Automated clinical analyzer with alternative substrate and method of use

Abstract

A method of performing an immunoassay with an automated clinical analyzer, includes, mixing an analyte from a patient sample with a first reagent having a plurality of particles configured to specifically bind to the analyte and a second reagent having an enzyme, incubating the assay mixture for a predetermined amount of time and then performing a wash cycle on the assay mixture in order to retain the plurality of particles while removing the remaining portion of the assay mixture not bound the plurality of particles, and selecting a first substrate material or a second substrate material, each housing with the automated clinical analyzer. 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, incubating the signal generating mixture, and analyzing the signal generating mixture with a luminometer.

Description

AUTOMATED CLINICAL ANALYZER WITH ALTERNATIVE SUBSTRATE ANDMETHOD OF USEPRIORITY

[0001] This application claims priority to U.S. Provisional Application 63 / 621,360, entitled “Automated Clinical Analyzer with Alternative Substrate and Method of Use,” filed on January 16, 2024, hereby incorporated by reference in its entirety.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 at least 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 bound 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.

[0006] In some embodiments, the automated assay mixes and incubates the sample with one or more reagents in order to bind a targeted analyte of the sample to a particle of a plurality of particles, and also bind reporter(s) to a respective particle of the plurality of particles and / or a respective analyte of the targeted sample analyte. Further, in some embodiments, the automated assay selects a first substrate or a second substrate and subsequently adds and mixes the selected substrate with a composition that includes the bound reporter(s) in order to generate a signal indicative of the amount of targeted patient analyte within the composition.

[0007] In some embodiments, when the first substrate is the selected substrate, the first substrate is added and mixed with the composition at a wash wheel of the clinical analyzer. In some embodiments, when the second substrate is the selected substrate, the second substrate is added and mixed with the composition at the wash wheel of the clinical analyzer. In some embodiments, when the second substrate is the selected substrate, the second substrate is added a mixed with the composition at a designated second substrate dispensing system. In some embodiments, the designated second substrate dispensing system is located at an incubation station. In some embodiments, the designated second substrate dispensing system is located adjacent to a reagent mixing station. In some embodiments, the designated second substrate dispensing system includes its own mixing station.

[0008] In some embodiments, the automated clinical analyzer includes a reagent mixing station, an incubation station, a wash wheel assembly, a substrate application assembly, and a luminometer. The reagent mixing station is configured to mix a composition comprising a patient sample with at least one reagent to form an assay mixture. The incubation station is configured to incubate the mixture containing at least a portion of the assay mixture. The wash wheel assembly is configured to isolate a plurality of particles from the assay mixture, some particles of the isolated plurality of particles are bound to a respective analyte of the patient sample. The substrate application assembly is configured to select a first substrate or a second substrate to be mixed with the isolated plurality of particles to from a signal generating mixture. The second substrate being located within the removable pack of the reagent station. The luminometer is configured to analyze light emitted from the signal generating mixture.BRIEF DESCRIPTION OF THE DRAWINGS

[0009] 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:

[0010] FIG. 1 depicts a schematic view of an illustrative assay;

[0011] FIG. 2 A depicts a cross-sectional view of an illustrative luminometer, taken along a centerline axis of the luminometer;

[0012] FIG. 2B depicts a sectional view of the luminometer of FIG. 2 A;

[0013] FIG. 3 A depicts a perspective view of an illustrative wash wheel;

[0014] FIG. 3B depicts a schematic view of the wash wheel of FIG. 3 A;

[0015] FIG. 4 depicts a schematic view of an illustrative pipetting system;

[0016] FIG. 5 depicts a schematic view of an illustrative probe washing arrangement;

[0017] FIG. 6 depict a schematic view of an illustrative computer system;

[0018] 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, an incubation station, a reagent storage station, a substrate storage station, 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;

[0019] FIG. 8 depicts a flowchart of an illustrative method of use of the automated clinical analyzer of FIG. 7;

[0020] FIG. 9 depicts a schematic view of a modified automated clinical analyzer similar to FIG. 7, with a second substrate dispensing system associated with a wash wheel, an example of which is shown in FIG. 3 A;

[0021] FIG. 10 depicts a schematic view of a modified automated clinical analyzer similar to FIG. 7, with a second substrate dispensing system;

[0022] FIG. 11 depicts a schematic view of a modified automated clinical analyzer similar to FIG. 7, with a second substrate dispensing system associated with an incubation station, an example of which is shown in FIG. 7;

[0023] FIG. 12 depicts a schematic view of a modified automated clinical analyzer similar to FIG. 7, with a second substrate dispensing system associated with a reagent mixing station, an example of which is shown in FIG. 7; and

[0024] FIG. 13 depicts a flowchart of an illustrative method of use of the analyzer of any of FIGS. 9, 10, 11, and 12.

[0025] The drawings are not intended to be limiting in any way, and 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

[0026] 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.

[0027] 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 readilyapparent 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.

[0028] 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 both reporting reagents are housed within the automated clinical analyzer. As further described herein, it may be desirable to have the option of 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.

[0029] I. Illustrative Assay and Automated Clinical Analyzer Configured to Perform Assay

[0030] A. Illustrative Assay

[0031] 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.

[0032] As shown in FIG.l, at stage 1, 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.

[0033] 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 pipette 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.

[0034] At stage 3, the reaction vessel 105, containing the patient sample 165 and the first reagent 115 (including the particles 120) mix 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 reaction 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 particles 120, 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.

[0035] 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 (orattempted 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.

[0036] 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 ten 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.

[0037] Thus, in an exemplary 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.

[0038] At stage 5, the pipette 110 may place a second reagent 155, including a reporter, in the reaction vessel 105 with the particles 120 and the bound analyte of the patient sample 165. Thereporter may be, for example, a suitable enzyme such as alkaline phosphatase (“ALP”) (for example, at a concentration of between O.Olmg / L and 2.0 mg / L) or 0-galactosidase, a fluorescent protein, or luciferase.

[0039] 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.

[0040] 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 heat source (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.

[0041] 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.

[0042] 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 agent150 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.

[0043] 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 pluralityof particles 120, some of which are bound to a respective analyte of the patent sample, within the reaction vessel 105.

[0044] 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.

[0045] 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. 9-12) 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. 9-12) is utilized to serve as the reporting reagent, a designated second substrate dispensing system 730, 740, 760, 770 (FIGS. 9-12) may be utilized in order to suitably dispense second substrate material 182 into reaction vessel 105. The selected reporting reagent (e.g., first substrate material 180 or second substrate material 182 (FIGS. 9-12) 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). In 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 toproduce light (i.e., photons) is proportional to the amount of patient analyte present within the reaction vessel 105.

[0046] 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.

[0047] 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.

[0048] 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 when the first substrate material 180 is added. In 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.

[0049] B. Illustrative Luminometer of Automated Clinical Analyzer

[0050] 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.

[0051] FIG. 2A provides a view of a chamber opening 430, which provides access to a 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 in detail) 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 regulatedinternal 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.

[0052] 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.

[0053] C. Illustrative Wash Assembly of Automated Clinical Analyzer

[0054] 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). The vessel and the holder 272 are axisymmetric with each other, when mated (e.g., when the vessel is located within the holder 272).

[0055] 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.

[0056] 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 labeledstations 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.

[0057] 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 and 3B), and then the contents of the vessel have been mixed or incubated (stage 3) (e.g., in an incubation wheel, also not shown in FIGS. 3 A 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.

[0058] 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 be rotated 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 will be spin mixed. This may then be repeated for stations S11-S18 to perform another illustrative wash cycle (i.e., the vessel may bemagnetized in positions SI 1 -SI 6, 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.

[0059] 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.

[0060] 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.

[0061] 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 clinical analyzer 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 in order 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)).

[0062] 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 may be 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. 9-12) 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. 9-12) is utilized to serve as the reporting reagent, a designated second substrate dispensing system 730, 740, 760, 770 (FIGS. 9-12) may be utilized to suitably dispense second substrate material 182 into reaction vessel 105.

[0063] 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.

[0064] D. Illustrative Pipetting System of Automated Clinical Analyzer

[0065] FIG. 4 and International Patent Application Publication No. WO 2020 / 139989 illustrate an example 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 ofperforming 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.

[0066] 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.

[0067] 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). 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- are defined for displacement dl.

[0068] 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 opposite second 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.).

[0069] 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 theinstrument) 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.

