, and these languages ​​refer to ranges, which can be subsequently decomposed into the above-mentioned sub-ranges. Finally, those skilled in the art will understand that ranges include each individual member. Therefore, a group with 1-3 items refers to a group with 1, 2, or 3 items. Similarly, a group with 1-5 items refers to a group with 1, 2, 3, 4, or 5 items, and so on.

[0367] Although the above invention has been described in some detail through illustration and example for the purpose of clarity of understanding, it is obvious to those skilled in the art that certain changes and modifications may be made thereto in accordance with the teachings of the present invention without departing from the spirit or scope of the appended claims.

[0368] Therefore, the foregoing only illustrates the principles of the present invention. It should be understood that those skilled in the art will be able to design various arrangements, which, although not explicitly described or shown in the present invention, embody the principles of the present invention and are included in its spirit and scope. In addition, all embodiments and conditional language described in the present invention are mainly intended to help readers understand the principles of the present invention and the concepts provided by the inventors for deepening the field, and should be interpreted as not being limited to these specifically listed examples and conditions. In addition, all statements recording the principles, aspects and embodiments of the present invention and their specific examples in the present invention are intended to cover their structural and functional equivalents. In addition, it is intended that such equivalents include currently known equivalents and equivalents developed in the future, i.e. any part (regardless of its structure) that performs the same function developed. In addition, any content disclosed by the present invention, whether or not it is explicitly listed in the claims, is not intended to be open to the public exclusively.

[0369] Therefore, the scope of the present invention is not intended to be limited to the exemplary embodiments shown and described herein. On the contrary, the appended claims embody the scope and spirit of the present invention. In the claims, 35 U.S. Patent Act Section 112(f) or 35 U.S. Patent Act Section 112(6) is expressly defined as being cited as a limitation in a claim only if such limitation in a claim begins with the exact phrase "manner" or the exact phrase "step"; if such exact phrase is not used in a claim, 35 U.S. Patent Act Section 112(f) or 35 U.S. Patent Act Section 112(6) is not cited.

Claims

1. A system for reconstitution of a dry reagent composition, the system comprising: a liquid container comprising an open end and a bottom separated by a wall, wherein an inner surface of the wall comprises a dry reagent composition; A solid volume displacer is placed inside the liquid container to occupy a majority of the liquid container volume below the top of the dry reagent composition.

2. A system according to any one of the preceding claims, wherein: The dry reagent composition includes dyes and / or microspheres.

3. The system according to claim 2, wherein: The dry reagent composition includes a dye.

4. The system according to claim 3, wherein: The dye is a polymer dye.

5. The system according to claim 4, wherein: The polymer dye includes a coupled polymer.

6. A system according to any one of the preceding claims, wherein: The liquid container contains a liquid volume that is greater than or equal to the volume of the liquid container below the top of the dry reagent composition that is not occupied by the solid volume displacer.

7. A system according to any one of the preceding claims, wherein: The inner surface of the liquid container wall includes two or more separately disposed dry reagent compositions.

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

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

10. A system according to any one of the preceding claims, wherein: The solid volume displacer includes a first end and a second end and a body therebetween, wherein the second end and the body are positionable inside a liquid container.

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

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

13. A method for reconstituted dry reagent composition, the method comprising: introducing a volume of liquid into a liquid container, the liquid container comprising an open end and a bottom separated by a wall, wherein an inner surface of the wall comprises a dry reagent composition; The solid volume displacer is introduced into the liquid container in a manner sufficient to displace liquid upwardly along the interior surface of the liquid container wall, thereby reconstituting the dry reagent composition.

14. A component comprising: a liquid container comprising an open end and a bottom separated by a wall, wherein an inner surface of the wall comprises a dry reagent composition; A certain volume of liquid is introduced into the liquid container; The solid volume displacer is placed inside the liquid container to displace the certain volume of liquid upward along the inner surface of the liquid container wall, thereby contacting with the dry reagent composition.

15. A kit comprising: a liquid container comprising an open end and a bottom separated by a wall, wherein an inner surface of the wall comprises a dry reagent composition; a solid volume displacer, positioned within the liquid container to occupy a majority of the liquid container volume below the top of the dry reagent composition; A package can accommodate the liquid container and the solid volume displacer.

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