Alternative substrate for automated clinical analyzer and methods of use

Abstract

A method of performing an immunoassay with an automated clinical analyzer, includes, creating an assay mixture utilizing an analyte from a patient sample, a first reagent having a plurality of particles configured to specifically bind to the analyte and a second reagent having an enzyme. The method further includes incubating the assay mixture and then performing a wash cycle on the assay mixture to retain the plurality of particles while removing the remaining portion of the assay mixture not bound the plurality of particles. The method further includes selecting a first substrate material or a second substrate material. The method further includes obtaining the selected substrate material and adding the selected substrate material with the plurality of particles remaining from the wash cycle to create a signal generating mixture. The method further includes incubating the signal generating mixture and analyzing the signal generating mixture with a luminometer.

Description

ALTERNATIVE SUBSTRATE FOR AUTOMATED CLINICAL ANALYZER ANDMETHODS OF USEPRIORITY

[0001] This application claims priority to U.S. Provisional Application 63 / 668,455, entitled “Alternative Substrate for Automated Clinical Analyzer and Methods of Use,” filed on July 8, 2024 and U.S. Provisional Application 63 / 746,0999, entitled “Alternative Substrate for Automated Clinical Analyzer and Methods of Use,” filed on January 16, 2025, hereby incorporated by reference in their entireties.BACKGROUND

[0002] Automated clinical analyzers are well known in the art and are generally used for the automated or semi-automated analysis of patient samples. Typically, prepared patient samples, such as blood, urine, spinal fluid, and the like are placed onto such an analyzer in sample containers such as test tubes or cups. The analyzer pipettes a portion of a patient sample and one or more reagents to a reaction cell (e.g., a reaction vessel, cuvette or flow cell) where an analysis of the sample is conducted, usually for a particular analyte of interest, and results of the analysis are reported.

[0003] In some instances, during an illustrative analysis of a patient sample, an enzyme is specifically bound, either directly or indirectly, to a respective analyte of interest within the reaction cell. A substrate is then added to the reaction cell, which is configured to chemically react with the bound enzyme to thereby produce light. The intensity of the produced light may be measured and the output utilized to ascertain the amount of the patient analyte within the reaction cell.

[0004] While various kinds of automated clinical analyzer have been made and used, it is believed that no one prior to the inventor(s) has made or used the invention described in the appended claims.SUMMARY

[0005] Described herein are devices, systems, and methods for performing an automated assay on a sample with a clinical analyzer configured to generate and measure an output signal. In some aspects, this disclosure describes devices, systems, and methods for providing a clinical analyzer system with multiple substrate materials that can be used for analyte detection and reporting. These substrates may be used alternatively to provide increased flexibility for the analyzer system. For example, a particular substrate may provide advantages in detecting certain analytes or in the context of specific assays (for example, different substrates could exhibit different sensitivities, different detection times, different effects on specificities, etc., that could be advantageous in some or all of the assay performed by the analyzer system). In some exemplary aspects, one substrate may be housed on the analyzer system, while an alternative substrate may be loaded onto the analyzer system (for example, via a sample rack or a sample carriage).

[0006] In some embodiments, the method of performing the automated assay includes mixing and incubation of the sample with one or more reagents to bind a targeted analyte of the sample to a respective particle of a plurality of particles, and also bind at least one reporter to a respective particle of the plurality of particles and / or a respective analyte of the targeted sample analyte. Further, the method of performing the automated assay includes selection of a first substrate or a second substrate and subsequently mixing of the selected substrate with a composition that includes the bound reporter(s) to generate a signal indicative of the amount of targeted patient analyte within the composition. In some embodiments, at least one of the first substrate or the second substrate is obtained via a pipette of the clinical analyzer from a sample rack that is processed by the clinical analyzer and subsequently ejected from the clinical analyzer. In some embodiments, the first substrate or the second substrate is obtained from an adjacent automated machine.

[0007] In some embodiments, where the first substrate or the second substrate is obtained via a pipette of the clinical analyzer from a sample rack, the sample rack includes a base configured to be received by the clinical analyzer, and a plurality of vessel holders configured to selectively receive a corresponding vessel such that the corresponding vessel is accessible by the pipette of the clinical analyzer while the base of the sample rack is received by the clinical analyzer. In some embodiments, the sample rack includes a substrate container holder that is configured toselectively receive a substrate such that the substrate container is accessible by the pipette of the clinical analyzer while the base of the sample rack is received by the clinical analyzer.

[0008] In some embodiments, the method of performing the automated assay includes mixing and incubating the sample with one or more reagents to bind a targeted analyte of the sample to a respective particle of a plurality of particles and also to bind at least one reporter to a respective particle of the plurality of particles and / or a respective analyte of the targeted sample analyte. Further, the method of performing the automated assay includes addition and mixing of a substrate with a composition that includes the bound reporter(s) to generate a signal indicator of the amount of targeted patient analyte within the composition, where the substrate is added and mixed with the composition with a sample aliquoting pipette and a disposable tip.

[0009] In some embodiments, the clinical analyzer uses the sample aliquoting pipette and a first disposable tip to transfer the substrate material from a vessel on the sample rack into a portable vessel at a mixing station and to transport the portable vessel within the clinical analyzer into a climate-controlled storage area. In some embodiments, when the substrate material is to be utilized, the clinical analyzer may transport the portable vessel back to the mixing station, and the sample aliquoting pipette may use a second disposable tip to add and mix the substrate with a composition that includes the bound reporter(s) to generate a signal indicator of the amount of targeted patient analyte within the composition. In other embodiments, when the substrate material is to be utilized, the clinical analyzer may transport the portable vessel to a wash wheel, which may then add and mix the substrate with a composition that includes the bound reporter(s) to generate a signal indicator of the amount of targeted patient analyte within the composition.BRIEF DESCRIPTION OF THE DRAWINGS

[0010] While the specification concludes with claims which particularly point out and distinctly claim the invention, it is believed the present invention will be better understood from the following description of certain examples taken in conjunction with the accompanying drawings, in which like reference numerals identify the same elements and in which:

[0011] FIG. 1 depicts a schematic view of an illustrative assay;

[0012] FIG. 2A depicts a cross-sectional view of an illustrative luminometer, taken along a centerline axis of the luminometer;

[0013] FIG. 2B depicts a sectional view of the luminometer of FIG. 2A;

[0014] FIG. 3A depicts a perspective view of an illustrative wash wheel;

[0015] FIG. 3B depicts a schematic top-down view of the wash wheel of FIG. 3 A;

[0016] FIG. 4 depicts a schematic view of an illustrative pipetting system;

[0017] FIG. 5 depicts a schematic view of an illustrative probe washing arrangement;

[0018] FIG. 6 depict a schematic view of an illustrative computer system for an automated clinical analyzer;

[0019] FIG. 7 depicts a schematic view of an illustrative automated clinical analyzer configured to perform an assay (including, for example, the illustrative assay of FIG. 1), where the automated clinical analyzer can include one or more of the luminometer of FIG. 2A, the wash wheel of FIG. 3 A, at least one pipetting system of FIG. 4, the computer system of FIG. 6, a reagent storage station, a substrate storage station housing a first substrate material, a reaction vessel carriage, and a reaction vessel transportation unit configured to pick and place a reaction vessel to various stations of the automated clinical analyzer;

[0020] FIG. 8 depicts a flowchart of an illustrative method of use of the automated clinical analyzer of FIG. 7;

[0021] FIG. 9 depicts a perspective view of an illustrative sample rack that may be used in conjunction with an automated clinical analyzer (including, for example, the automated clinical analyzer of FIG. 7);

[0022] FIG. 10 depicts a cross-sectional view of the sample rack of FIG. 9, with the vessels omitted for clarity;

[0023] FIG. 11 depicts a perspective view of an illustrative sample rack that may be used in conjunction with an automated clinical analyzer (including, for example, the automated clinical analyzer of FIG. 7);

[0024] FIG. 12 depicts a perspective view of an illustrative sample rack that may be used in conjunction with an automated clinical analyzer (including, for example, the automated clinical analyzer of FIG. 7);

[0025] FIG. 13 depicts a flowchart of an illustrative method of use of the automated clinical analyzer of FIG. 7 in conjunction with the sample rack of FIG. 9, FIG. 11, or FIG. 12;

[0026] FIG. 14A depicts a schematic view of a pipette and disposable tip of the clinical analyzer of FIG. 7 aligned and above a substrate container of the sample rack of FIG. 9, FIG. 11, or FIG. 12 where the substrate container houses a reservoir of substrate material;

[0027] FIG. 14B depicts a schematic view of the pipette and disposable tip of FIG. 14A, where the disposable tip is within the substrate container of FIG. 14A;

[0028] FIG. 14C depicts a schematic view of the pipette and disposable tip of FIG. 14A, where the disposable tip is aligned and above the substrate container of FIG. 14A, where the disposable tip contains the substrate material of FIG. 14A;

[0029] FIG. 14D depicts a schematic view of the pipette and disposable tip of FIG. 14A aligned and above a reaction vessel;

[0030] FIG. 14E depicts a schematic view of the pipette and disposable tip of FIG. 14A, where the disposable tip is within the reaction vessel of FIG. 14D;

[0031] FIG. 14F depicts a schematic view of the pipette and disposable tip of FIG. 14A, where the disposable tip is aligned and above the reaction vessel of FIG. 14D, where the reaction vessel contains a signal generating mixture;

[0032] FIG. 15 depicts a flowchart of an illustrative method of transferring a substrate material from a sample rack (for example, the sample rack of FIG. 9, FIG. 11, or FIG. 12) into the automated clinical analyzer of FIG. 7;

[0033] FIG. 16A depicts a schematic view of the pipette and the disposable tip of FIG. 14A are aligned and above a portable vessel, where the disposable tip contains a substrate material;

[0034] FIG. 16B depicts a schematic view of the pipette and the disposable tip of FIG. 14A, where the disposable tip is within the portable vessel of FIG. 16A;

[0035] FIG. 16C depicts a schematic view of the pipette and the disposable tip of FIG. 14A, where the pipette and the disposable tip are aligned and above the portable vessel of FIG. 16A, where the portable vessel contains the substrate material of FIG. 16A;

[0036] FIG. 17 depicts a flowchart of an illustrative method of transferring a substrate material from an adjacent automated machine into the automated clinical analyzer of FIG. 7;

[0037] FIG. 18 depicts a flowchart of an illustrative method of use of the automated clinical analyzer of FIG. 7 in conjunction with the portable vessel of FIG. 16A;

[0038] FIG. 19A depicts a schematic view of a pipette and disposable tip of FIG. 14A aligned and above the portable vessel of FIG. 16A, where the portable vessel houses a reservoir of substrate material;

[0039] FIG. 19B depicts a schematic view of the pipette and disposable tip of FIG. 14A, where the disposable tip is within the portable vessel of FIG. 16A;

[0040] FIG. 19C depicts a schematic view of the pipette and disposable tip of FIG. 14A, where the disposable tip is aligned and above the portable vessel of FIG. 16A, where the disposable tip contains the substrate material of FIG. 19A;

[0041] FIG. 19D depicts a schematic view of the pipette and disposable tip of FIG. 14A aligned and above a reaction vessel;

[0042] FIG. 19E depicts a schematic view of the pipette and disposable tip of FIG. 14A, where the disposable tip is within the reaction vessel of FIG. 19D;

[0043] FIG. 19F depicts a schematic view of the pipette and disposable tip of FIG. 14A, where the disposable tip is aligned and above the reaction vessel of FIG. 19D, where the reaction vessel contains a signal generating mixture;

[0044] FIG. 20 depicts another flowchart of an illustrative method of use of the automated clinical analyzer of FIG. 7 in conjunction with the portable vessel of FIG. 16A; and

[0045] FIG. 21 depicts a flowchart of an illustrative method of transferring a substrate material from an adjacent automated machine into the automated clinical analyzer of FIG. 7.

[0046] The drawings are not intended to be limiting in any way, and it is contemplated that various embodiments of the invention may be carried out in a variety of other ways, including those not necessarily depicted in the drawings. The accompanying drawings incorporated in and forming a part of the specification illustrate several aspects of the present invention, and together with the description serve to explain the principles of the invention; it being understood, however, that this invention is not limited to the precise arrangements shown.DETAILED DESCRIPTION

[0047] In some aspects, this disclosure describes devices, systems, and methods for providing a clinical analyzer system with multiple substrate materials that can be used for analyte detection and reporting. These substrates may be used alternatively to provide increased flexibility for the analyzer system. For example, a particular substrate may provide advantages in detecting certainanalytes or in the context of specific assays (for example, different substrates could exhibit different sensitivities, different detection times, different effects on specificities, etc., that could be advantageous in some or all of the assay performed by the analyzer system). In some exemplary aspects, one substrate may be housed on the analyzer system, while an alternative substrate may be loaded onto the analyzer system (for example, via a sample rack or a sample carriage).

[0048] The following description of certain examples of the invention should not be used to limit the scope of the present invention. Other examples, features, aspects, embodiments, and advantages of the invention will become apparent to those skilled in the art from the following description, which is by way of illustration, one of the best modes contemplated for carrying out the invention. As will be realized, the invention is capable of other different and obvious aspects, all without departing from the invention. Accordingly, the drawings and descriptions should be regarded as illustrative in nature and not restrictive.

[0049] It will be appreciated that any one or more of the teachings, expressions, versions, examples, etc. described herein may be combined with any one or more of the other teachings, expressions, versions, examples, etc. that are described herein. The following-described teachings, expressions, versions, examples, etc. should therefore not be viewed in isolation relative to each other. Various suitable ways in which the teachings herein may be combined will be readily apparent to those of ordinary skill in the art in view of the teachings herein. Such modifications and variations are intended to be included within the scope of the claims.

[0050] In one aspect, this disclosure describes a method of performing an immunoassay with an automated clinical analyzer wherein the automated clinical analyzer can select between two reporting reagents used to detect a reporter (for example, a first substrate material and a second substrate material), wherein at least one reporting reagent is housed within the automated clinical analyzer. As further described herein, it may be desirable to have an option of selecting between different reporting reagents, as different reporting reagents may have different resulting effects on desired parameters including amount of signal (for example, light) generated, background, signal- to-noise ratio, etc.

[0051] I. Illustrative Assay and Automated Clinical Analyzer Configured to Perform Assay

[0052] A. Illustrative Assay

[0053] Turning now to FIG. 1 , which illustrates an example assay 100 that may be performed by an automated clinical analyzer. Other types of assays (including other types of immunoassays) may also be performed by the same analyzer.

[0054] As shown in FIG.l a pipette 110 dispenses a first reagent 115 including a plurality of particles 120 (for example, iron particles) at a suitable concentration (for example, a concentration of between 0.3 mg / mL and 2.0 mg / mL) into a reaction vessel 105 (for example, a cuvette, test tube, or any other suitable receptacle). The first reagent 115 will also include antibodies or antigens that are tailored to bind specifically to an analyte of interest. The analyte of interest may be present in a patient sample 165 that the assay 100 is meant to measure. In assay 100, the particles 120 may be coated with the antibodies or antigens. Additionally, or alternatively, the particles 120 may be modified so that they specifically interact with antibodies or antigens which are also present in the first reagent 115. Thus, such binding between a particle and the antibodies or antigens with which the particle is coated may occur directly or indirectly.

[0055] At stage 2, the pipette 110 adds a sample including, for example, a patient sample 165 is added to the reaction vessel 105. The pipette 110 may be the same pipette used in stage 1, but which has been cleaned; a new, separate pipette from the pipette used in stage 1 ; or the same pipette used in stage 1, but with a new tip at each stage. Additionally, for some analyzers, different pipettes may be used in different stages (e.g., a first pipette for dispensing of the patient sample, a second pipette for dispensing of the reagent, a third pipette for aspiration and / or washing, etc.). Additionally, or alternatively, different pipette tips may be used with the different pipettes or different pipette tips may be used with the same pipette. It will be understood by one having skill in the art that stage 1 and stage 2 may be reversed; that is, the patient sample 165 may be added to the reaction vessel 105 before the first reagent 115. Furthermore, the order of reagents added in stages 1, 2, and 5 may be modified depending on assay design.

[0056] At stage 3, the reaction vessel 105, containing the patient sample 165 and the first reagent 115 (including the particles 120) are mixed to create a first mixture 170. In some instances, the first mixture may be mixed (for example, by additional pipetting) to ensure even mixture of the patient sample 165 and the first reagent 115. Additionally, or alternatively, in some methods and / or analyzers, the contents (i.e., first mixture 170) of the vessel 105 may be optionally subjected to a heat source (i.e., incubation). During incubation, the antibodies or antigens of the first reagent 115 bind with an analyte of interest in the patient sample 165. If not already bound to the particles120, the antibodies or antigens of the first reagent 115 may also bind to the particles 120 during incubation. The binding process can result in the analyte from the sample (for example, the patient sample 165) binding to the particles 120.

[0057] The first mixture may optionally be washed before additional reagent is added. With reference to FIG. 1, at stage 4, the reaction vessel 105 moves near one or more magnets 130, which attracts the particles 120 (e.g., iron particles) to one or more sides (e.g., perimeter portions) of the reaction vessel 105. Pipette 110 aspirates the fluid in the reaction vessel 105 and adds a washing agent 150 to the reaction vessel 105 to wash the particles 120. During aspiration, the magnet(s) 130 retain the plurality of particles 120 at the one or more sides of the reaction vessel 105. The particles 120 and the bound analyte of the patient sample 165 remain in the reaction vessel 105 after the washing is complete by virtue of the magnet(s) 130 retaining the particles 120 and the binding between the analyte of the patient sample 165 and the particles 120 via the antigens or antibodies of the first reagent. Therefore, at least one particle of the plurality of particles 120 is bound to a respective analyte from the patient sample 165. Other components of the first mixture 170 may be absent from the reaction vessel 105 after the washing is complete, having been washed away by the washing agent 150. Additionally, or alternatively, components other than particles 120 and bound analyte may be removed from the reaction vessel 105 via other methods of retaining the particles and aspiration, for example, with pipette 110 or other suitable means. Removal (or attempted removal) of remaining portions of a suitable mixture not bound to a plurality of particles (e.g., particles 120), while retaining the plurality of particles 120 themselves, may be referred to as a wash, a wash cycle, washing cycle, etc.

[0058] A wash cycle may include adding a washing agent 150 to the reaction vessel 105 and subsequently aspirating the added washing agent 150 any suitable number of times. For example, a wash cycle may include adding a washing agent 150 to the reaction vessel 105 at least one time, at least two times, at least three times, at least five times, at least seven times, at least nine times, at least ten times, and / or up to one hundred times. Further, a wash cycle may include aspirating the added washing agent 150 from the reaction vessel 105 at least one time, at least two times, at least three times, at least five times, at least seven times, at least nine times, at least 10 times, and / or up to one hundred times. Additionally, a wash cycle may include additional steps inbetween, before, and / or after adding a washing agent 150 and subsequent aspiration of the washing agent 150.

[0059] Thus, in an illustrative embodiment where the particles 120 are iron particles (that is, the particles include iron), performing the wash cycle can include (i) attracting the plurality of iron particles toward a magnet, and (ii) aspirating the mixture (for example, the first mixture 170 or an assay mixture 175) from the reaction vessel 105 while the plurality of iron particles remains attracted toward the magnet 130. In another illustrative embodiment, performing the wash cycle can include (i) adding a washing agent, (ii) attracting the plurality of iron particles toward a magnet, and (iii) aspirating the mixture and washing agent from the reaction vessel while the plurality of iron particles remains attracted toward the magnet. In a further illustrative embodiment, performing the wash cycle can include (i) adding a washing agent, (ii) attracting the plurality of iron particles toward a magnet, (iii) aspirating the mixture and washing agent from the reaction vessel while the plurality of iron particles remains attracted toward the magnet, (iii) adding additional washing agent, while the plurality of iron particles remains attracted toward the magnet, and (iv) aspirating washing agent from the reaction vessel while the plurality of iron particles remains attracted toward the magnet. As noted above, any of these illustrative wash cycles may further be repeated a suitable number of times.

[0060] At stage 5, the pipette 110 may place a second reagent 155 including a reporter can be placed in the reaction vessel 105 with the particles 120 and the bound analyte of the patient sample 165. The reporter may be, for example, a suitable enzyme such as alkaline phosphatase (“ALP”) (for example, at a concentration of between 0.01 mg / L and 2.0 mg / L) or P-galactosidase, a fluorescent protein, or luciferase.

[0061] At stage 5b, a composition including the second reagent 155, the first reagent (including the particles 120), and analyte from a patient sample (typically bound analyte associated with the particles 120) can be mixed in reaction vessel 105 to form an assay mixture (175).

[0062] The second reagent 155 can include an antibody attached to the reporter (e.g., ALP) that binds with the analyte of the patient sample 165 (while the analyte is attached to the particles 120). Such binding may occur directly or indirectly. Therefore, in the current example, the assay mixture 175 includes the analyte of the patient sample, bound to a particle 120 (from the first reagent 115) and a reporter (e.g., ALP) (from the second reagent 155). In some instances, the assay mixture 175 may be mixed (for example, by additional pipetting) after the addition of the second reagent 155 to ensure even mixture of the components. Additionally (or alternatively), in some analyzers, the contents of the vessel 105 (that is, the assay mixture 175) may be subjected to a heatsource (i.e., incubation). An incubation can speed or facilitate binding between the antibody attached to the reporter, present in the second reagent 155 with the analyte of interest from the patient sample 165 (which is already bound to a particle 120). That is, after incubation, the assay mixture 175 includes the analyte of the patient sample 165 bound to both a particle 120 (of the first reagent 115) and to the reporter present in the second reagent 155.

[0063] In the current example, assay mixture 175 is formed with incubation and washing steps occurring between the addition of first reagent 115 and second reagent 155. However, the first reagent 115, the patient sample 165, and the second reagent 155 may be combined to form assay mixture 175 using any suitable combination of steps. For example, in some instances, the first reagent 115, the second reagent 155, and the patient sample 165 may all be mixed together to form assay mixture 175 and then incubated.

