MICROFLUIDIC REACTION STRUCTURES

Abstract

A rotatable device includes a plurality of reaction structures arranged circumferentially over at least a portion of the device. Each reaction structure in the plurality of reaction structures includes an aliquot chamber and a reaction chamber positioned radially outwards of the aliquot chamber. The aliquot chamber has an outlet and the reaction chamber has an inlet connected to the outlet of the aliquot chamber. The rotatable device also includes one or more connecting siphons radially leveled with each other and radially inwards of the outlets of the aliquot chambers of the plurality of reaction structures. Each respective connecting siphon in the one or more connecting siphons connects the aliquot chambers of corresponding adjacent reaction structures in the plurality of reaction structures.

Description

1 / 45 MICROFLUIDIC REACTION STRUCTURES CROSS-REFERENCE TO RELATED REQUESTS

[0001] This application claims priority over U.S. Provisional Patent Application No. 63 / 489,422, filed March 9, 2023, and U.S. Provisional Patent Application No. 63 / 489,677, filed March 10, 2023. The disclosure of each application is incorporated herein by reference in its entirety for all purposes. TECHNICAL FIELD

[0002] This disclosure relates to devices and methods that allow parallel reactions to be performed in a microfluidic centrifuge. FUNDAMENTALS

[0003] Currently, 70% of all medical decisions depend on laboratory diagnoses, but today the diagnostic process is disconnected from how care is delivered. The primary care system requires patients to travel to external phlebotomy sites to have blood drawn, which is sent to laboratories by mail and processed overnight. This means that laboratory test results reach healthcare professionals long after the patient has left. This friction in the distribution of care and disease management leads to tremendous waste in the healthcare system: a. Patients frequently delay undergoing laboratory tests or fail to adhere to laboratory tests or subsequent care recommendations. b. The gap in the diagnostic process leads to a lack of testing, incorrect diagnoses, a lack of intervention, and ultimately, unsatisfactory results. c. Healthcare professionals waste time tracking down laboratory test orders and patient consultation notes. When intervention is needed, even more time is wasted contacting patients and conducting subsequent steps in the care pathway. Petition 870250101629, dated 06 / 11 / 2025, page 5 / 50 2 / 45 of the patient.

[0004] These problems are even more serious when it comes to rural populations or patients belonging to groups facing adverse social determinants of health, where there are many challenges to ensuring successful follow-up from the first encounter with the patient.

[0005] Several companies have created point-of-care instruments to close this gap. However, these instruments are limited to single types of tests and do not fully meet the workflow needs of primary care providers for a single system that produces simple, comprehensive, and rapid test results. A product to address these needs is currently under development. This is made possible through a highly automated workflow, enabled by the use of centrifugal microfluidic discs.

[0006] Microfluidic centrifuges are used in clinical chemistry, immunoassays, hematology, medicine, biomedical research, and other fields. Many of these applications involve a large number of reactions. To allow for this large number of reactions, existing microfluidics are often configured with multiple structures arranged sequentially to avoid contamination. Such configurations require radial displacement between each structure, which leads to at least two problems: limitation in the number of structures per cartridge and slightly different operating parameters for each structure.

[0007] Consequently, there is still a need for improved devices and methods to perform a large number of reactions in microfluidic centrifugation. SUMMARY

[0008] This disclosure addresses these and other needs in the art, providing devices and methods that can perform reactions in parallel and, in many cases, with controlled reconstitution of dry reagents and light-based intermediate reading. Petition 870250101629, dated 06 / 11 / 2025, page 6 / 50 3 / 45

[0009] In one aspect, the present disclosure provides a rotating device about a rotational axis. The device includes a reaction chamber with an inlet to receive a fluid and a first well to serve as both a reagent and reading well. A first reagent is disposed in the first well when the device is made. The radial position of the fluid meniscus in the reaction chamber depends, at least in part, on the rotational speed of the device. The first well has a substantially flat wall perpendicular to the axis of rotation to allow for reproducible light transmission.

[0010] In some embodiments, the reaction chamber also includes a second well and an intermediate chamber between the first and second wells. In some of these embodiments, a second reagent is disposed or placed in the second well when the device is made.

[0011] In some embodiments, the first and / or second reagent is lyophilized. In some embodiments, the first and / or second reagent is in granular form. In some embodiments, the second reagent is different from the first reagent.

[0012] In some embodiments, the reaction chamber is not vented to promote homogeneous mixing of the first reagent with the fluid. For example, in some of these embodiments, the reaction chamber includes an outlet connected to an air chamber. In one embodiment, the first well is connected to the air chamber. In some embodiments, the second well is connected to the air chamber.

[0013] In some embodiments, the device also includes an upstream chamber positioned radially inside the reaction chamber and connected to the inlet of the reaction chamber. In some embodiments, the inlet of the reaction chamber is located in the first well, and the second well is positioned radially inside the first well. In some embodiments, the outlet is located in the second well.

[0014] In some embodiments, the reaction chamber, the upstream chamber, the air chamber, or a combination thereof are configured to Petition 870250101629, dated 06 / 11 / 2025, page 7 / 50 4 / 45 allows the initial filling of the first well with fluid without wetting the second well.

[0015] In another aspect, the present disclosure provides a rotating device about a rotational axis. The device includes a plurality of reaction structures arranged circumferentially on at least one part of the device. Each reaction structure in the plurality of reaction structures includes an aliquot chamber and a reaction chamber positioned radially outward from the aliquot chamber. The aliquot chamber has an outlet and the reaction chamber has an inlet connected to the outlet of the aliquot chamber. In some embodiments, the device also includes one or more connecting siphons that are radially level with each other and radially inward to the outlets of the aliquot chambers of the plurality of reaction structures. Each respective connecting siphon in the one or more connecting siphons connects the corresponding adjacent aliquot chambers of reaction structures in the plurality of reaction structures.

