Multi-channel liquid dispenser compatible reagent consumables and workflow

Abstract

Consumables for use with multi-channel liquid dispensing devices and multi-channel test devices, as well as workflows for their use, are described. The consumables of the present disclosure can include multiple containers pre-loaded with reagents, such as known dilutions of reagents, such as standards, proteins, chemicals, or other desired materials. The multi-channel liquid dispensing device can be used to simultaneously withdraw a fixed amount of reagent from all of the containers in the consumable and simultaneously deposit the reagents into rows of a multi-well device (such as a microwell plate) for further downstream reactions and / or tests. The downstream tests can be in a testing machine, such as a spectrophotometer or fluorometer. The consumables of the present disclosure offer advantages, such as time savings, over individually preparing dilutions of reagents and individually measuring and dispensing the reagents / dilutions into individual wells of a multi-well device for downstream reactions / tests. The consumables of the present disclosure can include multiple rows, with each row including a consumable having multiple containers or tubes.

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 434,099, filed December 21, 2022, the entire disclosure of which is incorporated herein.

[0002] FIELD OF THE INVENTION FIELD OF THE DISCLOSURE The present disclosure relates generally to consumables compatible with multi-channel liquid dispensing devices and workflows using these consumables. [Background technology]

[0003] Related technologies Protein assays such as ELISA (enzyme-linked immunosorbent assay), BCA (bicinchoninic acid assay), and Bradford assay require specific standards to determine the relative protein concentration of a sample. For ELISA, the standard used is compatible with the protein being assayed. Common protein detection assays, such as the BCA assay, use a common protein standard, which for most purposes is bovine serum albumin (BSA). These standards must be run alongside the sample to obtain quantitative protein concentration values ​​(e.g., mg / mL). The relative concentration of protein in a sample is calculated / determined by generating a standard curve graph for the protein standard (e.g., BSA) and then plotting the assay-derived protein concentration in the sample on the protein standard curve graph. Protein standards typically must be created by the user or purchased from a commercial supplier and prepared in a dilution series to determine the assay's valid range and sensitivity. The precision with which standards are produced is crucial to the overall accuracy of a protein assay, and customer survey feedback has shown that preparation of standards is one of the common pain points when performing quantitative protein assays.

[0004] Users often prepare standards inaccurately and / or make mistakes when transferring standards to the appropriate assay containers. Typically, users use an 8 x 12-well (96-well) microplate format and only need to add 5–25 microliters of standard to the plate in duplicate or triplicate. A standard curve typically includes seven points representing seven different concentrations of protein standards and a sample blank. Without considering the pipetting steps required to accurately prepare protein standards, users must accurately dispense 21–24 standards into the correct wells of the microplate. This is typically done with a single-channel pipettor and involves retrieving standards from seven or eight different containers, potentially increasing the opportunity for error by selecting the wrong standard. Often, these assays are performed in a high-throughput format, which can pose ergonomic challenges when pipetting each of these standards into multiple plates. Manually dispensing standards into plates using a single-channel pipettor can be time-consuming, especially when multiple plates are used. Automation of these assays is often desirable to save time and reduce the ergonomic burden on the person performing these assays when large numbers of samples require many plates.

[0005] Protein standards are commercially available for ELISA and BCA and Bradford protein assays. In most cases, users purchase these standards and prepare appropriate dilutions of the protein standard appropriate for their assay. Commercial suppliers take the extra step of providing some protein standards, such as BSA and bovine gamma globulin (BGG), prediluted to the appropriate concentration required for each assay. These commercially available prediluted standards are provided in 1-3 mL screw-cap plastic vials, which are individually labeled with the concentration, but still require each plastic vial to be opened and closed after use. These prediluted protein standards also require manual pipetting by the user into microwell plates and are not compatible with multichannel pipettes.

[0006] Attempts have been made to address the issues of accuracy and error by using electronic repeat pipettors, automated instruments, and plate guides that instruct users to dispense the correct pre-diluted standards into the correct corresponding wells. However, there is still room for user error. Furthermore, it has been found that some users take shortcuts each time they run an assay, do not run it in the standard manner, and use previous standard curve data to determine the protein concentration of their own samples. Thus, the art still lacks an efficient and accurate solution for creating and dispensing diluted reagents, such as protein standards. Summary of the Invention

[0007] The present disclosure provides multichannel pipette compatible reagent consumables and corresponding workflows that address one or more of the problems described in the sections above.

[0008] One embodiment of the present disclosure includes a consumable for use with a multi-channel liquid dispensing device, the consumable including a plurality of containers coupled together, each of the plurality of containers containing a different dilution of a reagent, each of the plurality of containers configured to receive a tip of the multi-channel liquid dispensing device to dispense the reagent, and one or more orientation indicia configured to indicate which orientation to use when positioning the plurality of containers relative to the multi-channel liquid dispensing device.

[0009] Another embodiment of the present disclosure includes a consumable including multiple rows, each of the multiple rows including a plurality of containers coupled together, each of the plurality of containers containing a different dilution of a reagent, each of the plurality of containers configured to receive a tip of a multi-channel liquid dispensing device to dispense the reagent, and one or more orientation indicia configured to indicate which orientation to use when placing the plurality of containers relative to the multi-channel liquid dispensing device. The consumable of the present disclosure can include one or more covers, including seals, caps, or lids. The plurality of containers of the consumable can include tubes, vials, wells, trays, or any receptacle or container with dividing walls or surfaces to create multiple containers.

[0010] Another embodiment of the present disclosure includes a method of using a consumable compatible with a multichannel pipette device. The method includes: 1) removing one or more covers from a plurality of containers comprising the consumable, each of the plurality of containers containing a different dilution of a reagent, each of the plurality of containers including an upper portion adapted to receive a multichannel pipette tip of a multichannel liquid dispenser, the consumable further including one or more orientation markings configured to indicate which orientation to use when placing the plurality of containers relative to the multichannel liquid dispenser; and 2) placing each multichannel pipette tip of the multichannel liquid dispenser on each of the plurality of containers to draw a different dilution of the reagent into the multichannel pipette tip according to the one or more orientation markings. The method further includes dispensing the different dilutions of the reagent from the multichannel liquid dispenser into one or more wells of a multiwell container (and may include dispensing the reagent into one or more rows of wells of the multiwell container).