[0064] At stage 6, a suitable wash cycle is performed on the assay mixture 175. The reaction vessel 105 is moved near one or more magnet(s) 130, which attracts the particles 120 e.g., iron particles) to one or more sides e.g., perimeter portions) of the reaction vessel 105. At this moment, the patient analyte is bound to both a particle 120 and a reporter (e.g., ALP). Pipette 110 is used to wash the particles 120 by aspirating the fluid in the reaction vessel 105 and adding a washing agent 150 to the reaction vessel 105. For example, a wash cycle may include adding a washing agent 150 to the reaction vessel 105 at least once, at least two times, at least three times, at least five times, at least seven times, at least nine times, at least 10 times, and / or up to one hundred times. Further, a wash cycle may include aspirating the added washing agent 150 from the reaction vessel at least once, at least two times, at least three times, at least five times, at least seven times, at least nine times, at least 10 times, and / or up to one hundred times. During aspiration, the magnet(s) 130 retain the plurality of particles 120 at the one or more sides of the reaction vessel 105. The particles 120, the bound analyte of the patient sample 165, and the bound reporter of the second reagent 155 remain in the reaction vessel 105 after the washing is complete by virtue of the magnet(s) 130 and the binding between the analyte of the patient sample 165 and the antigens or antibodies that are coated on the particles 120 and the reporter. Other components of the assay mixture 175 may be absent from the reaction vessel 105 after the washing is complete, having been washed away by the washing agent 150. Additionally, or alternatively, components other than particles 120, the bounded analyte, and the bounded reporter may be removed from the reaction vessel 105 via aspiration, for example, with pipette 110 or other suitable means. Removal (or attempted removal)of remaining portions of a suitable mixture not bound to the plurality of particles (e.g., particles 120), while retaining the plurality of particles 120 themselves, may be referred to as a wash, a wash cycle, washing cycle, etc. Additionally, as discussed above, a wash cycle may include additional steps in-between, before, and / or after adding a washing agent 150 and subsequent aspiration of the washing agent 150.

[0065] In the current example, portions of the second reagent 155 not bound to the patient analyte may be removed from the reaction vessel 105 via a suitable wash cycle. In examples where first reagent 115 and / or the patient sample 165 are still housed within reaction vessel 105 (e.g., first reagent 115 and / or patient sample 165 form part of the assay mixture 175), portions of those materials not bound to the particles 120 may be removed from the reaction vessel 105 via the wash cycle at stage 6. Therefore, the plurality of particles 120 are retained within the reaction vessel 105 such that at least one particle of the plurality of particles is bound to a respective analyte from the patient sample 165, while the remaining portion of the assay mixture 175 not bound to the plurality of particles 120 is removed from the reaction vessel 105. Such a wash cycle may leave the plurality of particles 120, some of which are bound to a respective analyte of the patent sample, within the reaction vessel 105.

[0066] While the wash cycle at stage 4 is described as optional, the wash cycle at stage 6 is utilized to remove undesirable components of the assay mixture 175 that may otherwise undesirably affect the subsequent signal generated at stage 8 in accordance with description herein. Moreover, the wash cycle at stage 6 may be optimized (for example, by altering number of washes, amount or number of additions of washing agent, amount or number of aspirations, etc.) by a person having skill in the art to maximize removal of undesirable components while retaining the analyte of interest.

[0067] At stage 7, a reporting reagent such as a first substrate material 180 is added to the reaction vessel 105 with pipette 110 to create a signal generating mixture 177 (stage 8). As also described herein, in some instances, instead of selecting and adding first substrate material 180, a different reporting reagent such as a second substrate material 182 (FIGS. 14A-14F, 16A-16C, and 19A-19F) may be selected and added to reaction vessel 105 to create signal generating mixture 177. As will also be described in greater detail below, in some instances where a second substrate material 182 (FIGS. 14A-14F, 16A-16C, and 19A-19F) is utilized to serve as the reporting reagent, a sample rack 800, 830, 850 (FIGS. 9-12) housing a second substrate material 182 may be accessedby a suitable pipetting unit 706 (FIGS. 14A-14F and 16A-16C) to suitably dispense second substrate material 182 into reaction vessel 105. The selected reporting reagent (e.g., first substrate material 180, second substrate material 182, etc.) may be mixed and may be incubated using a heat source 125 (e.g., the reaction vessel 105 may be placed in an incubator). As shown in stage 7, in the current example, first substrate material 180 is the utilized reporting agent. However, as mentioned above, second substrate material 182 may be utilized as an alternative to first substrate material 180. Therefore, the below discussion related to first substrate material 180 in the context of assay 100 may also encompass suitable characteristics of second substrate material 182. When the first substrate material 180 is an ALP substrate, for example, the first substrate material 180 reacts with the ALP enzyme and thereby produces light 135 (that is, photons). The current example, the molecules of reporter (e.g., ALP enzymes) within reaction vessel at stage 7 are bound (directly or indirectly) to a respective patient analyte. Therefore, the amount of reporter (e.g., ALP) within reaction vessel 105 at stage 7 available to react with the first substrate material 180 to produce light (i.e., photons) is proportional to the amount of patient analyte present within the reaction vessel 105.

[0068] At stage 8, the signal (in this example, light 135), emitted by signal generating mixture 177 (e.g., the reaction of the reagent (e.g., the first substrate material 180) and the reporter (e.g., ALP) bound to the iron particles 120), can be measured (e.g., using a luminometer such as luminometer 600 of FIG. 2A) to generate an output signal that can be processed to generate an output response (e.g., a relative light unit (“RLU”) value), indicating a result of the assay 100. For example, in illustrative assay 100, a larger RLU value indicates more light, which indicates a larger amount of the analyte in the patient sample 165 as compared to a smaller RLU value.

[0069] Although particles 120 coated with antibodies or antigens are used to bind to a desired analyte, other suitable particle may be used in conjunction with a suitable antibody or antigen. Additionally, although a magnet 130 is used in the current example to retain particles within reaction vessel 105 during a wash cycle, any other suitable means of retaining particles within reaction vessel 105 during a wash cycle may be used.

[0070] Additionally, in the current illustrative assay 100, the enzyme (e.g., ALP) within second reagent 155 is configured to be bound (indirectly) with an analyte from a patient sample. Therefore, the more patient analyte that is present within reaction vessel 105, the more enzyme that is present within reaction vessel 105; which in turn leads to more light being produced whenthe first substrate material 180 is added. Tn other words, in the current illustrative assay 100, the amount of light produced at stage 8 is directly correlated to the amount of patient analyte present within a patient sample 165. However, in some instances, a “competing analyte” approach may be utilized, where the enzyme within the second reagent 155 is already bound to a “competing analyte” prior to being introduced within reaction vessel 105 at stage 5. In such instances, the enzyme in a “competing analyte” approach will only bind to particles 120 that are not already bound to a patient analyte. Therefore, in the “competing analyte” approach, the amount of light produced at stage 8 would be inversely correlated to the amount of patient analyte present within patient sample 165.

[0071] B. Illustrative Luminometer of Automated Clinical Analyzer

[0072] As described above, the exemplary immunoassay 100 generates an output response that indicates the results of the assay. The output response may be detected by any suitable detector. When the output response is a relative light unit (RLU) the detector may be a light detecting device such as a luminometer, including a photomultiplier tube (“PMT”). An exemplary luminometer 600 is described in FIGS. 2A and 2B and in International Patent Application Publication No. WO 2019 / 060375, titled “SYSTEM FOR ANALOG LIGHT MEASURING AND PHOTON COUNTING IN CHEMILUMINESCENCE MEASUREMENTS,” hereby incorporated by reference in its entirety. FIG. 2A illustrates a cross-sectional perspective view of a luminometer 600 for performing a portion of the assay 100 (that is, the stage of the assay that includes measuring the light generated, for example, stage 8 of FIG. 1), and FIG. 2B provides an enlarged view of a portion of FIG. 2A, as shown by the dashed circle in FIG. 2A. The cut portions of the cross- sectional perspective view are shown by cross-hatching. The cross-sectional perspective view illustrates a cap 415, a chassis 405, a luminometer computer system compartment 435, a PMT 630, a PMT cover 450, a stand 440, a motor 425, a thermal barrier 445, a reaction vessel chamber 610, and a calibration unit 460. Also shown is a luminometer output signal socket 465a and a luminometer output signal socket 465b.

[0073] FIG. 2A provides a view of a chamber opening 430, which provides access to reaction vessel chamber 610. The reaction vessel 105 is shown seated within the reaction vessel chamber 610. A light passage 640 intersects with the reaction vessel chamber 610 near the bottom of the reaction vessel chamber 610. The PMT 630, shown in FIGS. 2A and 2B, can be any other suitable light detecting device or light detector. The PMT 630 can include a sensing element (not shown indetail) that detects light from the light passage 640 and / or the reaction vessel chamber 610. The PMT 630 is adjacent an aperture 635 that is aligned with the light passage 640 and past an intersection of the light passage 640 and the reaction vessel chamber 610. The aperture 635 allows light to enter the PMT 630 and the sensing element to receive the light. The reaction vessel chamber 610 intersects with the light passage 640 such that when the reaction vessel 105 is placed in the reaction vessel chamber 610, the substance or sample within the reaction vessel 105 can emit photons viewable in the light passage 640 and to the aperture 635. The aperture 635 can be limited in size, for example to 8.5 centimeters in diameter, to limit the view of a meniscus within the reaction vessel 105. On the other end of the light passage 640, a calibration unit aperture 645 can align with the light passage 640. The calibration unit 460 can include a light emitting diode (“LED”) 620 and a photodiode 625. The LED 620 and photodiode 625 can provide a regulated internal light source used to calibrate PMT 630. The reaction vessel 105 is not needed in the luminometer, for example during calibration. While the luminometer 600 is described as including the reaction vessel 105, this is an optional component of the system that may not necessarily be part of the luminometer.

[0074] The luminometer 600 is an illustrative example of a suitable detector that may be used in accordance with the teaching herein and other detectors may be employed, and / or more or fewer components may be included in the luminometer 600. Therefore, the luminometer 600, and the individual structural elements of luminometer 600, are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0075] C. Illustrative Wash Assembly of Automated Clinical Analyzer

[0076] As described above, the immunoassay may include one or more wash cycles. The wash cycle(s) may be performed by any suitable instrumentation. FIGS. 3A and 3B and International Patent Application Publication No. WO 2020 / 139989, titled “CLINICAL ANALYZER AUTOMATED SYSTEM DIAGNOSTICS,” hereby incorporated by reference in its entirety, illustrate an exemplary wash assembly in the form of a wash wheel 270 that can be used in performing the wash steps of an assay 100, as exemplified in FIG. 1. The wash wheel 270 includes a plurality of holders 272 (e.g., holes, etc.). As depicted, the wash wheel 270 includes 27 holders 272. In other embodiments, the wash wheel 270 may include less than or more than 27 holders 272. The holders 272 are each configured to receive a vessel (e.g., a reaction vessel 105). Thevessel and the holder 272 are axisymmetric with each other, when mated (e.g., when the vessel is located within the holder 272).

[0077] In the example of FIGS. 3A and 3B, 27 stations S attached to the frame 262 of the clinical analyzer are defined, about which the wash wheel 270 moves the holders 272. In particular, the wash wheel 270 rotates about a rotational axis RAI and thereby moves the holders 272 from station to station about a rotational displacement Rl. In the illustrative wash wheel 270 of FIGS. 3 A and 3B, the wash wheel 270 is indexed 13 1 / 3 degrees per cycle and thereby advances each of the 27 holders 272 one station forward per cycle.

[0078] In FIG. 3A, the stations S are labeled with respect to the wash wheel 270 at a given position, with individual stations being designated using the letter “S” followed by a station number. Not all stations S are labeled, but can be determined by counting between the labeled stations S. In FIG. 3B, the station designations using the letter “S’ followed by a station number are omitted. However, correspondence between the figures can be established by mapping the in / out station in FIG. 3B to station SI in FIG. 3 A, and mapping the station labeled QS in FIG. 3B to station S2 in FIG. 3A. Descriptions of the various stations and roles they can play in an assay 100 (for example, as shown in FIG. 1) are set forth below.

[0079] As mentioned above, and as will be described in greater detail below, wash wheel 270 may be configured to perform one or more washing cycles on a suitable mixture within reaction vessel 105. In some clinical analyzers, station SO may be a no-function station, but may transfer a vessel 105 between neighboring stations. Station SI may be an entrance / exit station. The vessel 105 is introduced to one of the holders 272 of the wash wheel 270 at station SI . This may be done, for example, after the first reagent 115 (stage 1) and sample 165 (stage 2) have been added to the reaction vessel (e.g., in a reaction build carriage, not pictured in FIGS. 3A and3B), and then the contents of the vessel have been mixed and / or incubated (stage 3) (e.g., in an incubation wheel, also not shown in FIGS. 3A and 3B). Additionally, or alternatively, vessel 105 may be introduced to one of the holders 272 of the wash wheel 270 at station SI after assay mixture (175) has been suitably created (stage 5b). From station SI, the vessel 105 is rotated to the other stations S and eventually returns to the station SI where it is removed from the holder 272 of the wash wheel 270.

[0080] After a vessel 105 has been added to the wash wheel 270 at station S 1, it will be rotated to station S2, where a wash fluid 150 is dispensed. The contents of the vessel 105 will then berotated through stations S3-S8 where the particles 120 in the reaction vessel 105 will be drawn to the side of the vessel 105 via magnets in the current example. The vessel 105 will then be rotated to station S9 (labeled as station Al in FIG. 3B) where the contents of the vessel that are not bound to a particle 120 that is attracted to the side of the vessel by a magnet will be aspirated, thereby completing an illustrative wash cycle. Therefore, wash assembly (in the current form of wash wheel 270) is configured to isolate a plurality of particles 120 from a mixture, where some particles 120 of the plurality of particles 120 are bound to a respective analyte of the patient sample 165. The vessel 105 will then be moved to station S10 (labeled as DI in FIG. 3B) where wash buffer will be added to the vessel 105 and the contents of the vessel 105 will be spin mixed. This process may then be repeated for stations S11-S18 to perform another illustrative wash cycle (i.e., the vessel may be magnetized in positions S11-S16, have its contents aspirated in position S17 / A2, then have additional buffer added and be mixed in position S18 / D2). The vessel 105 may then be subjected to another magnetization / aspiration procedure in positions S19-S25.

[0081] In the current example, wash wheel 270 adds wash fluid 150 to the vessel 105, aspirates contents not bound to particles 120, subsequently adds more wash fluid 150 to the vessel 105, and spin mixes the contents within vessel 105. However, wash wheel 270 may be configured to add wash fluid 150, aspirate, add more wash fluid 150, and spin mix contents of vessel 105 in any suitable order and / or pattern. For example, in one instance, wash wheel 270 may be configured to add wash fluid 150 to the vessel 105, aspirate contents not bound to particles 120, spin mix the contents within the vessel 105, and then add wash fluid 150 for a subsequent wash cycle. As another example, wash wheel 270 may be configured to add wash fluid 150 to vessel 105, aspirate contents not bound to particles 120, add more wash fluid 150 to the vessel, re-aspirate the contents not bound to particles 120, spin mix the contents within the vessel 105, and repeat.

[0082] While in one embodiment, the current illustrative example contemplated wash wheel 270 performing three wash cycles, wash wheel 270 may perform any suitable number of wash cycles. For example, vessel 105 may be moved about wash wheel 270 multiple times such that additional wash cycles are performed.

[0083] In some exemplary instances where wash wheel 270 is utilized in the context of assay 100 described above, after stage 4, the vessel 105 could be moved to position SI, from which it could be removed (e.g., using a pick and place device) from wash wheel 270 to continue with assay 100. For example, in instances where wash wheel 270 is incorporated with automated clinicalanalyzer 700, described in greater detail below, reaction vessel 105 may be moved from wash wheel 270 and transferred to a reaction vessel carriage (e.g., reaction vessel carriage 712 of FIG. 7) for dispensing of the second reagent 155 (stage 5) via reagent pipetting stations 714 ( FIG. 7) and reagent storage unit 716 (FIG. 7) in accordance with the description herein, thereby creating the assay mixture 175. As mentioned above, wash wheel 270 may receive reaction vessel 105 housing assay mixture 175 to perform one or more wash cycles (e.g., magnetization + aspiration cycle described as stage 6), either directly or after being subjected to additional mixing and / or incubation (e.g., in an incubation wheel 750 (FIG. 7)).

[0084] In some exemplary instances where wash wheel 270 is utilized in the context of assay 100 described above, the additional magnetization + aspiration cycle of stage 6 would be performed in the same manner as described above for stage 4 (i.e., wash buffer would be dispensed at positions QS, DI and D2, the contents of the buffer would be aspirated in positions Al, A2 and A3, etc.). However, in instances where the first substrate material 180 is utilized, at the conclusion of stage 6, the reaction vessel 105, rather than being moved directly to the In / Out position SI and removed from the wash wheel 270, may be moved to station S26 where the first substrate material 180 would be dispensed (i.e., the beginning of stage 7) and the contents of the vessel 105 would be mixed to form signal generating mixture 177. A designated substrate dispensing pipette system 724 (FIG. 7) may be used to dispense the first substrate material 180 into vessel at station S26. From there, the vessel could be rotated to the In / Out position SI, from which it could be moved to another portion of the analyzer (e g., incubation wheel 750 (FIG. 7)) to further advance the ALP / substrate reaction, thereby completing stage 7. Finally, at the end of stage 7, the reaction vessel maybe moved to a luminometer 600 such as shown in FIGS. 2A and 2B for measurement of the light generated by the ALP / substrate reaction as shown in stage 8 of the assay 100 from FIG. 1. As also described herein, in some instances, instead of selecting and adding first substrate material 180, a different reporting reagent such as a second substrate material 182 (FIGS. 14A- 14F, 16A-16C, and 19A-19F) may be selected and added to reaction vessel 105 to create signal generating mixture 177. As will also be described in greater detail below, in instances where a second substrate material 182 (FIGS. 14A-14F, 16A-16C, and 19A-19F) is utilized to serve as the reporting reagent, a sample rack 800, 830, 850 (FIGS. 9-12) housing a second substrate material 182 may be accessed by a suitable pipetting unit 706 (FIGS. 14A-14F and 16A-16C) to suitably dispense second substrate material 182 into reaction vessel 105. Alternatively, in someembodiments, an alternate substrate may be obtained from an adjacent automated machine to suitably dispense second substrate material 182 into reaction vessel 105.

[0085] The wash wheel 270 is an illustrative example of a suitable wash assembly that may be used in accordance with the teaching herein. Therefore, the wash wheel 270, and the individual structural elements of wash wheel 270, are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0086] D. Illustrative Pipetting System of Automated Clinical Analyzer

[0087] FIG. 4, and International Patent Application Publication No. WO 2020 / 139989 an exemplary pipetting system 510 that may move pipettes between various probe receiving stations for dispensing and / or aspirating various fluids as described previously in the context of performing an assay 100. As an example, suitable portions of pipetting system 510 may be incorporated into sample pipetting unit 706 (FIG. 7), reagent pipetting stations 714 (FIG. 7), designated substrate dispensing pipette system 724 (FIG. 7), other suitable portions of wash wheel 270 configured to aspirate and / or deliver wash buffer, etc.

[0088] The pipetting system 510 is an illustrative example of a suitable pipetting system that may be used in accordance with the teachings herein. Therefore, the pipetting system 510, and the individual structural elements of pipetting system 510 are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0089] In FIG. 4, the example pipetting system 510 is configured to transfer fluids between a first probe receiving station PSI (e.g., a sample rack, a reagent pack, a sample vessel, a substrate vessel, etc.) and a second probe receiving station PS2 (e.g., a reaction vessel, a substrate vessel, a station along the periphery of wash wheel 270, etc.). This may be done in part using first actuator 514 mounted to a first frame 512 mounted to the frame of the instrument. In the example of FIG. 4, the first actuator 514 is a linear actuator that provides movement along displacement dl. A sign convention is defined with respect to the displacement dl. In particular, a first direction dl+ and an opposite second direction dl- is defined for displacement dl.

[0090] In addition to the first frame 512, the example pipetting system 510 of FIG. 4 also includes a second frame 516. The second frame 516 may be mounted to the first actuator 514, and a second actuator 518 may be mounted to the second frame 516. As depicted, the second actuator 518 is a linear actuator that provides movement along displacement d2. A sign convention is defined with respect to the displacement d2. In particular, a first direction d2+ and an oppositesecond direction d2- are defined for displacement d2. As depicted, the displacements dl and d2 are perpendicular. In other embodiments, the displacements dl and d2 may be non-perpendicular (e.g., skew, parallel, etc.).

[0091] As depicted in FIG. 4, a probe P, including a probe tip PT, may be mounted to the second actuator 518. The probe P may be a hollow probe. Accordingly, in the example pipetting system 510 of FIG. 4, by actuating the first and second actuators 514 and 518, the probe P and the probe tip PT can be moved to a plurality of locations within a two-dimensional space including the probe receiving stations PSI and PS2. In other embodiments, an additional frame and / or an additional actuator may be provided (e.g., between the first frame 512 and the frame of the instrument) thereby allowing the probe P and the probe tip PT to be moved to a plurality of locations within a three-dimensional space. In other embodiments, pipetting system 510 may include only one actuator 514, 518 such that probe P and the probe tip PT maybe moved along a single linear direction.

[0092] The probe P may define an axis A. The probe receiving station PS may define an axis A0. The probe P may be aligned with the corresponding probe receiving station PS when the axes A and A0 are aligned within an acceptable tolerance.

[0093] In typical use, such as in dispensing and aspiration of fluids as described in the context of FIGS. 1, 3A, and 3B, the first actuator 514 axially aligns the probe P with the desired probe receiving station PS, PSI and thereby aligns the axes A and A0. As illustrated at FIG. 4, the probe P and the probe receiving station PSI of the example pipetting system 510 are aligned when the first actuator 514 is at an actuated position dpi. Upon alignment between the probe P and the probe receiving station PS, PSI, the second actuator 518 may move the probe P along its axis A and thereby along a probe path 300 (e.g., away from an actuated position apl of the second actuator 518). Upon the probe P dispensing and / or aspirating fluid at an actuated position in a probe receiving station, the probe P may retract along the probe path 300 and the first actuator 514 may then move the second frame 516 and thereby move the probe P, the probe tip PT and the probe path 300 to an additional receiving station within the range of the pipetting system.

[0094] In practice, a clinical analyzer may incorporate multiple pipetting systems, for purposes such as allowing specialization of various assemblies. For example, in some cases, pipetting systems used to transfer reagents from reagent packs to a reaction vessel may be different from pipetting systems used to transfer samples from a sample vessel to a reaction vessel. In thistype of system, the pipettor used for transferring reagents may have additional specialization to aid in this task. For instance, a reagent pipettor may be outfitted with a tip that allows the reagent pipettor to perform ultrasonic mixing of a reagent in a reagent pack before aspirating the reagent for transport to a reaction vessel, thereby ensuring that the aspirated reagent would not be impacted by any settling that may have taken place in the reagent pack. Sample pipettors may similarly be specialized. For instance, there may be multiple sample pipettors adapted to move portions of a sample either directly to a particular test (which would be done by a sample precision pipettor), or (via a sample aliquot pipettor) to a holding area (e.g., a sample wheel) in which the portion of the sample may be held for use in a later test (including, in some cases, a reflex test). Multiple pipetting systems may also be incorporated for reasons besides supporting multiple workflows. For example, some instruments may be provided with multiple pipetting systems to avoid individual pipetting systems becoming bottlenecks.