[0016] In some embodiments, for each reaction structure in the plurality of reaction structures, the reaction chamber includes a first well. In some embodiments, for each reaction structure in the plurality of reaction structures, a first reagent is disposed in the first well when the device is made. In some embodiments, for each reaction structure in the plurality of reaction structures, the first well serves as both a reagent and a readout well. In some embodiments, the first wells of the plurality of reaction structures are radially leveled with respect to each other.

[0017] In some embodiments, for each reaction structure in at least one subset of the plurality of reaction structures, the reaction chamber includes a second well and an intermediate chamber between the first and second wells. In some embodiments, for each reaction structure in at least one subset of the plurality of reaction structures, a second reagent is disposed in the second well when the device is made. In some embodiments, the second wells in at least one subset of the Petition 870250101629, dated 06 / 11 / 2025, page 8 / 50 5 / 45 A plurality of reaction structures are radially leveled relative to each other. In some embodiments, the inlet of the reaction chamber is located in the first well, and the second well is positioned radially into the first well.

[0018] In some embodiments, each respective reaction structure in the plurality of reaction structures includes an air chamber, and the reaction chamber of the respective reaction structure has an outlet connected to the air chamber.

[0019] In some embodiments, the device further includes an overflow chamber and an overflow channel connecting the aliquot chamber of a final reaction structure in the plurality of reaction structures to the overflow chamber. In some of these embodiments, the overflow channel includes a siphon portion, and the siphon portion includes an inlet and a crest. In some embodiments, the inlet of the siphon portion of the overflow channel is radially level with an inlet and an outlet of each connecting siphon in one or more siphons. In some embodiments, the crest of the siphon portion of the overflow channel is level with or radially outward from a crest of each connecting siphon in one or more siphons. In some embodiments, the overflow channel further includes a U-shaped channel with an outlet connected to the overflow chamber. In some embodiments, a portion of the U-shaped channel extends beyond the outlet of the overflow channel.

[0020] In a further aspect, the present disclosure provides a method that includes (A) obtaining a rotating device about a rotational axis. The device includes a plurality of reaction structures and one or more connecting siphons. Reaction structures in the plurality of reaction structures are arranged circumferentially on at least one portion of the device. Each reaction structure in the plurality of reaction structures includes an aliquot chamber and a reaction chamber positioned radially outward from the aliquot chamber. The aliquot chamber has an outlet and the chamber Petition 870250101629, dated 06 / 11 / 2025, p. 9 / 50 6 / 45 of the reaction has an inlet connected to the outlet of the aliquot chamber. One or more connecting siphons are radially leveled relative to each other and radially into the outlets of the aliquot chambers of the plurality of reaction structures. Each respective connecting siphon in the one or more connecting siphons connects the corresponding adjacent reaction structure aliquot chambers in the plurality of reaction structures. The method also includes (B) rotating the device at a speed to fill a first portion of the reaction chamber of each reaction structure in the plurality of reaction structures with a fluid. The method further includes (C) increasing the speed to empty the fluid from each corresponding connecting siphon into one or more connecting siphons.Furthermore, the method includes (D) slowing down to create a volume of trapped gas within each corresponding connecting siphon in one or more connecting siphons, thereby disrupting the fluid connection between the plurality of reaction structures.

[0021] In some embodiments, the increase (C) moves at least a portion of the fluid out of the aliquot chamber of each respective reaction structure in the plurality of reaction structures, so that the fluid contained in the aliquot chamber of each respective reaction structure in the plurality of reaction structures is positioned out of the inlet and outlet of each corresponding connection siphon in one or more connection siphons, thus emptying the fluid from each corresponding connection siphon in one or more connection siphons.

[0022] In some embodiments, the decrease (D) moves at least a portion of the fluid contained in the reaction chamber of each respective reaction structure in the plurality of reaction structures to the aliquot chamber of each respective reaction structure in the plurality of reaction structures, so that the fluid contained in the aliquot chamber of each respective reaction structure in the plurality of reaction structures is positioned into the inlet and outlet of each corresponding connecting siphon in the one or more connecting siphons, thereby wetting the inlet and the Petition 870250101629, dated 06 / 11 / 2025, page 10 / 50 7 / 45 outlet of each corresponding connection siphon in one or more connection siphons with the volume of gas trapped inside each corresponding connection siphon.

[0023] In some embodiments where the first portion of the reaction chamber of each reaction structure in the plurality of reaction structures includes a first well with a first reagent, the method further includes (E) repeating the increase (C) and decrease (D) one or more times to promote mixing of the fluid with the first reagent. In some embodiments, the method further includes (F) detecting, for each reaction structure in the plurality of reaction structures, light transmitted through the first well or a wall of the first well of the reaction chamber. In some embodiments, the detection (F) is performed without repetition (E) (e.g., without mixing).

[0024] In some embodiments, the method further includes (G) increasing the rate to fill a second portion of the reaction chamber of each reaction structure in at least one subset of the plurality of reaction structures, and (H) performing the decrease (D). In some embodiments where the second portion of the reaction chamber of each reaction structure in at least the subset of the plurality of reaction structures includes a second well with a second reagent, the method further includes (I) repeating the increase (G) and performing (H) one or more times to promote mixing of the fluid with the second reagent. In some embodiments, the method further includes (J) detecting, for each reaction structure in at least the subset of the plurality of reaction structures, light transmitted through the first well or a wall of the first well of the reaction chamber.

[0025] The devices, systems and methods of the present disclosure have other features and advantages which will be apparent or are presented in more detail in the accompanying drawings, which are incorporated herein, and in the Detailed Description below, which together serve to explain certain principles of exemplary embodiments of the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS Petition 870250101629, dated 06 / 11 / 2025, p. 11 / 50 8 / 45

[0026] The accompanying drawings, which are incorporated into and form part of this specification, illustrate one or more exemplary embodiments of the present disclosure and, together with the Detailed Description, serve to explain the principles and implementations of exemplary embodiments of the invention. The accompanying drawings are not necessarily to scale. The specific design features of the present invention, as disclosed herein, including, for example, specific dimensions, orientations, locations, and shapes, will be determined in part by the intended application and the specific environment of use. Furthermore, the components illustrated in the figures can be combined in any useful number and combination.