[0070] The probe P may define an axis A. The probe receiving station PS may define an axis AO. The probe P may be aligned with the corresponding probe receiving station PS when the axes A and AO are aligned within an acceptable tolerance.

[0071] In typical use, such as in dispensing and aspiration of fluids as described in the context of FIGS. 1, 3 A 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.

[0072] 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 this type 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 to 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 thesample 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.

[0073] 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.

[0074] FIG. 5 illustrates an exemplary 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 position pw2 (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.

[0075] 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 thethird 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.

[0076] 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.

[0077] 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.

[0078] E. Illustrative Computer System for Automated Clinical Analyzer

[0079] Any suitable computer system can be integrated into, or connected with, a clinical analyzer in order to control various actions of the analyzer such as by sending commands to thewash 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 may 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 (I / O) interface 57, and a Human Machine Interface (HMI) 59. Optionally, computer system 49 may be operatively coupled to one or more external resources 61 via a network 63 or I / O interface 57. External resources 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.

[0080] 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.

[0081] 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.

[0082] 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. Application67 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.

[0083] 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.

[0084] 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. The 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.

[0085] 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.

[0086] F. Illustrative Automated Clinical Analyzer

[0087] 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 a 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.

[0088] Additionally, clinical analyzer 700 includes a patient 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.

[0089] 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.

[0090] 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.

[0091] 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.

[0092] 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 a sample rack is aligned with the 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 for testing. 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 or around 4 degrees Celsius. The patient sample wheel 710 may be configured to maintain any other suitable internal temperature.

[0093] The sample pipetting unit 706 may actuate a sample pipette 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 707in accordance with the description herein. Sample aliquoting pipette 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 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.

[0094] 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.

[0095] 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 use of 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.

[0096] 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 patient sample 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 sample 165 may be transported from sample wheel 710 and / or sample rack carriage 704 onto reaction vessel carriage 712 using any suitable means.

[0097] Reaction vessel carriage 712 is configured to actuate at least one reaction vessel 105 along an axis Aciin order 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, reagentmixing 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 Acito 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.

[0098] 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. Further, pipettes of reagent pipetting station 714 may then actuate along their respective axis (A3, A4, A5, A6) to disburse the recently aspirated reagent into reaction vessel 105. In the current example, there are four reagent pipetting stations 714. However, any suitable number of reagent pipetting station 714 may be incorporated.

[0099] 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 mixthe 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.

[0100] 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.

[0101] 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.

[0102] 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 in order to perform one or more washing cycles in accordance with the description herein, luminometer 600, or another suitable station.

[0103] 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 incubation wheel 750, are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0104] As mentioned above, clinical analyzer 700 may include a substrate dispensing system725. As further discussed herein, when first substrate material 180 is the selected reporting agent,a 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 a first substrate material 180 within reaction vessel 105. 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. 9-12) 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. 9-12) is utilized to serve as the reporting reagent, a designated second substrate dispensing system 730, 740, 760, 770 (FIGS. 9-12) may be utilized to suitably dispense second substrate material 182 into reaction vessel 105.

[0105] 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.

[0106] 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 washwheel 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.

[0107] The substrate 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 structural elements of substrate dispensing system 725, are optional and may be substituted for other suitable elements, and even omitted when appropriate.

[0108] Similar to reaction vessel carriage 712, wash wheel 270 may be configured to suitably vibrate reaction vessel 105 in order 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.

[0109] G. Illustrative Workflow of Automated Clinical Analyzer

[0110] 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. The automated clinical analyzer 700 may perform at least 50 assays per hour, at least 100 assays per hour, at least 200 assays per hour, at least 300 assays per hour, at least 400 assays per hour, or at least 450 assay per hour. In some examples, the automated clinical analyzer 700 may perform at least 50 assays per hour and up to 450 assays per hour, at least 100 assays per hour and up to 450 assays per hour, at least 200 assays per hour and up to 450 assays per hour, at least 300 assays per hour and up to 450 assays per hour, or at least 400 assays per hour and up to 450 assays per hour, Therefore, the analyzer 700, as well as other similar analyzers with similar throughputs of at least 50 tests per hour, may be considered a “high throughput” clinical analyzer. 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 thedescription 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.[oni] 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.

[0112] 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.

[0113] Optionally, next, the reaction vessel 105 may be transferred 1010 to the incubation wheel to 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 vessel 105 is transported 1012 to the wash wheel 270 to suitably perform one or more wash cycles in accordance with the description herein.

[0114] 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 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 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.

[0115] 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, clinical analyzer 700 performing assay workflow 1000 moves directly to the adding1022 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 as would be apparent to one skilled in the art in view of the teaching herein.

[0116] 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.

[0117] 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 where first 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. 3 A and 3B). If first substrate 180 is added by substrate dispensing pipette system 724, 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., predetermined amount of time). Next, the reaction vessel 105 is transferred to the luminometer 600 to measure 1026 a RLU 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. As also described herein, in some instances, instead of selecting and adding first substrate material 180, a different reporting agent such as a second substrate material 182 (FIGS. 9-12) 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. 9-12) is utilized to serve as the reporting reagent, a designated second substrate dispensing system 730, 740, 760, 770 ( FIGS. 9-12) may be utilized in order to suitably dispense second substrate material 182 into reaction vessel 105. In some instances, the assay performed includes a competing analyte, such that less RLU detected by luminometer 600 is indicative of the presence of more patient analyte. In some instances, the assay performed does not include a competing analyte, such that more RLU detected by luminometer 600 is indicative of the presence of more patient analyte. The assay workflow 1000 applies to both of the aforementioned instances.

[0118] II. Illustrative Automated Clinical Analyzer with Alternative Substrate and Method of Use

[0119] As mentioned above, a reporting reagent (e.g., first substrate material 180 or second substrate material 182) is added to a reaction vessel 105 to chemically react with a bound reporter (e.g., an enzyme) to generate a signal (e.g., light) to 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 RLU via luminometer 600, is 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) 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 fiirther optimize desired parameter(s), the clinical analyzer 700 may be configured to select from multiple reporting 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 parameters(s).

[0120] 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 substrateand 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).

[0121] 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 second substrate 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 in order 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.

[0122] Further, in instances where clinical analyzer 700 can select a reporting reagent from multiple reporting reagents (e.g., first substrate material 180 or second substrate material 182 shown in FIGS 9-12) for use in a particular assay, it may be undesirable to utilize the same designated substrate dispensing pipette system 724 to deliver both the first substrate material 180 and the second substrate material 182 into a reaction vessel 105. Utilizing the same designatedsubstrate dispensing pipette system 724 to deliver both the first substate material 180 and the second substrate material 182 may require suitably cleaning of pipette system 724 between assays utilizing a different substrate material (e.g., first substrate material 180 and second substrate material 182). Therefore, it may be desirable to have a designated substrate dispensing system for each type of reporting agent (including, for example, the first substrate material 180 and the second substrate material 182) available within clinical analyzer 700, as described in more detail to follow.

[0123] FIGS. 9-12 show three modified automated clinical analyzers 700’, 700”, 700”’, 700””, respectively, that may be used to perform suitable assays in a similar manner to clinical analyzer 700 described above, with differences elaborated herein. Therefore, clinical analyzers 700’, 700”, 700’”, 700”” may be substantially similar to clinical analyzer 700, but with differences elaborated herein. As will be described in greater detail below, each clinical analyzer 700’, 700”, 700’”, 700”” contains a respective second substrate dispensing system 730, 740, 760, 770, each having a suitable second substrate material 182 that may be used during an illustrative assay 1100 (FIG. 13) as an alternative to first substrate material 180 located within substrate container 722 and housing 720. Therefore, at least two substrate materials (a first substrate material 180 and a second substrate material 182) are both housed within the automated clinical analyzer 700’, 700”, 700’”, 700””. Additionally, substrate dispensing system 725 in combination with a respective second substrate dispensing system 730, 740, 760, 770 of clinical analyzer 700’, 700”, 700’”, 700””, together form a substrate application assembly 735 configured to select the first substrate material 180 or the second substrate material 182 to be mixed with isolated plurality of particles 120 (some of which being bound to a respective analyte of the patient sample), to form the signal generating mixture 177.

[0124] Turning to FIG. 9, clinical analyzer 700’ includes a second substrate dispensing system 730 having a second substrate reservoir in the form of a second substrate container 732, a designated second substrate dispensing pipette system 734, and a fluid line 736 providing fluid communication between container 732 and pipette system 734. Since clinical analyzer 700’ is a modified version of clinical analyzer 700 described above, clinical analyzer 700’ may include luminometer 600, wash wheel 270, computer system 49, sample aliquot system 705, reagent mixing station 715, incubation wheel 750, and substrate dispensing system 725. Of course, suchcomponents of clinical analyzer 700’ may be substituted for other suitable elements, and even omitted when appropriate.