[0095] Although one or more pipetting system(s) such as shown in FIG. 4 may be present in clinical analyzers that are implemented based on this disclosure, such pipetting systems are not a requirement, and other types of pipetting arrangements, either in combination with or as alternatives to systems such as shown in FIG. 4 may also be present. For example, in some embodiments, various vessel positions (e.g., positions SO to S26 from FIG. 3A) may have dedicated pipettors that may move up and down to interact with (e.g., dispense fluid into, aspirate fluid from) the vessels at their respective positions (e.g., wash buffer dispensing positions in a wash wheel such as shown in FIGS. 3A and 3B), but would not have the additional degrees of freedom illustrated in FIG. 4. Accordingly, the above discussion of variations, like the discussion of the pipetting system 510 of FIG. 4, should be understood as being illustrative only, and should not be treated as limiting.

[0096] FIG. 5 illustrates an illustrative arrangement that may be employed for washing probes in a pipetting system 510 such as shown in FIG. 4. The probe washing arrangement includes the probe P, the frame 516, the probe actuator 518, a probe washer 530, and a probe washer actuator 520, also referred to herein as a third actuator 520. The probe actuator 518 actuates the probe P relative to the frame 516. The probe washer 530 cleans the probe P. The probe washer 530 includes a cleaning cavity 532, having a longitudinal, centerline cavity axis Ac, that is adapted to receive at least a portion of the probe P when the probe washer 530 is positioned at a deployed position pw2, intersects the probe path 300 when the probe washer 530 is positioned at the deployed positionpw2 (shown in dashed line), and clears the probe path 300 when the probe washer 530 is positioned at a stowed position pwl. The probe washer actuator 520 moves the probe washer 530 between the deployed position pw2 and the stowed position pwl. The probe washer actuator 520 actuates the probe washer 530 relative to the frame 516.

[0097] In some embodiments, the probe actuator 520 is adapted to move the probe P along displacement d2 between a stowed probe position and a probe washing position. The probe washer 530 may correspondingly be moved along displacement d3 relative to the probe path 300 by the third actuator 520 (e.g., to an actuated position pw2) such that the probe washer 530 (e.g., the cleaning cavity 532 of the probe washer 530 and / or a wall 534 at a bottom of the cleaning cavity 532) intersects the probe path 300 when cleaning or preparing to clean the probe P and thereby allows the probe P to pass into and out of the cleaning cavity 532 of the probe washer 530. The probe washer 530 could also be moved relative to the probe path 300 by the third actuator 520 (e.g., to an actuated position pwl) such that the probe washer 530 clears the probe path 300 when the probe P dispenses, aspirates, prepares for dispensing, and / or prepares for aspirating and thereby allows the probe P to pass by the probe washer 530.

[0098] Upon the axis A and the cavity axis Ac being aligned, the second actuator 518 may advance the probe P to a washing position in which at least a portion of the probe P is within the cleaning cavity 532 of the probe washer 530. Upon the probe P or a portion thereof entering the cleaning cavity 532, the probe P may be internally and / or externally cleaned. Upon the probe P being cleaned, the second actuator 518 may retract the probe P to a stowed position and thereby remove the probe P or portion thereof from the cleaning cavity 532 of the probe washer 530.

[0099] The probe washing arrangement of FIG. 5 is illustrative and should not be treated as limiting. For instance, in some analyzers, a probe washing arrangement may include a cleaning fluid supply, a pump for transferring cleaning fluid into and / or out of a probe washer 530, and one or more valves for configuring fluid flow through the probe washer. The fact that these additional components are not explicitly illustrated in FIG. 5 should not be treated as implying that analyzers implemented based on this disclosure will necessarily lack such features. Similarly, in some cases, analyzers may be equipped with wash stations that are separate from pipetting assemblies, either as alternatives to, or in addition to, washing arrangements such as shown in FIG. 5. Such wash stations may include, for example, wash towers into which probes could be inserted for cleaning and wash dispensing pumps for dispensing fluid into and / or inside of a wash tower. Accordingly,while the mobile washing arrangement of FIG. 5 may be present in some analyzers implemented to include functionality described in this document, the mobile washing arrangements are intended to be illustrative only.

[0100] E. Illustrative Computer System for Automated Clinical Analyzer

[0101] Any suitable computer system can be integrated into, or connected with, a clinical analyzer to control various actions of the analyzer such as by sending commands to the wash wheel 270, the pipette 110, and / or other components. Turning now to FIG. 6, which illustrates an exemplary computer system 49 that can be integrated into, or connected with, a clinical analyzer, and that could control various actions of the analyzer such as by sending commands to the wash wheel 270, the pipette 110 and / or other components. As shown in FIG. 6, such a computer system 49 may include a processor 51, a memory 53, a mass storage memory device 55, an input / output (VO) interface 57, and a Human Machine Interface (HMI) 59. Optionally, computer system 49 may also be operatively coupled to one or more external resources 61 via a network 63 and / or VO interface 57. External resources 61 may include, but are not limited to, servers, databases, mass storage devices, peripheral devices, cloud-based network services, or any other suitable computer resource that may be employed by computer system 49.

[0102] Processor 51 may include one or more devices selected from microprocessors, microcontrollers, digital signal processors, microcomputers, central processing units, field programmable gate arrays, programmable logic devices, state machines, logic circuits, analog circuits, digital circuits, or any other devices that manipulate signals (analog or digital) based on operational instructions that are stored in memory 53. Memory 53 may include a single memory device or a plurality of memory devices including, but not limited, to read-only memory (ROM), random access memory (RAM), volatile memory, non-volatile memory, static random access memory (SRAM), dynamic random access memory (DRAM), flash memory, cache memory, or any other device capable of storing information. Mass storage memory device 55 may include data storage devices such as a hard drive, optical drive, tape drive, non-volatile solid state device, or any other device capable of storing information.

[0103] Processor 51 may operate under the control of an operating system 65 that resides in memory 53. Operating system 65 may manage computer resources so that computer program code embodied as one or more computer software applications, such as an application 67 residing in memory 53, may have instructions executed by the processor 51. In an alternative embodiment,processor 51 may execute application 67 directly, in which case the operating system 65 may be omitted. One or more data structures 69 may also reside in memory 53, and may be used by processor 51, operating system 65, and / or application 67 to store and / or manipulate data.

[0104] The I / O interface 57 may provide a machine interface that operatively couples processor 51 to other devices and systems, such as network 63 or external resource 61. Application 67 may thereby work cooperatively with network 63 or external resource 61 by communicating via I / O interface 57 to provide the various features, functions, applications, processes, or modules comprising embodiments of the invention. Application 67 may also have program code that is executed by one or more external resources 61, or otherwise rely on functions or signals provided by other system or network components external to computer system 49. Indeed, given the nearly endless hardware and software configurations possible, persons having ordinary skill in the art will understand that different versions of the invention may include applications that are located externally to computer system 49, distributed among multiple computers or other external resources 61, or provided by computing resources (hardware and software) that are provided as a service over network 63, such as a cloud computing service.

[0105] HMI 59 may be operatively coupled to processor 51 of computer system 49 in a known manner to allow a user to interact directly with the computer system 49. HMI 59 may include video or alphanumeric displays, a touch screen, a speaker, and any other suitable audio and visual indicators capable of providing data to the user. HMI 59 may also include input devices and controls such as an alphanumeric keyboard, a pointing device, keypads, pushbuttons, control knobs, microphones, etc., capable of accepting commands or input from the user and transmitting the entered input to the processor 51.

[0106] A database 71 may reside on mass storage memory device 55 and may be used to collect and organize data used by the various systems and modules described herein. Database 71 may include data and supporting data structures that store and organize the data. In particular, database 71 may be arranged with any database organization or structure including, but not limited to, a relational database, a hierarchical database, a network database, or combinations thereof. A database management system in the form of a computer software application executing as instructions on processor 51 may be used to access the information or data stored in records of the database 71 in response to a query, where a query may be dynamically determined and executed by operating system 65, other applications 67, or one or more modules.

[0107] The computer system 49 is an illustrative example of a suitable computer system that may be used in accordance with the teachings herein. Therefore, the computer system 49, and the individual structural elements of computer system 49 are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0108] F. Illustrative Automated Clinical Analyzer

[0109] FIG. 7 shows an illustrative automated clinical analyzer 700. The automated clinical analyzer 700 is configured to perform a variety of suitable assays, some of which may be similar to assay 100 described above. The various types of assays which may be performed by automated clinical analyzer 700 will be apparent to one skilled in the art in view of the teachings herein. The automated clinical analyzer 700 includes a detector in the form of luminometer 600, wash wheel 270, and computer system 49 described above, however, the luminometer 600, wash wheel 270, and computer system 49, and the individual structural elements thereof, are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0110] Additionally, clinical analyzer 700 includes a sample aliquot system 705, a reagent mixing station 715, and a substrate dispensing system 725. As will be described in greater detail below, sample aliquot system 705 is configured to aliquot patient samples from a rack including vessels containing at least one patient sample into a plurality of suitable reaction vessels 105. As will also be described in greater detail below, reagent mixing station 715 is configured to add a suitable amount of a desired reagent(s) into reaction vessels 105 and to suitably mix a composition of the added reagent(s) with a patient sample 165 (or patient analytes bounded to a respective particle of a plurality of particles 120) within reaction vessel 105. Further, substrate dispensing system 725 is configured to dispense first substrate material 180 into a reaction vessel 105 at wash wheel 270. The sample aliquot system 705, reagent mixing station 715, and substrate dispensing system 725 are illustrative examples of a suitable sample aliquot system, mixing station, and substrate dispensing system, respectively, that may be used within the context of automated clinical analyzer 700. Therefore, the sample aliquot system 705, reagent mixing station 715, and substrate dispensing system 725, and the individual structural elements thereof, are optional and may be substituted for other suitable elements, and even omitted when appropriate. For example, the substrate dispensing system 725 may be omitted when the substrate (e.g., substrate 182) is provided by other systems, as described herein, or, in some examples, both the substrate dispensingsystem 725 and the alternative systems for providing the second substrate are both included in the automated clinical analyzer 700.

[0111] Computer system 49 is in suitable communication with various components of automated clinical analyzer 700 such that computer system 49 may operatively control automated clinical analyzer 700. Such control of automated clinical analyzer 700 may be based on a combination of user input, software, suitable feedback components of clinical analyzer 700, such as sensors, motors, etc., and any other suitable information.

[0112] Automated clinical analyzer 700 includes a frame station 701. Frame station 701 acts as a mechanical ground and / or housing for various components of clinical analyzer 700. Further, automated clinical analyzer 700 includes suitable pick and place devices configured to suitably transport reaction vessels 105 (FIG. 1) and any other suitable containers between suitable stations of automated clinical analyzer 700. Such pick and place devices may have any suitable component(s).

[0113] An exemplary sample aliquot system 705 includes a sample loading station 702, a sample rack carriage 704, a sample pipetting unit 706, a disposable tip housing 708, and a sample wheel 710. Sample loading station 702 is configured to receive one or more patient samples 165 via a sample rack. Therefore, a user may simply place a sample rack onto sample loading station 702 to initiate the patient sample aliquoting process. Sample loading station 702 may include a suitable drive belt that may receive and actuate the sample rack to a suitable loading zone for further processing. In the current example, sample loading station 702 may actuate sample rack to a location that is adjacent to sample rack carriage 704.

[0114] Sample rack carriage 704 is configured to actuate a sample rack along an axis ASR between sample loading station 702 and sample pipetting unit 706. Once sample rack is aligned with sample pipetting unit 706, the sample pipetting unit 706 is configured to aliquot suitable amounts of patient samples from patient sample rack and dispense the aliquoted patient samples 165 into suitable containers, such as reaction vessels 105. Suitable containers housing aliquoted patient samples 165 may be housed within sample wheel 710 for a suitable amount of time thereby providing access to the aliquoted patient sample 165 when ready fortesting. Patient sample wheel 710 may be climate controlled and contain any suitable structures and components . Therefore, patient sample wheel 710 may act as a climate-controlled sample housing. As one illustrative example, patient sample wheel 710 may be configured to maintain an internal temperature at oraround 4 degrees Celsius. The patient sample wheel 710 may be configured to maintain any other suitable internal temperature.

[0115] The sample pipetting unit 706 may actuate a sample pipette 707 (FIGS. 14A-14F, 16A-16C, and 19A-19F) along axis ASPI and / or ASP2 and a vertical axis perpendicular to axis ASPI and / or ASP2 (e.g., into and out of the page) to access disposable tip housing 708, sample wheel 410, a sample rack located on sample rack carriage 704, and / or suitable sections of reaction vessel carriage 712. The sample pipetting unit 706 may include any of the suitable features of pipetting system 510 to actuate sample pipette 707 in accordance with the description herein. Sample aliquoting pipette 707 (FIGS. 14A-14F, 16A-16C, and 19A-19F) may reach disposable tip housing 708 to suitably obtain an unused disposable tip to suitably aliquot different patient samples 165. Therefore, during illustrative use, sample aliquoting pipette 707 (FIGS. 14A-14F, 16A-16C, and 19A-19F) may use a first disposable tip to aliquot a first patient sample 165 into a suitable container, dispose of the first disposable tip, obtain a new disposable tip from tip housing 708, and then aliquot a second patient sample into another suitable container. The sample aliquoting pipette 707 may dispense of a first disposable tip and obtain a new disposable tip for any suitable purpose.

[0116] Once samples from the sample rack are suitably aliquoted, the sample rack may be returned to sample loading station 702 via sample rack carriage 704 such that a new patient sample rack may be loaded and aliquoted in accordance with the teachings herein.

[0117] The sample aliquot system 705 is an illustrative example of a sample aliquot system 705 that may be used within the context of automated clinical analyzer 700 in accordance with the teachings herein. Therefore, the sample aliquot system 705, and the individual structural elements of sample aliquot system 705 are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0118] Once an aliquot of patient sample 165 (FIG. 1) has been removed from the sample rack by the sample aliquot system 705, the aliquot of patient sample can be placed in sample wheel 710, and from there may be transferred to the reaction vessel carriage 712. Alternatively, in some instances, an aliquot of patient sample 165 may be aliquoted from the sample rack and directly dispensed into a reaction vessel 105 already placed on reaction vessel carriage 712, effectively bypassing use of sample wheel 710. In some instances, a pick and place device obtains the container or vessel housing the patient sample 165 within sample wheel 710 and places the container or vessel on reaction vessel carriage 712. In some instances, a pipette aliquots the patientsample 165 from the sample wheel 710 and / or the sample rack and distributes the patient sample 165 into a reaction vessel 105 already placed on reaction vessel carriage 712. The patient samples 165 may be transported from sample wheel 710 and / or sample rack carriage 704 onto reaction vessel carriage 712 using any suitable means.

[0119] Reaction vessel carriage 712 is configured to actuate at least one reaction vessel 105 along an axis Aci to suitably transport a reaction vessel 105 between sample aliquot system 705 and a reagent station in the form of reagent mixing station 715. In the current example, reagent mixing station 715 includes a plurality of reagent pipetting stations 714, and a reagent storage unit 716. Therefore, reaction vessel carriages 712 may actuate a reaction vessel 105 along axis Aci to align a reaction vessel 105 with a pipette from a respective reagent pipetting station 714. Each reagent pipetting station 714 is configured to actuate a respective pipette along a respective axis (A3, A4, A5, A6) and a vertical axis perpendicular to their respective axis (A3, A4, A5, A6) (e.g., into and out of the page) to (A) obtain a suitable reagent(s) housed within reagent storage unit 716, and (B) add the obtained reagent into the aligned reaction vessel 105. Therefore, reagent storage unit 716 is accessible via at least one reagent pipetting station 714.

[0120] Reagent storage unit 716 houses at least one reagent, which may include reagents 115, 155. Reagent storage unit 716 may be climate controlled. Reagents housed within reagent storage unit 716 may be housed within a reagent pack. Additionally, reagent storage unit defines a plurality of access openings 718A, 718B, 718C, 718D; which may be suitably aligned with chambers defined by reagent pack housing reagent. The access openings 718A, 718B, 718C, 718D are aligned with a respective reagent pipetting station 714 such that a pipette of the respective pipetting station 714 may actuate within a respecting access opening 718A, 718B, 718C, 718D to aspirate reagent from a respective reagent pack within reagent storage unit 716. That is, a first plurality of access openings 718A are aligned with reagent pipetting station 714 configured to actuate a respective pipette along axis A3, a second plurality of access openings 718B are aligned with reagent pipetting station 714 configured to actuate a respective pipette along axis A4, a third plurality of access openings 718C are aligned with reagent pipetting station 714 configured to actuate a respective pipette along axis A5, and a fourth plurality of access openings 718D are aligned with reagent pipetting station 714 configured to actuate a respective pipette along axis A6. In the current example, there are four reagent pipetting stations 714. However, any suitable number of reagent pipetting stations 714 may be incorporated.

[0121] Reagent mixing station 715 is configured to mix reagent within reaction vessel 105. In some instances, pipettes of reagent pipetting station 714 are configured to mix reagent within reaction vessel 105. In some instances, reaction vessel carriage 712 may be configured to shake and / or otherwise suitably vibrate a reaction vessel 105 housed within carriage 712 to suitably mix the components (e.g., patient sample, reagents, etc.) housed within such a reaction vessel 105. Therefore, reaction vessel carriage 712 may have any suitable components to therefore suitably mix compositions housed within reaction vessel 105.

[0122] The reagent mixing station 715 is an illustrative example of a reagent mixing station that may be used within the context of automated clinical analyzer 700 in accordance with the teachings herein. Therefore, the reagent mixing station 715, and the individual structural elements of reagent mixing station 715 are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0123] As mentioned above, clinical analyzer 700 includes suitable pick and place devices configured to move a reaction vessel 105 between suitable stations. Therefore, clinical analyzer 700 includes a pick and place device that is configured to actuate a reaction vessel 105 from reaction vessel carriage 712 to incubation wheel 750, wash wheel 270, back to reaction vessel carriage 712, into luminometer 600, and any other suitable path.

[0124] Clinical analyzer 700 includes an incubation container in the form of an incubation wheel 750. The incubation container is configured to suitably house and incubate one or more reaction vessels 105 and / or suitable mixtures (or suitable portions of mixtures) containing a patient sample 165 and one or more reagents 115, 155. As one illustrative example, incubation container may be configured to maintain an internal temperature at or around 37 degrees Celsius. Of course, incubation container may be configured to maintain any other suitable internal temperature. Incubation wheel 750 may include any suitable components. Once a mixture is suitably incubated, the pick and place device may transport reaction vessel 105 either to wash wheel 270 to perform one or more washing cycles in accordance with the description herein, luminometer 600, or another suitable station.

[0125] The incubation wheel 750 is an illustrative example of an incubation container that may be used within the context of automated clinical analyzer 700 in accordance with the teachings herein. Therefore, the incubation wheel 750, and the individual structural elements of incubationwheel 750 are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0126] As mentioned above, clinical analyzer 700 may include a substrate dispensing system 725. As further discussed herein, when first substrate material 180 is the selected reporting agent, substrate dispensing system 725 is configured to add first substrate material 180 into reaction vessel 105 while the reaction vessel 105 is located at the wash wheel assembly 270. Wash wheel assembly 270 may further be configured to suitably mix first substrate material 180 within reaction vessel 105. In some examples, the first substrate material 180 and / or a second substrate material 182 may be added into the reaction vessel 105 from an alternative location than the substrate dispensing system 725. The present disclosure provides disclosure of the alternative locations. Although the following description provides for adding of the second substrate material 182 at the alternative locations, the first substrate material 180 may be added from the alternative locations. Furthermore, the alternative locations may be in addition to the substrate dispensing system 725 or in lieu of the substrate dispensing system 725. As described herein, the alternative locations may include a sample rack and / or an adjacent automated machine (e.g., an automation line). The substrate material may be obtained directly from the sample rack or adjacent automated machine to a reaction vessel (e.g., pipetted directly into a reaction vessel) or may be obtained from the sample rack or adjacent automated machine to be temporarily stored on the instrument in a vessel, before being provided to a reaction vessel (e.g., pipetted into an intermediate vessel prior to a reaction vessel). Further details of the alternative locations are provided in the description to follow.

[0127] As noted, in some instances, instead of selecting and adding first substrate material 180, another reporting reagent such as a second substrate material 182 (FIGS. 14A-14F, 16A-16C, and 19A-19F) may be selected and added to reaction vessel 105 to create signal generating mixture 177. The second substrate material 182 may be the same as or different from the first substrate material 180. As will also be described in greater detail below, in instances where a second substrate material 182 (FIGS. 14A-14F, 16A-16C, and 19A-19F) is utilized to serve as the reporting reagent (or where the first substrate material 180 is loaded from an alternate location from substrate dispensing system 725), the second substrate material 182 may be loaded onto the clinical analyzer (for example, via a sample rack 800, 830, 850 (FIGS. 9-12) housing a second substrate material 182; or via an adjacent automatic machine 1700) and may be accessed by asuitable pipetting unit 706 (FIGS. 14A-14F and 16A-16C) to dispense second substrate material 182 into reaction vessel 105.

[0128] Substrate dispensing system 725 includes a substrate housing 720, a substrate reservoir in the form of a substrate container 722 located within housing 720, a designated substrate dispensing pipette system 724, and a fluid line 726 providing fluid communication between container 722 and pipette system 724. Substrate dispensing system 725 may include suitable temperature control systems configured to suitably control the temperature of first substrate material 180 while contained within both container 722 and housing 720, as well as when transferred to dispensing pipette system 724. Such temperature control systems may have any suitable components.