[0027] In the drawings:

[0028] FIG. 1A is a schematic diagram illustrating a device according to some exemplary embodiments of the present disclosure;

[0029] FIG. 1B is a schematic cross-sectional view taken along the dotted line in FIG. 1A according to an exemplary embodiment of the present disclosure;

[0030] FIG. 1C is a schematic diagram illustrating a reaction structure according to an alternative exemplary embodiment of the present disclosure;

[0031] FIG. 1D is a schematic diagram illustrating a device according to another alternative exemplary embodiment of the present disclosure;

[0032] FIG. 2A is a schematic diagram illustrating a device according to exemplary embodiments of this disclosure;

[0033] FIG. 2B is a partially enlarged view of FIG. 2A;

[0034] FIGS. 3A and 3B are flowcharts that collectively illustrate a method according to some exemplary embodiments of this disclosure;

[0035] FIGS. 4A, 4B, 4C, 4D, 4E and 4F are schematic diagrams Petition 870250101629, dated 06 / 11 / 2025, p. 12 / 50 9 / 45 which collectively illustrate a process carried out using the device in FIG. 2A according to some exemplary embodiments of the present disclosure;

[0036] FIG. 5A is a schematic diagram illustrating a device (e.g., a disk) according to some exemplary embodiments of the present disclosure;

[0037] FIG. 5B is a schematic diagram illustrating a loading process according to some exemplary embodiments of the present disclosure;

[0038] FIG. 5C is a schematic diagram illustrating a separation and dosing process according to some exemplary embodiments of the present disclosure;

[0039] FIG. 5D is a schematic diagram illustrating a dosing process according to some exemplary embodiments of the present disclosure;

[0040] FIG. 5E-1 is a schematic diagram illustrating a dilution process according to some exemplary embodiments of the present disclosure;

[0041] FIG. 5E-2 is a partially enlarged view of FIG. 5E-1;

[0042] FIG. 5F-1 is a schematic diagram illustrating a transfer and dissolution process according to some exemplary embodiments of the present disclosure;

[0043] FIG. 5F-2 is a partially enlarged view of FIG. 5F-1;

[0044] FIG. 5G is a schematic diagram illustrating a mixing process according to some exemplary embodiments of the present disclosure;

[0045] FIG. 5H is a schematic diagram illustrating a dissolution process according to exemplary embodiments of the present disclosure;

[0046] FIG. 5I is a schematic diagram illustrating a mixing and detection process according to some exemplary embodiments of Petition 870250101629, dated 06 / 11 / 2025, p. 13 / 50 10 / 45 present disclosure;

[0047] FIG. 6 is a block diagram illustrating a workflow according to some exemplary embodiments of the present disclosure; and

[0048] FIG. 7 is a schematic diagram illustrating a device (e.g., a disk) according to some exemplary embodiments of the present disclosure. DETAILED DESCRIPTION

[0049] This disclosure provides systems and methods for performing a large number of reactions in parallel. In various embodiments, the systems and methods of this disclosure allow for controlled reconstitution of dry reagents and light-based intermediate reading, such as in the first step of two reagent reactions. This can be achieved by controlling reaction times and volumes, minimizing dead volume (e.g., the volume of liquid not being read by detection methods). This is critical because a small increase in dead volume can lead to a large amount of space (e.g., cartridge space) being allocated to non-critical functions and limiting the number of reactions that can be performed in a single device (e.g., cartridge).

[0050] Referring now to the drawings, where similar reference numbers indicate similar elements throughout the drawing, an exemplary device 100 is shown in FIGS. 1A-1C according to some embodiments of this disclosure. The device 100 is rotatable about a rotational axis 102. The device 100 includes a reaction structure 104 which is configured to allow the use of pneumatic pressure and centrifugal force to control the liquid position and allow the reconstitution of the reagent in the same chamber where incubation and detection occur. This eliminates the dead volume and variability associated with transferring liquid from a reconstitution chamber to a separate detection chamber.

[0051] Reaction structure 104 includes a reaction chamber 110 Petition 870250101629, dated 06 / 11 / 2025, p. 14 / 50 11 / 45 having an inlet 112 to receive a fluid, for example, from an upstream chamber or from a pipetting port or similar. The reaction chamber 110 is configured so that the radial position 116 of a fluid meniscus in the reaction chamber depends at least in part on the rotational speed of the device. The reaction chamber 110 includes one or more wells. As a non-limiting example, FIG. 1A illustrates the reaction chamber 110 having a first well 121 and a second well 122 connected by an intermediate chamber 123 between the first and second wells. In some embodiments, the inlet is located in the first well, and the second well is positioned radially into the first well. As another non-limiting example, FIG. 1C illustrates the reaction chamber 110 having a single well, such as the first well 121. It is worth noting that the reaction chamber may have more than two, more than three, more than four, or more than five wells.

[0052] At least one or more wells are configured to serve as both reagent and readout wells. For example, in some embodiments, as illustrated in FIG. 1B, the first well 121 is configured to serve as both reagent and readout wells. In some of these embodiments, as illustrated in FIG. 1B, a first reagent 151 is disposed in the first well when the device is made. The first well 121 is made with a substantially flat wall 124 perpendicular to the axis of rotation to allow reproducible light transmission. Alternatively, in some embodiments, the second well 122 is configured to serve as both reagent and readout wells, or both the first and second wells are configured to serve as both reagent and readout wells. In some embodiments, a second reagent 152 is disposed in the second well 122 when the device is made. The second reagent may be the same as or different from the first reagent.

[0053] In some embodiments, the first and / or second reagents are lyophilized. In some embodiments, the first and / or second reagents are in granular form. In some embodiments, the device or at least a portion of the device that includes the reaction chamber is made Petition 870250101629, dated 06 / 11 / 2025, p. 15 / 50 12 / 45 of an injection-molded thermoplastic part with one or more cavities, where the first and / or second reagents (e.g., lyophilized granules) are placed, followed by a bonding step with a pressure-sensitive adhesive-coated laminate to define the upper surface of the chamber, as illustrated in FIG. 1B.