[0125] Similar to substrate container 722 described above, substrate housing 720 houses second substrate container 732 and provides access to second substrate container 732, thereby allowing replacement of second substrate container 732 when appropriate. Second substrate container 732 may be formed of any suitable material, such as aplastic. Further, second substrate container 732 may include any suitable size and shape as would be apparent to one skilled in the art in view of teachings herein. Second substrate container 732 includes a suitable amount of second substrate material 182. Second substrate material 182 may be preloaded into second substrate container 732 and sealed prior to being suitably inserted into substrate housing 720. Therefore, in the current example, substrate housing 720 houses both substrate containers 722, 732, each of which house a respective substrate material 180, 182. Additionally, second substrate container 732 is suitably attached to second fluid line 736. Second fluid line 736 is in fluid communication with both an interior of substrate container 732 and designated second substrate dispensing pipette system 734.

[0126] In some instances, substrate containers 722, 732 incorporate features that inhibit a user from accidentally coupling substrate container 732 (which contains second substrate material 182) with fluid line 726 of first substrate dispensing system 725 (which is intended to receive first substrate material 180) and also accidentally coupling substrate container 722 (which contains first substrate material 180) with second fluid line 736 of second substrate dispensing system 730 (which is intended to receives second substrate material 182). Therefore, substrate containers 722, 732 may be configured to inhibit inadvertent loading errors when coupling containers 722, 732 to their respective dispensing systems 725, 730. Substrate containers 722, 732 may include any suitable components configured to inhibit such loading errors. For example, substrate containers 722, 732 may include unique bottle shapes for each container 722, 732 that only mate with corresponding features of housing 720 dimensioned to receive such containers 722, 732 (e.g., the feature that provides fluid communication between containers 722, 732 and their respective fluid line 726, 736). As another example, substrate container 722, 732 may include different fluid coupling ports that are specifically designed to only fluidly couple to their intended fluid line 726, 736, respectively.

[0127] In some examples, the substrate application assembly 735 may include one or more readers to read identifiers on the substrate containers 722, 732, such as described in International Patent Application Publication No. WO 2024 / 006490, titled “CONSUMABLES CONTAINER LOADING / UNLOADING SYSTEM,” hereby incorporated by reference in its entirety. For example, at least one reader of the one or more readers may be located at an operator accessible container station. The reader may be configured to read identifiers on the substrate containers 722, 732, that uniquely represent the respective substrate containers 722, 732. That is, the identifiers may be unique to a specific substrate material such that the automated analyzer is configured to determine what substrate material is loaded into the analyzer. In some examples, an error may present if the reader identifies the substrate container is not mated with the correct dispensing system. The reader may be a scanner, for example, a barcode reader, a QR code reader, or an RFID reader which may read out information from the identifiers of the substrate containers 722, 732. The identifiers may include information related to the substrate material (e.g., the first substrate material 180 and the second substrate material 182) contained in the substrate container. The information may include one or more analyses in which the substrate material contained in the substrate containers may be used, the name of the substrate material, the expiration date of the substrate material, the volume of the substrate material, lot information, container information, and the like. In some embodiments, the identifiers may include a bar code, which may include encoded information and is optically read, as well as an RFID tag that may transmit the information stored via radio waves.

[0128] Additionally, substrate container 732 is suitably attached to second fluid line 736. Second fluid line 736 is in fluid communication with both an interior of second substrate container 732 and designated second substrate dispensing pipette system 734. Designated second substrate dispensing pipette system 734 is configured to selectively remove the second substrate material 182 from second substrate container 732 and dispense second substrate material 182 into a reaction vessel 105 suitably located on wash wheel 270. Second designated substrate dispensing pipette system 734 may include a suitable pump, pipette, temperature controls, and any other suitable components. In some instances, one or more of the features of second designated substrate dispensing pipette system 734 may be optionally incorporated into second substrate container 732 and / or incorporated as discrete components in communication with second fluid line 736.

[0129] Designated second substrate dispensing pipette system 734 of the current example is associated with wash wheel 270. Designated second substrate dispensing pipette system 734 may be mounted onto and / or relative to wash wheel 270 such that designated second substrate dispensing pipette system 734 is a component of wash wheel 270. Designated second substrate dispensing pipette system 734 may be located adjacent to wash wheel 270 but mounted to other suitable structure of clinical analyzer 700 (e.g., frame station 701). Second substrate dispensing pipette system 734 may be located at a suitable location on wash wheel 270 to dispense second substrate material 182 into reaction vessel 105. In one example, second substrate dispensing pipette system 734 is located at station SO (FIGS. 3 A and 3B); which is directly adjacent to (A) the station S26 that first substrate dispensing system 725 is associated with, as well as (B) in / out station SI. Therefore, during illustrative use, if the second substrate material 182 is selected in accordance with decision herein, wash wheel 270 may cycle reaction vessel 105 through station S26 associated with first substrate dispensing system 225, place reaction vessel 105 in station SO, and add second substrate material 182 via designated second substrate dispensing pipette system 734. Additionally, as mentioned above, wash wheel 270 is configured to suitably vibrate reaction vessel 105. In the current example, station SO of wash wheel 270 is configured to vibrate in order to suitably mix second substrate material 182 with other components housed within reaction vessel 105 in accordance with the description herein. Once the second substrate material 182 is suitably mixed within reaction vessel 105, the pick and place device may move reaction vessel 105 to incubation wheel 750 to incubate the mixture for a suitable amount of time (e.g., a predetermined amount of time) in accordance with the description herein. For example, reaction vessel 105 may be incubated within incubation wheel 750 for 64 seconds, 192 seconds, etc.

[0130] In some examples, the second substrate dispensing pipette system 734 may be located at other positions of the wash wheel 270 (e.g., any of positions SO to S26). For example, the second substrate dispensing pipette system 734 may be dispensed at the same location (e.g., S26) as the first substrate material 180. In such examples, a device may be operable to align either the first substrate dispensing pipette system 724 or the second substrate dispensing pipette system 734 with the location (e.g., S26).

[0131] Therefore, in the current example, reaction vessel 105 may receive either first or second substrate material 180, 182 at wash wheel 270 such that wash wheel 270 is also configuredto suitably mix first or second substrate material 180 in reaction vessel 105 in accordance with the description herein.

[0132] In FIG. 9, the designated second substrate dispensing pipette system 734 is associated with wash wheel 270, however, the designated second substrate dispensing pipette system 734 may be located at any other suitable location, as described with respect to FIGS. 10 to 12. Accordingly, the aforementioned options apply to FIGS. 10 to 12 (e.g., the reader, unique bottle, etc.) For example, designated second substrate dispensing pipette system 734 may be associated with, or be located adjacent to, reagent mixing station 715, as described with respect to FIG. 12. Additionally, there may be more than one second substrate dispensing pipette system 734 located at various locations on wash wheel 270 and / or other locations such as those of second substrate dispensing pipette system 744, 764, 774 described herein. In such instances, each substrate dispensing pipette system 734 may be configured to deliver the same substrate material and / or different substrate materials. Therefore, in some instances, clinical analyzer 700’ may have two, three, four, or five substrate dispensing pipette systems 734, each located at a suitable location within clinical analyzer 700’. Additionally, second substrate material 182, when selected, may be mixed in reaction vessel 105 at any suitable location. For example, second substrate material 182 may be mixed in reaction vessel 105 with reaction vessel carriage 712.

[0133] Turning to FIG. 10, clinical analyzer 700” includes a second substrate dispensing system 740 having a second substrate reservoir in the form of a second substrate container 742, a designated second substrate dispensing pipette system 744, and a second fluid line 746 providing fluid communication between second substrate container 742 and second substrate dispensing pipette system 744. In the current example, designated second substrate dispensing pipette system 744 is located at a designated mixing station 748 attached to frame 701. As will be described in greater detail below, a pick and place system of analyzer 700”, when the second substrate material 182 is the selected substrate, is configured to transport a reaction vessel 105 from wash wheel 270 to mixing station 748 such that (A) second substrate dispensing pipette system 744 delivers second substrate material 182 into reaction vessel 105, and (B) mixing station 748 suitably mixes the second substrate material 182 within reaction vessel 105.