[0129] Substrate housing 720 houses substrate container 722 and provides access to substrate container 722, thereby allowing replacement of substrate container 722 when appropriate. Substrate container 722 includes a suitable amount of first substrate material 180. Additionally, substrate container 722 is suitably attached to fluid line 726. Fluid line 726 is in fluid communication with both an interior of substrate container 722 and designated substrate dispensing pipette system 724. Therefore, substrate dispensing pipette system 724 is in fluid communication with first substrate material 180. Designated substrate dispensing pipette system 724 is configured to selectively remove first substrate material 180 from container 722 and dispense first substrate material 180 into a reaction vessel 105 suitably located on wash wheel 270. Designated substrate dispensing pipette system 724 may be mounted onto and / or relative to wash wheel 270 such that designated substrate dispensing pipette system 724 is a component of wash wheel 270. Designated substrate dispensing pipette system 724 may be located adjacent to wash wheel 270 but mounted to other suitable structure of clinical analyzer 700 (e.g. frame station 701). Designated substrate dispensing pipette system 724 may include a suitable pump, pipette, temperature controls, and any other suitable components. Therefore, when a reaction vessel 105 located within wash wheel 270 requires addition of first substrate material 180, designated substrate dispensing pipette system 724 may suitably dispense first substrate material 180 into reaction vessel 105 in accordance with the description herein.

[0130] The dispensing system 725 is an illustrative example of a substrate dispensing system that may be used within the context of automated clinical analyzer 700 in accordance with the teachings herein. Therefore, the substrate dispensing system 725, and the individual structuralelements of substrate dispensing system 725 are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0131] Similar to reaction vessel carriage 712, wash wheel 270 may be configured to suitably vibrate reaction vessel 105 to suitably mix first substrate material 180 with other components housed within reaction vessel 105. Additionally, or alternatively, designated substrate dispensing pipette system 724 (or any other suitable components of clinical analyzer 700) may be configured to suitably mix first substrate material 180 within reaction vessel 105.

[0132] G. Illustrative Workflow of Automated Clinical Analyzer

[0133] FIG. 8 shows an illustrative workflow 1000 of automated clinical analyzer 700 that may be used to perform a suitable assay, which may be the assay 100, an assay similar to assay 100, or another assay. First, analyzer 700 may suitably receive 1002 a patient sample 165, and then aliquot 1004 the patient sample 165 into a suitable vessel. Receiving 1002 and aliquoting 1004 of the patient sample 165 may be performed by sample aliquot system 705 in accordance with the description herein. Alternatively, the analyzer 700 may receive 1002 and aliquot 1004 the patient sample 165 utilizing any suitable structures and techniques. For example, in some instances, the received patient sample 165 may already be aliquoted 1004 an appropriate amount.

[0134] Next, with the patient sample 165 suitably aliquoted 1004, reagent pipetting station 714 may obtain 1006 at least the first reagent 115 in accordance with the description herein. Next, at least the first reagent 115 and the patient sample may be mixed 1008 within a reaction vessel 105 to create a composition of a first mixture. As described in greater detail herein, in some assays, the second reagent 155 is also added to and mixed within reaction vessel 105 at this time to create the assay mixture 175.

[0135] In some instances, the first reagent 115 is added to the reaction vessel 105 first, while the patient sample is added to the reaction vessel 105 subsequently. In some instances, the patient sample is added to the reaction vessel 105 first, while the first reagent 115 is added to the reaction vessel 105 subsequently. Therefore, the patient sample and the first reagent 115, (and when appropriate, the second reagent 155) may be added to reaction vessel 105 using any suitable order.

[0136] Optionally, next, the reaction vessel 105 may be transferred 1010 to the incubation wheel 750 to optionally incubate the first mixture for a suitable amount of time (e.g., a predetermined amount of time). Any suitable incubation time may be utilized. The reaction vessel105 is transported 1012 to the wash wheel 270 to suitably perform one or more wash cycles in accordance with the description herein.

[0137] Next, automated clinical analyzer 700 performing the assay workflow 1000 checks 1014 to see if the assay being performed requires a secondary reagent addition and wash cycle. In the illustrative assay 100 described above, an optional wash cycle(s) was (were) performed between the addition of the first reagent 115 into reaction vessel 105 and the addition of the second reagent 155 into reaction vessel 105, and a subsequent wash cycle(s) was (were) performed after the addition of the second reagent 155 into reaction vessel 105. In such instances, when the clinical analyzer 700 performing assay workflow checks 1014 to see if the assay requires use of a secondary reagent requiring additional wash cycle(s), the answer is yes.

[0138] However, in some instances, the first reagent and the second reagent may be added to the patient sample prior to the first wash cycle such that a second wash cycle is not necessary. In such instances, the clinical analyzer 700 performing assay workflow 1000 moves directly to the adding 1022 substrate to reaction vessel step, bypassing steps 1016, 1018, and 1020, described below. In such instances, the first reagent 115, second reagent 155, and patient sample 165 may be added to reaction vessel 105 in any suitable order.

[0139] If the assay requires use of a secondary reagent requiring additional wash cycle(s), clinical analyzer 700 transfers 1016 reaction vessel 105 to reagent pipetting station 714, obtains second reagent 155, and mixes the second reagent 155 within reaction vessel 105 to create the assay mixture 175 in accordance with the description herein. With the assay mixture 175 created, clinical analyzer 700 then transfers the reaction vessel 105 with the assay mixture 175 to the incubation wheel 750 to incubate 1018 the assay mixture 175 for a suitable amount of time (e.g., a predetermined amount of time) in accordance with the description herein. Once the incubation 1018 step is completed, the reaction vessel 105 is then transferred 1020 to the wash wheel 270 to perform a wash cycle(s) on the assay mixture 175.

[0140] Next, irrespective of whether a second reagent wash cycle is required (e.g., steps 1016, 1018, 1020), a suitable reporting reagent (e.g., first substrate material 180, second substrate material 182, etc.) is added 1022 to the reaction vessel 105 at a suitable time during the assay in accordance with the description herein to create the signal generating mixture 177. As described elsewhere herein, in some instances, first substrate material 180 may be added to reaction vessel 105 at wash wheel 270 via designated substrate dispensing pipette system 724. In instances wherefirst substrate material 180 is selected, wash wheel 270 is configured to add the first substrate material 180 into reaction vessel 105 (e.g., at station S26 in FIGS. 3A and 3B).

[0141] As also described herein, in some instances, instead of selecting and adding first substrate material 180, a different reporting reagent such as a second substrate material 182 (FIGS. 14A-14F, 16A-16C, and 19A-19F) may be selected and added to reaction vessel 105 to create signal generating mixture 177. Various non-limiting means of receiving, aliquoting, dispensing, and sometimes storing second substrate material 182 are described below. Such a second substrate material 182 may be loaded into clinical analyzer 700 in variety of ways. Additionally, such a second substrate material 182 may be obtained from a variety of locations within clinical analyzer 700. As will also be described in greater detail below, in some instances where a second substrate material 182 (FIGS. 14A-14F, 16A-16C, and 19A-19F) is utilized to serve as the reporting reagent, a sample rack 800, 830, 850 (FIGS. 9-12) housing a second substrate material 182 may be accessed by a suitable pipetting unit 706 (FIGS. 14A-14F and 16A-16C) to suitably dispense second substrate material 182 into reaction vessel 105. In some instances where a second substrate material 182 (FIGS. 14A-14F, 16A-16C, and 19A-19F) is utilized to serve as the reporting reagent, the second substrate may be obtained from an adjacent automated machine. As will also be described in greater detail below, in some instances, regardless of the source of the second substrate material 182, the second substrate material may be stored on the analyzer 700 between being accessed or obtained and being dispensed into the reaction vessel 105.

[0142] Regardless of which substrate material 180, 182 is selected, added, and mixed to reaction vessel 105, the reaction vessel 105 is then transferred to the incubation wheel 750 and is suitably incubated 1024 for a suitable amount of time (e.g., a predetermined amount of time). Next, the reaction vessel 105 is transferred to the luminometer 600 to measure 1026 RLUs generated by the chemical reaction between the added first substrate material 180 and the enzyme within the reaction vessel 105 in accordance with the description herein. In some instances, for example, when the assay performed includes a competing analyte, fewer RLUs detected by luminometer 600 is indicative of the presence of more patient analyte. In some instances, more RLUs detected by luminometer 600 is indicative of the presence of more patient analyte. The assay workflow 1000 applies to both of the aforementioned instances.

[0143] II. Illustrative Means of Introducing A Second Substrate Material into an Automated Clinical Analyzer and Method of Use

[0144] As mentioned above, a reporting reagent (e.g., first substrate material 180 or second substrate material 182) is added to a reaction vessel 105, when the substrate material chemically reacts with a bound reporter (e.g., an enzyme), a signal (e.g., light) is generated, and the signal can be detected by a detector (e.g., a luminometer 600). Further, in an exemplary embodiment, the strength of signal (e g., light) generated from the chemical reaction, and the measured RLUs via luminometer 600, may be indicative of the amount of patient analyte within the reaction vessel 105. The type of reporting reagent (e.g., first substrate material 180 or second substrate material 182) selected for an assay may affect various parameters of the generated signal (e.g., light) to be measured by the detector (e.g., luminometer 600). To further optimize desired parameter(s), for the clinical analyzer 700 may be configured to select from multiple reagents (e.g., first substrate material 180 or second substrate material 182), as different reporting reagents or substrates materials may have different resulting effects on the desired parameter(s).

[0145] Various parameters may be considered in determining which reporting reagent or substrate material to select, such that one reporting reagent or substrate may have known advantages and disadvantages compared to a second reporting reagent or substrate; and such advantages and disadvantages may be considered when deciding which reagent or substrate to use in an assay. For example, reagents or substrates may vary in strength of signal generated, or time required during incubation to generate a suitable or maximal signal to be measured in an assay. The selection of reporting reagent (e.g., a first substrate material 180 or a second substrate material 182) may be determined by any suitable means. For example, the analyte being detected may be used to select the reporting reagent (that is, with some analytes being detected by a first substrate and other analytes by a second substrate); the results of a previous assay for the same analyte may be used to select the reporting reagent (that is, when the results of an assay for a particular analyte with a first substrate fall within certain parameters, performing the assay with the second substrate may be indicated), or a user of the clinical analyzer may select the reporting reagent (that is, the clinical analyzer may be configured such that the user of the analyzer may be able to select the use of particular substrate for a particular assay).

[0146] In an exemplary embodiment, a first substrate material 180 may, after chemically reacting with a bound enzyme, generate light with a stronger signal (i.e., light with higher RLU measurement) as compared to a second substrate material 182; while the first substrate material 180 may take longer to generate a suitable signal to be measured as compared to the secondsubstrate material 182. Therefore, the first substrate material 180 may be better suited for an assay that may anticipate having fewer bound enzymes once a substrate is added; as use of the first substrate material 180 may thereby generate an appropriate RLU value to be detected luminometer 600; which, if the second substrate material 182 was selected, would otherwise have been undetectable (or at least measured in an undesirable RLU range) by luminometer 600. Accordingly, the second substrate material 182 may be better suitable for an assay that may have more bound enzymes once a substrate is added; as use of the second substrate material 182 may generate an appropriate RLU value to be detected by luminometer 600; which, if the first substrate material 180 was selected, would otherwise have too strong a signal (e.g., too much noise) for luminometer 600 to meaningfully measure. Further, in instances where strength of light to be generated is not anticipated to be an issue, it may be desirable to use the second substrate material 182 to efficiently perform an assay. Of course, the characteristics of first substrate material 180 and second substrate material 182 in the current exemplary embodiment are merely illustrative such that first substrate material 180 and second substrate material 182 may have any suitable characteristics, respectively.

[0147] This disclosure describes different ways of loading the second substrate material 182 onto the clinical analyzer 700, storing (or in some instances not storing) the second substrate material 182 within the clinical analyzer 700, and utilizing the second substrate material 182 in an assay, when appropriate.

[0148] In some aspects, the second substrate material 182 is obtained by the analyzer 700 (that is, loaded and / or introduced into the analyzer 700) from a rack (as described in Section II-A- i); in other aspects, the second substrate material 182 is obtained by the analyzer 700 (that is, loaded and / or introduced into the analyzer 700) from an adjacent automated machine (as described in Section II-A-ii).

[0149] The second substrate material 182 may be added to a reaction directly from a substrate container or vessel on a rack (e.g., sometimes referred to herein as a sample rack) or a substrate container or vessel loaded from an adjacent automated machine (as described in Section II-B). Adding the second substrate material 182 directly may mean, for example, adding without transferring the second substrate material 182 into an additional vessel or container or otherwise storing the second substrate material 182 on the analyzer 700. Alternatively, the second substrate material 182 may be in a portable vessel 106 (FIGS. 16A-16C and 19A-19F) that can be stored onthe analyzer (e.g., on the sample wheel or the incubation wheel) prior to being added to a reaction. The second substrate material 182 may be loaded directly onto the machine in a portable vessel 106, where the portable vessel 106 can be loaded onto the analyzer 700 via an adjacent automated machine (as described in Section II-A-ii); or the second substrate may be aliquoted into a portable vessel 106 (FIGS. 16A-16C and 19A-19F) that can be stored on the analyzer 700, where the second substrate aliquoted into the portable vessel 106 was obtained from either (A) a substrate container on a rack (as described in Section II-C), or (B) a substrate container introduced via the adjacent automated machine (as described in Section II-A-ii).

[0150] The second substrate material 182 can be dispensed into reaction vessel 105 at the reaction vessel carriage 712 of the reagent mixing station 715 (as described in Section II-D); or the second substrate material 182 can be dispensed into reaction vessel 105 at the wash wheel (as described in Section II-E).

[0151] A. Illustrative Loading of Second Substrate Material into Automated Clinical Analyzer

[0152] The following describe non-limiting, illustrative examples of devices that may be used to load the second substrate material into automatic clinical analyzer 700, as well as illustrative methods of loading the second substrate material into automated clinical analyzer 700.

[0153] i. Introducing Second Substrate Material into Automated Clinical Analyzer Using a Sample Rack

[0154] In some aspects, where a second substrate material 182 (FIGS. 14A-14F, 16A-16C, and 19A-19F) is used as the reporting reagent, a rack, also referred to herein as a sample rack, housing a second substrate material 182 may be used to introduce the second substrate material 182 to the clinical analyzer 700. The second substrate material 182 may be introduced via vessel, tube, cup, or container a sample rack, these terms are used interchangeably in the following disclosure. Additionally, or alternatively, a portion of the sample rack may be configured to accommodate a specific container intended to house the second substrate. FIGS. 9-12 illustrate exemplary sample racks for providing substrate to the clinical analyzer. The sample racks 800, 830, and 850 of FIGS. 9-12 may be loaded into the clinical analyzer 700 at the sample loading station 702 (FIG. 7) and / or presented to the clinical analyzer 700 by the adjacent automated machine 1700 (FIG. 7).

[0155] FIGS. 9 and 10 show an exemplary rack 800, also referred to herein as sample rack 800, that may be readily incorporated into clinical analyzer 700 described above that includes a configuration to accommodate a specific container intended to house the second substrate. Sample rack 800 may be configured to be readily loaded on sample loading station 702 of automated clinical analyzer 700 such that sample rack carriage 704 and sample pipetting unit 706 may suitably process vessels loaded onto sample rack 800 in accordance with the description herein.

[0156] As will be described in greater detail below, sample rack 800 may be configured to house a suitable substrate container 806 housing a second substrate material 182. As is described in greater detail below, sample pipetting unit 706 may suitably access the interior of substrate container 806 while sample rack 800 is loaded onto sample rack carriage 704 to obtain the second substrate material 182, dispense the second substrate material 182 into reaction vessel 105, mix the second substrate material 182 into reaction vessel 105, and / or dispense the second substrate material 182 into a portable vessel 106 (FIGS. 16A-16C and 19A-19F). In some embodiments, automated clinical analyzer 700 may temporarily store the acquired second substrate material 182 prior to use in a suitable assay in accordance with the description herein. Alternatively, in some embodiments, automated clinical analyzer 700 may use the acquired second substrate material in a suitable assay in accordance with the description herein without a storage step.

[0157] The sample rack 800 includes a base 801 having a plurality of vessel holders 802 extending upwardly from base 801, and at least one substrate container holder 804 also extending upwardly from base 801. While sample rack 800 in the current example is configured to hold five vessels 816, which may be patient sample vessels, and one substrate container 806, the sample rack 800 may be configured to hold any suitable number of substrate containers 806 and / or vessels. Two such examples are described in more detail with respect to FIGS. 11 and 12. In some examples, sample rack 800 may be configured to only hold a substrate container 806 (including just a single substrate container 806 or multiple substrate containers 806). In some examples, sample rack 800 may be configured to hold two or more substrate containers 806. In some instances, a vessel on a vessel holder 802 may house a suitable substrate material such that substrate container holder 804 is not present (e.g., FIG. 12). In some instances, each substrate container 806 housed by sample rack 800 may include a different substrate material. Therefore, while in the current example, use of second substrate material 182 is described in conjunction with sample rack 800, sample rack 800 may provide access to a variety of substrate materials, whereone of those substrate materials may be chosen over the other substrate materials (including first substrate material 180 housed within analyzer 700) to be the reporting agent for a particular assay.

[0158] Base 801 is configured to be received by sample rack carriage 704 of automated clinical analyzer 700. In the current example, a user may load sample rack 800 onto sample loading station by placing base 801 on the conveyor belt of sample loading station 702. Sample loading station 702 may then guide sample rack 800 toward sample rack carriage 704 such that sample rack carriage 704 receives sample rack 800 via suitable contact / engagement with base 801.

[0159] As shown in FIG. 9, each vessel holder 802 of the plurality of vessel holders 802 are configured to selectively receive a corresponding vessel. When sample rack 800 is loaded onto sample rack carriage 704 in accordance with the description herein, each loaded vessel is accessible by a pipette of automated clinical analyzer 700 (e.g., pipette 707 (FIGS. 14A-14F) of sample pipetting unit 706) while base 801 is received by sample rack carriage 704.

[0160] Each vessel holder 802 of the plurality of vessel holders 802 may include a pair of arms 807 that are resiliently biased toward each other. The biased nature of arms 807 may allow arms 807 to suitably receive a vessel such that arms 807 inhibit inadvertent removal of a vessel once suitably attached in vessel holder 802. Arms 807 may also suitably engage a received vessel to properly align to open of vessels with pipette 707 (see FIGS. 14A-14F) of sample pipetting unit 706. Arms 807 may be formed of any suitable material and may be biased via any suitable means.

[0161] Substrate container holder 804 is configured to selectively receive substrate container 806 that houses a suitable amount of second substrate material 182. When sample rack 800 is loaded onto sample rack carriage 704 in accordance with the description herein, substrate container 806 is accessible by a pipette of automated clinical analyzer 700 (e.g., pipette 707 (FIGS. 14A- 14F) of sample pipetting unit 706) while base 801 is received by sample rack carriage 704. In some instances, substrate container holder 804 may also include a pair of resilient arms that function in substantially similar manner of arms 807 described above but are configured for use with substrate container 806.

[0162] Sample rack 800 also includes a plurality of upwardly extending columns 805 that extend upwardly from base 801. Columns 805 may help structurally support vessels and / or substrate container 806 received by sample rack 800. Columns 805 may also act as structural support for arms 807. An exemplary rack and arms is described in International Patent ApplicationPublication No Wo 2018 / 232364, titled “APPARATUS AND METHOD FOR HANDLING SAMPLE CONTAINERS,” hereby incorporated by reference in its entirety.

[0163] As mentioned, substrate container holder 804 houses a substrate container 806. Substrate container 806 suitably houses second substrate material 182. When the automated clinical analyzer 700 receives rack 800, sample rack carriage 704 may align substrate container 806 with sample aliquoting pipette 707 of sample pipetting unit 706, thereby allowing sample pipetting unit 706 to obtain a desired amount of second substrate material 182 from container 806. As described in greater detail, sample pipetting unit 706 may directly add the obtained second substrate material 182 into a reaction vessel 105 to form signal generating mixture 177 (as described, for example, in Section II-B), or sample pipetting unit 706 may aliquot the second substrate material 182 into a portable vessel 106 (FIGS. 16A-16C and 19A-19F) that can be stored on the analyzer (as described, for example, in Section ILC). In instances where the second substrate material 182 is temporarily stored within suitable components of analyzer 700 prior to being used in an appropriate assay, analyzer 700 may use suitable mechanisms, such as pick and place devices, to transfer and store the portable vessel 106 to a suitable climate-controlled sample housing (such as sample wheel 710) or an incubation container / station (e.g., incubation wheel 750). Therefore, in instances where a particular assay requires use of second substrate material 182 to form signal generating mixture 177, clinical analyzer 700 may suitably access second substrate material 182 from the stored portable vessel 106 in accordance with Section II-D.

[0164] Substrate container 806 may also have a removable cap 808 configured to selectively attach to substrate container 806. Cap 808 may be removed from substrate container 806 to provide access to an interior defined by container 806 housing second substrate material 182. In some instances, before rack 800 is suitably loaded into sample rack carriage 704, a user may remove cap 808 from container 806 to allow sample pipetting unit 706 to suitably access second substrate material 182. In some instances, before rack 800 is suitably loaded into sample rack carriage 704, another instrument (on the analyzer 700, on the adjacent automated machine 1700, and / or on another adjacent device) may be used to remove cap 808 from container 806 to allow sample pipetting unit 706 to suitably access second substrate material 182.

[0165] Although illustrated and described in FIGS. 9 and 10 with the substrate container holder 804 located at one end of the sample rack 800, the substrate holder 804, and thus, the substrate container 806, may be located at any position of the sample rack 800. For example, thesubstrate holder 804 may be located at the opposing end of the sample rack 800 or anywhere in between the opposing end and the location depicted in FIGS. 9 and 10. Although illustrated and described in FIGS. 9 and 10 as a substrate container 806 having a cap 808, the cap 808 may be omitted. Furthermore, the substrate container 806 may have the same appearance (e.g., same size and shape) as the vessels. In such an example, the size and shape of the substrate container holder 804 may be the same or similar to the vessel holder 802. The size, shape, and dimensions of the vessels and the substrate container 806 are not limited to the depiction of FIGS. 9 and 10.

[0166] The analyzer 700 may include more than one sample rack 800. In examples with more than one sample rack 800, any number of the sample racks 800 may accommodate the substrate container 806. In some examples, only a single sample rack 800 includes the substrate container holder 804 and the substrate container 806. In some examples, all of the sample racks 800 include the substrate container holder 804 and the substrate container 806. In some examples, more than one but fewer than all of the sample racks 800 include the substrate container 804 and the substrate container 806.

[0167] FIGS. 11 and 12 illustrate alternative sample racks 830 and 850, respectively. For the sake of brevity, the above description is not repeated herein, but all features, functions, and optionality of the sample rack 800 is applicable to the sample racks 800 and 850. The only difference between the sample rack 800 and the sample racks 830 and 850 are the differences described below.

[0168] FIG. 11 illustrates the exemplary sample rack 830 arranged to accommodate only the substrate container 806 and not to accommodate the vessels. In the example of FIG. 11, three substrate holders 804 are depicted, but more or fewer may be provided.