[0011] In some embodiments of the disclosed methods, one or more wells of the multi-well container can include one or more additional materials (such as substances, chemicals, biochemicals, or additional reagents) that react with a reagent included in the consumable to form a detectable reactant, and the method can further include detecting the reactant formed in the one or more wells of the multi-well container.

[0012] In some alternative embodiments of the methods of the present disclosure, the method can further include adding one or more additional materials (such as substances, chemicals, biochemicals, or additional reagents) to one or more wells of the multi-well container, where the materials can react with the reagents to form a detectable reactant. The method can further include detecting the reactant formed in the one or more wells of the multi-well container.

[0013] Further embodiments under the present disclosure include a method for testing one or more substances or reagents in a multi-well plate. The method includes using a multi-channel liquid dispensing device to dispense a quantity of reagent into one or more wells of a multi-well container, the reagent being contained in a consumable including a plurality of containers configured to receive a tip of the multi-channel liquid dispensing device, each of the plurality of containers containing a different dilution of the reagent, and one or more orientation indicia configured to indicate which orientation to use when positioning the plurality of containers of the consumable relative to the multi-channel liquid dispensing device. The method further includes adding one or more substances to one or more wells of the multi-well container to form a reaction between the reagent and the one or more substances, inserting the multi-well container into a receptacle of a testing machine, and exciting the reaction in the plurality of wells of the multi-well container with one or more light sources. A further step can include measuring the output of the excitation. The measurement can include, for example, measuring light quenching, colorimetric properties, fluorescence levels, luminescence, or chemiluminescent properties.

[0014] Further embodiments under the present disclosure include a method for testing one or more substances or reagents in a multiwell plate. The method includes dispensing a reagent from a consumable into a plurality of wells comprising the multiwell plate, the consumable including a plurality of containers joined together, each containing a different dilution of the reagent, each configured to receive a tip of a multichannel liquid dispensing device for dispensing the reagent, and one or more orientation indicia configured to indicate which orientation to use when positioning the plurality of containers relative to the multichannel liquid dispensing device. The method may further include adding one or more substances to the multiwell plate to form a reaction between the reagent and the one or more substances, inserting the multiwell plate into a receptacle of a testing machine, exciting the reactants in the plurality of wells with one or more light sources, and measuring the excitation output. The excitation output can be used to detect or quantify the one or more substances or reagents. Measuring the excitation output may include measuring one or more of absorbance, light quenching, colorimetric properties, fluorescence levels, luminescence, and chemiluminescent properties. The tester may include measuring at least one of a fluorometer, a plate reader, a luminometer, a colorimeter, a chemiluminescence reader, and a spectrophotometer. In some embodiments of the method, testing the one or more substances or reagents includes detecting or quantifying the one or more substances or reagents.

[0015] Another embodiment of the present disclosure includes a method for testing one or more substances in a multiwell plate. The method includes dispensing a reagent from a consumable into a plurality of wells of the multiwell plate, the consumable including a plurality of containers joined together, each containing a different dilution of the reagent, each configured to receive a tip of a multichannel liquid dispensing device to dispense the reagent, and one or more orientation indicia configured to indicate which orientation to use when positioning the plurality of containers relative to the multichannel liquid dispensing device. The method also includes adding one or more substances to the multiwell plate to form a reaction between the reagent and the one or more substances, inserting the multiwell plate into a receptacle of a testing machine, subjecting the reaction in the plurality of wells to one or more light sources, and measuring the output of the reaction. In some examples, the measuring may include measuring one or more of absorbance, light quenching, colorimetric properties, fluorescence levels, luminescence, and chemiluminescent properties. An additional step may include obtaining a standard curve corresponding to different dilutions of the reagent by plotting different concentrations of the reactant and reagent on a graph axis. Other additional steps may include dispensing a sample containing an unknown concentration of a reagent into one or more empty rows of a multiwell plate, mixing the sample with one or more substances in the one or more empty rows to promote multiple detectable reactants, and plotting the multiple detectable reactants on a standard curve to extrapolate a value for the sample with unknown concentration. Thus, reagent concentrations may be detected and quantified by the methods described herein, facilitated by the consumables of the present disclosure.

[0016] This Summary is provided to introduce a selection of concepts in a simplified form that are further described below in the Detailed Description. This Summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used as an index of the scope of the claimed subject matter.

[0017] The foregoing has outlined rather broadly the features and technical advantages of the present invention in order that the detailed description of the embodiments that follow may be better understood. Additional features and advantages of the present disclosure will be set forth in the description that follows, and in part will be obvious from the description, or may be learned by the practice of the present disclosure. The features and advantages of the present disclosure may be realized and obtained by means of the instruments and combinations particularly pointed out in the appended claims. These and other features of the present disclosure will become more fully apparent from the following description and the appended claims, or may be learned by the practice of the present disclosure as set forth hereinafter. It should be appreciated by those skilled in the art that the conception and specific embodiments disclosed may be readily utilized as a basis for modifying or designing other structures for carrying out the same purposes of the present invention. It should also be appreciated by those skilled in the art that such equivalent constructions do not depart from the spirit and scope of the invention as set forth in the appended claims. The novel features believed characteristic of the present invention, both as to its organization and method of operation, together with further objects and advantages, will be better understood from the following description when considered in conjunction with the accompanying figures. It is to be expressly understood, however, that each of the figures is provided for the purpose of illustration and description only and is not intended as a definition of the limits of the present invention.

[0018] To explain how the above-mentioned and other advantages and features of the present disclosure can be obtained, a more particular description of the present disclosure, briefly described above, will be rendered by reference to specific embodiments thereof which are illustrated in the accompanying drawings. It will be understood that these drawings depict only typical embodiments of the disclosure and therefore should not be considered as limiting its scope. The present disclosure will be described and explained with additional specificity and detail using the accompanying drawings. [Brief explanation of the drawings]