[0054] In some embodiments, as illustrated in FIG. 1A, the reaction chamber 110 is non-vented to promote homogeneous mixing of the first reagent and / or the second reagent with the fluid. For example, in some embodiments, the reaction structure 104 includes a pressure chamber (e.g., an air chamber) 130 and the reaction chamber includes an outlet 114 connected to the pressure chamber. The outlet may be located in the second well, as illustrated in FIG. 1A, in the intermediate chamber, as illustrated in FIG. 1C, or in any other suitable location. This allows control of the liquid meniscus position within the reaction chamber using the centrifugation speed and ensures homogeneous mixing of the first or second reagent with the fluid in the reaction chamber. For example, changing the centrifugation speed moves the fluid in or out, thus promoting homogeneous mixing of the reagents.This has the added benefit of allowing precise control of the liquid position in all reaction chambers of multiple reaction structures simultaneously, as will be explained in more detail. The only factors influencing the liquid position are the rotation speed, the chamber geometry, and the reaction volume.

[0055] In some embodiments, the reaction structure 104 includes an upstream chamber 140 (e.g., an aliquot chamber) positioned radially inside the reaction chamber 110 and having an outlet (e.g., outlet 145, outlet 146, or both) connected to the inlet of the reaction chamber to supply fluid to the reaction chamber. In some embodiments, the reaction chamber, the upstream chamber, the pressure chamber (e.g., air chamber), or a combination thereof are configured to allow initial filling of the first well 121 with fluid without wetting the Petition 870250101629, dated 06 / 11 / 2025, page 16 / 50 13 / 45 second well 122, if present. For example, in some embodiments, the upstream chamber 140 is configured with a first deeper portion 141, a second deeper portion 142, and a shallow portion 143 between the first and second deeper portions. In some embodiments, the intermediate chamber 123 of the reaction chamber is configured to have a relatively small volume but a relatively larger radial length. This can ensure minimal variability in the reaction volume during dosing, even with a large volume variation in the upstream chamber. This can ensure minimal variability in the reaction volume during dosing, even with a large variation in the volume of the upstream chamber, ensuring that a variation in the radial position of the liquid meniscus 116 within the reaction chamber 110 does not correspond to a large variation in volume within the reaction chamber 110.

[0056] The reaction structure 104 can be configured to include additional, optional, or alternative components. For example, as a non-limiting example, FIG. 1D illustrates the reaction structure 104 which includes a vent port 160, the first well 121 is connected to the vent port, and the second well 122 is connected to the first well 121, the air chamber 130, and the upstream chamber 140. In this embodiment, the first well 121 is filled during the initial filling, leaving a minimum volume in the upstream chamber 140. By decreasing the rotation speed, the liquid meniscus will move inward and wet the siphon crest, filling the second reagent well 122.

[0057] Referring to FIGS. 2A and 2B, an exemplary device 200 is shown according to some embodiments of the present disclosure. The device 200 is rotatable about a rotational axis 202. The device 200 includes a plurality of reaction structures, such as the reaction structure 104 disclosed herein. Reaction structures in the plurality of reaction structures are arranged circumferentially on at least one portion of the device. The device 200 may include any suitable number of reaction structures. For example, the device may include 2, 3, 4, 5, 6, 7, 8, 9, 10 or more than 10. Petition 870250101629, dated 06 / 11 / 2025, page 17 / 50 14 / 45 reaction structures. As a non-limiting example, FIGS. 2A and 2B illustrate the device with four reaction structures, for example, reaction structures 104-1, 104-2, 104-3 and 104-4.

[0058] A reaction structure in a plurality of reaction structures may be configured in the same way or differently from another reaction structure in a plurality of reaction structures. In some embodiments, each reaction structure in a plurality of reaction structures is configured substantially the same way as other reaction structures in a plurality of reaction structures. In some embodiments, at least one reaction structure in a plurality of reaction structures is configured differently from one or more other reaction structures in a plurality of reaction structures. By way of example, FIGS. 2A and 2B illustrate that reaction structures 104-2, 104-3, and 104-4 are configured substantially the same or identical to each other, and reaction structure 104-1 is configured differently from reaction structures 104-2, 104-3, and 104-4.

[0059] Each reaction structure in the plurality of reaction structures includes an aliquot chamber, such as the upstream chamber 140 disclosed herein, and a reaction chamber, such as the reaction chamber 110 disclosed herein. The aliquot chamber has an outlet (e.g., outlet 145, outlet 146, or both) and the reaction chamber has an inlet (e.g., inlet 112) connected to the outlet of the aliquot chamber. The reaction chamber is positioned radially outward from the aliquot chamber. In some embodiments, each reaction structure in the plurality of reaction structures includes an air chamber, such as the air chamber 130 disclosed herein. For each respective reaction structure, the reaction chamber has an outlet (e.g., outlet 114) that is connected to the air chamber.

[0060] In some embodiments, for each reaction structure in the plurality of reaction structures, the reaction chamber includes a first well, such as the first well 121 disclosed in this document. In some embodiments, for each reaction structure in the plurality of reaction structures Petition 870250101629, dated 06 / 11 / 2025, page 18 / 50 15 / 45 reaction, a first reagent, such as the first reagent 151, is placed in the first well when the device is made. In some embodiments, for each reaction structure in the plurality of reaction structures, the first well serves as both a reagent and a readout well. In some embodiments, the first wells of the plurality of reaction structures are radially leveled (e.g., in the same radial position) relative to each other. In some embodiments, for any and all reaction structures in the plurality of reaction structures, the reaction chamber consists of a single well, such as the first well 121 disclosed in this document. In some embodiments, for each reaction structure in at least one subset of the plurality of reaction structures, the reaction chamber includes a second well, such as the second well 122 disclosed herein, and an intermediate chamber, such as the intermediate chamber 123 disclosed herein, between the first and second wells.In some embodiments, for each reaction structure in at least the subset of the plurality of reaction structures, a second reagent, such as the second reagent 152, is disposed in the second well when the device is made. In some embodiments, for each reaction structure in the plurality of reaction structures, the reaction chamber includes the first and second wells. By way of example, FIGS. 2A and 2B illustrate that the reaction chamber of reaction structure 104-1 includes a single well (e.g., the first well 121) and the reaction chamber of each of reaction structures 104-2, 104-3, and 104-4 includes the first and second wells.