[0169] FIG. 12 illustrates the exemplary sample rack 850 arranged to accommodate only the vessels 816 and not to accommodate the substrate container 806. In the example of FIG. 12, seven vessel holders 802 are depicted, but more or fewer may be provided. In some examples, only vessels 816 housing substrate (e.g., second substrate material 182) are provided in the rack 850, either completely or partially filling the rack 850. In some examples, one or more vessels 816 house the substrate and one or more vessels 816 contain a patient sample. The clinical analyzer is configured to detect whether the vessels 816 contain substrate (and which substrate) or patient sample.

[0170] ii. Introducing Second Substrate Material into Clinical Analyzer via Adjacent Automated Machine

[0171] Second substrate material 182 may be suitably loaded into analyzer 700 using other suitable means instead of, or in addition to, the use of rack 800 described above, such as an adjacent automated machine 1700. In some instances, as shown in FIG. 7, suitable portions of analyzer 700 may be in communication with other adjacent automated machines 1700. Adjacent automated machines 1700 may be configured to perform other tasks that are complementary to the tasks performed by analyzer 700 and / or identical to the tasks performed by analyzer 700. In the current example, adjacent automated machine 1700 includes vessel transfer station 1702 that may suitably transfer a vessel (e.g., reaction vessel 105, portable vessel 106, vessel 816 (FIGS. 16A-16C and 19A-19F), or a container substantially similar to substrate container 806) into the possession of clinical analyzer 700 via reaction vessel carriage 712. As referred to herein, a portable vessel is a vessel that may be transported through the clinical analyzer 700, for example, with a pick and place device. In some examples, the portable vessel is the reaction vessel 105 or has the same or similar structure as the reaction vessel 105. In some examples, the portable vessel 106 may by the vessel 816 provided in the sample rack 800, 830, 850.

[0172] FIG. 17 shows an illustrative method 1300 of analyzer 700 receiving a portable vessel 106 already filled with second substrate materials 182. Portable vessel 106 may be appropriately sized and configured for analyzer 700 to suitably store portable vessel 106 (such as within sample wheel 710 and / or incubation wheel 750). First, analyzer 700 may receive 1302 a portable vessel 106 already housing second substrate material 182 from adjacent automated machine 1700. Portable vessel 106 may obtain second substrate material 182 through any suitable means. After portable vessel 106 is suitably received by analyzer 700, analyzer 700 may directly utilize substrate material 182 within a suitable assay (as further described, for example, in Section II-B); or analyzer 700 may store second substrate material 182 for a suitable amount of time prior to utilizing second substrate material 182 in a suitable assay (as described, for example, in Section II-C).

[0173] In instances where the received portable vessel 106 is temporarily stored within suitable components of analyzer 700 prior to being used in an appropriate assay, analyzer 700 may use suitable mechanisms, such as pick and place devices, to transfer and store 1304 the received portable vessel 106 to a suitable climate-controlled sample housing (such as sample wheel 710) or an incubation container / station (e g., incubation wheel 750). Therefore, in instances where aparticular assay requires use of second substrate material 182 to form signal generating mixture 177, clinical analyzer 700 may suitably access second substrate material 182 from the stored portable vessel 106 in accordance with Section II-D.

[0174] In instances where the second substrate material 182 housed within portable vessel 106 is directly added into a reaction vessel 105 to form signal generating mixture 177, analyzer 700 may aliquot 1306 second substrate material 182 from the received portable vessel 106 directly into the reaction vessel 105 to create the signal generating mixture 177 without storing the portable vessel 106 prior to the creation of the signal generator mixture. Adding the second substrate material 182 directly may mean, for example, adding without transferring the second substrate material 182 into an additional vessel or container or otherwise storing the second substrate material 182 on the analyzer 700. Directly adding second substrate material 182 into reaction vessel 105 without storing the received portable vessel 106 may be substantially similar to the method described in Section II-B below, except the second substrate material 182 is obtained from the portable vessel 106 received from adjacent automated machine 1700 rather than rack 800.

[0175] Analyzer 700 may use any suitable means (e.g., programming, operator instructions, etc.) to decide whether to directly use or store second substrate material 182 received from adjacent automated machine 1700.

[0176] FIG. 21 shows an illustrative method 1310 of analyzer 700 receiving a container filled with second substrate material 182, where such a container may not be compatible for storage within clinical analyzer 700. In such cases, the second substrate material 182 may be obtained from the substrate container filled with second substrate material 182, and obtaining the second substrate material can include accessing a reservoir of the substrate container. First, analyzer 700 may receive 1312 a container already housing second substrate material 182 from adjacent automated machine 1700. Next, analyzer 700 may aliquot 1314 second substrate material 182 from the received container. Analyzer 700 may suitably aliquot 1314 second substrate material 182 from the received container in substantially similar manner that analyzer 700 aliquots second substrate material 182 from rack 800, as described in Section II-B and Section II-C described below; except that second substrate material 182 is within the container received by adjacent automated machine 1700 rather than rack 800. After second substrate material 182 is suitably aliquoted by analyzer 700, analyzer 700 may directly utilize substrate material 182 within a suitable assay; or analyzer700 may store second substrate material 182 for a suitable amount of time prior to utilizing second substrate material 182 in a suitable assay.

[0177] In instances where second substrate material 182 is to be stored within suitable components of analyzer 700 prior to being used in an appropriate assay, analyzer 700 may dispense the aliquoted second substrate material into a portable vessel 106 and transfer 1316 the portable vessel 106 to incubation wheel 750 or the sample wheel 720 for storage. Such dispensing and transferring 1316 may be substantially similar to the dispensing and transferring described in Section II-C, except the second substrate material 182 is taken from the container received by adjacent automated machine 1700 rather than rack 800.

[0178] In instances where the aliquoted second substrate material 182 may be directly added into a reaction vessel 105 to form signal generating mixture 177, analyzer 700 may dispense 1318 the recently aliquoted second substrate material 182 directly into the reaction vessel 105 to create the signal generating mixture 177 without storing the portable vessel 106 prior to the creation of the signal generator mixture 177. Adding the second substrate material 182 directly may mean, for example, adding without transferring the second substrate material 182 into an additional vessel or container or otherwise storing the second substrate material 182 on the analyzer 700. Directly adding second substrate material 182 in reaction vessel 105 without storing the received portable vessel 106 may be substantially similar to the method described in Section II-B below, except the second substrate material 182 is obtained from the container received from adjacent automated machine 1700 rather than rack 800.

[0179] Analyzer 700 may use any suitable means (e.g., programming, operator instructions, etc.) to decide whether to directly use or store second substrate material 182 received from adjacent automated machine 1700.

[0180] B. Illustrative Direct Use of Loaded Second Substrate Material by Automated Clinical Analyzer

[0181] As mentioned above, in some instances when a particular assay is designed to use second substrate material 182 as the reporting reagent, instead of the first substrate material 180, sample pipetting unit 706 may suitably access the interior of substrate container 806 or the interior of vessel 816 while sample rack 800, 830, 850 is loaded on sample rack carriage 704 to (A) obtain the second substrate material 182, and (B) dispense and / or mix the second substrate material 182 into the reaction vessel 105 (which contains particles 120, some of which are bound to both ananalyte of interest and the bound reporter of the second reagent 155) to create the signal generating mixture 177 (FIG. 1, stage 8). Such use is considered direct use of the second substrate material 182 when the second substrate material 182 is added to the reaction vessel 105 to create a signal generating mixture without being stored on the automated clinical analyzer 700. Adding the second substrate material 182 directly may mean, for example, adding without transferring the second substrate material 182 into an additional vessel or container or otherwise storing the second substrate material 182 on the analyzer 700. For example, the second substrate material 182 may be added to the reaction vessel 105 within four minutes of being loaded onto the automated clinical analyzer, within two minutes of being loaded onto the automated clinical analyzer, within one minute of being loaded onto the automated clinical analyzer, or within thirty seconds of being loaded onto the automated clinical analyzer. In another example, the second substrate material 182 may be added to the reaction vessel 105 at least eight seconds, at least ten seconds, at least twenty second, or at least thirty seconds after being loaded onto the automated clinical analyzer. In one exemplary embodiment, the second substrate material 182 may be added to the reaction vessel 105 to create a signal generating mixture at a time in a range of eight seconds to one minute after being loaded onto the automated clinical analyzer. In another exemplary embodiment, the second substrate material 182 may be added to the reaction vessel 105 to create a signal generating mixture at a time in a range of eight seconds to two minutes after being loaded onto the automated clinical analyzer.

[0182] FIG. 13 shows an illustrative method 1100 of using an automated clinical analyzer 700 in conjunction with sample rack 800, 830, 850 such that clinical analyzer 700 may select between using a first substrate material 180 or a second substrate material 182 for a particular assay. Further, FIGS. 14A-14F show an illustrative examples of sample pipetting unit 706 obtaining the second substrate material 182 from rack 800, 830, 850 and mixing the second substrate material 182 within reaction vessel 105 to create a signal generating mixture 177 (when second substrate material 182 is the selected reporting agent over first substrate material 180). Although the current example, the second substrate material 182 is obtained from rack 800, 830, 850, second substrate material 182 may alternatively be obtained from portable vessel 106 or another container which analyzer 700 received from adjacent automated machine 1700, as described in Section II-A-ii above.

[0183] Although not illustrated, the clinical analyzer 700 may receive and aliquot patient samples in substantially similar manner as steps 1002, 1004 of workflow 1000 described above,or utilizing any other suitable manner. Tn some instances, the analyte of interest from the patient sample may be obtained from sample aliquoting pipette 707 (FIGS. 14A-14F, 16A-16C, and 18A- 18F) of a patient sample aliquot system (i.e., sample pipetting unit 706). Sample aliquot pipette 707 may utilize a first disposable tip 709 to obtain the analyte from the patient sample, and then discard of the first disposable tip 709. Therefore, the manner in which an aliquot of a patient sample 165 is received may vary.

[0184] Next, clinical analyzer 700 may mix patient samples 165 with reagent(s) 1102, incubate mixtures 1104, and perform wash cycle(s) 1106 to prepare to create a mixture by adding a reporting agent (for example, first substrate material 180 or second substrate material 182) in accordance with the description herein (e.g., the assay 100 of FIG. 1). For sake of simplicity, the steps 1102, 1104, and 1106 of the current workflow may, in the aggregate, be substantially similar to steps 1006, 1008, 1010, 1012, 1014, 1016, 1018, 1020, in the aggregate, described above.

[0185] Therefore, analyzer 700 may perform steps 1102, 1104, 1106 to (A) suitably create an assay mixture 175 (FIG. 1) in accordance with the description herein, and then (B) wash the assay mixture 175 such that the particles 120, the bound analyte of the patient sample 165, and the bound reporter of the second reagent 155 remain in the reaction vessel 105 after washing is complete while other components of the assay mixture 175 may be absent from the reaction vessel 105 after the washing is complete. Therefore, steps 1102, 1104, 1106 may include (a) creating an assay mixture within a reaction vessel utilizing an analyte from a patient sample, a first reagent, and a second reagent, wherein the first reagent comprises a plurality of particles configured to specifically bind to the analyte from the patient sample, wherein the second reagent comprises a reporter; (b) incubating the assay mixture; and (c) performing a wash cycle on the assay mixture, wherein performing the wash cycle comprises retaining the plurality of particles while removing the remaining potions of the assay mixture not bound to the plurality of particles, wherein at least one particle of the plurality of particles is bound to a respective analyte from the patient sample.

[0186] Once steps 1102, 1104, 1106 are suitably completed in accordance with the description herein, reaction vessel 105 may be optionally located at the portion of wash wheel 270 configured to receive the first substrate material 180 via substrate dispensing system 725. Alternatively, reaction vessel 105 may be located at any other suitable location.

[0187] Once the contents of a reaction vessel 105 are configured to receive a reporting agent (e.g. either first substrate material 180 or second substrate material 182) in accordance with thedescription herein, the analyzer 700, based on provided programming or operator instructions, selects a substrate material 1108 (e.g., selecting either first substrate material 180 or second substrate material 182) to be used for the particular assay. Analyzer 700 may select 1108 between a first substrate material 180 and a second substrate material 182, where the first substrate material 180 may be housed within the clinical analyzer 700 (e.g., within substrate housing 720) and the second substrate material 182 may be located on sample rack 800, which may or may not be currently housed within sample rack carriage 704 at the time the selection is made.

[0188] Analyzer 700 may use any suitable means (e.g., programming, operator instructions, etc.) to decide which reporting agent (including, for example, first substrate material 180 or second substrate material 182) to utilize during a particular assay. For illustrative purposes, analyzer 700 may select reporting agent (including between at least first substrate material 180 and second substrate material 182) based on known parameter(s) of the patient sample, based on results of assays performed on another aliquot of the same patient sample, based on user input, suitable empirical data, etc. As one illustrative example, clinical analyzer 700 may perform a first assay with a first aliquot of a patient sample 165 with first substrate material 180 in accordance with the description herein; then clinical analyzer 700 may reflex to a second assay performed with a second aliquot from the same patient sample 165, yet with the second substrate material 182. It is important to understand that clinical analyzer 700 does not use both the first substrate material 180 and the second substrate material 182 in the same assay.

[0189] If the first substrate material 180 is selected 1110, clinical analyzer 700 adds and mixes 1112 the selected substate material (i.e., the first substrate material 180) into the reaction vessel 105 with the plurality of particles 120 at wash wheel 270 in accordance with the description herein to create the signal generating mixture 177. Next, clinical analyzer 700 transfers 1114 the reaction vessel 105 to the incubation wheel 750 and suitably incubates the signal generating mixture in accordance with the description herein. Next, clinical analyzer 700 transfers the reaction vessel to a detector (e.g., the luminometer 600) to analyze and / or measure 1116 light generated by the signal generating mixture within reaction vessel 105 in accordance with the description herein.

[0190] However, if the second substrate material 182 is selected 1120, clinical analyzer 700 transfers 1122 the reaction vessel 105 back to reaction vessel carriage 712, which then moves reaction vessel 105 to a suitable location to receive second substrate material 182 in accordancewith the description herein. Reaction vessel 105 may be transferred from wash wheel 270 toward reaction vessel carriage 712 via suitable pick and place device.

[0191] In the current example, as shown in FIGS. 14A-14F, reaction vessel carriage 712 includes an actuating vessel holder 713. Actuating vessel holder 713 is configured to actuate along a predetermined path between sample pipetting unit 706 and reagent mixing station 715. Additionally, actuating vessel holder 713 is configured to suitably house vessels (e.g., reaction vessels 105 and / or other portable vessels 106 as shown in FIGS. 16A-16C and 19A-19F) while actuating between various locations in the predetermined path to suitably align such vessels with various pipettes (e.g., sample aliquoting pipette 707 shown in FIGS. 14A-14F). Therefore, once reaction vessel 105 is transferred 1122 back to reaction vessel carriage 712, actuating vessel holder 713 may then move reaction vessel 105 to the location shown in FIG. 14A to prepare reaction vessel 105 to receive second substrate material 182.

[0192] In the current example, actuating vessel holder 713 may simultaneously house three vessels. However, actuating vessel holder 713 may be configured to simultaneously house and suitable number of vessels.

[0193] Before, during, or after reaction vessel 105 being transferred 1122 to reaction vessel carriage 712 and suitably positioned to receive second substrate material 182, sample aliquoting pipette 707 of sample pipetting unit 706 may couple 1124 to a suitable un-used disposable tip 709. Disposable tip 709 may be initially housed within disposable tip housing 708 and accessed via sample aliquoting pipette 707 via any suitable means and / or structures. Disposable tip 709 may be configured to selectively attach and detach from aliquoting pipette 707 such that aliquoting pipette 707 may utilize and discard disposable tip 709. While disposable tip 709 is coupled to sample aliquoting pipette 707, disposable tip 709 may be configured to aspirate / deliver suitable fluids in accordance with the description herein. Disposable tip 709 may be formed of any suitable material, such as a suitable plastic material.

[0194] The aliquoting pipette 707 of sample pipetting unit 706 may include suitable components of pipetting system 510 described above to function in accordance with the description herein. Aliquoting pipette 707 may be coupled to various suitable actuators and / or frames to move along at least two axes (e.g., ASPI and / or ASP2 and the vertical axis (VA)).

[0195] With disposable tip 709 coupled 1124 to sample aliquoting pipette 707, next, aliquoting pipette 707 may align 1126 disposable tip 709 with substrate container 806 on samplerack 800, as shown in FIG. 14A. Therefore, disposable tip 709 is located above substrate container 806 such that if disposable tip 709 were to be actuated toward container 806, tip 709 would be dipped into second substrate material 182.

[0196] As mentioned above, when sample rack 800, 830, 850 is suitably loaded into automated clinical analyzer 700, sample rack carriage 704 suitably actuates sample rack 800, 830, 580 and substrate container 806 or vessel 816 to the position shown in FIG. 14A. The sample rack 800, 830, 850 and substrate container 806 or vessel 816 may be loaded onto sample rack carriage 704 at any suitable time during and / or before the assay 1100. For example, sample rack 800, 830, 850 may be loaded into clinical analyzer 700 prior to the beginning as assay 1100 or during assay 1100. Further, removable cap 808 (FIG. 9) of container 806 or vessel 816 (which may also include a removable cap) may be removed such that the reservoir of second substrate material 182 is accessible by sample pipetting unit 706. In some instances, when sample rack 800, 830, 850 is not loaded into sample rack carriage 704 during assay 1100 when second substrate material 182 is the selected reporting agent, automated clinical analyzer 700 may generate a suitable alert signal, thereby indicating to a user that a sample rack 800 housing second substrate material 182 is required to assay 1100 to continue.

[0197] Next, with disposable tip 709 suitably aligned 1126 with substrate container 806 or vessel 816 housing a reservoir of the second substrate material 182, sample aliquoting pipette 707 may utilize disposable tip 709 to access the reservoir of the substrate container 806 or vessel 816 and aliquot 1128 second substrate material 182 from substrate container 806 or vessel 816. Therefore, sample pipetting unit 706 may be utilized to obtain the second substate material from the sample rack 800 on sample rack carriage 704. As shown between FIGS. 14A-14B, sample pipetting unit 706 may actuate sample aliquoting pipette 707 and disposable tip 709 in a direction parallel with vertical axis (VA) until disposable tip 709 is suitably dipped into second substrate material 182. Sample aliquoting pipette 707 may utilize disposable tip 709 to aliquot 1128 a suitable amount of substrate material 182. Sample aliquoting pipette 707 may aliquoted 1128 any suitable amount of substrate material 182 from container 806. Some non-limiting examples include at least 50 microliters, or at least 100 microliters, and up to 100 microliters, up to 200 microliters, up to 300 microliters, or up to 500 microliters.

[0198] Next, as shown in FIG. 14C, sample pipetting unit 706 may vertically actuate sample aliquoting pipette 707 and disposable tip 709 vertically away from container 806. Therefore, thesecond substrate material 182 is obtained from a rack 800 on a sample rack carriage 704 of the automated clinical analyzer 700. Although in the current example, the second substrate material 182 is obtained from container 806 on rack 800, second substrate material 182 may alternatively be obtained from portable vessel 106 or another container which analyzer 700 received from adjacent automated machine 1700 as described in Section II-A-ii . Further, the sample rack carriage 704 is configured to selectively receive and eject the rack 800.

[0199] Next as shown between FIGS. 14C-14D, sample pipetting unit 706 may actuate sample aliquoting pipette 707, disposable tip 709, and the second substrate material 182 contained within disposable tip 709, in a direction parallel with axis Aspl until disposable tip 709 and sample aliquoting pipette 707 are aligned with reaction vessel 105. Although the time shown in FIGS. 14A-14D, reaction vessel 105 contains a plurality of particles 120, some of which are bound to an analyte of interest as well as a suitable reporter, that are ready to receive a reporting reagent (e.g., second substrate material 182) to form a signal generating mixture 177.

[0200] Once sample aliquoting pipette 707 and disposable tip 709 are suitably aligned with reaction vessel 105 (whose position is controlled via actuating vessel holder 713 of reaction vessel carriage 712), pipette 707 and disposable tip 709 may be actuated vertically toward reaction vessel 105 in a direction parallel with the vertical axis (VA) to add and mix 1130 the second substrate material 182 with the particles 120 of reaction vessel 105 to form signal generating mixture 177 in accordance with the description herein. Sample aliquoting pipette 707 may add 1130 any suitable amount of substrate material 182 into reaction vessel 105. Some non-limiting examples include at least 50 microliters, or at least 100 microliters, and up to 100 microliters, up to 200 microliters, up to 300 microliters, or up to 500 microliters.

[0201] In the example shown in FIG. 14E, disposable tip 709 is actuated into the interior reaction vessel 105 to add and mix 1130 second substrate material 182 with the plurality of particles 120 (some of which are bound to an analyte of interest as well as a suitable reporter). Therefore, sample pipetting unit 706 may be used to directly add the second substrate material 182 into the reaction vessel 105 to create the signal generating mixture 177. Disposable tip 709 and sample aliquoting pipette 707 may use any suitable technique to suitably mix the contents of reaction vessel 105 to form signal generating mixture 177. For example, disposable tip 709 and sample aliquoting pipette 707 may utilize a push-pull motion technique as described in U.S. Patent Application Publication No. 2020 / 0264207, entitled, “Substance Dispense System for BiologicalSample Analysis Instrument,” published on August 20, 2020, hereby incorporated by reference in its entirety.

[0202] In instances where second substrate material 182 is directly added to create signal generating mixture 177, second substrate material 182 may be within analyzer 700 for less than one minute, less than two minutes, and / or less than five minutes before second substrate material 182 is added to reaction vessel 105.

[0203] In some instances, disposable tip 709 and sample aliquoting pipette 707 add second substrate material 182, but do not mix second substrate material 182 within reaction vessel 105 to form signal generating mixture 177. In such instances, other suitable components of analyzer 700 may be utilized to mix the content of reaction vessel 105 to form signal generating mixture 177.

[0204] After signal generating mixture 177 is formed, disposable tip 709 and aliquoting pipette 707 may be moved vertically out of reaction vessel 105, as shown in FIG. 14F. Subsequently, disposable tip 709 may be discarded. Therefore, in some instances, disposable tip 709 may be considered a single use disposable tip 709. Therefore, in some instances, a first distal tip 709 may be utilized to obtain the patient sample, while a second disposal tip 709 may be utilized to obtain the second substrate material. The disposable tip 709 is formed of a suitable material that does not interfere with the properties of second substrate material 182 after coming into contact with second substrate material 182. Therefore, exposure of second substrate material 182 to disposable tip 709 does not interfere with the ability of second substrate material’s 182 to suitably form signal generating mixture 177.