[0134] Since clinical analyzer 700” is a modified version of clinical analyzer 700 described above, clinical analyzer 700” may include luminometer 600, wash wheel 270, computer system49, sample aliquot system 705, reagent mixing station 715, incubation wheel 750, and substrate dispensing system 725. Of course, such components of clinical analyzer 700” may be substituted for other suitable elements, and even omitted when appropriate.

[0135] Similar to substrate container 722 described above, substrate housing 720 houses second substrate container 742 and provides access to second substrate container 742, thereby allowing replacement of second substrate container 742 when appropriate. Second substrate container 742 may be formed of any suitable material, such as a plastic. Further, second substrate container 742 may include any suitable size and shape as would be apparent to one skilled in the art in view of teachings herein. Second substrate container 742 includes a suitable amount of second substrate material 182. Second substrate material 182 may be preloaded into second substrate container 742 and sealed prior to being suitably inserted into substrate housing 720. Therefore, in the current example substrate housing 720 houses both substrate containers 722, 742, each of which house a respective substrate material 180, 182. Additionally, second substrate container 742 is suitably attached to second fluid line 746. Second fluid line 746 is in fluid communication with both an interior of second substrate container 742 and designated second substrate dispensing pipette system 744.

[0136] In some instances, substrate containers 722, 742 incorporate features that inhibit a user from accidentally coupling second substrate container 742 (which contains second substrate material 182) with fluid line 726 of first substrate dispensing system 725 (which is intended to receive first substrate material 180) and also accidentally coupling substrate container 722 (which contains first substrate material 180) with second fluid line 746 of second substrate dispensing system 740 (which is intended to receive second substrate material 182). Therefore, substrate containers 722, 742 may be configured to inhibit inadvertent loading errors when coupling containers 722, 742 to their respective dispensing systems 725, 740. Substrate containers 722, 742 may include any suitable components configured to inhibit such loading errors. For example, substrate containers 722, 742 may include unique bottle shapes for each container 722, 742 that only mate with corresponding features of housing 720 dimensioned to receive such containers 722, 742 (e.g., the feature that provides fluid communication between containers 722, 742 and their respective fluid line 726, 746). As another example, substrate container 722, 742 may include different fluid coupling ports that are specifically designed to only fluidly couple to their intendedfluid line 726, 746, respectively. Additionally, or alternatively, the substrate application assembly 735 may include a reader and the substrate containers 722, 742 may include identifiers as described above and in WO 2024 / 006490.

[0137] Additionally, second substrate container 742 is suitably attached to second fluid line 746. Second fluid line 746 is in fluid communication with both an interior of second substrate container 742 and designated second substrate dispensing pipette system 744. Designated second substrate dispensing pipette system 744 is configured to selectively remove the second substrate material 182 from second substrate container 742 and dispense second substrate material 182 into a reaction vessel 105 suitably located at mixing station 748. Second designated substrate dispensing pipette system 744 may include a suitable pump, pipette, temperature controls, and any other suitable components. In some instances, one or more of the features of designated second substrate dispensing pipette system 744 may be optionally incorporated into second substrate container 742 and / or incorporated as discrete components in communication with second fluid line 746.

[0138] Designated second substrate dispensing pipette system 744 of the current example is associated with its own mixing station 748 configured to suitably receive a reaction vessel 105 from wash wheel 270 when the second substrate material 182 is the selected substrate. As mentioned above, mixing station 748 is configured to suitably mix second substate 182 once dispensed within reaction vessel 105 in accordance with the description herein.

[0139] Second substrate dispensing pipette system 744 and mixing station 748 may be located at a suitable location on frame 701. For example, designated second substrate dispensing pipette system 744 may associated with, or be located adjacent to, reagent mixing station 715. Additionally, there may be more than one second substrate dispensing pipette system 744 located at various locations on wash wheel 270 and / or other locations such as those of second substrate dispensing pipette system 734, 764, 774 described herein. In such instances, each second substrate dispensing pipette system 744 may be configured to deliver the same substrate material and / or different substrate materials, as would be apparent to one skilled in the art. Therefore, in some instances, clinical analyzer 700” may have two, three, four, or five substrate dispensing pipette systems 744, each located at a suitable location within clinical analyzer 700”.

[0140] During illustrative use, if the second substrate material 182 is selected in accordance with decision herein, wash wheel 270 may cycle reaction vessel 105 through station S26 associated with first substrate dispensing system 225, place reaction vessel 105 at the in / out station SI, and transfer reaction vessel 105 to mixing station 748 in order to add and mix the second substrate material 182 via designated second substrate dispensing pipette system 744. Once the second substrate material 182 is suitably mixed within reaction vessel 105, the pick and place device may move reaction vessel 105 to incubation wheel 750 in order to incubate the mixture for a suitable amount of time (e.g., a predetermined amount of time) in accordance with the description herein. For example, reaction vessel 105 may be incubated within incubation wheel 750 for 64 seconds, 192 seconds, etc.

[0141] Turning to FIG. 11, clinical analyzer 700”’ includes a second substrate dispensing system 760 having a second substrate reservoir in the form of a second substrate container 762, a designated second substrate dispensing pipette system 764, and a second fluid line 766 providing fluid communication between second substrate container 762 and second substrate dispensing pipette system 764. In the current example, designated second substrate dispensing pipette system 764 is located at incubation wheel 750 such that second substrate dispensing pipette system 764 may distribute second substrate material 182 into a reaction vessel 105 housed in incubation wheel 750. As will be described in greater detail below, a pick and place system of analyzer 700” ’, when the second substrate material 182 is the selected substrate, is configured to transport a reaction vessel 105 from wash wheel 270 to incubation wheel 750 such that (A) second substrate dispensing pipette system 764 delivers second substrate material 182 into reaction vessel 105, and (B) incubation wheel 750 suitably mixes the second substrate material 182 within reaction vessel 105.

[0142] Since clinical analyzer 700’” is a modified version of clinical analyzer 700 described above, clinical analyzer 700’” may include luminometer 600, wash wheel 270, computer system 49, sample aliquot system 705, reagent mixing station 715, incubation wheel 750, and substrate dispensing system 725. Of course, such components of clinical analyzer 700’” may be substituted for other suitable elements, and even omitted when appropriate.

[0143] Similar to substrate container 722 described above, substrate housing 720 houses second substrate container 762 and provides access to second substrate container 762, thereby allowing replacement of second substrate container 762 when appropriate. Second substratecontainer 762 may be formed of any suitable material, such as aplastic. Further, second substrate container 762 may include any suitable size and shape as would be apparent to one skilled in the art in view of teachings herein. Second substrate container 762 includes a suitable amount of second substrate material 182. Second substrate material 182 may be preloaded into second substrate container 762 and sealed prior to being suitably inserted into substrate housing 720. Therefore, in the current example, substrate housing 720 houses both substrate containers 722, 762, each of which house a respective substrate material 180, 182. Additionally, second substrate container 762 is suitably attached to second fluid line 766. Second fluid line 766 is in fluid communication with both an interior of second substrate container 762 and designated second substrate dispensing pipette system 764.

[0144] In some instances, substrate containers 722, 762 incorporate features that inhibit a user from accidentally coupling second substrate container 762 (which contains second substrate material 182) with fluid line 726 of first substrate dispensing system 725 (which is intended to receive first substrate material 180) and also accidentally coupling substrate container 722 (which contains first substrate material 180) with second fluid line 766 of second substrate dispensing system 760 (which is intended to receive second substrate material 182). Therefore, substrate containers 722, 762 may be configured to inhibit inadvertent loading errors when coupling containers 722, 762 to their respective dispensing systems 725, 760. Substrate containers 722, 762 may include any suitable components configured to inhibit such loading errors. For example, substrate containers 722, 762 may include unique bottle shapes for each container 722, 762 that only mate with corresponding features of housing 720 dimensioned to receive such containers 722, 762 (e.g., the feature that provides fluid communication between containers 722, 762 and their respective fluid line 726, 766). As another example, substrate container 722, 762 may include different fluid coupling ports that are specifically designed to only fluidly couple to their intended fluid line 726, 746, respectively. Additionally, or alternatively, the substrate application assembly 735 may include a reader and the substrate containers 722, 762 may include identifiers as described above and in WO 2024 / 006490.