[0061] In some embodiments, the second wells of at least the subset of the plurality of reaction structures are radially leveled to each other. In some embodiments, for each of the pluralities of reaction structures, the inlet of the reaction chamber is located in the first well, and the second well (if present) is positioned radially into the first well, as illustrated in FIGS. 1A and 2A.

[0062] In some embodiments, reaction structures in the plurality of reaction structures are positioned within a range of Petition 870250101629, dated 06 / 11 / 2025, page 19 / 50 16 / 45 radius. For example, in some embodiments, reaction structures in the plurality of reaction structures may be positioned between a first radius and a second radius relative to the axis of rotation. The first radius may be less than about 30 mm, less than about 35 mm, less than about 40 mm, less than about 45 mm, or less than 50 mm. The second radius may be greater than about 35 mm, greater than about 40 mm, greater than about 45 mm, greater than 50 mm, greater than 55 mm, or greater than 60 mm. In some specific implementations, the plurality of reaction structures may be positioned within a radius range that is between about 25 mm and about 45 mm, between about 30 mm and about 50 mm, between about 35 mm and about 55 mm, or between about 40 mm and about 60 mm. In one specific implementation, the plurality of reaction structures can be positioned within a radius range that is between approximately 43 mm and approximately 50 mm.

[0063] The volumes of the first wells (e.g., reaction volumes for a first reaction 1R in the reaction structures) may be equal to or different from the volumes of the second wells (e.g., reaction volumes for a second reaction 2R in the reaction structures). “1R” denotes a reaction occurring in the first wells or with the first reactants arranged in the first wells. Similarly, “2R” denotes a reaction occurring in the second wells or with the second reactants arranged in the second wells. In some embodiments, the reaction volumes for the 1R reaction are smaller than the reaction volumes for the 2R reaction.For example, in some specific implementations, the reaction volumes for reaction 1R may be less than about 10 μL, less than about 15 μL, less than about 20 μL, less than about 25 μL, or less than about 30 μL, and the reaction volumes for reaction 1R may be greater than about 20 μL, greater than about 25 μL, greater than about 30 μL, greater than about 35 μL, or greater than about 40 μL. In one specific implementation, the reaction volumes for reaction 1R may be about 15 μL and the volumes... Petition 870250101629, dated 06 / 11 / 2025, p. 20 / 50 17 / 45 of the reaction volume for reaction 2R could be 29 μE. In another specific implementation, the reaction volumes for reaction 1R could be about half the reaction volumes for reaction 2R.

[0064] In some embodiments, a channel (for example, the channel connecting the upstream chamber to the reaction chamber) may have a width between about 100 μm and 150 μm, between about 150 μm and 200 μm, between about 200 μm and 250 μm, or between about 250 μm and 300 μm, and a depth between about 50 μm and 100 μm, 100 μm and 150 μm, between about 150 μm and 200 μm, or between about 150 μm and 250 μm. In one specific implementation, a channel might have a width between approximately 120 μm and 175 μm and a depth between approximately 100 μm and 175 μm.

[0065] A chamber may have constant or variable depth. For example, a reaction chamber may have a depth in the intermediate chamber that is different from that in the first and / or second wells. Similarly, the upstream chamber may have a depth in the shallow portion that is different from that in the first and / or second deeper portions. In some embodiments, a chamber may have a depth between about 0.2 mm and about 5 mm. In some embodiments, a chamber may have a depth between about 0.4 mm and about 3 mm.

[0066] The device 200 also includes one or more radially leveled (e.g., in the same radial positions) connecting siphons 220 relative to each other and radially into the outlets of the aliquot chambers of the plurality of reaction structures. Each respective connecting siphon in the one or more connecting siphons connects the corresponding adjacent reaction structure aliquot chambers in the plurality of reaction structures. For example, in some embodiments, the device includes two reaction structures and a connecting siphon connecting the two reaction structures. In some embodiments, the device includes three reaction structures and two connecting siphons, each connecting two adjacent reaction structures. In the embodiment illustrated in FIG. 2A, where there are four Petition 870250101629, dated 06 / 11 / 2025, page 21 / 50 18 / 45 reaction structures, the device includes three connecting siphons, for example, connecting siphons 220-1, 220-2 and 220-3. Connecting siphon 220-1 connects reaction structure 104-1 and reaction structure 104-2. Connecting siphon 220-2 connects reaction structure 104-2 and reaction structure 104-3. Connecting siphon 220-3 connects reaction structure 104-3 and reaction structure 104-4. The connecting siphons 220-1, 220-2 and 220-3 are radially leveled relative to each other (i.e., the radial positions of the siphons are the same with respect to the rotational axis 202) and radially into the outlets of the aliquot chambers of the plurality of reaction structures.

[0067] In some embodiments, the reaction structures (e.g., the reaction chamber, the air chamber, and / or the aliquot chamber) and / or the connecting siphons are adjusted to meet the reaction requirements and minimize dead volume. For example, in some embodiments, the reaction structures and / or connecting siphons are adjusted to maintain the reaction volume within a desired range (e.g., within 1% to 10% or within 5% to 15%) according to the reaction kinetics, minimizing dead volume. In some embodiments, the reaction structures and / or connecting siphons are adjusted to ensure that the liquid position within the reaction chamber of each reaction structure completely fills the reading / reconstitution chamber (e.g., the first well) during the initial filling step without wetting the second reagent chamber (e.g., the second well, if present).