[0205] Next, clinical analyzer 700 transfers 1132 the reaction vessel 105 to the incubation wheel 750 and suitably incubates the mixture in accordance with the description herein. Reaction vessel 105 may be incubated within incubation wheel 750 for any suitable amount of time. For example, reaction vessel 105 may be incubated within incubation wheel 750 for 64 seconds, 192 seconds, etc. Clinical analyzer 700 may utilize a suitable pick and place device to obtain reaction vessel 105 from reaction vessel carriage 712 to transfer reaction vessel 105 to incubation wheel 750. Next, clinical analyzer 700 transfers the reaction vessel 105 to the luminometer 600 (or any other suitable detector) and analyzes 1134 the mixture containing the selected substrate with a detector (e.g., measures the light generated by the mixture within reaction vessel 105 in accordance with the description herein).

[0206] Therefore, the workflow 1100 utilizing clinical analyzer 700 in conjunction with sample rack 800 and / or substrate container 806 may allow for selective use of either a first substrate material 180 or a second substrate material 182 for a particular assay without requiring any type of manual adjustments of clinical analyzer 700.

[0207] C. Illustrative Loading of Second Substrate Material for Temporary Storage by Automated Clinical Analyzer

[0208] In some instances, it may be desirable to obtain second substrate material 182 from sample rack 800, 830, 850 without directly adding second substrate material 182 from sample rack 800, 830, 850 into a reaction vessel 105 to form signal generating mixture 177. For instance, in some examples, it may be desirable to aliquot second substrate material 182 into a dedicated portable vessel 106 (or otherwise receive a second substrate material 182 housed within a dedicated portable vessel 106), and then temporarily store the dedicated portable vessel 106 within a suitable container of analyzer (e.g., sample wheel 710 or incubation wheel 750). Subsequently in such instances, once needed, the stored second substrate material 182 may be acquired from the dedicated portion vessel 106 and dispensed / mixed into the reaction vessel 105 to form signal generating mixture 177.

[0209] FIG. 15 shows an illustrative method 1200 of using sample rack 800, 830, 850 sample pipetting unit 706, and reaction vessel carriage 712 to transfer second substrate material 182 from substrate container 806 of sample rack 800 (for the sake of simplicity, the description refers only to container 806 and sample rack 800, but the description also covers vessel 816 and sample racks 830, 850) into a dedicated substrate housing vessel in the form of portable vessel 106; while FIGS. 16A-16C show an illustrative example of sample pipetting unit 706 depositing an aliquot of second substrate material 182 (acquired from sample rack 800) into dedicated portable vessel 106. The portable vessel 106 may be substantially similar to reaction vessel 105 described above, except portable vessel 106 is intended to act as a dedicated housing for second substrate material 182 rather than be utilized to form a signal generating mixture 177.

[0210] As shown in FIGS. 16A-16C, portable vessel 106 is suitably coupled to actuating vessel holder 713 of reaction vessel carriage 712. A suitable pick and place device may be utilized to place an empty portable vessel 106 on actuating vessel holder 713. Analyzer 700 may obtain and place an empty portable vessel 106 onto actuating vessel holder 713 at any suitable time during or prior to method 1200.

[0211] Method 1200 includes analyzer 700 receiving 1202 sample rack 800 and substrate container 806 utilizing sample rack carriage 704 in accordance with the description herein. Sample rack carriage 704 may actuate sample rack 800 and substrate container 806 to a location accessible by sample pipetting unit 706 in accordance with the description herein. As mentioned above, container 806 houses a reservoir of second substrate material 182. Further, removable cap 808 of container 806 may be removed such that the reservoir of second substrate material 182 is accessible by sample pipetting unit 706.

[0212] Before, during, or after sample rack 800 being received 1202 by sample rack carriage 704 of an analyzer 700, sample aliquoting pipette 707 of sample pipetting unit 706 may couple 1204 to a suitable un-used disposable tip 709. Coupling of sample aliquoting pipette 707 and disposable tip 709 may be substantially similar to the coupling of such two components described above with reference to method 1100. Therefore, once coupled together, sample aliquoting pipette 707 may utilize disposable tip 709 to aliquot and dispense second substrate material 182 in accordance with the description herein.

[0213] With disposable tip 709 coupled 1204 to sample aliquoting pipette 707, next, aliquoting pipette 707 may align 1206 disposable tip 709 with substrate container 806 on sample rack 800. Therefore, disposable tip 709 is located above substrate container 806 such that if disposable tip 709 were to be actuated toward container 806, tip 709 would be dipped into second substrate material 182.

[0214] Next, with disposable tip 709 suitably aligned 1206 with substrate container 806, sample aliquoting pipette 707 may utilize disposable tip 709 to aliquot 1208 second substrate material 182 from substrate container 806. Sample aliquoting pipette 707 may aliquoted 1208 any suitable amount of substrate material 182 from container 806. Some non-limiting examples include at least 50 microliters, or at least 100 microliters, and up to 100 microliters, up to 200 microliters, up to 300 microliters, or up to 500 microliters.

[0215] Once second substrate material 182 is suitable aliquoted, sample pipetting unit 706 may vertically actuate sample aliquoting pipette 707 and disposable tip 709 vertically away from container 806. Therefore, the second substrate material 182 is obtained from a rack 800 on a sample rack carriage 704 of the automated clinical analyzer 700. Further, the sample rack carriage 704 is configured to selectively receive and eject the rack 800. Next as shown in FIG. 16A, sample pipetting unit 706 may actuate sample aliquoting pipette 707, disposable tip 709, and the secondsubstrate material 182 contained within disposable tip 709, in a direction parallel with axis Aspl until disposable tip 709 and sample aliquoting pipette 707 are aligned with portable vessel 106.

[0216] Aligning 1206 of disposable tip 709 with substrate container 806, inserting disposable tip 709 into substrate container 806, aliquoting 1208 of second substrate material 182 from substrate container 806, and subsequent removing of disposable tip 709 from substrate container 806 is not shown in the FIG. 16 series, as such functionality may be substantially similar to that shown in FIGS. 14A-14C shown above. Although in the current example, the second substrate material 182 is obtained from container 806 on rack 800, second substrate material 182 may alternatively be obtained from portable vessel 106 or another container which analyzer 700 received from adjacent automated machine 1700 as described in Section II-A-ii.

[0217] As shown between FIG. 16A-16B, once sample aliquoting pipette 707 and disposable tip 709 are suitably aligned with portable vessel 106 (whose position is controlled via actuating vessel holder 713 of reaction vessel carriage 712), pipette 707 and disposable tip 709 may be actuated vertically toward portable vessel 106 in a direction parallel with the vertical axis (VA) such that disposable tip 709 is within an interior of portable vessel 106. Once suitably positioned, such as at the position shown in FIG. 16B, pipette 707 and disposable tip 709 may, in conjunction with each other, dispense and / or add 1210 the aliquoted second substrate 182 into portable vessel 106, and then vertically actuate away from portable vessel 106 as shown in FIG. 16C. Sample aliquoting pipette 707 may dispense 1210 any suitable amount of substrate material 182 into portable vessel 106. Some non-limiting examples include at least 50 microliters, or at least 100 microliters, and up to 100 microliters, up to 200 microliters, up to 300 microliters, or up to 500 microliters .

[0218] Subsequently, disposable tip 709 may be discarded. Therefore, in some instance, disposable tip 709 may be considered a single use disposable tip 709. Therefore, in some instances, a first distal tip 709 may be utilized to obtain the patient sample, while a second disposal tip 709 may be utilized to obtain the second substrate material. In some instances, the second substrate material is obtained prior to the patient analyte being obtained. The disposable tip 709 is formed of a suitable material that does not interfere with the properties of second substrate material 182 after coming into contact with second substrate material 182. Therefore, exposure of second substrate material 182 to disposable tip 709 does not interfere with the ability of second substrate material’s 182 to suitably form signal generating mixture.

[0219] Next, after portable vessel 106 has suitably received second substrate material 182, the second substrate material 182 may be used directly, or the portable vessel 106 may be transferred 1212 and stored in a suitable climate-controlled sample housing (such as sample wheel 710) or an incubation container / station (e.g., incubation wheel 750). Adding the second substrate material 182 directly may mean, for example, adding without transferring the second substrate material 182 into an additional vessel or container or otherwise storing the second substrate material 182 on the analyzer 700. In some examples, adding the second substrate material 182 directly may include proceeding with steps 1128, 1130, 1132, and 1134 as described in FIG. 11 to add the second substrate material 182 to the reaction vessel 105 and measure the output response. As illustrative examples, portable vessel 106 and second substrate material 182 may be stored at 4 degrees Celsius (e.g., within sample wheel 710), 37 degrees Celsius (e.g., within incubation wheel 750), or any other suitable temperature.

[0220] As will be described in greater detail below, when second substrate material 182 is the selected substrate material for an illustrative assay, portable vessel 106 housing second substrate material 182 may be transported to a suitable location such that automated clinical analyzer 700 may aliquot second substrate material 182 from portable vessel 106 and deposit the aliquoted second substrate material 182 into reaction vessel 105 to create signal generating mixture 177 in accordance with the description herein. Alternatively, if substrate material 182 has been stored for longer than a predetermined amount of time, portable vessel 106 and second substrate material 182 may be discarded. In some instances, second substrate material 182 may be stored on machine for at least five minutes, at least ten minutes, at least fifteen minutes, at least twenty minutes, at least one hour, at least five hours, at least ten hours, or at least twelve hours. In some instances, second substrate material 182 may be stored on machine for up to ten minutes, up to fifteen minutes, up to twenty minutes, up to one hour, up to five hours, up to ten hours, up to twelve hours, or up to twenty -four hours.

[0221] D. Illustrative Use of Temporarily Stored Second Substrate Material by Automated Clinical Analyzer at Reaction Vessel Carriage

[0222] FIG. 18 depicts an illustrative method 1400 of using an automated clinical analyzer 700 in conjunction with portable vessel 106 housing second substrate material 182 such that clinical analyzer 700 may select between using a first substrate material 180 or a second substrate material 182 for a particular assay. Further, FIGS. 19A-19F show an illustrative example of samplepipetting unit 706 obtaining the second substrate material 182 from portable vessel 106 and mixing the second substrate material 182 within reaction vessel 105 to create a signal generating mixture 177 (when second substrate material 182 is the selected reporting agent over first substrate material 180). The second substrate material 182 stored within portable vessel 106 may have been obtained by analyzer 700 (that is, loaded and / or introduced into the analyzer 700) from a rack (as described in Section II-A-i), from an adjacent automated machine (as described in Section II-A-ii), or any other suitable means.

[0223] First, while not shown, the clinical analyzer 700 may receive and aliquot patient samples in substantially similar manner as steps 1002, 1004 of workflow 1000 described above, or utilizing any other suitable manner. In some instances, the analyte of interest from the patient sample may be obtained from sample aliquoting pipette 707 (FIGS. 14A-14F, 16A-16C, and 19A- 19F) of a patient sample aliquot system (i.e., sample pipetting unit 706). Sample aliquot pipette 707 may utilize a first disposable tip 709 to obtain the analyte from the patient sample, and then discard of the first disposable tip 709. Therefore, the manner in which an aliquot of a patient sample 165 is received may vary.

[0224] Next, clinical analyzer 700 may mix patient samples with reagent(s) 1402, incubate mixtures 1404, and perform wash cycle(s) 1406 to prepare to create a mixture by adding a reporting agent (for example, first substrate material 180 or second substrate material 182) in accordance with the description herein. For sake of simplicity, the steps 1402, 1404, and 1406 of the current workflow may, in the aggregate, be substantially similar to steps 1006, 1008, 1010, 1012, 1014, 1016, 1018, 1020, in the aggregate, described above.

[0225] Therefore, analyzer 700 may perform steps 1402, 1404, 1406 to (A) suitably create an assay mixture 175 in accordance with the description herein, and then (B) wash the assay mixture 175 such that the particles 120, the bound analyte of the patient sample 165, and the bound reporter of the second reagent 155 remain in the reaction vessel 105 after washing is complete while other components of the assay mixture 175 may be absent from the reaction vessel 105 after the washing is complete. Therefore, steps 1402, 1404, 1406 may include (a) creating an assay mixture within a reaction vessel utilizing an analyte from a patient sample, a first reagent, and a second reagent, wherein the first reagent comprises a plurality of particles configured to specifically bind to the analyte from the patient sample, wherein the second reagent comprises a reporter; (b) incubating the assay mixture; and (c) performing a wash cycle on the assay mixture, wherein performing thewash cycle comprises retaining the plurality of particles while removing the remaining potions of the assay mixture not bound to the plurality of particles, wherein at least one particle of the plurality of particles is bound to a respective analyte from the patient sample.

[0226] Once steps 1402, 1404, 1406 are suitably completed in accordance with the description herein, reaction vessel 105 may be optionally located at the portion of wash wheel 270 configured to receive the first substrate material 180 via substrate dispensing system 725. Alternatively, reaction vessel 105 may be located at any other suitable location.

[0227] Once the contents of a reaction vessel 105 are configured to receive a reporting agent (e.g. either first substrate material 180 or second substrate material 182) in accordance with the description herein, the analyzer 700, based on provided programming or operator instructions, selects a substrate material 1408 (e.g., selecting either first substrate material 180 or second substrate material 182) to be used for the particular assay. Analyzer 700 may select 1408 between a first substrate material 180 and a second substrate material 182, where the first substrate material 180 may be housed within the clinical analyzer 700 (e.g., within substrate housing 720) and the second substrate material 182 may be located in portable vessel 106 that is housed within suitable storage areas (e.g., sample wheel 710 or incubation wheel 750) of analyzer 700 or other suitable areas (e.g., adjacent automated machines 1700).

[0228] Analyzer 700 may use any suitable means (e.g., programming, operator instructions, etc.) to decide which reporting agent (including, for example, first substrate material 180 or second substrate material 182) to utilize during a particular assay. For illustrative purposes, analyzer 700 may select reporting agent (including between at least first substrate material 180 and second substrate material 182) based on known parameter(s) of the patient sample, based on results of assays performed on another aliquot of the same patient sample, based on user input, suitable empirical data, etc. As one illustrative example, clinically analyzer 700 may perform a first assay with a first aliquot of a patient sample 165 with first substrate material 180 in accordance with the description herein; then clinical analyzer 700 may reflex to a second assay performed with a second aliquot from the same patient sample 165, yet with the second substrate material 182. It is important to understand that clinical analyzer 700 does not use both the first substrate material 180 and the second substrate material 182 in the same assay.

[0229] If the first substrate material 180 is selected 1410, clinical analyzer 700 adds and mixes 1412 the selected substate material (i.e., the first substrate material 180) into the reaction vessel105 with the plurality of particles 120 at wash wheel 270 in accordance with the description herein to create the signal generating mixture 177. Next, clinical analyzer 700 transfers 1414 the reaction vessel 105 to the incubation wheel 750 and suitably incubates the signal generating mixture in accordance with the description herein. Next, clinical analyzer 700 transfers the reaction vessel to a detector (e.g., the luminometer 600) to analyze and / or measure 1416 light generated by the signal generating mixture within reaction vessel 105 in accordance with the description herein.

[0230] However, if the second substrate material 182 is selected 1420, clinical analyzer 700 transfers 1422 the reaction vessel 105 back to reaction vessel carriage 712, which then moves reaction vessel 105 to a suitable location to receive second substrate material 182 in accordance with the description herein. Further, clinical analyzer 700 also transfers 1422 portable vessel 106 containing the second substrate material 182 back to reaction vessel carriage 712. Reaction vessel 105 may be transferred from wash wheel 270 toward reaction vessel carriage 712 via suitable pick and place devices; while portable vessel 106 may be transferred toward reaction carriage 712 via suitable pick and place devices as well. As mentioned above, in the current example, actuating vessel holder 713 is configured to hold three different vessels. Therefore, actuating vessel holder 713 may simultaneously hold both portable vessel 106 and reaction vessel 105 and shown in FIGS. 19A-19F. Actuating vessel holder 713 holds both reaction vessel 106 and portable vessel 106 such that sample pipetting unit 706 may suitably access each vessel 105, 106 in accordance with the description herein.

[0231] Before, during, or after reaction vessel 105 and portable vessel 106 being transferred 1422 to reaction vessel carriage 712 and suitably positioned to be accessed by sample pipetting unit 706, sample aliquoting pipette 707 of sample pipetting unit 706 may couple 1424 to a suitable un-used disposable tip 709. Disposable tip 709 may be initially housed within disposable tip housing 708 and accessed via sample aliquoting pipette 707 via any suitable means and / or structures.

[0232] With disposable tip 709 coupled 1424 to sample aliquoting pipette 707, next, aliquoting pipette 707 may align 1426 disposable tip 709 with portable vessel 106 on actuating vessel holder 713, as shown in FIG. 19A. Therefore, disposable tip 709 is located above aliquoting pipette 707 such that if disposable tip 709 were to be actuated toward portable vessel 106, tip 709 would be dipped into second substrate material 182.

[0233] Next, with disposable tip 709 suitably aligned 1426 with portable vessel 106, sample aliquoting pipette 707 may utilize disposable tip 709 to aliquot 1428 second substrate material 182 from portable vessel 106. Therefore, the second substate material 182 may be obtained from portable vessel 106. As shown between FIGS. 19A-19B, sample pipetting unit 706 may actuate sample aliquoting pipette 707 and disposable tip 709 in a direction parallel with vertical axis (VA) until disposable tip 709 is suitably dipped into second substrate material 182. Sample aliquoting pipette 707 may utilize disposable tip 709 to aliquot 1428 a suitable amount of substrate material 182, and then, as shown in FIG. 16C, sample pipetting unit 706 may vertically actuate sample aliquoting pipette 707 and disposable tip 709 vertically away from portable vessel 106. Therefore, the second substrate material 182 is obtained from portable vessel 106. Sample aliquoting pipette 707 may aliquot 1428 any suitable amount of substrate material 182 from portable vessel 106. Some non-limiting examples include at least 50 microliters, or at least 100 microliters, and up to 100 microliters, up to 200 microliters, up to 300 microliters, or up to 500 microliters.

[0234] Next as shown between FIGS. 19C-19D, sample pipetting unit 706 may actuate sample aliquoting pipette 707, disposable tip 709, and the second substrate material 182 contained within disposable tip 709, in a direction parallel with axis Al until disposable tip 709 and sample aliquoting pipette 707 are aligned with reaction vessel 105. At the time shown in FIGS. 19A-19D, reaction vessel 105 contains a plurality of particles 120, some of which are bound to an analyte of interest as well as a suitable reporter, that are ready to receive a reporting reagent (e.g., second substrate material) to form a signal generating mixture 177.

[0235] Once sample aliquoting pipette 707 and disposable tip 709 are suitably aligned with reaction vessel 105 (whose position is controlled via actuating vessel holder 713 of reaction vessel carriage 712), pipette 707 and disposable tip 709 may be actuated vertically toward reaction vessel 105 in a direction parallel with the vertical axis (VA) to add and mix 1430 the second substrate material 182 with the particles 120 of reaction vessel 105 to form signal generating mixture 177 in accordance with the description herein. Therefore, the obtained second substrate material 1872 may be dispensed into the reaction vessel 105 to create the signal generating mixture 177. Sample aliquoting pipette 707 may add 1430 any suitable amount of substrate material 182 into reaction vessel 105. Some non-limiting examples include at least 50 microliters, or at least 100 microliters, and up to 100 microliters, up to 200 microliters, up to 300 microliters, or up to 500 microliters.

[0236] Tn the example shown in FIG. 19E, disposable tip 709 is actuated into the interior reaction vessel 105 to add / dispense and mix 1430 second substrate material 182 with the plurality of particles 120 (some of which are bound to an analyte of interest as well as a suitable reporter). In the current example, the portable vessel 106 and the reaction vessel 105 are possessed at the reaction build station (i.e., the reaction vessel carriage 712) while the second substrate material 182 is dispensed into the reaction vessel 105. Disposable tip 709 and sample aliquoting pipette 707 may use any suitable technique to suitably mix the contents of reaction vessel 105 to form signal generating mixture 177. For example, disposable tip 709 and sample aliquoting pipette 707 may utilize a push-pull motion technique as described in U.S. Patent Application Publication 2020 / 0264207, entitled, “Substance Dispense System for Biological Sample Analysis Instrument,” published on August 20, 2020, hereby incorporated by reference in its entirety.

[0237] In some instances, disposable tip 709 and sample aliquoting pipette 707 add second substrate material 182, but do not mix second substrate material 182 within reaction vessel 105 to form signal generating mixture 177. In such instances, other suitable components of analyzer 700 may be utilized to mix the content of reaction vessel 105 to form signal generating mixture 177.

[0238] After signal generating mixture 177 is formed, disposable tip 709 and aliquoting pipette 707 may be moved vertically out of reaction vessel 105, as shown in FIG. 19F. Subsequently, disposable tip 709 may be discarded. Therefore, in some instances, disposable tip 709 may be considered a single use disposable tip 709. Therefore, in some instances, a first distal tip 709 may be utilized to obtain the patient sample, while a second disposal tip 709 may be utilized to obtain the second substrate material. The disposable tip 709 is formed of a suitable material that does not interfere with the properties of second substrate material 182 after coming into contact with second substrate material 182. Therefore, exposure of second substrate material 182 to disposable tip 709 does not interfere with the ability of second substrate material’s 182 to suitably form signal generating mixture 177.

[0239] Next, clinical analyzer 700 transfers 1432 the reaction vessel 105 to the incubation wheel 750 and suitably incubates the mixture in accordance with the description herein. Reaction vessel 105 may be incubated within incubation wheel 750 for any suitable amount of time. For example, reaction vessel 105 may be incubated within incubation wheel 750 for 64 seconds, 192 seconds, etc. Clinical analyzer 700 may utilize a suitable pick and place device to obtain reaction vessel 105 from reaction vessel carriage 712 to transfer 1432 reaction vessel 105 to incubationwheel 750. Next, clinical analyzer 700 transfers the reaction vessel 105 to the luminometer 600 (or any other suitable detector) and analyzes 1434 the mixture containing the selected substrate with a detector (e.g., measures the light generated by the mixture within reaction vessel 105 in accordance with the description herein).

[0240] Therefore, the workflow 1400 utilizing clinical analyzer 700 in conjunction with portable vessel 106 housing second substrate material 182 may allow for selective use of either a first substrate material 180 or a second substrate material 182 for a particular assay without requiring any type of manual adjustments of clinical analyzer 700.