[0019] [Figure 1A] 1 illustrates an exemplary consumable embodiment of the present disclosure. [Figure 1B] 1 illustrates an exemplary consumable embodiment of the present disclosure. [Figure 2A]1 illustrates an exemplary consumable embodiment of the present disclosure. [Figure 2B] 1 illustrates an exemplary consumable embodiment of the present disclosure. [Figure 2C] 1 illustrates an exemplary consumable embodiment of the present disclosure. [Figure 2D] 1 illustrates an exemplary consumable embodiment of the present disclosure. [Figure 2E] 1 illustrates an exemplary consumable embodiment of the present disclosure. [Figure 2F] 1 illustrates an exemplary consumable embodiment of the present disclosure. [Figure 3] 1 illustrates an exemplary heat seal embodiment of the present disclosure. [Figure 4] 1 illustrates an exemplary embodiment for pipetting consumables into a multi-well plate of the present disclosure. [Figure 5] 1 illustrates an exemplary spectrophotometer embodiment of the present disclosure. [Figure 6] 1 illustrates an exemplary workflow for using consumables compatible with the multi-channel dispensing device of the present disclosure. [Figure 7] 1 illustrates an exemplary workflow for testing one or more substances in a multi-well plate of the present disclosure. [Figure 8] 1 shows a comparison of the time spent generating a standard curve by existing methods compared to an exemplary workflow of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0020] Before describing various embodiments of the present disclosure in detail, it should be understood that the present disclosure is not limited to particularly exemplified systems, methods, apparatus, products, processes, kits, and / or examples, which may, of course, vary. Thus, while particular embodiments of the present disclosure will be described in detail with reference to particular configurations, parameters, components, elements, etc., the description is illustrative and should not be construed as limiting the scope of the claimed invention. Additionally, the terminology used herein is for the purpose of describing embodiments and is not necessarily intended to limit the scope of the claimed invention.

[0021] Consumables for use with multi-channel liquid dispensing devices and multi-channel test devices, as well as workflows for their use, are described. The consumables of the present disclosure include multiple containers pre-loaded with reagents, such as known dilutions of any reagent, standard, protein, chemical, biochemical, nucleic acid, or other desired material. A multi-channel liquid dispensing device can be used to simultaneously withdraw a quantity of reagent from all of the containers of the consumables of the present disclosure and simultaneously deposit the reagents into multiple wells and / or rows of a multi-well container device (such as a microwell plate) for further downstream reactions and / or tests. The downstream tests can be performed in a testing machine such as a spectrophotometer or fluorometer. The consumables of the present disclosure offer advantages such as time savings over individually preparing dilutions of reagents and individually measuring and dispensing the reagents / dilutions into individual wells of a multi-well device for downstream reactions / tests. The consumables of the present disclosure can be configured in multiple rows, with each row containing a consumable having multiple containers or tubes.

[0022] Embodiments under the present disclosure include multichannel pipette compatible (MCPC) tube format consumables containing pre-diluted reagents such as protein standards of interest for microplate-based protein assays, ELISAs, or nucleic acid standards for determining nucleic acid concentrations, or any other chemicals or biochemicals, including other test standards, that can be used in a variety of assays and systems. Such embodiments can solve several problems present in the prior art. Features of certain embodiments can include one or more of the following: The consumable and its multiple containers can be appropriately spaced for direct pick-up with a multi-channel pipette and for direct dispensing of samples into standard microplates (typically containing 96 wells). The consumable containers of the present invention can be spaced for easy use in automated liquid handlers. The consumable may have one or more orientation indicia configured to indicate which orientation to use when placing multiple containers relative to the multi-channel liquid dispensing device. The containers of the consumables of the present invention can be connected in such a way that the order of concentrations of the reagents, such as pre-diluted reagents (e.g., protein standards or nucleic acid standards) contained therein, is not disturbed. The container of the consumable of the present invention can be thermally sealed with foil to prevent leakage and evaporation. A foil tab may be included on the seal to facilitate opening the consumable and to add an additional layer of orientation to the loading order of reagents included in consumables, including reagent dilutions. Individual containers of consumables can be easily opened and resealed with a single strip cap for multiple uses. Consumables can be supplied in holders for convenient dispensing or pipetting of reagents from the consumable's individual containers, improving ease of dispensing and pipetting.

[0023] Embodiments include consumables that include multiple containers (e.g., tubes) that can facilitate the process of adding reagents or other materials to wells or other containers of a multi-well type device (e.g., a multi-well plate). The multi-well device can then be inserted into a multi-channel test device for further downstream processing of the reagents and any other materials added thereto.

[0024] Consumable embodiments described herein may be compatible with multichannel pipette devices (either manual or automated). Embodiments also include groups of consumables, for example, groups of 10, 12, 16, 20, or more consumables joined together, each containing multiple containers. Such groups of consumables are also referred to as consumables. Embodiments further include removable covers or caps for the consumables.

[0025] Embodiments also include methods of using the consumables of the present disclosure.

[0026] The multiple container embodiments described herein can include consumables with, for example, eight containers, or ten containers, or twelve or more containers prepackaged with reagents or materials, such as various dilutions of a reagent or other material. Such embodiments allow a multichannel liquid dispensing device, such as a multichannel pipette, to withdraw a reagent or other material from a tube and dispense it into another multiwell container (such as a multiwell plate) in a single step. Currently available pre-diluted standards are available in individual screw-cap vials and tubes and are not available in a multichannel dispenser-compatible format. The multichannel dispenser-compatible format of the consumables disclosed herein provides users with one or more advantages, such as consistent pipetting of diluted reagents / materials from the consumable into a multiwell container for further downstream reaction with the reagents / materials; the ability to automate or semi-automate test assays where reagents / materials / dilutions of reagents must be pipetted / dispensed into multiple containers; reduced time spent dispensing reagents; and reduced error rates when pipetting the correct dilution of a reagent / material into multiple containers.

[0027] 1A and 1B illustrate an embodiment of a consumable under the present disclosure. The consumable 100 includes multiple containers, in this embodiment, tubes 110. In this embodiment, the tubes 110 are joined together in eight rows. Other numbers of tubes 110 (2, 4, 6, 10, 12, 16, 20, etc.), container types, and combinations (single row, 4x2, 10x7, other arrangements) are possible. Each tube 110 has a different dilution of reagent 130 therein. Orientation indicia 150 are optional. The indicia 150 may include letters, numbers, shapes, arrows, colors, or other markings, or physical components or devices that indicate how the consumable should be oriented with a multichannel liquid dispensing device or how a user should insert a manual multichannel pipetting device into the tubes 110. The indicia 150 may be located on one tube 110, all tubes 110, some of the tubes 110, or on a cap or seal (not shown here) attached to the consumable 100. The indicia 150 may include tabs, holes, or other protrusions or variations on or from the cap of the consumable 110. The tubes 110 are preferably joined together, for example, at or near the top surface. The tubes 110 have open ends to allow for dispensing (e.g., pipetting) of liquid.