[0145] Additionally, second substrate container 762 is suitably attached to second fluid line766. Second fluid line 766 is in fluid communication with both an interior of second substrate container 762 and designated second substrate dispensing pipette system 764. Designated secondsubstrate dispensing pipette system 764 is configured to selectively remove the second substrate material 182 from second substrate container 762 and dispense second substrate material 182 into a reaction vessel 105 suitably located at incubation wheel 750. Designated second substrate dispensing pipette system 764 may include a suitable pump, pipette, temperature controls, and any other suitable components. In some instances, one or more of the features of designated second substrate dispensing pipette system 764 may be optionally incorporated into second substrate container 762 and / or incorporated as discrete components in communication with fluid line 766.

[0146] Designated second substrate dispensing pipette system 764 of the current example is associated with incubation wheel 750, which is configured to suitable receive a reaction vessel 105 from wash wheel 270 when the second substrate material 182 is the selected substrate. In the current example, incubation wheel 750 is configured to suitably mix second substrate material 182 within reaction vessel 105 in accordance with the description herein. Second substrate dispensing pipette system 764 and incubation wheel 750 may be located at a suitable location with frame 701. For example, designated second substrate dispensing pipette system 764 may associated with, or be located adjacent to, reagent mixing station 715. Additionally, there may be more than one second substrate dispensing pipette system 764 located at various locations on wash wheel 270 and / or other locations such as those of second substrate dispensing pipette system 734, 744 described herein. In such instances, each substrate dispensing pipette system 764 may be configured to deliver the same substrate material 182 and / or different substrate materials, as would be apparent to one skilled in the art. Therefore, in some instances, clinical analyzer 700’” may have two, three, four, or five substrate dispensing pipette systems 764, each located at a suitable location within clinical analyzer 700”’.

[0147] During illustrative use, if the second substrate material 182 is selected in accordance with decision herein, wash wheel 270 may cycle reaction vessel 105 through station S26 associated with first substrate dispensing system 225, place reaction vessel 105 at the in / out station SI, and transfer reaction vessel 105 to incubation wheel 750 in order to add and mix the second substrate material 182 via designated second substrate dispensing pipette system 764 and incubation wheel 750. Once the second substrate material 182 is suitably mixed within reaction vessel 105, incubation wheel 750 may then suitably incubate the mixture for a suitable amount of time (e.g., apredetermined amount of time) in accordance with the description herein. For example, reaction vessel 105 may be incubated within incubation wheel 750 for 64 seconds, 192 seconds, etc.

[0148] FIG. 12 illustrates a clinical analyzer 700”” having a second substrate dispensing system 770. The second substrate dispensing system 770 has a second substrate reservoir in the form of a second substrate container 772, a designated second substrate dispensing pipette system 774, and a second fluid line 766 providing fluid communication between second substrate container 772 and second substrate dispensing pipette system 774. In the current example, designated second substrate dispensing pipette system 764 is located at or adjacent to a reagent station housing of the reagent mixing station 715 such that second substrate dispensing pipette system 774 may distribute second substrate material 182 into a reaction vessel 105 housed at a reaction vessel carriage 712. As will be described in greater detail below, a pick and place system of analyzer 700””, when the second substrate material 182 is the selected substrate, is configured to transport a reaction vessel 105 to the reaction vessel carriage 712 such that (A) second substrate dispensing pipette system 774 delivers second substrate material 182 into reaction vessel 105, and (B) the reaction mixing station 715 suitably mixes the second substrate material 182 within reaction vessel 105.

[0149] Since clinical analyzer 700”” is a modified version of clinical analyzer 700 described above, clinical analyzer 700”” may include luminometer 600, wash wheel 270, computer system 49, sample aliquot system 705, reagent mixing station 715, incubation wheel 750, and substrate dispensing system 725. Of course, such components of clinical analyzer 700’ ’ ’ ’ may be substituted for other suitable elements, and even omitted when appropriate.

[0150] Similar to substrate container 722 described above, substrate housing 720 houses second substrate container 772 and provides access to second substrate container 772, thereby allowing replacement of second substrate container 772 when appropriate. Second substrate container 772 may be formed of any suitable material, such as a plastic. Further, second substrate container 772 may include any suitable size and shape as would be apparent to one skilled in the art in view of teachings herein. Second substrate container 772 includes a suitable amount of second substrate material 182. Second substrate material 182 may be preloaded into second substrate container 772 and sealed prior to being suitably inserted into substrate housing 720. Therefore, in the current example, substrate housing 720 houses both substrate containers 722,772, each of which house a respective substrate material 180, 182. Additionally, second substrate container 772 is suitably attached to second fluid line 776. Second fluid line 776 is in fluid communication with both an interior of second substrate container 772 and designated second substrate dispensing pipette system 774.

[0151] In some instances, substrate containers 722, 772 incorporate features that inhibit a user from accidentally coupling second substrate container 772 (which contains second substrate material 182) with fluid line 726 of first substrate dispensing system 725 (which is intended to receive first substrate material 180) and also accidentally coupling substrate container 722 (which contains first substrate material 180) with second fluid line 776 of second substrate dispensing system 760 (which is intended to receive second substrate material 182). Therefore, substrate containers 722, 772 may be configured to inhibit inadvertent loading errors when coupling containers 722, 772 to their respective dispensing systems 725, 770. Substrate containers 722, 772 may include any suitable components configured to inhibit such loading errors. For example, substrate containers 722, 772 may include unique bottle shapes for each container 722, 772 that only mate with corresponding features of housing 720 dimensioned to receive such containers 722, 772 (e.g., the feature that provides fluid communication between containers 722, 772 and their respective fluid line 726, 776). As another example, substrate container 722, 772 may include different fluid coupling ports that are specifically designed to only fluidly couple to their intended fluid line 726, 776, respectively. Additionally, or alternatively, the substrate application assembly 735 may include a reader and the substrate containers 722, 772 may include identifiers as described above and in International Patent Application Publication No. WO 2024 / 006490.

[0152] Additionally, second substrate container 772 is suitably attached to second fluid line 776. Second fluid line 776 is in fluid communication with both an interior of second substrate container 772 and designated second substrate dispensing pipette system 774. Designated second substrate dispensing pipette system 774 is configured to selectively remove the second substrate material 182 from second substrate container 772 and dispense second substrate material 182 into a reaction vessel 105 suitably located at reagent mixing station 715. Designated second substrate dispensing pipette system 774 may include a suitable pump, pipette, temperature controls, and any other suitable components. In some instances, one or more of the features of designated secondsubstrate dispensing pipette system 774 may be optionally incorporated into second substrate container 772 and / or incorporated as discrete components in communication with fluid line 776.

[0153] Additionally, there may be more than one second substrate dispensing pipette system 774 located at various locations on wash wheel 270 and / or other locations such as those of second substrate dispensing pipette system 734, 744, 764 described herein. In such instances, each substrate dispensing pipette system 774 may be configured to deliver the same substrate material 182 and / or different substrate materials, as would be apparent to one skilled in the art. Therefore, in some instances, clinical analyzer 700”” may have two, three, four, or five substrate dispensing pipette systems 764, each located at a suitable location within clinical analyzer 700””.

[0154] During illustrative use, if the second substrate material 182 is selected in accordance with decision herein, wash wheel 270 may cycle reaction vessel 105 through station S26 associated with first substrate dispensing system 225, place reaction vessel 105 at the in / out station SI, and transfer reaction vessel 105 to the reagent mixing station 715 to add and mix the second substrate material 182. Once the second substrate material 182 is suitably mixed within reaction vessel 105, the pick and place device may move reaction vessel 105 to the incubation wheel 750 to incubate the mixture for a suitable amount of time (e.g., a predetermined amount of time) in accordance with the description herein. For example, reaction vessel 105 may be incubated within incubation wheel 750 for 64 seconds, 192 seconds, etc.

[0155] The second substrate dispensing pipette system 734, 744, 764, and 774 may be the same as or similar to the first substrate dispensing pipette system 724. For example, any of substrate dispensing pipette systems 724, 734, 744, 764, and 774 may be in accordance with the pipetting system 510 described with respect to FIGS. 4 and 5.