[0241] E. Illustrative Use of Second Substrate Material by Automated Clinical Analyzer at Wash Wheel

[0242] While reaction vessel carriage 712 and sample pipetting unit 706 were used in workflow 1400 described above to obtained second substrate material 182 from portable vessel 106 and dispend the second substrate material 182 into reaction vessel 105, when appropriate, this is merely optional. Any other suitable mechanism of analyzer 700 may be utilized to obtain second substrate material 182 from portable vessel 106 and dispense the second substrate material 182 into reaction vessel 105.

[0243] FIG. 20 an illustrative method 1500 of using an automated clinical analyzer 700 in conjunction with portable vessel 106 housing second substrate material 182 such that clinical analyzer 700 may select between using a first substrate material 180 or a second substrate material 182 for a particular assay. However, in instance where second substrate material 182 is the selective reporting reagent, analyzer 700 utilizes wash wheel 270 in accordance with the description herein to (A) obtain second substrate material 182 from portable vessel 106, and (B) dispense second substrate material 182 into reaction vessel 105 to create signal generating mixture 177.

[0244] Although not illustrated, the clinical analyzer 700 may receive and aliquot patient samples in substantially similar manner as steps 1002, 1004 of workflow 1000 described above, or utilizing any other suitable manner. Therefore, the manner in with an aliquot of a patient sample 165 is received may vary. Additionally, since the differences between method 1500 and method 1400 described above are when the second substrate material 182 is selected 1520; the steps 1502, 1504, 1506, 1508, 1510, 1512, 1514, 1516 may be substantially similar to steps 1402, 1404, 1406, 1408, 1410, 1412, 1414, 1416, described above, respectively.

[0245] Therefore, the description of method 1500 shown in FIG. 20 will start in instances where second substrate material 182 is selected 1520. If second substrate material 182 is the selected 1520 reporting agent, analyzer 700 transfers 1522 portable vessel 106 housing second substrate material 182 to wash wheel 270. The second substrate material 182 stored within portable vessel 106 may have been obtained by analyzer 700 (that is, loaded and / or introduced into the analyzer 70)0) from a rack (as described in Section II-A-i), from an adjacent automated machine (as described in Section II-A-ii), or any other suitable means.

[0246] Analyzer 700 may utilize any suitable combination of pick and place devices, as well as reaction vessel carriage 712, to transport portable vessel 106 from its housed location (e.g., sample wheel 710, incubation wheel 750, etc.) onto wash wheel 270. Next, analyzer 700 transfers 1524 reaction vessel 105 housing particles 120 ready to mix with a reporting agent (e.g., second substrate material 182) to a suitable station S on wash wheel 270 such that reaction vessel 105 trails portable vessel 106 housing second substrate material 182.

[0247] Next, wash wheel 270 uses a suitable pipetting station at a suitable station S to aliquot 1526 second substrate material 182 from portable vessel 106. Wash wheel 270 may aliquot 1526 any suitable amount of substrate material 182 from portable vessel 106. Some non-limiting examples include at least 50 microliters, or at least 100 microliters, and up to 100 microliters, up to 200 microliters, up to 300 microliters, or up to 500 microliters.

[0248] Next, wash wheel 270 rotates 1528 reaction vessel 105 into the station S of wash wheel 270 possessing the recently aliquoted second substrate material 182. Next, wash wheel 270 adds / dispenses and mixes 1530 the second substrate material 182 previous obtained from portable vessel 106 into reaction vessel 105 to create the signal generating mixture 177. Wash wheel 270 may add / dispense 1530 any suitable amount of substrate material 182 into reaction vessel 105. Some non-limiting examples include Some non-limiting examples include at least 50 microliters, or at least 100 microliters, and up to 100 microliters, up to 200 microliters, up to 300 microliters, or up to 500 microliters.

[0249] In the current example, portable vessel 106 and reaction vessel 105 are both located at the wash wheel 270 while the second substrate material 182 is dispensed into the reaction vessel 105. The wash wheel 270 may contain any suitable components to aliquot and dispense second substrate material 182 to create signal generating mixture 177.

[0250] Next, clinical analyzer 700 transfers 1532 the reaction vessel 105 to the incubation wheel 750 and suitably incubates the mixture in accordance with the description herein. Reaction vessel 105 may be incubated within incubation wheel 750 for any suitable amount of time. For example, reaction vessel 105 may be incubated within incubation wheel 750 for 64 seconds, 192 seconds, etc. Next, clinical analyzer 700 transfers the reaction vessel to the luminometer 600 and measures 1532 to light generated by the mixture within reaction vessel 105 in accordance with the description herein.

[0251] Accordingly, the present application describes different systems, apparatus, and methods for introducing a substrate into a clinical analyzer. Although described in the context of providing a second substrate material 182, the systems, apparatus, and methods described herein may provide the first substrate material 180 to the clinical analyzer in addition to the second substrate material 182 or instead of the second substrate material 182. As described herein, the substrate material may be provided to the clinical analyzer by way of a sample rack, housing either or both of a substrate container or a vessel containing substrate. In some examples, the substrate is provided to the clinical analyzer via an adjacent automated system (e.g., 1700), such as an automated sample presentation line that moves between multiple analyzers or instruments. In such examples, a pick and place within the clinical analyzer 700 may pipette the substrate directly from the adjacent automated system or may receive a vessel containing the substrate from the adjacent automated system.

[0252] III. Illustrative First and Second Substrate Materials

[0253] Second substrate material 182 has different properties compared to first substrate material 180. As mentioned above, second substrate material 182 may have various advantages and disadvantages compared to first substrate material 180 such that some assays will be better suited using first substrate material 180 as a reporting agent, while other assays will be better suited using second substrate material 182 as a reporting agent. Therefore, in certain examples, sample rack 800 may enable analyzer 700 to select between at least the first substrate material 180 or the second substrate material 182 to serve as the reporting agent for a particular assay, where the selected reporting agent may be better suited for a particular assay compared to the reporting agent not selected.

[0254] Reporting agents (including, for example, first substrate material 180 and second substrate material 182) may include any suitable characteristics that would be apparent to oneskilled in the art in view of the teachings herein. In one illustrative example, substrate material, for example, the first substrate material 180, may include(a) a compound comprising a compound of formula I or a salt thereof:whereinA is Ci-6haloalkyl, naphthyl, phenyl, substituted phenyl, or heteroaryl, wherein substituted phenyl comprises from 1 to 3 halo, Ci-6 alkyl, Ci-6 alkoxy, Ci-6 haloalkyl, C(0)Ri5, CN or NO2 substituents;Ri is selected from the group consisting of Cs-naryl, Ci-6 alkyl, Ci-6 haloalkyl, and C5- 14 aralkyl groups;R7-R14 are independently H, Ci-6 alkoxy, halo, Ci-4alkyl, or R7 -Rs or Rs -R9 or R9-R10 R11-R12 or R12-R13 or R13-R14, can be joined together as a carbocyclic or heterocyclic ring system comprising at least one 5 or 6-membered ring;Ri5 is Ci-6 alkyl; each M is independently selected from H, or an alkali metal, alkaline earth metal, transition metal, ammonium, phosphonium, organic amine salt, and an amino acid salt;Z is O or S; and n is 0, 1, or 2;(b) a cationic aromatic compound,(c) a background reducing agent, and(d) an ether-linked nonionic surfactant or a hydrophilic polymer or a combination thereof.Such a substrate may be further configured in accordance with the teachings of International Patent Application Publication No. WO 2018 / 006059, entitled “Chemiluminescent Substrates,” published on January 4, 2018, hereby incorporated by reference in its entirety.

[0255] Additionally, or alternatively, substrate material, for example, the second substrate material 182, may include a composition comprising a compound of Formula II and salts thereof:(Formula II) wherein each of R1and R2is independently C3-C10 alkyl, or R1and R2taken together with the carbon to which they are attached provide a C5-C10 cycloalkyl ring; R3is C1-C10 alkyl, Ce-Cio aryl, or heteroaryl;R4is C2-C10 alkenyl; R3is H or C1-C10 alkyl; X is a phosphate; and at least one surfactant. For example, the substrate material(s) may include a composition comprising(Formula II).Such a composition comprising a compound of Formula II may be further configured in accordance with the teachings of International Patent Application No. PCT / US2023 / 071231, entitled “CHEMILUMINESCENT REAGENTS FOR DETECTION OF ALKALINE PHOSPHATASES,” hereby incorporated by reference in its entirety. The composition comprising a compound of Formula II may further include a phosphonium surfactant. Examples of polymeric phosphonium surfactants include polyvinyl type polymers with pendant quaternary phosphonium groups, which are disclosed in U.S. Patent No. 5,393,469. compositions including formula II and a phosphonium surfactant are further described in PCT Application No. PCT / US2023 / 071231.

[0256] IV. Additional Examples

[0257] To further illustrate potential implementations and embodiments of the disclosed technology, exemplary systems and methods which could be practiced based on this disclosure are set forth below.

[0258] Example 1

[0259] A method of performing an immunoassay with an automated clinical analyzer, the method comprising: (a) creating an assay mixture within a reaction vessel, the assay mixture comprising an analyte from a patient sample, a first reagent, and a second reagent, wherein the first reagent comprises a plurality of particles configured to specifically bind to the analyte from the patient sample, wherein the second reagent comprises a reporter; (b) incubating the assay mixture; (c) performing a wash cycle on the assay mixture, wherein performing the wash cycle comprises retaining the plurality of particles while removing the remaining potions of the assay mixture not bound to the plurality of particles, wherein at least one particle of the plurality of particles is bound to a respective analyte from the patient sample; (d) selecting a first substrate material or a second substrate material; (e) adding and mixing the selected substrate material into the reaction vessel with the plurality of particles remaining from the wash cycle to create a signal generating mixture; (f) incubating the signal generating mixture; and (g) analyzing the signal generating mixture with a detector.

[0260] Example 2.

[0261] The method of Example 1, further comprising obtaining the second substrate material from a rack on a sample rack carriage of the automated clinical analyzer, wherein the sample rack carriage is configured to selectively receive and eject the rack.

[0262] Example s.

[0263] The method of Example 2, wherein the rack comprises a substrate container, wherein the substrate container houses a reservoir of the second substrate material, wherein obtaining the second substrate material comprises accessing the reservoir of the substrate container of the rack.

[0264] Example 4.

[0265] The method of Examples 2 or 3, further comprising utilizing a sample pipetting unit of the automated clinical analyzer to obtain the second substrate material from the rack on the sample carriage.

[0266] Example 5.

[0267] The method of Example 4, further comprising using a disposable pipette tip in conjunction with the sample pipetting unit to obtain the second substrate material from the rack on the sample carriage.

[0268] Example 6.

[0269] The method of Example 5, further comprising discarding the disposable pipette tip used to obtain the second substrate material.

[0270] Example 7.

[0271] The method of any one or more of Examples 4 through 6, wherein the second substrate material is the selected substrate material, wherein the method further comprises using the sample pipetting unit to directly add the second substrate material into the reaction vessel to create the signal generating mixture.

[0272] Example 8.

[0273] The method of Example 7, further comprising using the disposable pipette tip to mix the added second substrate material with the plurality of particles remaining from the wash cycle to create the signal generating mixture.

[0274] Example 9.

[0275] The method of any one or more of Examples 2 through 6, further comprising dispensing the obtained second substrate material into a portable vessel.

[0276] Example 10.

[0277] The method of Example 9, further comprising storing the portable vessel containing the second substrate material in a climate-controlled sample housing.

[0278] Example 11.

[0279] The method of Example 9 or 10, further comprising storing the portable vessel containing the second substrate material in an incubation container of the automated clinical analyzer.

[0280] Example I la.

[0281] The method of Example 10 or 11, wherein the portable vessel is stored for at least five minutes, at least ten minutes, at least fifteen minutes, at least twenty minutes, at least one hour, at least five hours, at least ten hours, or at least twelve hours, and / or up to ten minutes, up to fifteen minutes, up to twenty minutes, up to one hour, up to five hours, up to ten hours, up to twelve hours, or up to twenty -four hours.

[0282]

[0283] Example 12.

[0284] The method of any one or more of Examples 9 through I la, wherein the second substrate material is the selected substrate material, wherein the method further comprises obtaining the second substrate material from the portable vessel and dispensing the obtained second substrate material into the reaction vessel to create the signal generating mixture.

[0285] Example 13.

[0286] The method of Example 12, further comprising using a single use disposable tip and the sample pipetting unit to obtain the second substrate material from the portable vessel and add the second substrate into the reaction vessel.

[0287] Example 14.

[0288] The method of Example 13, further comprising utilizing the single-use disposable tip to mix the second substrate material with the plurality of particles remaining from the wash cycle to create the signal generating mixture.

[0289] Example 15.

[0290] The method of any one or more of Examples 12 through 14, further comprising possessing the portable vessel and the reaction vessel at a reaction build station while the second substrate material is dispensed into the reaction vessel.

[0291] Example 16.The method of Example 12, further comprising possessing the portable vessel and the reaction vessel at a wash wheel while the second substrate material is dispensed into the reaction vessel.

[0292] Example 17.

[0293] The method of Example 1, further comprising obtaining the second substrate material from an adjacent automated machine.

[0294] Example 17a.

[0295] The method of Example 17, wherein the second substrate material is obtained from a substrate container, and wherein obtaining the second substrate material comprises accessing a reservoir of the substrate container.

[0296] Example 17b.

[0297] The method of Example 17a, further comprising utilizing a sample pipetting unit of the automated clinical analyzer to obtain the second substrate material.

[0298] Example 17c.

[0299] The method of Example 17b, further comprising using a disposable pipette tip in conjunction with the sample pipetting unit to obtain the second substrate material.

[0300] Example 17d.

[0301] The method of Example 17c, further comprising discarding the disposable pipette tip used to obtain the second substrate material.

[0302] Example 17e.

[0303] The method of any one or more of Examples 17 through 17d, wherein the second substrate material is the selected substrate material, and wherein the method further comprises using the sample pipetting unit to directly add the second substrate material into the reaction vessel to create the signal generating mixture.

[0304] Example 17f.

[0305] The method of Example 17e, further comprising using the disposable pipette tip to mix the added second substrate material with the plurality of particles remaining from the wash cycle to create the signal generating mixture.

[0306] Example 18.

[0307] The method of Example 17, wherein the second substrate material is housed within a portable vessel while obtaining the second substrate material from an adjacent automated machine.

[0308] Example 18a.

[0309] The method of Example 18 further comprising storing the portable vessel containing the second substrate material in a climate-controlled sample housing.

[0310] Example 18b.

[0311] The method of Example 18 or 18a, further comprising storing the portable vessel containing the second substrate material in an incubation container of the automated clinical analyzer.

[0312] Example 18c.

[0313] The method of Example 18a or 18b, wherein the portable vessel is stored for at least five minutes, at least ten minutes, at least fifteen minutes, at least twenty minutes, at least one hour, at least five hours, at least ten hours, or at least twelve hours, and / or up to ten minutes, up to fifteenminutes, up to twenty minutes, up to one hour, up to five hours, up to ten hours, up to twelve hours, or up to twenty -four hours.

[0314] Example 19

[0315] The method of any one or more of Examples 18 to 18c, wherein the second substrate material is the chosen substrate material, the method further comprising utilizing a sample pipetting unit of the automated clinical analyzer in conjunction with a disposable pipette tip to obtain the second substrate material from the portable vessel and add the second substrate material into the reaction vessel to create the signal generating mixture.

[0316] Example 19a.

[0317] The method of Example 19, further comprising using a single use disposable pipette tip and the sample pipetting unit to obtain the second substrate material from the portable vessel and add the second substrate into the reaction vessel.

[0318] Example 20.

[0319] The method of Example 19, further comprising utilizing the disposable pipette tip to mix the added second substrate material with the plurality of particles remaining from the wash cycle to create the signal generating mixture.

[0320] Example 20a.

[0321] The method of any one or more of Examples 19 through 20, further comprising possessing the portable vessel and the reaction vessel at a reaction build station while the second substrate material is dispensed into the reaction vessel.

[0322] Example 20b.

[0323] The method of Example 19, further comprising possessing the portable vessel and the reaction vessel at a wash wheel while the second substrate material is dispensed into the reaction vessel.

[0324] Example 21.

[0325] The method of any one or more of the preceding Examples, wherein the second substrate material is the selected substrate material, wherein the amount of substrate material comprises at least 50 microliters, or at least 100 microliters, and up to 100 microliters, up to 200 microliters, up to 300 microliters, or up to 500 microliters.

[0326] Example 22.

[0327] The method of any one or more of Examples 1 through 21, wherein the reporter comprises an enzyme.

[0328] Example 23

[0329] The method of any one or more of Examples 1 through 22, wherein the detector comprises a luminometer.

[0330] Example 24

[0331] The method of any one or more of Examples 1 through 23, wherein creating the assay mixture comprises adding the first reagent and the second reagent to the reaction vessel sequentially, prior to incubation.

[0332] Example 25

[0333] The method of any one or more of Examples 1 through 23, wherein creating the assay mixture comprises adding the first reagent into the reaction vessel prior to adding the second reagent to the reaction vessel.

[0334] Example 26

[0335] The method of Example 25, further comprising incubating the reaction vessel prior to adding the second reagent to the reaction vessel.

[0336] Example 27

[0337] The method of either of Examples 25 or 26, further comprising performing a wash cycle prior to adding the second reagent to the reaction vessel.

[0338] Example 28

[0339] The method of any one or more of Examples 1 through 24, wherein creating the assay mixture comprises adding the second reagent into the reaction vessel prior to adding the first reagent to the reaction vessel.

[0340] Example 29

[0341] The method of any one or more of Examples 1 through 28, wherein the second reagent further comprises an antibody.

[0342] Example 30

[0343] The method of any one or more of Examples 1 through 29, wherein the second reagent further comprises an analyte.

[0344] Example 31

[0345] The method of Example 30, wherein the analyze comprises a competing analyte.

[0346] Example 32

[0347] The method of any one or more of Examples 1 through 31, wherein the first reagent comprises an antibody.

[0348] Example 33

[0349] The method of any one or more of Examples 1 through 32, wherein the plurality of particles comprises a plurality of iron particles.

[0350] Example 34

[0351] The method of Example 33, wherein performing the wash cycle further comprises (i) attracting the plurality of iron particles toward a magnet, and (ii) aspirating the assay mixture from the reaction vessel while the plurality of iron particles remains attracted toward the magnet.

[0352] Example 35

[0353] The method of any one or more of Examples 1 through 34, wherein performing the wash cycle further comprises adding a washing agent to the reaction vessel.

[0354] Example 36

[0355] The method of Example 35, wherein performing the wash cycle further comprises adding the washing agent to the reaction vessel at least three times, at least five times, at least seven times, at least nine times, at least 10 times.

[0356] Example 37

[0357] The method of Example 35 or Example 36, wherein performing the wash cycle further comprises adding the washing agent to the reaction vessel up to 100 times.

[0358] Example 38

[0359] The method of any one or more of Examples 33 through 37, wherein adding the washing agent is performed while the plurality of iron particles is attached toward the magnet.

[0360] Example 39

[0361] A method of performing an immunoassay with an automated clinical analyzer, the method comprising: (a) mixing a composition comprising an analyte from a patient sample with at least one reagent within a reaction vessel to create an assay mixture; (b) incubating the reaction vessel containing the assay mixture for a predetermined amount of time; (c) performing a wash cycle on the reaction vessel containing the assay mixture, thereby leaving a plurality of particles, some of which are bound to a respective analyte of the patient sample; (d) selecting a first substrate material or a second substrate material; (e) obtaining the selected substrate material; (f) adding andmixing the selected substrate material into the reaction vessel with the plurality of particles remaining from the wash cycle to create a signal generating mixture; (g) incubating the reaction vessel containing the signal generating mixture; and (h) analyzing the signal generating mixture with a luminometer.

[0362] Example 40

[0363] The method of Example 39, further comprising obtaining the analyte from the patient sample utilizing a pipette of a patient sample aliquot system of the automated clinical analyzer.

[0364] Example 41

[0365] The method of Example 40, further comprising obtaining the second substrate material utilizing the pipette of the patient sample aliquot system of the automated clinical analyzer.

[0366] Example 42

[0367] The method of Example 41, further comprising: (i) utilizing a first disposable tip with the pipette to obtain the analyte from the patient sample, and (ii) utilizing a second disposable tip with the pipette to obtain the second substrate material.

[0368] Example 43

[0369] The method of Example 42, wherein the analyte from the patient sample is obtained prior to obtaining the second substrate material.

[0370] Example 44

[0371] The method of Example 42, wherein the second substrate material is obtained prior to obtaining the analyte from the patient sample.

[0372] Example 45

[0373] The method of any one or more of Examples 41-44, further comprising dispensing the obtained second substrate material into a portable vessel and storing the portable vessel.

[0374] Example 46

[0375] The method of Example 45, further comprising storing the portable vessel at a climate controlled sample housing of the automated clinical analyzer or an incubation wheel of the automated clinical analyzer.

[0376] Example 47

[0377] A sample rack configured for use with an automated clinical analyzer, the sample rack comprising: (a) a base configured to be received by a sample rack carriage of the automated clinical analyzer; (b) a plurality of vessel holders extending upwardly from the base, wherein each vesselholder of the plurality of vessel holders is configured to selectively receive a corresponding vessel such that the corresponding vessel is accessible by a pipette of the automated clinical analyzer while the base is received by the sample rack carriage; and (c) a substrate container holder extending upwardly from the base, wherein the substrate container holder is configured to selectively receive a substrate container such that the substrate container is accessible by the pipette of the automatic clinical analyzer while the base is received by the sample rack carriage.

[0378] Example 48

[0379] The method or clinical analyzer of any one or more of the preceding Examples, wherein the substrate material comprises:(a) a compound comprising a compound of formula I or a salt thereof:whereinA is Ci-6haloalkyl, naphthyl, phenyl, substituted phenyl, or heteroaryl, wherein substituted phenyl comprises from 1 to 3 halo, Ci-6 alkyl, Ci-6 alkoxy, Ci-6 haloalkyl, C(0)Ri5, CN or NO2 substituents;Ri is selected from the group consisting of Cs-naryl, Ci-6 alkyl, Ci-6 haloalkyl, and C5- 14 aralkyl groups;R7-R14 are independently H, Ci-6 alkoxy, halo, Ci-4alkyl, or R7 -Rs or Rs -R9 or R9-R10 Rl 1-R12 or R12-R13 or R13-R14, can be joined together as a carbocyclic or heterocyclic ring system comprising at least one 5 or 6-membered ring;Ri5 is Ci-6 alkyl; each M is independently selected from H, or an alkali metal, alkaline earth metal, transition metal, ammonium, phosphonium, organic amine salt, and an amino acid salt;Z is O or S; and n is 0, 1, or 2;(b) a cationic aromatic compound,(c) a background reducing agent, and(d) an ether-linked nonionic surfactant or a hydrophilic polymer or a combination thereof.