[0028] 2A-2F show various possible embodiments of consumables under the present disclosure.

[0029] 2A shows a consumable product 210 having multiple rows 212, each row 212 containing a consumable product, such as the consumable product 100 of FIGS. 1A-1B. The rows 212 may preferably all be joined together such that individual rows 212 may be torn, cut, or otherwise easily removed from the other rows 212 by a user.

[0030] 2B shows a consumable product 220 in which each container 223 has its own removable cap 225. The tab 227 can include orientation markings.

[0031] 2C shows a consumable 230 having a single, one-piece cap 235 with multiple domes 233 configured to mate with each container 237. A tab 239 can include orientation indicia, such as a hole 238.

[0032] 2D shows consumable 240 with tab 245 extending therefrom. Hole 247 may include orientation markings. In consumable 240, individual tubes 243 are joined together by sheath 241, which joins tubes 243 together around the top half of tubes 243. Sheath 241 is physically integrated into consumable 240.

[0033] 2E shows a side view of consumable 250. In consumable 250, tubes 255 are joined together at a top surface 257 of each tube 255.

[0034] 2F shows consumable 260. Consumable 260 helps illustrate that container 265 need not include a tube as in other embodiments. Consumable 260 more closely resembles a tray with multiple slots.

[0035] 3 shows a heat seal 520 bonded to the top of a consumable (not shown) that includes eight containers 515. The heat seal 520 may include a protrusion 525 that can be gripped to facilitate removal of the heat seal 520 from the consumable. The heat seal 520 may include various markings 530. The markings 530 may indicate the contents of each container 515, orientation indicia, reagent type, or other information. The heat seal 520 may include an aluminum or other metal seal, or may include other materials. The heat seal 520 may be bonded to a single consumable (e.g., a row of eight tubes) or multiple consumables (e.g., a group of 5, 8, 10, or any number of consumables).

[0036] In a typical embodiment, a batch of consumables may be delivered to a customer with a heat seal attached. The consumables may include multiple containers pre-loaded with standard dilutions of reagents. After initial use (and removal of the heat seal), the user can attach a cover, such as cap 225 of FIG. 2B or cap 235 of FIG. 2C, to the consumable to protect the contents for later use.

[0037] FIG. 4 illustrates an embodiment of a consumable 660 and how it can be used with a multichannel liquid dispensing / pipette device 670 and a multiwell plate 600. The consumable 660 has multiple containers 610 and can have markings such as color coding, printed text or numbers or symbols, physical features (such as holes or tabs), or any other indicative markings to indicate different concentrations or dilutions of a given reagent. The markings can include indicia for orientation of the consumable. The multiwell plate 660 can include typical multiwell containers used in biological or chemical test assays. Such multiwell containers can be placed in spectrophotometers, fluorometers, luminometers, plate readers, and other types of testing instruments. The multiwell plate 600 includes rows 630 and columns 625, each of which has multiple wells 635 (in this case, and by way of example only, eight wells in the rows 630 and twelve wells in the columns 625). The wells 635 may already contain test materials, proteins, nucleic acids, biochemicals, chemicals, or other reagents or other substances that can react with pre-diluted reagents in the consumable to form detectable reactants. Alternatively, these materials may be added to the wells 635 of the multiwell plate 600 later. The multichannel pipette device 670 may be manual or part of an automated machine. A user may wish to pipette reagents (manually or via automated means, such as programming a machine) from the consumable 660 into each row 630 of the multiwell plate 600. In prior art approaches, this must be done one pipette and one container 610 at a time. Under the present disclosure, the multichannel pipette device 670 can simultaneously pipette reagents from each container 610 and then simultaneously deposit the reagents into the wells 635 of the rows 630 (or columns 625) of the multiwell plate 600.The multi-channel dispenser 670 compatible format of the consumable 660 of the present disclosure provides the user with one or more advantages, such as consistent pipetting of diluted reagents / materials from the consumable 660 into the multi-well container 600, which can be used for further downstream reactions with other test materials / other reagents / substances. This also enables the ability to automate or semi-automate test assays where reagents / materials / reagent dilutions need to be pipetted / dispensed into multiple containers, reducing the time spent dispensing reagents and reducing the error rate in pipetting the correct dilution of reagent / reagent dilution / material into multiple containers.

[0038] In some cases, if further downstream reactions are to be performed, some test rows 640 of the multiwell plate 600 may be reserved for standardization purposes. These test rows 640 may not be used with a second material and may simply receive reagents from the consumable 660. When the test rows 640 are tested in a spectrophotometer / fluorometer / etc. testing machine, the detected light / color / excitation change / characteristics should confirm the standard dilution of the reagent, as indicated by the orientation indicia. Test materials, such as proteins, other reagents, or other substances, may be added to each well 635 before or after pipetting the reagent from the consumable 660 to allow a reaction to form between the reagent from the consumable and the test material in the well to form a detectable reactant. Depending on the particular test desired, additional substances / reagents / chemicals / stimuli and incubation periods may be required to form the detectable reactant. Figure 4 illustrates the components and steps of a workflow using the multichannel liquid dispenser-compatible consumable of the present disclosure.

[0039] FIG. 5 shows an exemplary testing machine, depicted here as an exemplary spectrophotometer 800, capable of testing a multiwell plate 820. The multiwell plate 820 can be inserted into the spectrophotometer 800 after being loaded with reagents and test materials from a consumable or other desired substances. The spectrophotometer can excite or illuminate each well and detect an output. However, those skilled in the art will recognize, in light of the present disclosure, that other testing machines, such as a fluorometer, plate reader, luminometer, or chemiluminescence detector, can be used in place of the spectrophotometer 800 used herein as the exemplary testing machine to illustrate the workflow and methods of the present disclosure. Continuing from the exemplary workflow and method embodiments described in FIG. 4 and other sections above, after forming a reaction in one or more wells of the multiwell plate 820 between one or more test substances or materials added to or contained in the wells of the multiwell plate and a reagent from a consumable, the multiwell plate can be inserted into a receptacle of the testing machine 800. The tester 800 can be used to cause excitation of reactants in multiple wells using one or more light sources and then measure the excitation output of the reactants. The excitation output can be used to detect or quantify one or more substances or reagents. Measuring the excitation output can include measuring one or more of absorbance, light quenching, colorimetric properties, fluorescence levels, luminescence, and chemiluminescent properties. In some embodiments of the method, testing one or more substances or reagents includes detecting or quantifying one or more substances or reagents.