[0068] For example, referring to FIGS. 2A and 2B, in some embodiments, for each reaction structure in at least one subset of the plurality of reaction structures, the upstream / aliquot chamber includes a shallow portion, for example, shallow portion 143 as illustrated in FIG. 1B, adjacent to the corresponding connecting siphon. In some embodiments, the shallow portion may have a depth less than 1,000 μm, less than 900 μm, less than 800 μm, less than 700 μm, less than 600 μm, or less than 500 μm. In some embodiments, the shallow portion may have a depth between Petition 870250101629, dated 06 / 11 / 2025, p. 22 / 50 19 / 45 approximately 100 μm and approximately 500 μm. In some embodiments, the shallow portion may be positioned within approximately 2 mm, approximately 1.5 mm, approximately 1 mm, or approximately 0.5 mm radii of the connecting siphon radius. In some embodiments, to accommodate the entire required reaction volume, the aliquot chamber includes deeper portions, such as the first deeper portion 141 and the second deeper portion 142, in radial positions both outward and inward from the shallow portion. In some embodiments, the first and / or second deeper portions may have a depth of at least 1 mm, at least 1.5 mm, at least 2 mm, at least 2.5 mm, at least 3 mm, at least 3.5 mm, or at least 4 mm. In some modalities, the first and / or second deepest portions may have a depth of approximately 2 mm to approximately 3 mm.

[0069] In some embodiments, for each reaction structure in at least one subset of the plurality of reaction structures, the maximum angular length (e.g., arc length) of the upstream / aliquot chamber is defined by the total reaction volume and the need to minimize space with the minimum limit defined by the minimum angular length occupied by the connecting siphon and the downstream characteristics of the reaction structure. In some embodiments, for each reaction structure in at least one subset of the plurality of reaction structures, the angular length of the upstream / aliquot chamber may be between about 1 mm and about 5 mm, between about 2 mm and about 6 mm, between about 3 mm and 7 mm, or between about 4 mm and about 8 mm.

[0070] In some embodiments, the device 200 includes an overflow chamber 230 and an overflow channel 240. The overflow channel 240 connects the aliquot chamber of a reaction structure, for example, the last reaction structure in the plurality of reaction structures, to the overflow chamber. For example, in the illustrated embodiment, the overflow channel 240 connects the aliquot chamber of reaction structure 104-4 to the overflow chamber. The channel of Petition 870250101629, dated 06 / 11 / 2025, page 23 / 50 20 / 45 overflow allows for the dosing of reaction volumes within reaction structures by overflowing the excess volume. In some embodiments, these reaction structures can be loaded with an excess volume, and only the target volume can be retained, with the remainder overflowing.

[0071] In some embodiments, the overflow channel includes a siphon portion 241. The siphon portion 241 has an inlet 242 and a crest 243. The inlet 242 is radially level with the inlet 231 and outlet 232 of each connecting siphon in one or more siphons. The crest 243 is level with or slightly radially outward from the crest 243 of each connecting siphon in one or more siphons. In some embodiments, the overflow channel further includes a U-channel 244 having an outlet connected to the overflow chamber. In some embodiments, the U-channel 244 includes an outer segment and an inner segment, as illustrated in FIGS. 2A and 2B. In some embodiments, the U-channel retains the liquid after overflow. The 200 device allows precise control of the liquid positions in all reaction chambers simultaneously.In particular, it allows the use of pneumatic pressure and centrifugal force to control the liquid positions during each reaction step in parallel in all reaction chambers of the plurality of reaction structures.

[0072] For example, during the initial filling, each connecting siphon is filled and forms a connection with the next reaction structure; for example, it simply allows liquid to overflow from reaction structure (e.g., aliquot chamber) N to reaction structure N+1. After this initial filling, there is a known volume of liquid within each reaction structure. Increasing the rotation speed introduces more liquid volume into the reaction chamber and increases the radial position of the liquid in the upstream chamber. This ensures that the connecting siphons are emptied. Reducing the speed subsequently decreases the radial position of the liquid in the upstream channel, re-wetting the connecting siphons. How does this happen in Petition 870250101629, dated 06 / 11 / 2025, page 24 / 50 21 / 45 all reaction structures at the same time, both ends (e.g., the inlet and outlet channels) of the connecting siphons are filled simultaneously, and an air space is created within the connecting siphon. This air space prevents liquid movement between the reaction structures during the mixing process. It has the added benefit of improving mixing efficiency by allowing a wider velocity range during mixing: the lowest velocity can be lower than the initial filling velocity, and this would not be possible if there were a simple overflow connection between the chambers.

[0073] The overflow channel allows for the dosing of the reaction volume for each reaction structure. When all reaction structures are filled to a certain internal radius, and once that radius is inside the crest 243 of the siphon portion of the overflow channel, it can prepare the siphon portion of the overflow channel and empty all the liquid into the inlet 242 of the siphon portion of the overflow channel into the overflow chamber. By adding the U-channel 244 to the outlet portion of the overflow channel, while simultaneously defining the radial position of the crest 243 to be close to the desired radial position of the meniscus in the upstream chamber during filling, the need for additional dead volume to allow for initial emptying is minimized. This ensures that there is sufficient volume within the overflow channel when the siphon breaks, creating an air space.

[0074] Furthermore, the device may have different reaction structures for single and multiple reagent reactions, which operate at exactly the same operating rates and which can be used in parallel in the same fluidic system. For example, in some embodiments, such as that illustrated in FIG. 2A, the device has at least one structure (structure 1R) with a single well (e.g., the first well) for a single reagent reaction and at least one structure (structure 2R) with two wells (e.g., the first and second wells) for two reagent reactions in the Petition 870250101629, dated 06 / 11 / 2025, page 25 / 50 22 / 45 same device. This can minimize the reaction volume to get very close to the minimum volume required to reconstitute a lyophilized bead (e.g., structure 1R uses ~1Λ volume of structure 2R). Furthermore, this provides extreme flexibility when designing new cartridges. Referring to FIGS. 3A and 3B, a flowchart is shown illustrating an exemplary method for carrying out a series of reactions in parallel according to some embodiments of the present disclosure. In the flowchart, preferred parts of the method are shown in solid-line boxes, while additional, optional, or alternative parts of the method are shown in dashed-line boxes. It should be noted that the processes disclosed herein and exemplified in the flowchart can be, but do not have to be, carried out in their entirety or in the order in which they are presented.