[0156] FIG. 13 shows an illustrative method of using automated clinical analyzer 700’, 700”, 700’”, 700”” such that clinical analyzer 700’, 700”, 700’”, 700”” may select a first substrate material 180 or a second substrate material 182 for a particular assay. The clinical analyzer 700’, 700”, 700”’, 700”” may be programmed to select a first substrate material 180 or a second substrate material 182 for detection of a particular analyte or may be programmed to select a particular substrate material based on the results of a previous assay. Additionally, or alternatively, the operator of a clinical analyzer 700’, 700”, 700’”, 700”” may provide instructions to that clinical analyzer 700’, 700”, 700”’, 700”” indicating which substrate should be selected. First,although not shown, the clinical analyzer 700’, 700”, 700”’, 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. Next, clinical analyzer 700’, 700”, 700”’, 700”” may mix patient samples with reagent(s) 1102, incubate mixtures 1104, and perform wash cycle(s) 1106 in order to prepare to create a mixture by adding either first substrate material 180 or second substrate material 182 in accordance with the description herein. 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. Therefore, clinical analyzer 700’, 700”, 700’”, 700”” may perform steps 1102, 1104, 1106 in order to (A) suitably form 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. 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 substate material 180 via first substrate dispensing system 725. Alternatively, reaction vessel 105 may be located at any other suitable location.

[0157] Once the contents of a reaction vessel 105 are configured to receive a reporting agent in accordance with the description herein, the clinical analyzer 700’, 700”, 700’”, 700”” selects a substrate material 1108 to be used for the particular assay. Clinical analyzer 700’, 700”, 700’”, 700”” may select 1108 either a first substrate material 180 or a second substrate material 182, both of which are housed within automated clinical analyzer 700’, 700”, 700’”, 700””. Clinical analyzer 700’, 700”, 700’”, 700”” may use any suitable means to select either first substrate material 180 or second substrate material for a particular assay. For illustrative purposes, clinical analyzer 700’, 700”, 700”’, 700”” may select either first substrate material 180 or 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’, 700”, 700’”, 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’, 700”, 700’”, 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’, 700”, 700’”, 700”” does not use both the first substrate material 180 and the second substrate material 182 in the same assay.

[0158] If the first substrate material 180 is selected 1110, clinical analyzer 700’, 700”, 700’”, 700”” transfers 1112 the reaction vessel 105 to the substrate receiving station s26 of the wash wheel 270. The clinical analyzer 700’, 700”, 700”’, 700”” obtains, adds, and mixes 1114 the first substrate material 180 to reaction vessel 105 at wash wheel 270 in accordance with the description herein to create the signal generating mixture 177. Next, clinical analyzer 700’, 700”, 700’” transfers 1116 the reaction vessel 105 to the incubation wheel 750 and suitably incubates the mixture for a suitable amount of time (e.g., a predetermined amount of time) in accordance with the description herein. Next, clinical analyzer 700’, 700”, 700’”, 700”” transfers the reaction vessel to the luminometer 600 (or any other suitable detector) and analyzes the mixture containing the selected substrate with a detector (e.g., measures 1118 light generated by the mixture within reaction vessel 105) in accordance with the description herein.

[0159] However, if the second substrate material 182 is selected 1120, clinical analyzer 700’,700”, 700”’, 700””, 700”” transfers 1122 the reaction vessel 105 to the designated second substrate dispensing station 734, 744, 764, 774 in accordance with the description herein. Next, clinical analyzer 700’, 700”, 700’”, 700”” utilizes second substrate dispensing station 734, 744, 764, 774 to obtain and add 1124 the second substrate material 182 to reaction vessel 105. At this moment, suitable components of a respective second substrate dispensing station 734, 744, 764, 774 may mix the second substrate material 182 with the contents of reaction vessel 105 in accordance with the description herein to create a suitable signal generating mixture 177.

[0160] Next, in instances where reaction vessel 105 is not already located at incubation wheel 750, clinical analyzer 700’, 700”, 700’”, 700”” transfers 1126 the reaction vessel 105 to the incubation wheel 750. Incubation wheel 750 then suitably incubates the mixture for a suitable amount of time (e.g., a predetermined amount of time) in accordance with the description herein. For example, reaction vessel 105 may be incubated within incubation wheel 750 for 64 seconds,192 seconds, etc. Next, clinical analyzer 700’, 700”, 700”’, 700”” transfers the reaction vessel to the luminometer 600 and measures 1128 to light generated by the mixture within reaction vessel 105 in accordance with the description herein.

[0161] Therefore, the clinical analyzer 700’, 700”, 700’”, 700”” performing steps in workflow 1100 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 (e.g., changing the substrate container 722 of substrate dispensing system 725).

[0162] III. Illustrative First and Second Substrate Materials

[0163] 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, while other assays will be better suited using second substrate material 182. Therefore, when second substrate dispensing system 730, 740, 760, 770 is incorporated into clinical analyzer 700, analyzer 700 may select one of at least two substrate materials 180, 182 to perform an assay, where the selected substrate material 180, 182 may be better suited for a particular assay compared to the substrate material 180, 182 not selected.

[0164] Substrate materials 180, 182 may include any suitable substrate material that would be apparent to one skilled in the art in view of the teachings herein. In one illustrative example, substrate material(s) may include(d) 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. Such a substrate may be further configured in accordance with the teachings of PCT Publication No. WO 2018 / 006059, entitled “Chemiluminescent Substrates,” published on January 4, 2018.Additionally, or alternatively, substrate material(s) 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, C6-C10 aryl, or heteroaryl;R4is C2-C10 alkenyl; R5is 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.

[0165] IV. Additional Examples

[0166] 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.

[0167] Example 1

[0168] 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 a first reagent and a second reagent within a reaction vessel to create an assay mixture, the first reagent comprising a plurality of particles configured to specifically bind to the analyte from the patient sample, the second reagent comprising a reporter; (b) incubating the reaction vessel containing the assay mixture for a predetermined amount of time; (c) 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; (d) selecting a first substrate material or a second substrate material, wherein both the first substrate material and the second substrate material are housed within the automated clinical analyzer; (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 signal generating mixture; and (h) analyzing the signal generating mixture with a detector.

[0169] Example la.

[0170] The method of Example 1, wherein the reporter comprises an enzyme.

[0171] Example lb.

[0172] The method of Example 1 or la, wherein the detector comprises a luminometer.

[0173] Example 2.

[0174] The method of Example 1, la, or lb„ wherein the automated clinical analyzer comprises a wash assembly configured to perform the wash cycle, wherein the wash assembly comprises a substrate dispensing pipette system, wherein if the first substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the substrate dispensing pipette system while the reaction vessel is located at the wash assembly.

[0175] Example 3.

[0176] The method of Example 1, la, or lb, wherein the automated clinical analyzer comprises a wash assembly configured to perform the wash cycle, wherein the wash assembly comprises a substrate dispensing pipette system, wherein the first substrate material is the selected substrate material, wherein addition of the selected substrate material into the reaction vessel occurs with the substrate dispensing pipette system while the reaction vessel is located at the wash assembly.

[0177] Example 4.

[0178] The method of Example 2 or 3, wherein the substrate dispensing pipette system is in fluid communication with the first substrate material.

[0179] Example 5.

[0180] The method of any one or more of Examples 2 through 4, wherein the automated clinical analyzer comprises an incubation station configured to incubate the assay mixture and the signal generating mixture, wherein the incubation station comprises a second substrate dispensing pipette system.

[0181] Example 6.

[0182] The method of Example 5, wherein if the second substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at the incubation station.

[0183] Example 7.

[0184] The method of Example 5, wherein the second substrate material is the selected substrate material, wherein addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at the incubation station.

[0185] Example 8.

[0186] The method of any one or more of Examples 5 through 7, wherein the second substrate dispensing pipette system is in fluid communication with the second substrate material.

[0187] Example 9.

[0188] The method of any one or more of Examples 2 through 4, wherein the automated clinical analyzer comprises a second substrate dispensing pipette system located adjacent to or at the same position of the wash wheel, wherein if the second substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system.

[0189] Example 10.

[0190] The method of Example 9, wherein if the second substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at the wash assembly.

[0191] Example 11.

[0192] The method of any one or more of Examples 2 through 4, wherein the automated clinical analyzer comprises a second substrate dispensing pipette system located adjacent to or at the same position of the wash wheel, wherein the second substrate material is the selected substrate material, wherein addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system.

[0193] Example 12.

[0194] The method of Example 11, wherein addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at the wash assembly.

[0195] Example 13.

[0196] The method of any one or more of Examples 9 through 12, wherein the second substrate dispensing pipette system is in fluid communication with the second substrate material.

[0197] Example 14.

[0198] The method of any one or more of Examples 2 through 4, wherein the automated clinical analyzer comprises a second substrate dispensing pipette system located adjacent to a reagent station housing the first reagent, wherein if the second substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system.

[0199] Example 15.

[0200] The method of Example 14, wherein if the second substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at a reagent mixing station where the assay mixture is formed.