[0040] The embodiments described herein offer numerous advantages. One advantage is automation capability: the consumables of the present disclosure can be made with any number of vessels in any shape and in any combination of rows and columns and vessel numbers to match the format of any liquid handling device, any robotic liquid dispenser, and any multichannel liquid dispensing device, enabling automation of any microplate assay. Another advantage is time savings, reducing the number of liquid dispensing / pipetting steps and the number of opening and closing steps of each individual standard vessel or tube. A further advantage is precision: pre-diluted reagents (e.g., protein standards, nucleic acid standards, chemical standards, etc.) can be prepared at a manufacturing site with appropriate quality control specifications in place. Furthermore, loading errors can be substantially reduced; a multichannel pipette can simultaneously withdraw all of the pre-diluted reagents in the consumable in one step and, guided by the orientation indicia, deposit them into the corresponding wells of the multi-well container (e.g., all protein standards required for a protein standard curve can be withdrawn from a consumable of the present disclosure and deposited into the corresponding wells of the multi-well container in one step), reducing the chance of loading the wrong dilution into a well due to user error in 1) withdrawing, 2) depositing, and 3) picking a reagent with the wrong dilution, as well as reducing the number of pipetting steps. In one example where the pre-diluted reagents include protein standards, the protein standards can be marked with indicia indicating the highest (or lowest) protein concentration in the top container of the consumable to ensure that the loading orientation into the microplate is correct.

[0041] Exemplary consumables may be pre-loaded with reagents and / or standard solutions. The test standards used may be proteins, such as BSA, BGG, or IgG (immunoglobulin G) proteins for ELISA, enzymes, or other peptides, nucleic acids, and compounds. For example, the consumables may be loaded with pre-diluted BSA standards, such as, but not limited to, 2000 μg / mL, 1500 μg / mL, 1000 μg / mL, 750 μg / mL, 500 μg / mL, 250 μg / mL, and 125 μg / mL, for use with any protein concentration detection assay, such as, but not limited to, BCA, Lowry, and Bradford-based protein assays. These amounts may vary by + / - 5%.

[0042] Another possibility is a range of BSA standards such as, but not limited to, assays in the ranges of 10000 μg / mL, 5000 μg / mL, 2000 μg / mL, 1000 μg / mL, 500 μg / mL, 250 μg / mL, or 125 μg / mL.

[0043] A single row of consumables can be loaded with a wide variety of dilutions, for example, from picograms to milligrams or even gram levels per milliliter. In some embodiments, ELISA protein standards, such as intracellular or extracellular proteins, can be used. Examples of intracellular proteins can include phosphoproteins, cyclins, and nuclear transcription factors. Examples of extracellular proteins can include cytokines or biomarkers found in whole blood, serum, or plasma. Chemical standards used can be glutathione for use in Ellman assays or N-ethylmaleimide for maleimide concentration assays. Other exemplary pre-diluted reagents that can be loaded into consumables of the present disclosure include protein-fluorophore conjugates, DNA standards, RNA standards (DNA and RNA standards are common, for example, in fluorescence assays), chemical standards, or others.

[0044] 6 and 7 show flow charts of possible method embodiments under the present disclosure.

[0045] 6 illustrates a method for using a consumable compatible with a multichannel pipette device. Step 910 includes removing one or more covers from a plurality of containers of the consumable, each of which contains a different dilution of a reagent, each of which includes an upper portion adapted to receive a multichannel pipette tip of a multichannel liquid dispenser, and the consumable further includes one or more orientation indicia configured to indicate which orientation to use when placing the plurality of containers relative to the multichannel liquid dispenser. Step 920 includes placing each multichannel pipette tip of the multichannel liquid dispenser in accordance with the one or more orientation indicia on each of the plurality of containers to withdraw a different dilution of the reagent into the multichannel pipette tip. Step 930 includes dispensing the different dilutions from the multichannel liquid dispenser into one or more rows of multiwell containers.

[0046] Method 900 can include various variations and additional or alternative steps. For example, one or more rows can contain one or more additional materials configured to facilitate a detectable reaction with the reagent. In some embodiments, the method can further include depositing one or more additional materials in one or more rows configured to facilitate a detectable reaction with the reagent. The one or more additional materials can include one or more of a chemical, another reagent, a protein, or other substance. An additional step can include detecting a reactant in one or more rows / wells. In some embodiments, the detecting can include one or more of colorimetry, absorbance, fluorescence, luminescence, and chemiluminescence. These methods can be used to detect the presence of a reagent or substance / material.

[0047] The method can optionally further include generating a standard curve by plotting the detectable reactants corresponding to different dilutions of the reagent on a graph.

[0048] Some embodiments may further include dispensing a sample containing an unknown concentration of a reagent into one or more empty rows of a multi-well container, mixing the sample with one or more substances in the one or more empty rows to promote a plurality of detectable reactants, and plotting the plurality of detectable reactants on a standard curve to extrapolate a value for the sample with unknown reagent / substance concentration. These methods may be used to quantify a reagent or substance.

[0049] In some cases, the method can include placing one or more caps on the plurality of containers, the one or more caps configured to be coupled together and coupled to the plurality of containers after the one or more covers are removed. In some cases, the detecting can be performed by one or more of a fluorometer, a plate reader, a luminometer, a colorimeter, a chemiluminescence reader, and a spectrophotometer.

[0050] Method 1100 of FIG. 7 is a method for testing one or more substances in a multiwell plate. Step 1110 involves using a multichannel liquid dispensing device to dispense a quantity of reagent into one or more wells of a multiwell container, the reagent being contained in a consumable including: 1) a plurality of containers configured to receive tips of the multichannel liquid dispensing device, each of the plurality of containers containing a different dilution of the reagent; and 2) one or more orientation indicia configured to indicate which orientation to use when placing the plurality of containers of the consumable relative to the multichannel liquid dispensing device. Step 1120 involves adding one or more substances to one or more wells of the multiwell container to form a reaction between the reagent and the one or more substances. Step 1130 involves inserting the multiwell container into a receptacle of a testing machine. Step 1140 involves exposing or exciting the reactants in the plurality of wells of the multiwell container to one or more light sources. Step 1150 involves measuring the output of the excitation (or exposure). Examples of measuring the output of exposure or excitation to light include measuring absorbance, light quenching, colorimetric properties, fluorescence levels, luminescence, or chemiluminescent properties.