[0075] Referring to block 302, in some embodiments, method 300 includes (A) obtaining a device like device 200 disclosed herein (for example, as illustrated in FIG. 2A). The device is rotatable about an axis of rotation. In some embodiments, the device includes a plurality of reaction structures and one or more connecting siphons. Reaction structures in the plurality of reaction structures are arranged circumferentially on at least one portion of the device. Each reaction structure in the plurality of reaction structures includes an aliquot chamber with an outlet and a reaction chamber with an inlet connected to the outlet of the aliquot chamber. The reaction chamber is positioned radially outward from the aliquot chamber. The one or more connecting siphons are radially leveled relative to each other and radially inward to the outlets of the aliquot chambers of the plurality of reaction structures.Each respective connecting siphon in one or more connecting siphons connects the aliquot chambers of the corresponding adjacent reaction structures in the plurality of reaction structures.

[0076] Referring to block 304, in some embodiments, method 300 includes (B) rotating the device at a speed to fill a Petition 870250101629, dated 06 / 11 / 2025, p. 26 / 50 23 / 45 first portion (e.g., the first well) of the reaction chamber of each reaction structure in the plurality of reaction structures with a fluid. In some embodiments, the device is rotated at a relatively low speed (e.g., about 2,000 rpm to about 2,500 rpm, or any other suitable speed depending on the implementation). On the one hand, this speed is slow enough not to wet the R2 wells prematurely and fast enough to ensure the filling of the first wells of all reaction structures within the time allowed in the workflow. On the other hand, this speed is high enough to avoid surface effects and ensure a repeatable meniscus position during overflow.

[0077] This process is illustrated in FIGS. 4A-4C. Initially, rotation of the device causes the fluid to fill the first well of the first reaction structure (e.g., reaction structure 104-1), as shown in FIG. 4A.

[0078] As more fluid flows into the chamber upstream of the first reaction structure, it overflows into the next reaction structure (e.g., reaction structure 104-2 through connecting siphon 220-1) and so on (e.g., reaction structure 104-3 through connecting siphon 220-2 and then reaction structure 104-4 through connecting siphon 220-3), as shown in FIG. 4B. The time to fill the first wells of all reaction structures depends, at least in part, on the number of reaction structures, the upstream flow rate, the rotation speed, or any combination thereof. It can be long or short. In some embodiments, it may take a few seconds, tens of seconds, a minute, or more than a minute.During this period, the fluid positions within the reaction structures may fluctuate; for example, the radial position of the fluid meniscus in the upstream chamber may fluctuate between radial positions 402-1 and 402-2, and the radial position of the fluid meniscus in the reaction chamber may fluctuate between radial positions 404-1 and 404-2. It should be noted that the radial positions illustrated in FIG. 4B are merely illustrative and not limiting. Petition 870250101629, dated 06 / 11 / 2025, page 27 / 50 24 / 45 actual radial positions will depend, at least in part, on the application (e.g., fluid volume, applied velocity) and the configuration of the reaction structure (e.g., size and shape of the first well and / or intermediate chamber).

[0079] As the last reaction structure (e.g., reaction structure 104-4) is filled with fluid, the position of the liquid in the aliquot chambers (e.g., upstream chamber 140) becomes inward to the crest of the overflow siphon 406 in FIG. 4C (e.g., the radial position of the crest 243 of the siphon portion of the overflow channel 240). Thus, the fluid overflows and empties all excess volume from all reaction structures into the overflow chamber (e.g., overflow chamber 230). This leaves a measured reaction volume within each reaction structure, defined by the connecting siphons and the overflow channel.

[0080] With reference to block 306, in some embodiments, method 300 includes (C) increasing the speed to empty the fluid from each corresponding connection siphon into one or more connection siphons. In some embodiments, the increase (C) moves at least a portion of the fluid out of the aliquot chamber of each respective reaction structure in the plurality of reaction structures, so that the fluid contained in the aliquot chamber of each respective reaction structure in the plurality of reaction structures is positioned out of the inlet and outlet of each corresponding connection siphon into one or more connection siphons, thus emptying the fluid from each corresponding connection siphon into one or more connection siphons.

[0081] For example, as a non-limiting example, FIG. 4D illustrates the increase in rotational speed (e.g., from between 2,000 rpm and about 2,500 rpm to between 2,500 rpm and about 3,000 rpm, or any other suitable speed depending on the implementation) that moves radially into the position of the fluid meniscus 404 in each reaction chamber and moves radially out of the position of the meniscus 402 in each chamber in Petition 870250101629, dated 06 / 11 / 2025, p. 28 / 50 25 / 45 upstream (e.g., the aliquot chamber). In some embodiments, the position of meniscus 402 and / or the position of meniscus 404 are moved to avoid wetting chambers R2 (e.g., the second wells).

[0082] With reference to block 308, in some embodiments, method 300 includes (D) slowing down to create a volume of trapped gas within each corresponding connecting siphon in one or more connecting siphons, thereby interrupting the fluid connection between the plurality of reaction structures.In some embodiments, the decrease (D) moves at least a portion of the fluid contained in the reaction chamber of each respective reaction structure in the plurality of reaction structures to the aliquot chamber of each respective reaction structure in the plurality of reaction structures, so that the fluid contained in the aliquot chamber of each respective reaction structure in the plurality of reaction structures is positioned into the inlet and outlet of each corresponding connection siphon in the one or more connection siphons, thereby wetting the inlet and outlet of each corresponding connection siphon in the one or more connection siphons with the volume of gas trapped within each corresponding connection siphon.