[0201] Example 16.

[0202] The method of any one or more of Examples 2 through 4, wherein the automated clinical analyzer comprises a second substrate dispensing pipette system located adjacent to a reagent station housing the first reagent, wherein the second substrate material is the selected substrate material, wherein addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system.

[0203] Example 17.

[0204] The method of Example 16, wherein the second substrate material is the selected substrate material, wherein addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at a reagent mixing station where the assay mixture is formed.

[0205] Example 18.

[0206] The method of any one or more of Examples 1 through 17, wherein mixing the composition with the first reagent and the second reagent within the reaction vessel further comprises adding the first reagent and the second reagent to the reaction vessel sequentially, prior to incubation.

[0207] Example 19.

[0208] The method of any one or more of Examples 1 through 18, wherein mixing the composition with the first reagent and the second reagent within the reaction vessel further comprises adding the first reagent into the reaction vessel prior to adding the second reagent to the reaction vessel.

[0209] Example 20.

[0210] The method of Example 19, further comprising incubating the reaction vessel prior to adding the second reagent to the reaction vessel.

[0211] Example 21.

[0212] The method of either of Examples 19 or 20, further comprising performing a wash cycle prior to adding the second reagent to the reaction vessel.

[0213] Example 22.

[0214] The method of any one or more of Examples 1 through 18, wherein mixing the composition with the first reagent and the second reagent within the reaction vessel further comprises adding the second reagent into the reaction vessel prior to adding the first reagent to the reaction vessel.

[0215] Example 23.

[0216] The method of any one or more of Examples 1 through 22, wherein the second reagent further comprises an antibody.

[0217] Example 24.

[0218] The method of any one or more of Examples 1 through 23, wherein the second reagent further comprises a competing analyte.

[0219] Example 25.

[0220] The method of any one or more of Examples 1 through 24, wherein the first reagent comprises an antibody.

[0221] Example 25a.

[0222] The method of any one or more of Examples 1 through 25, wherein the second reagent comprises an analyte.

[0223] Example 26.

[0224] The method of any one or more of Examples 1 through 25, wherein either the first reagent, the second reagent, or both the first reagent and the second reagent comprise an antibody.

[0225] Example 27.

[0226] The method of any one or more of Examples 1 through 26, wherein the plurality of particles further comprises a plurality of iron particles.

[0227] Example 28.

[0228] The method of Example 27, 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.

[0229] Example 29.

[0230] The method of any one or more of Examples 1 through 28, wherein performing the wash cycle further comprises adding a washing agent to the reaction vessel.

[0231] Example 30.

[0232] The method of Example 29, 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, and up to 100 times.

[0233] Example 31.

[0234] The method of Example 29, wherein adding the washing agent is performed while the plurality of iron particles is attached toward the magnet.

[0235] Example 32.

[0236] 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 a 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 first 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 material, wherein both the first substrate material and the second substrate material are housed within the automated clinical analyzer; (e) obtaining the selected substrate material; (f) adding and mixing the selected substrate material into the reaction vessel with the plurality of particles to create a signal generating mixture; (g) incubating the signal generating mixture; and (h) analyzing the signal generating mixture with a luminometer.

[0237] Example 33.

[0238] The method of Example 32, wherein the automated clinical analyzer comprises a substrate housing, wherein the first substrate material and the second substrate material are both located within the substrate housing.

[0239] Example 34.

[0240] The method of either Example 32 or Example 33, wherein the first substrate material is contained in a first bottle having a first unique bottle shape, wherein the second substrate material is contained in a second bottle having a second unique bottle shape.

[0241] Example 35.

[0242] The method of any one or more of Examples 32 through 34, wherein the automated clinical analyzer comprises a wash assembly configured to perform the first wash cycle, wherein the automated clinical analyzer comprises an incubation station configured to incubate the assay mixture and the signal generating mixture.

[0243] Example 36.

[0244] The method of Example 35, wherein if the second substrate material is the selected substrate material, the second substrate material is added to the reaction vessel while the reaction vessel is housed within the incubation station.

[0245] Example 37.

[0246] The method of Example 35, wherein if the second substrate material is the selected substrate material, the second substrate material is added to the reaction vessel while the reaction vessel is housed within the wash assembly.

[0247] Example 38.

[0248] The method of Example 35, wherein the second substrate material is the selected substrate material, wherein the second substrate material is added to the reaction vessel while the reaction vessel is housed within the incubation station.

[0249] Example 39.

[0250] The method of Example 35, wherein the second substrate material is the selected substrate material, wherein the second substrate material is added to the reaction vessel while the reaction vessel is housed within the wash assembly.

[0251] Example 40.

[0252] An automated clinical analyzer, comprising: (a) a reagent mixing station configured to mix a composition comprising a patient sample with at least one reagent to form an assay mixture; (b) an incubation station configured to incubate the mixture containing at least a portion of the assay mixture; (c) a wash assembly configured to isolate a plurality of particles from the assay mixture, some particles of the isolated plurality of particles are bound to a respective analyte of the patient sample; (d)a substrate application assembly configured to select a first substrate or a second substrate to be mixed with the isolated plurality of particles to form a signal generating mixture; and (e) a luminometer configured to analyze light emitted from the signal generating mixture.

[0253] Example 41.

[0254] The automated clinical analyzer of Example 40, wherein the substrate application assembly comprises a first substrate dispensing pipette system configured to deliver the first substrate, and a second substrate dispensing pipette system configured to deliver the second substrate.

[0255] Example 42.

[0256] The automated clinical analyzer of Example 41, wherein the first substrate dispensing pipette system is associated with the wash assembly, wherein the second substrate dispensing pipette system is associated with the incubation station.

[0257] Example 43.

[0258] The automated clinical analyzer of Example 41 , wherein the first substrate dispensing pipette system and the second substrate dispensing pipette system are associated with the wash assembly.

[0259] Example 44.

[0260] The automated clinical analyzer of any one or more of Examples 41 through 43, wherein the substrate application assembly comprises a first substrate reservoir and a second substrate reservoir, wherein the first substrate reservoir is in communication with the first substrate dispensing pipette system, wherein the second substrate reservoir is in communication with the second substrate dispensing pipette system.

[0261] Example 45.

[0262] The method or clinical analyzer of any one or more of the preceding Examples, wherein the first 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.

[0263] Example 46

[0264] The method or clinical analyzer of any one or more of the preceding Examples, wherein 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, C6-C10 aryl, or heteroaryl;R4is C2-C10 alkenyl; R5is H or C1-C10 alkyl; X is a phosphate; and at least one surfactant.

[0265] Example 47

[0266] The method of Example 44, wherein the second substrate material comprisescompound having a structure:

[0267] IV. Interpretation

[0268] 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.

[0269] 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”).

[0270] 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.

[0271] 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.

[0272] 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 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.

[0273] 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.

[0274] 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.

[0275] 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.

[0276] 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.

[0277] 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.

[0278] 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.

[0279] 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.

[0280] 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.

[0281] 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.

[0282] 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.

[0283] 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 embodiment of the disclosure. Furthermore, the particular features, configurations, compositions, or characteristics may be combined in any suitable manner in one or more embodiments.

[0284] 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.

[0285] 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 documentincorporated 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) mixing a composition comprising an analyte from a patient sample with a first reagent and a second reagent within a reaction vessel to create an assay mixture, the first reagent comprising a plurality of particles configured to specifically bind to the analyte from the patient sample, the second reagent comprising a reporter;(b) incubating the reaction vessel containing the assay mixture for a predetermined amount of time;(c) 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;(d) selecting a first substrate material or a second substrate material, wherein both the first substrate material and the second substrate material are housed within the automated clinical analyzer;(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 signal generating mixture; and(h) analyzing the signal generating mixture with a detector.

2. The method of claim 1, wherein the automated clinical analyzer comprises an incubation station configured to incubate the assay mixture and the signal generating mixture, wherein the incubation station comprises a second substrate dispensing pipette system.

3. The method of claim 2, wherein if the second substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at the incubation station.

4. The method of claim 2, wherein the second substrate material is the selected substrate material, wherein addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at the incubation station.

5. The method of any one or more of claims 2 through 4, wherein the second substrate dispensing pipette system is in fluid communication with the second substrate material.

6. The method of claim 1 , wherein the automated clinical analyzer comprises a second substrate dispensing pipette system located adjacent to or at the same position of the wash wheel, wherein if the second substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system.

7. The method of claim 6, wherein if the second substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at the wash assembly.