[0051] Method 1100 can include various variations and additional or alternative steps. In some embodiments, the measurement can be performed by analyzing one or more of absorbance, colorimetric properties, fluorescence levels, luminescence, chemiluminescent properties, or another property. The method can also include obtaining a standard curve corresponding to different dilutions of the reagent by plotting different concentrations of the reactant and reagent on the axes of a graph. This can further include dispensing a sample containing an unknown concentration of the reagent into one or more empty rows of a multiwell plate, mixing the sample with one or more substances in the one or more empty rows to promote multiple detectable reactants, and plotting the multiple detectable reactants on the standard curve to extrapolate values ​​for samples with unknown concentrations. In some embodiments, the testing machine can include at least one of a fluorometer, a plate reader, a luminometer, a spectrophotometer, or another type of testing machine.

[0052] A summary list of defined terms To aid in understanding the scope and content of this written description and the appended claims, a selection of terms are directly defined below. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0053] As used herein, the terms "approximately," "about," and "substantially" refer to an amount or condition that is close to a particular stated amount or condition that still performs a desired function or achieves a desired result. For example, the terms "approximately," "about," and "substantially" can refer to an amount or condition that deviates from the specifically stated amount or condition by less than 10%, or less than 5%, or less than 1%, or less than 0.1%, or less than 0.01%.

[0054] Various aspects of the present disclosure, including devices, systems, and methods, may be illustrated with reference to one or more embodiments or implementations that are exemplary in nature. As used herein, the term "exemplary" means "serving as an example, instance, or illustration" and should not necessarily be construed as preferred or advantageous over other embodiments disclosed herein. Additionally, references to "implementations" of the disclosure or invention include specific references to one or more embodiments thereof, and vice versa, and are intended to provide examples without limiting the scope of the invention, which is indicated by the appended claims, rather than by the following description.

[0055] As used herein, unless otherwise implicitly or explicitly understood or stated, words appearing in the singular include their plural equivalents, and words appearing in the plural include their singular equivalents. Accordingly, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, a reference to a single referent (e.g., "a widget") includes one, two, or more referents unless otherwise implicitly or explicitly understood or stated. Similarly, a reference to a plural referent should be interpreted as including a single referent and / or multiple referents unless the content and / or context clearly dictates otherwise. For example, a reference to a plural referent (e.g., "widgets") does not necessarily require that a plurality of such referents exist. Instead, it will be understood that one or more referents are contemplated herein, regardless of the presumed number of referents, unless otherwise specified.

[0056] As used herein, directional terms such as "top," "bottom," "left," "right," "upper," "lower," "superior," "inferior," "proximal," "distal," "adjacent," and the like are used herein for relative orientation purposes only and are not intended to otherwise limit the scope of the present disclosure and / or claimed invention(s).

[0057] Unless otherwise implicitly or explicitly understood or stated, it is understood that for any given component or embodiment described herein, any of the possible candidates or alternatives listed for that component may generally be used individually or in combination with each other. Further, unless otherwise implicitly or explicitly understood or stated, it will be understood that any listing of such candidates or alternatives is merely exemplary and not limiting.

[0058] Additionally, unless otherwise indicated, numbers expressing quantities, components, distances, or other measurements used in the specification and claims are to be understood as modified by the term "about," as defined herein. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and appended claims are approximations that may vary depending upon the desired properties sought to be obtained by the subject matter presented herein. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should be construed at least in light of the number of reported significant digits and by applying ordinary rounding approaches. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the subject matter presented herein are approximations, the numerical values ​​set forth in the specific examples are reported as precisely as possible. However, any numerical value inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements.

[0059] The headings and sub-headings used herein are for organizational purposes only and are not meant to be used to limit the scope of the description or the claims.

[0060] The terms and expressions used in this specification are used as terms of description and not of limitation, and the use of such terms and expressions is not intended to exclude equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention described in the accompanying sections. Thus, while the present invention has been specifically disclosed, in part, by preferred embodiments, exemplary embodiments, and optional features, it should be understood that modifications and variations of the concepts disclosed herein may be resorted to by those skilled in the art, and such modifications and variations are deemed to be within the scope of the invention as defined by the accompanying sections. The specific embodiments provided herein are examples of useful embodiments of the present invention, and various modifications and / or variations of the features of the invention exemplified herein, as well as further applications of the principles exemplified herein that occur to those skilled in the art in possession of this disclosure, can be made to the exemplified embodiments without departing from the spirit and scope of the invention as defined by the accompanying sections, and should be deemed to be within the scope of this disclosure.

[0061] It will also be understood that systems, devices, articles of manufacture, kits, methods, and / or processes according to certain embodiments of the present disclosure may include, incorporate, or otherwise comprise properties or features (e.g., components, members, elements, parts, and / or portions) described in other embodiments disclosed and / or described herein. Thus, various features of particular embodiments may be compatible with, combined with, included in, and / or incorporated into other embodiments of the present disclosure. Thus, the disclosure of a particular feature in connection with a particular embodiment of the present disclosure should not be construed as limiting the application or inclusion of that feature to the particular embodiment. Rather, it will be understood that other embodiments may also include such features, members, elements, parts, and / or portions without necessarily departing from the scope of the present disclosure.

[0062] Furthermore, unless a feature is described as requiring another feature in combination with it, any feature herein can be combined with any other feature of the same or different embodiments disclosed herein. Moreover, various well-known aspects of example systems, methods, devices, etc. have not been described in particular detail herein to avoid obscuring aspects of the example embodiments. However, such aspects are also contemplated herein.

[0063] All references cited in this application are incorporated herein by reference in their entirety to the extent they do not contradict the disclosure of this application. It will be apparent to those skilled in the art that methods, devices, device elements, materials, procedures, and techniques other than those specifically described herein can be adapted to practice the invention broadly disclosed herein without resort to undue experimentation. All art-known functional equivalents of the methods, devices, device elements, materials, procedures, and techniques specifically described herein are intended to be encompassed by this invention.