[0380] Example 49

[0381] The method or clinical analyzer of any one or more of the preceding Examples, wherein the substrate material comprises: a compound of Formula II and salts thereof:(Formula II) wherein each of R1and R2is independently C3-C10 alkyl, or R1and R2taken together with the carbon to which they are attached provide a C5-C10 cycloalkyl ring; R3is C1-C10 alkyl, Ce-Cio aryl, or heteroaryl;R4is C2-C10 alkenyl; R5is H or Ci-C 10 alkyl; X is a phosphate; and at least one surfactant.

[0382] Example 50

[0383] The method of Example 49, wherein the second substrate material comprises compound having a structure:

[0384] Example 51A method of performing an immunoassay with an automated clinical analyzer, the method comprising:(a) creating an assay mixture within a reaction vessel, the assay mixture comprising an analyte from a patient sample, a first reagent, and a second reagent, wherein the first reagent comprises a plurality of particles configured to specifically bind to the analyte from the patient sample, wherein the second reagent comprises a reporter;(b) incubating the assay mixture;(c) performing a wash cycle on the assay mixture, wherein performing the wash cycle comprises retaining the plurality of particles while removing the remaining potions of the assay mixture not bound to the plurality of particles, wherein at least one particle of the plurality of particles is bound to a respective analyte from the patient sample;(d) obtaining a substrate material from a sample rack loaded into the automated clinical analyzer;(e) adding and mixing the substrate material into the reaction vessel with the plurality of particles remaining from the wash cycle to create a signal generating mixture;(f) incubating the signal generating mixture; and(g) analyzing the signal generating mixture with a detector.

[0385] Example 52

[0386] The method of Example 51, further comprising loading the sample rack into the automated clinical analyzer using a sample loading station of the automated clinical analyzer.

[0387] Example 53

[0388] The method of Example 51, further comprising loading the sample rack into the automated clinical analyzer using an adjacent automated machine.

[0389] Example 54

[0390] The method of any one or more of Examples 51 through 53, wherein the sample rack, while loaded into the automated clinical analyzer, holds a substrate container.

[0391] Example 55

[0392] The method of any one or more of Examples 51 through 54, wherein the sample rack, while loaded into the automated clinical analyzer, holds a patient sample vessel.

[0393] Example 56

[0394] The method of any one or more of Examples 51 through 55, wherein the sample rack is configured to accommodate a substrate container.

[0395] Example 57

[0396] The method of any one or more of Examples 51 through 55, wherein the sample rack is configured to accommodate three substrate holders.

[0397] Example 58

[0398] The method of any one or more of Examples 51 through 55, wherein the sample rack is configured to accommodate only vessels.

[0399] Example 59

[0400] The method of Example 58, wherein the sample rack comprises seven vessel holders.

[0401] V. Interpretation

[0402] It should be understood that, in the above examples and the claims, a statement that something is “based on” something else should be understood to mean that it is determined at least in part by the thing that it is indicated as being based on. To indicate that something must be completely determined based on something else, it is described as being “based EXCLUSIVELY on” whatever it must be completely determined by.

[0403] It should be understood that a statement that “one or more” or “at least one” of a type of item have a characteristic indicates that the items in the indicated group collectively have the characteristic. To indicate that each item in a group has a characteristic, the phrase “each of’ will be used with the group identifier (e.g., “one or more” or “at least one”).

[0404] It should be understood that, in the claims, “set” should be understood as referring to one or more thing of similar nature, design or function.

[0405] It should be understood that any of the examples described herein may include various other features in addition to or in lieu of those described above. By way of example only, any of the examples described herein may also include one or more of the various features disclosed in any of the various references that are incorporated by reference herein.

[0406] It should be understood that any one or more of the teachings, expressions, embodiments, examples, etc. described herein may be combined with any one or more of the other teachings, expressions, embodiments, examples, etc. that are described herein. The abovedescribed teachings, expressions, embodiments, examples, etc. should therefore not be viewed in isolation relative to each other. Various suitable ways in which the teachings herein may becombined will be readily apparent to those of ordinary skill in the art in view of the teachings herein. Such modifications and variations are intended to be included within the scope of the claims.

[0407] It should be appreciated that any patent, publication, or other disclosure material, in whole or in part, that is said to be incorporated by reference herein is incorporated herein only to the extent that the incorporated material does not conflict with existing definitions, statements, or other disclosure material set forth in this disclosure. As such, and to the extent necessary, the disclosure as explicitly set forth herein supersedes any conflicting material incorporated herein by reference. Any material, or portion thereof, that is said to be incorporated by reference herein, but which conflicts with existing definitions, statements, or other disclosure material set forth herein will only be incorporated to the extent that no conflict arises between that incorporated material and the existing disclosure material.

[0408] Having shown and described various versions of the present invention, further adaptations of the methods and systems described herein may be accomplished by appropriate modifications by one of ordinary skill in the art without departing from the scope of the present invention. Several of such potential modifications have been mentioned, and others will be apparent to those skilled in the art. For instance, the examples, versions, geometries, materials, dimensions, ratios, steps, and the like discussed above are illustrative and are not required. Accordingly, the scope of the present invention should be considered in terms of the following claims and is understood not to be limited to the details of structure and operation shown and described in the specification and drawings.

[0409] The words “preferred” and “preferably” refer to embodiments of the invention that may afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful and is not intended to exclude other embodiments from the scope of the invention.

[0410] The term “comprises” and variations thereof do not have a limiting meaning where these terms appear in the description and claims. Such terms will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements.

[0411] By ‘ ‘consisting of’ is meant including, and limited to, whatever follows the phrase “consisting of.” Thus, the phrase “consisting of’ indicates that the listed elements are required or mandatory, and that no other elements may be present. By “consisting essentially of’ is meant including any elements listed after the phrase, and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase “consisting essentially of’ indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present depending upon whether or not they materially affect the activity or action of the listed elements.

[0412] Unless otherwise specified, “a,” “an,” “the,” and “at least one” are used interchangeably and mean one or more than one. As used herein, the term “or” is generally employed in its usual sense including “and / or” unless the content clearly dictates otherwise. The term “and / or” means one or all of the listed elements or a combination of any two or more of the listed elements.

[0413] Any reference to standard methods (e.g., ASTM, TAPPI, AATCC, etc.) refer to the most recent available version of the method at the time of filing of this disclosure unless otherwise indicated.

[0414] Also herein, the recitations of numerical ranges by endpoints include all numbers subsumed within that range (e.g., 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.80, 4, 5, etc.). Herein, “up to” a number (for example, up to 50) includes the number (for example, 50). The term “in the range” or “within a range” (and similar statements) includes the endpoints of the stated range.

[0415] For any method disclosed herein that includes discrete steps, the steps may be conducted in any feasible order. And, as appropriate, any combination of two or more steps may be conducted simultaneously.

[0416] All headings are for the convenience of the reader and should not be used to limit the meaning of the text that follows the heading, unless so specified.

[0417] Reference throughout this specification to “one embodiment,” “an embodiment,” “certain embodiments,” or “some embodiments,” etc., means that a particular feature, configuration, composition, or characteristic described in connection with the embodiment is included in at least one embodiment of the disclosure. Thus, the appearances of such phrases in various places throughout this specification are not necessarily referring to the same embodimentof the disclosure. Furthermore, the particular features, configurations, compositions, or characteristics may be combined in any suitable manner in one or more embodiments.

[0418] Unless otherwise indicated, all numbers expressing quantities of components, molecular weights, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about.” As used herein in connection with a measured quantity, the term “about” refers to that variation in the measured quantity as would be expected by the skilled artisan making the measurement and exercising a level of care commensurate with the objective of the measurement and the precision of the measuring equipment used. Accordingly, unless otherwise indicated to the contrary, the numerical parameters set forth in the specification and claims are approximations that may vary depending upon the desired properties sought to be obtained by the present invention. At the very least, and not as an attempt to limit the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

[0419] Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. All numerical values, however, inherently contain a range necessarily resulting from the standard deviation found in their respective testing measurements. The complete disclosure of all patents, patent applications, and publications, and electronically available material cited herein are incorporated by reference. In the event that any inconsistency exists between the disclosure of the present application and the disclosure(s) of any document incorporated herein by reference, the disclosure of the present application shall govern. The foregoing detailed description and examples have been given for clarity of understanding only. No unnecessary limitations are to be understood therefrom. The invention is not limited to the exact details shown and described, for variations obvious to one skilled in the art will be included within the invention defined by the claims.

Claims

WHAT IS CLAIMED IS:

1. A method of performing an immunoassay with an automated clinical analyzer, the method comprising:(a) providing a first substrate material and a second substrate material;(b) creating an assay mixture within a reaction vessel, the assay mixture comprising an analyte from a patient sample, a first reagent, and a second reagent, wherein the first reagent comprises a plurality of particles configured to specifically bind to the analyte from the patient sample, wherein the second reagent comprises a reporter;(c) incubating the assay mixture;(d) performing a wash cycle on the assay mixture, wherein performing the wash cycle comprises retaining the plurality of particles while removing the remaining portions of the assay mixture not bound to the plurality of particles, wherein at least one particle of the plurality of particles is bound to a respective analyte from the patient sample;(e) selecting the first substrate material or the second substrate material;(f) adding and mixing the selected substrate material into the reaction vessel with the plurality of particles remaining from the w ash cycle to create a signal generating mixture;(g) incubating the signal generating mixture; and(h) analyzing the signal generating mixture with a detector.

2. The method of claim 1, further comprising selecting the second substrate material and obtaining the second substrate material from a rack on a sample rack camage of the automated clinical analyzer, wherein the sample rack carriage is configured to selectively receive and ej ect the rack.

3. The method of claim 2, wherein the rack comprises a substrate container, wherein the substrate container houses a reservoir of the second substrate material, wherein obtaining the second substrate material comprises accessing the reservoir of the substrate container of the rack.

4. The method of claims 2 or 3, further comprising utilizing a sample pipetting unit of the automated clinical analyzer to obtain the second substrate material from the rack on the sample rack carriage.

5. The method of claim 4, further comprising using a disposable pipette tip in conjunction with the sample pipetting unit to obtain the second substrate matenal from the rack on the sample rack carriage.

6. The method of claim 5, further comprising discarding the disposable pipette tip used to obtain the second substrate material.

7. The method of any one or more of claims 4 through 6, wherein the second substrate material is the selected substrate material, wherein the method further comprises using the sample pipetting unit to directly add the second substrate material into the reaction vessel to create the signal generating mixture.

8. The method of claim 7, further comprising using the disposable pipette tip to mix the added second substrate material with the plurality of particles remaining from the wash cycle to create the signal generating mixture.

9. The method of any one or more of claims 2 through 6, further comprising dispensing the obtained second substrate material into a portable vessel.

10. The method of claim 9, further comprising storing the portable vessel containing the second substrate material in a climate-controlled sample housing.

11. The method of claim 9 or 10. further comprising storing the portable vessel containing the second substrate material in an incubation container of the automated clinical analyzer.

12. The method of claim 10 or 11, wherein the portable vessel is stored forat least five minutes, at least ten minutes, at least fifteen minutes, at least twenty minutes, at least one hour, at least five hours, at least ten hours, or at least twelve hours, and / or up to ten minutes, up to fifteen minutes, up to twenty minutes, up to one hour, up to five hours, up to ten hours, up to twelve hours, or up to twenty-four hours.

13. The method of any one or more of claims 9 through 12. wherein the second substrate material is the selected substrate material, wherein the method further comprises obtaining the second substrate material from the portable vessel and dispensing the obtained second substrate material into the reaction vessel to create the signal generating mixture.

14. The method of claim 13. further comprising using a single use disposable tip and the sample pipetting unit to obtain the second substrate material from the portable vessel and add the second substrate into the reaction vessel.

15. The method of claim 14, further comprising utilizing the single-use disposable tip to mix the second substrate material with the plurality of particles remaining from the wash cycle to create the signal generating mixture.

16. The method of any one or more of claims 13 through 15, further comprising possessing the portable vessel and the reaction vessel at a reaction build station while the second substrate material is dispensed into the reaction vessel.

17. The method of claim 13, further comprising possessing the portable vessel and the reaction vessel at a wash wheel while the second substrate material is dispensed into the reaction vessel.

18. The method of claim 1, further comprising obtaining the second substrate material from an adjacent automated machine.

19. The method of claim 18, wherein the second substrate material is obtained from a substrate container, and wherein obtaining the second substrate material comprises accessing a reservoir of the substrate container.

20. The method of claim 19, further comprising utilizing a sample pipetting unit of the automated clinical analyzer to obtain the second substrate material.

21. The method of claim 20, further comprising using a disposable pipette tip in conjunction with the sample pipetting unit to obtain the second substrate material.

22. The method of claim 21. further comprising discarding the disposable pipette tip used to obtain the second substrate material.

23. The method of any one or more of claims 17 through 22, wherein the second substrate material is the selected substrate material, and wherein the method further comprises using the sample pipetting unit to directly add the second substrate material into the reaction vessel to create the signal generating mixture.

24. The method of claim 23, further comprising using the disposable pipette tip to mix the added second substrate material with the plurality of particles remaining from the wash cycle to create the signal generating mixture.

25. The method of claim 18, wherein the second substrate material is housed within a portable vessel while obtaining the second substrate material from an adjacent automated machine.

26. The method of claim 25 further comprising storing the portable vessel containing the second substrate material in a climate-controlled sample housing.

27. The method of claim 25 or 26, further comprising storing the portable vessel containing the second substrate material in an incubation container of the automated clinical analyzer.

28. The method of claim 26 or 27, wherein the portable vessel is stored for at least five minutes, at least ten minutes, at least fifteen minutes, at least twenty minutes, at least one hour, at least five hours, at least ten hours, or at least twelve hours, and / orup to ten minutes, up to fifteen minutes, up to twenty minutes, up to one hour, up to five hours, up to ten hours, up to twelve hours, or up to twenty-four hours.

29. The method of any one of claims 25to 28, wherein the second substrate material is the chosen substrate material, the method further comprising utilizing a sample pipetting unit of the automated clinical analyzer in conjunction with a disposable pipette tip to obtain the second substrate material from the portable vessel and add the second substrate material into the reaction vessel to create the signal generating mixture.

30. The method of claim 29, further comprising using a single use disposable pipette tip and the sample pipetting unit to obtain the second substrate material from the portable vessel and add the second substrate into the reaction vessel.

31. The method of claim 29 or 30, further comprising utilizing the disposable pipette tip to mix the added second substrate material with the plurality of particles remaining from the wash cycle to create the signal generating mixture.

32. The method of any one or more of claims 29 through 31, further comprising possessing the portable vessel and the reaction vessel at a reaction build station while the second substrate material is dispensed into the reaction vessel.

33. The method of claim 29, further comprising possessing the portable vessel and the reaction vessel at a wash wheel while the second substrate material is dispensed into the reaction vessel.

34. The method of claim 1, wherein the selected substrate material is added directly to the reaction vessel.

35. The method of claim 1, wherein the selected substrate material is temporarily stored on the automated clinical analyzer prior to being added to the reaction vessel.

36. The method of claim 35, wherein the selected substrate material is temporarily stored on the automated clinical analyzer in a vessel.

37. A method of performing an immunoassay with an automated clinical analyzer, the method comprising:(a) mixing a composition comprising an analyte from a patient sample with at least one reagent within a reaction vessel to create an assay mixture;(b) incubating the reaction vessel containing the assay mixture for a predetermined amount of time;(c) performing a wash cycle on the reaction vessel containing the assay mixture, thereby leaving a plurality of particles, some of which are bound to a respective analyte of the patient sample;(d) selecting a first substrate material or a second substrate material based on the immunoassay;(e) obtaining the selected substrate material;(f) adding and mixing the selected substrate material into the reaction vessel with the plurality of particles remaining from the wash cycle to create a signal generating mixture;(g) incubating the reaction vessel containing the signal generating mixture; and(h) analyzing the signal generating mixture with a luminometer.

38. The method of claim 37, further comprising obtaining the analyte from the patient sample utilizing a pipette of a patient sample aliquot system of the automated clinical analyzer.

39. The method of claim 38, further comprising obtaining the second substrate material utilizing the pipette of the patient sample aliquot system of the automated clinical analyzer.

40. The method of claim 39, further comprising:(i) utilizing a first disposable tip with the pipette to obtain the analyte from the patient sample, and(ii) utilizing a second disposable tip with the pipette to obtain the second substrate material.

41. The method of claim 40, wherein the analyte from the patient sample is obtained prior to obtaining the second substrate material.

42. The method of claim 40, wherein the second substrate material is obtained prior to obtaining the analyte from the patient sample.

43. The method of any one or more of claims 39-42, further comprising dispensing the obtained second substrate material into a portable vessel and storing the portable vessel.

44. The method of claim 43, further comprises storing the portable vessel at a climate controlled sample housing of the automated clinical analyzer or an incubation wheel of the automated clinical analyzer.

45. The method of any one or more of the preceding claims, wherein the second substrate material is the selected substrate material, wherein the amount of substrate material comprises at least 50 microliters, or at least 100 microliters, and up to 100 microliters, up to 200 microliters, up to 300 microliters, or up to 500 microliters.

46. The method of any one or more of the preceding claims, wherein the reporter comprises an enzyme.

47. The method of any one or more of the preceding claims, wherein the detector comprises a luminometer.

48. The method of any one or more of the preceding claims, wherein creating the assay mixture comprises adding the first reagent and the second reagent to the reaction vessel sequentially, prior to incubation.

49. The method of any one or more of the preceding claims, wherein creating the assay mixture comprises adding the first reagent into the reaction vessel prior to adding the second reagent to the reaction vessel.

50. The method of any one or more of the preceding claims, further comprising incubating the reaction vessel prior to adding the second reagent to the reaction vessel.

51. The method of any one or more of the preceding claims, further comprising performing a wash cycle prior to adding the second reagent to the reaction vessel.

52. The method of any one or more of the preceding claims, wherein creating the assay mixture comprises adding the second reagent into the reaction vessel prior to adding the first reagent to the reaction vessel.

53. The method of any one or more of the preceding claims, wherein the second reagent further comprises an antibody.

54. The method of any one or more of the preceding claims, wherein the second reagent further comprises an analyte.

55. The method of any one or more of the preceding claims, wherein the analyte comprises a competing analyte.

56. The method of any one or more of the preceding claims, wherein the first reagent comprises an antibody.

57. The method of any one or more of the preceding claims, wherein the plurality of particles comprises a plurality of iron particles.

58. The method of any one or more of the preceding claims, wherein performing the wash cycle further comprises (i) attracting the plurality of iron particles toward a magnet, and (ii) aspirating the assay mixture from the reaction vessel while the plurality of iron particles remains attracted toward the magnet.

59. The method of any one or more of the preceding claims, wherein performing the wash cycle further comprises adding a washing agent to the reaction vessel.

60. The method of any one or more of the preceding claims, wherein performing the wash cycle further comprises adding the washing agent to the reaction vessel at least three times, at least five times, at least seven times, at least nine times, at least 10 times.

61. The method of any one or more of the preceding claims, wherein performing the wash cycle further comprises adding the washing agent to the reaction vessel up to 100 times.

62. The method of any one or more of the preceding claims, wherein adding the washing agent is performed while the pl urali ty of iron particles is attached toward the magnet.

63. The method of any one or more of the preceding claims, wherein either the first substrate material or the second substrate material comprises:(a) a compound comprising a compound of formula I or a salt thereof:whereinA is Ci-6haloalkyl, naphthyl, phenyl, substituted phenyl, or heteroaryl, wherein substituted phenyl comprises from 1 to 3 halo, Ci-6 alkyl. Ci-6 alkoxy. Ci-6 haloalkyl. C(0)Ri5, CN or NO2 substituents;Ri is selected from the group consisting of Cs-naryl. Ci-6 alky l, Ci-6 haloalkyl, and C5-14 aralkyl groups;R7-R14 are independently H, Ci-6 alkoxy, halo. Ci-4alkyl, or R7 -Rs or Rs -R9 or R9-R10 R11-R12 or R12-R13 or R13-R14, can be joined together as a carbocyclic or heterocyclic ring system comprising at least one 5 or 6-membered ring;Ri5 is Ci-6 alkyl; each M is independently selected from H, or an alkali metal, alkaline earth metal, transition metal, ammonium, phosphonium, organic amine salt, and an amino acid salt;Z is O or S: and n is 0, 1, or 2;(b) a cationic aromatic compound.(c) a background reducing agent, and(d) an ether-linked nonionic surfactant or a hydrophilic polymer or a combination thereof.

64. The method of any one or more of the preceding claims, wherein either the first substrate material or the second substrate material comprises: a compound of Formula II and salts thereof:(Formula II) wherein each of R1and R2is independently C3-C10 alkyl, or R1and R2taken together with the carbon to which they are attached provide a C5-C10 cycloalkyl ring; R3is C1-C10 alkyl, Ce-Cio aryl, or heteroaryl;R4is C2-C10 alkenyl; R5is H or C1-C10 alkyl; X is a phosphate; and at least one surfactant.

65. The method of claim 52, wherein the second substrate material comprises compound having a structure:

66. The method of claim 37, wherein the selected substrate material is added directly to the reaction vessel.

67. The method of claim 37, wherein the selected substrate material is temporarily stored on the automated clinical analyzer prior to being added to the reaction vessel.

68. The method of claim 67, wherein the selected substrate material is temporarily stored on the automated clinical analyzer in a vessel.

69. The method of any preceding claim, further comprising providing both the first substrate material and the second substrate material and selecting only one substrate material for the immunoassay.

70. A sample rack configured for use with an automated clinical analyzer, the sample rack comprising:(a) a base configured to be received by a sample rack carriage of the automated clinical analyzer;(b) a plurality of vessel holders extending upwardly from the base, wherein each vessel holder of the plurality of vessel holders is configured to selectively receive a corresponding vessel such that the corresponding vessel is accessible by a pipette of the automated clinical analyzer while the base is received by the sample rack carriage; and(c) a substrate container holder extending upwardly from the base, wherein the substrate container holder is configured to selectively receive a substrate container such that the substrate container is accessible by thepipete of the automatic clinical analyzer while the base is received by the sample rack carriage.

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