8. The method of claim 1 , wherein the automated clinical analyzer comprises a second substrate dispensing pipette system located adjacent to or at the same position of the wash wheel, wherein the second substrate material is the selected substrate material, wherein addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system.

9. The method of claim 8, wherein addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at the wash assembly.

10. The method of any one or more of claims 6 through 9, wherein the second substrate dispensing pipette system is in fluid communication with the second substrate material.

11. The method of claim 1 , wherein the automated clinical analyzer comprises a second substrate dispensing pipette system located adjacent to a reagent station housing the first reagent, wherein if the second substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system.

12. The method of claim 11, wherein if the second substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at a reagent mixing station where the assay mixture is formed.

13. The method of claim 1 , wherein the automated clinical analyzer comprises a second substrate dispensing pipette system located adjacent to a reagent station housing the first reagent, wherein the second substrate material is the selected substrate material, wherein addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system.

14. The method of claim 13, wherein the second substrate material is the selected substrate material, wherein addition of the selected substrate material into the reaction vessel occurs with the second substrate dispensing pipette system while the reaction vessel is located at a reagent mixing station where the assay mixture is formed.

15. The method of any one or more of claims 1 through 14, wherein mixing the composition with the first reagent and the second reagent within the reaction vessel further comprises adding the first reagent and the second reagent to the reaction vessel sequentially, prior to incubation.

16. The method of any one or more of claims 1 through 15, wherein mixing the composition with the first reagent and the second reagent within the reaction vessel further comprises adding the first reagent into the reaction vessel prior to adding the second reagent to the reaction vessel.

17. The method of claim 16, further comprising incubating the reaction vessel prior to adding the second reagent to the reaction vessel.

18. The method of either of claims 16 or 17, further comprising performing a wash cycle prior to adding the second reagent to the reaction vessel.

19. The method of any one or more of claims 1 through 15, wherein mixing the composition with the first reagent and the second reagent within the reaction vessel further comprises adding the second reagent into the reaction vessel prior to adding the first reagent to the reaction vessel.

20. The method of any one or more of claims 1 through 19, wherein the second reagent further comprises an antibody.

21. The method of any one or more of claims 1 through 20, wherein the second reagent further comprises a competing analyte.

22. The method of any one or more of claims 1 through 21, wherein the first reagent comprises an antibody.

23. The method of any one or more of claims 1 through 22, wherein the second reagent comprises an analyte.

24. The method of any one or more of claims 1 through 23, wherein either the first reagent, the second reagent, or both the first reagent and the second reagent comprise an antibody.

25. The method of any one or more of claims 1 through 24, wherein the plurality of particles further comprises a plurality of iron particles.

26. The method of claim 25, 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.

27. The method of any one or more of claims 1 through 26, wherein performing the wash cycle further comprises adding a washing agent to the reaction vessel.

28. The method of claim 27, 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, and up to 100 times.

29. The method of claim 27, wherein adding the washing agent is performed while the plurality of iron particles is attached toward the magnet.

30. The method of any one or more of claims 1 through 29, wherein the reporter comprises an enzyme.

31. The method of any one or more of claims 1 through 30, wherein the detector comprises a luminometer.

32. The method of any one or more of claims 1 through 31, wherein the automated clinical analyzer comprises a wash assembly configured to perform the wash cycle, wherein the wash assembly comprises a substrate dispensing pipette system, wherein if the first substrate material is the selected substrate material, addition of the selected substrate material into the reaction vessel occurs with the substrate dispensing pipette system while the reaction vessel is located at the wash assembly.

33. The method of any one or more of claims 1 through 31, wherein the automated clinical analyzer comprises a wash assembly configured to perform the wash cycle, wherein the wash assembly comprises a substrate dispensing pipette system, wherein the first substrate material is the selected substrate material, wherein addition of the selected substrate material into the reaction vessel occurs with the substrate dispensing pipette system while the reaction vessel is located at the wash assembly.

34. The method of claim 32 or 33, wherein the substrate dispensing pipette system is in fluid communication with the first substrate material.

35. 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 a 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 first 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 material, wherein both the first substrate material and the second substrate material are housed within the automated clinical analyzer;(e) obtaining the selected substrate material;(f) adding and mixing the selected substrate material into the reaction vessel with the plurality of particles to create a signal generating mixture;(g) incubating the signal generating mixture; and(h) analyzing the signal generating mixture with a luminometer.

36. The method of claim 35, wherein the automated clinical analyzer comprises a substrate housing, wherein the first substrate material and the second substrate material are both located within the substrate housing.

37. The method of either claim 35 or claim 36, wherein the first substrate material is contained in a first bottle having a first unique bottle shape, wherein the second substrate material is contained in a second bottle having a second unique bottle shape.

38. The method of any one or more of claims 35 through 37, wherein the automated clinical analyzer comprises a wash assembly configured to perform the first wash cycle, wherein the automated clinical analyzer comprises an incubation station configured to incubate the assay mixture and the signal generating mixture.

39. The method of claim 38, wherein if the second substrate material is the selected substrate material, the second substrate material is added to the reaction vessel while the reaction vessel is housed within the incubation station.

40. The method of claim 38, wherein if the second substrate material is the selected substrate material, the second substrate material is added to the reaction vessel while the reaction vessel is housed within the wash assembly.

41. The method of claim 38, wherein the second substrate material is the selected substrate material, wherein the second substrate material is added to the reaction vessel while the reaction vessel is housed within the incubation station.

42. The method of claim 38, wherein the second substrate material is the selected substrate material, wherein the second substrate material is added to the reaction vessel while the reaction vessel is housed within the wash assembly.

43. An automated clinical analyzer, comprising:(a) a reagent mixing station configured to mix a composition comprising a patient sample with at least one reagent to form an assay mixture;(b) an incubation station configured to incubate the mixture containing at least a portion of the assay mixture;(c) a wash assembly configured to isolate a plurality of particles from the assay mixture, some particles of the isolated plurality of particles are bound to a respective analyte of the patient sample;(d) a substrate application assembly configured to select a first substrate or a second substrate to be mixed with the isolated plurality of particles to form a signal generating mixture; and(e) a luminometer configured to analyze light emitted from the signal generating mixture.

44. The automated clinical analyzer of claim 43, wherein the substrate application assembly comprises a first substrate dispensing pipette system configured to deliver the first substrate, and a second substrate dispensing pipette system configured to deliver the second substrate.

45. The automated clinical analyzer of claim 44, wherein the first substrate dispensing pipette system is associated with the wash assembly, wherein the second substrate dispensing pipette system is associated with the incubation station.

46. The automated clinical analyzer of claim 44, wherein the first substrate dispensing pipette system and the second substrate dispensing pipette system are associated with the wash assembly.

47. The automated clinical analyzer of any one or more of claims 44 through 46, wherein the substrate application assembly comprises a first substrate reservoir and a second substrate reservoir, wherein the first substrate reservoir is in communication with the first substrate dispensing pipette system, wherein the second substrate reservoir is in communication with the second substrate dispensing pipette system.

48. The method or automated clinical analyzer of any one or more of the preceding claims, wherein the first 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-RIO 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.

49. The method or automated clinical analyzer of any one or more of the preceding claims, wherein 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, C6-C10 aryl, or heteroaryl;R4is C2-C10 alkenyl; R5is H or C1-C10 alkyl; X is a phosphate; and at least one surfactant.

50. The method or automated clinical analyzer of any one or more of the preceding claims, wherein the second substrate material comprises compound having a structure:

51. A method of performing an immunoassay with an automated clinical analyzer, the method comprising:(a) mixing an analyte from a patient sample with a first reagent and a second reagent within a reaction vessel to create an assay mixture, the first reagent comprising a plurality of particles configured to specifically bind to the analyte from the patient sample, the second reagent comprising a reporter;(b) incubating the reaction vessel containing the assay mixture for a predetermined amount of time;(c) 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;(d) adding a substrate material into the reaction vessel with the plurality of particles remaining from the wash cycle, wherein the substrate material is added into the reaction vessel by a substrate dispensing pipette system located at one of the following locations:(i) a designated mixing station attached to a frame of the automated clinical analyzer,(ii) an incubation wheel of the automated clinical analyzer, or(iii) a reagent mixing station of the automated clinical analyzer;(e) mixing the substrate material with the plurality of particles remaining from the wash cycle to create a signal generating mixture;(g) incubating the signal generating mixture; and(h) analyzing the signal generating mixture with a detector.

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