[0064] When a group of materials, compositions, components, or compounds is disclosed herein, it is understood that all individual members of that group and all subgroups thereof are separately disclosed. When a Markush group or other group is used herein, all individual members of the group and all possible combinations and subcombinations of the group are intended to be individually included in the disclosure. All formulations or combinations of components described or exemplified herein can be used to practice the invention, unless otherwise specified. Whenever a range, e.g., a temperature range, a time range, or a composition range, is given in the specification, all intermediate ranges and subranges, as well as all individual values ​​included in the given range, are intended to be included in the disclosure. All variations that fall within the meaning and range of equivalents of the terms are intended to be encompassed within those ranges. [Example]

[0065] Example 1 One exemplary embodiment demonstrates advantageous reductions in setup time using the present workflow compared to other commercially available protein assays.

[0066] The Pierce Dilution-Free Rapid Gold BCA assay, including the disclosed workflow for generating a BSA standard curve, reduces setup time by up to 80% compared to other protein assays. Various assays, including the Bradford assay and the Pierce BCA protein assay, were compared with the Pierce Dilution-Free Rapid Gold BCA assay (including the disclosed dilution workflow) and performed according to the manufacturer's protocol in a microplate format. Five cell lysates and five pure proteins were prepared, resulting in a total of 10 samples. Standard curves for the BCA and Bradford assays were generated through serial dilutions of a 2 mg / mL bovine serum albumin (BSA) standard. The standard curve for the Pierce Dilution-Free Rapid Gold BCA assay was generated using the Pierce Dilution-Free BSA protein standard (a version of the present invention) packaged in a multichannel, pipette-friendly tube strip. Four of the 10 samples were expected to have starting concentrations >2 mg / mL, which required a sample dilution step for the BCA and Bradford assays, but no sample dilution was required for the undiluted Rapid Gold BCA assay. Samples were then mixed with working reagent from each assay and incubated according to the manufacturer's instructions.

[0067] Table 1 and Figure 8 show the results of the above experiments, demonstrating that the setup time for the Pierce No-Dilution Rapid Gold BCA Assay, including the disclosed workflow, was significantly reduced compared to other commercially available protein assays. In contrast to the disclosed automated dilution workflow used in the Pierce No-Dilution Rapid Gold BCA Assay, standard curves were generated through serial dilutions of a 2 mg / mL bovine serum albumin (BSA) standard for the Pierce BCA Protein Assay and Bradford Assay. [Table 1]

[0068] While the present invention and its advantages have been described in detail, it should be understood that various changes, substitutions, and alterations can be made therein without departing from the spirit and scope of the invention, as defined by the appended claims. Moreover, the scope of this application is not intended to be limited to the particular embodiments of the processes, machines, manufacture, compositions of matter, means, methods, and steps described herein. As those skilled in the art will readily appreciate from this disclosure, any now-existing or later-developed process, machine, manufacture, composition of matter, means, method, or step that performs substantially the same function or achieves substantially the same result as the corresponding embodiment described herein can be utilized in accordance with the present invention. Accordingly, it is intended that the appended claims include within their scope such processes, machines, manufacture, compositions of matter, means, methods, or steps.

Claims

1. 1. A consumable for use with a multi-channel liquid dispensing device, comprising: a plurality of containers coupled together, each of the plurality of containers containing a different dilution of a reagent, each of the plurality of containers configured to receive a tip of a multi-channel liquid dispensing device to dispense the reagent; one or more orientation indicia configured to indicate which orientation to use when placing the plurality of containers relative to the multi-channel liquid dispensing device.

2. The consumable product of claim 1 , wherein the plurality of containers of the consumable product are joined by being physically joined on their exterior surfaces.

3. The consumable product of claim 1 , further comprising one or more covers configured to couple to top surfaces of the plurality of containers.

4. The consumable product of claim 3 , wherein the one or more covers include a plurality of lids or caps joined together in a row.

5. 4. The consumable product of claim 3, further comprising heat seals coupled to the top surfaces of the plurality of containers, the one or more covers configured to be coupled to the plurality of tubes after the heat seals are removed.

6. The consumable product of claim 3 , wherein the one or more covers are removable from one another.

7. The consumable product of any one of claims 1 to 6, wherein the one or more orientation indicia include one or more of letters, text, numbers, symbols, shapes, arrows, colors, other markings, or physical components.

8. The consumable product of any one of claims 1 to 7, wherein the one or more orientation indicia include one or more of the following: a tab, an indentation, and a hole.

9. The consumable product of claim 7 or 8, wherein the one or more orientation indicia are coupled to one or more of the plurality of containers, the one or more covers, and the heat seal.

10. The consumable product of claim 1 , wherein the plurality of containers includes eight tubes and the multi-channel liquid dispensing device includes eight channels.

11. The consumable product of claim 1 , wherein the plurality of containers includes 12 tubes and the multi-channel liquid dispensing device includes 12 channels.

12. 10. The consumable of claim 1, wherein the reagents comprise one or more of a test standard, a protein-fluorophore conjugate, a DNA standard, an RNA standard, a protein standard, and a chemical standard.

13. 13. The consumable of claim 12, wherein the test standard is a protein test standard.

14. 14. The consumable of claim 13, wherein the protein test standard is at least one of bovine serum albumin (BSA), bovine gamma globulin (BGG), enzyme-linked immunosorbent assay (ELISA) protein standard, and immunoglobulin G (IgG) standard.

15. 15. The consumable of claim 14, wherein the ELISA protein standard comprises an intracellular protein or an extracellular protein.

16. 16. The consumable product of claim 15, wherein the intracellular proteins include one or more of a phosphoprotein, a cyclin, and a nuclear transcription factor.

17. 16. The consumable of claim 15, wherein the extracellular proteins include one or more of cytokines and biomarkers found in whole blood, serum, and plasma.

18. 13. The consumable of claim 12, wherein the chemical standards include one of glutathione for use in an Ellman assay or N-ethylmaleimide for a maleimide concentration assay.

19. 15. The consumable of claim 14, wherein the plurality of containers contain pre-diluted BSA standards ranging from about 10,000 μg / mL, 5,000 μg / mL, 2,000 μg / mL, 1,500 μg / mL, 1,000 μg / mL, 750 μg / mL, 500 μg / mL, 250 μg / mL, to about 125 μg / mL.

20. 15. The consumable of claim 14, wherein the plurality of tubes contain pre-diluted BSA or BGG standards at approximately 2000 μg / mL, 1000 μg / mL, 500 μg / mL, 250 μg / mL, 125 μg / mL.

21. 15. The consumable of claim 14, wherein the plurality of containers are configured for use with one or more of a BCA protein assay, a Bradford protein assay, a Lowry protein assay, or other protein assays.

22. The consumable of claim 1 , wherein the plurality of containers comprises at least one of receptacles, tubes, test tubes, microtubes, vials, multi-well tubes, and multi-well vials.

23. The consumable of claim 1 , wherein the plurality of tubes contain dilutions in one or more of the following ranges: picograms / milliliter, milligrams / milliliter, and grams / milliliter.

24. The consumable of claim 1 , wherein the multi-channel liquid dispensing device comprises at least one of a multi-channel pipette, an automated multi-channel pipette, and a manual multi-channel pipette.

25. Consumables, A plurality of rows are configured, and each of the plurality of rows is a plurality of containers coupled together, each of the plurality of containers containing a different dilution of a reagent, each of the plurality of containers configured to receive a tip of a multi-channel liquid dispensing device to dispense the reagent; one or more orientation indicia configured to indicate which orientation to use when placing the plurality of containers relative to the multi-channel liquid dispensing device.

26. 26. The consumable product of claim 25, wherein the one or more orientation indicia include one or more of a letter, text, number, symbol, shape, arrow, color, tab, indentation, or hole.

27. 26. The consumable product of claim 25, further comprising one or more covers configured to cover open ends of the plurality of containers.

28. 30. The consumable product of claim 27, wherein the one or more covers include one or more of the following: one or more heat seals; one or more caps.

29. The consumable product of any one of claims 25 to 28, wherein each of the plurality of containers is color-coded according to its dilution.

30. 28. The consumable product of claim 26 or 27, wherein the one or more covers are color-coded according to the dilutions of each of the plurality of containers.

31. The consumable product of any one of claims 25 to 30, wherein the plurality of rows includes two or more.

32. The consumable product of any one of claims 25 to 31, wherein the plurality of containers comprises 8 tubes, 10 tubes, 12 tubes, 16 tubes, or 20 tubes.

33. 33. The consumable product of any one of claims 25 to 32, wherein each of the plurality of rows contains the same set of different dilutions in the same location.

34. 1. A method of using a consumable compatible with a multi-channel pipette device, comprising: removing one or more covers from a plurality of containers comprising a consumable, each of the plurality of containers of the consumable containing a different dilution of a reagent, each of the plurality of containers including an upper portion adapted to receive a multi-channel pipette tip of a multi-channel liquid dispenser, the consumable further including one or more orientation indicia configured to indicate which orientation to use when placing the plurality of containers relative to the multi-channel liquid dispenser; placing each multi-channel pipette tip of the multi-channel liquid dispenser in a respective one of the plurality of containers of the consumable product according to the one or more orientation indicia to draw the different dilutions of the reagent into the multi-channel pipette tip; dispensing the different dilutions of the reagent from the multi-channel liquid dispenser into one or more rows of a multi-well container.

35. 35. The method of claim 34, further comprising forming a detectable reactant in the one or more rows of the multi-well container by reacting the reagent with a substance, material, or additional reagent in the multi-well container.

36. 36. The method of claim 35, further comprising detecting the reactants in the one or more rows of the multi-well vessel.

37. 35. The method of claim 34, wherein the one or more rows of the multi-well container contain one or more additional materials that react with the reagents contained in the consumable to form a detectable reactant.

38. 35. The method of claim 34, further comprising depositing one or more additional materials in the one or more rows of the multi-well container configured to react with the reagent to form a detectable reactant.

39. 39. The method of claim 37 or 38, wherein the one or more additional materials comprise one or more of a chemical, another reagent, or a protein.

40. 37. The method of claim 36, wherein the detecting comprises the use of one or more of colorimetry, absorbance, quenching, fluorescence, luminescence, and chemiluminescence.

41. 41. The method of any one of claims 34 to 40, further comprising generating a standard curve by plotting the detectable reactant corresponding to the different dilutions of the reagent on a graph.

42. dispensing a sample containing an unknown concentration of said reagent into one or more empty rows of said multi-well container; mixing the sample with one or more substances in the one or more empty rows to promote multiple detectable reactants; 42. The method of claim 41, further comprising plotting the plurality of detectable reactants on the standard curve to extrapolate values ​​for the samples of unknown concentration.

43. 43. The method of any one of claims 34-42, further comprising placing one or more caps on the plurality of containers, the one or more caps being coupled together and configured to be coupled to the plurality of containers after the one or more covers are removed.

44. 37. The method of claim 36, wherein said detecting is performed by one or more of a fluorometer, a plate reader, a luminometer, and a spectrophotometer.

45. 1. A method for testing one or more substances in a multi-well plate, comprising:

1. Use of a multi-channel liquid dispensing device to dispense a volume of a reagent into one or more wells of a multi-well container, the reagent being contained in a consumable product comprising: a plurality of vessels configured to receive tips of the multi-channel liquid dispensing device, each vessel containing a different dilution of the reagent; one or more orientation indicia configured to indicate which orientation to use when placing the plurality of containers of the consumable product relative to the multi-channel liquid dispensing device; adding one or more substances to the one or more wells of the multi-well container to form a reaction between the reagent and the one or more substances; inserting the multi-well vessel into a receptacle of a testing machine; exposing the reactants in the plurality of wells to one or more light sources; and measuring the output of said exposure.

46. 46. ​​The method of claim 45, wherein said measuring comprises measuring one or more of absorbance, light quenching, colorimetric properties, fluorescence levels, luminescence, and chemiluminescent properties.

47. 47. The method of claim 45 or 46, further comprising obtaining a standard curve corresponding to the different dilutions of the reagent by plotting the different concentrations of the reactant and the reagent on the axes of a graph.

48. dispensing a sample containing an unknown concentration of said reagent into one or more empty rows of said multi-well plate; mixing the sample with one or more substances in the one or more empty rows to promote multiple detectable reactants; 48. The method of any one of claims 45 to 47, further comprising plotting the plurality of detectable reactants on the standard curve to extrapolate values ​​for the samples of unknown concentration.

49. 49. The method of any one of claims 45 to 48, wherein the test instrument comprises at least one of a fluorometer, a plate reader, a colorimeter, a chemiluminescence reader, a luminometer, and a spectrophotometer.

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