[0083] For example, as a non-limiting example, FIG. 4E illustrates that, due to the decrease in rotational speed (for example, from between 2,500 rpm and about 3,000 rpm to about 900 rpm, or any other suitable speed depending on the implementation), the fluid exits the reaction chambers and enters the aliquot chambers. In some embodiments, the entire volume of fluid in the reaction chambers is moved out of the reaction chambers. In some embodiments, only a portion of the volume of fluid in the reaction chambers is moved out of the reaction chambers. For example, in some specific implementations, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the volume of fluid in the reaction chambers is moved out of the reaction chambers. In some other specific implementations, at most 90%, at most 80%, or at least 90% of the fluid volume in the reaction chambers is moved out of the reaction chambers. Petition 870250101629, dated 06 / 11 / 2025, p. 29 / 50 26 / 45 maximum: 70% of the fluid volume in the reaction chambers is moved out of the reaction chambers. In some modes, gear shifting promotes mixing. Typically, the greater the proportion of volume that enters and exits, the more efficient the mixing.

[0084] As the fluid returns to the aliquot chambers, it wets the inlets of one or more connecting siphons and the overflow channel, forming air spaces between the reaction volumes and between the last reaction volume and the liquid trapped in the overflow channel. The air spaces separate the reaction structures from each other and from the overflow chamber, as illustrated in FIGS. 2A and 2B.

[0085] Referring to block 310, in some embodiments, method 300 includes (E) repeating the increase (C) and decrease (D) one or more times. For example, in some embodiments, the first portion of the reaction chamber of each reaction structure in the plurality of reaction structures includes a first well with a first reagent, the method includes repeating the increase (C) and decrease (D) one or more times to promote mixing of the fluid with the first reagent. In some embodiments, to promote mixing, the increase (C) and decrease (D) are repeated at least once, at least 5 times, at least 10 times, at least 15 times, at least 20 times, or any number of times suitable to meet the requirements of the applications. In some embodiments, the method repeats the mixing step(s) after detection, which may be performed any number of times.

[0086] Referring to block 312, in some embodiments, method 300 includes (F) detecting, for each reaction structure in the plurality of reaction structures, light transmitted through the first well or a wall of the first well of the reaction chamber. For example, in some embodiments, the device is rotated at a speed after homogenization R1, as illustrated in FIG. 4F, or at any desirable time or stage during the process. Light absorption at one or more wavelengths is read through the reading wells (e.g., the first wells). This may take less Petition 870250101629, dated 06 / 11 / 2025, page 30 / 50 27 / 45 of a minute, about a minute, several minutes, or any period of time, depending on the reactions used and their kinetics.

[0087] Referring to block 312, in some embodiments, method 300 includes (G) increasing the speed to fill a second portion (e.g., the second well) of the reaction chamber of each reaction structure in at least one subset of the plurality of reaction structures. For example, as a non-limiting example, FIG. 4G illustrates increasing the rotational speed (e.g., to about 7500 ~ 8000 rpm or any other suitable speed depending on the applications) to completely fill the second wells and initiate the second incubation.

[0088] Referring to block 314 and block 316, in some embodiments, the subsequent method 300 includes increasing (H), performing the decrease (D) and (I), repeating the increase (G) and performing (H) one or more times. For example, in some embodiments where the second portion of the reaction chamber of each reaction structure in at least the subset of the plurality of reaction structures includes a second well with a second reagent, the method includes repeating the increase (G) and performing (H) one or more times to promote mixing of the fluid with the second reagent.

[0089] In some embodiments, to promote mixing, the increase (G) and execution (H) are repeated at least once, at least 5 times, at least 10 times, at least 15 times, at least 20 times, or any number of times suitable to meet the application requirements. It should be noted that in execution (H), the velocity may be reduced to the same velocity or different from the velocity to promote fluid mixing in the first wells.

[0090] Referring to block 318, in some embodiments, method 300 includes (J) detecting, for each reaction structure in at least the subset of the plurality of reaction structures, light transmitted through the first well or a wall of the first well of the reaction chamber. For example, in some embodiments, the device is rotated at a speed Petition 870250101629, dated 06 / 11 / 2025, p. 31 / 50 28 / 45 after homogenization R2 or at any desired time or stage during the process. Light absorption at one or more wavelengths is read through the reading wells (e.g., the first or second wells). This can take less than a minute, about a minute, several minutes, or any length of time, depending on the reactions used and their kinetics.

[0091] The devices and methods of this disclosure may have additional, optional, or alternative components. For example, in some embodiments, the device may include more than two reagents per reaction chamber (e.g., including one or more additional wells in addition to the first and second wells). The reaction wells may contain more than one bead each. There is no hard limit. It is primarily a trade-off between the available radial distance, the maximum rotation speed, and the available space for volume aliquoting. In some embodiments, both the first and second wells may be configured as reading wells to allow readings in both reagent wells in the 2R reaction structure. In some embodiments, emptying may be controlled with an additional channel after 2R reconstitution to allow reading while the device is not rotating.In some embodiments, the device may not include the connecting siphon, and the reaction chamber of each reaction structure is individually loaded with a controlled volume. In some embodiments, certain components are arranged differently to further minimize the reaction volume.

[0092] With reference to FIG. 7, a device 700 (e.g., a disk) is shown according to some exemplary embodiments of the present disclosure. The device 700 (e.g., the disk) includes a plurality of units, such as units 710-1, 710-2, 710-3, arranged circumferentially. In some embodiments, the device 700 includes 2, 3, 4, 5, 6, 7, 8, 9, 10 or more than 10 units. In some embodiments, a unit 710 includes one or more features / components / devices (e.g., Petition 870250101629, dated 06 / 11 / 2025, p. 32 / 50 29 / 45 the reaction structure 104) disclosed herein. In some embodiments, each unit 710 includes one or more features / components / devices (e.g., the reaction structure 104) disclosed herein. In some embodiments, each unit is identical to another unit in the plurality of units. In some embodiments, at least one unit is different from the other units in the plurality of units. The device 700 can be used in various applications, including but not limited to clinical chemistry.

[0093] Example Workflow(s).

[0094] Figures 5A-5I collectively illustrate an exemplary workflow according to some exemplary embodiments of the present disclosure. Although specific specimens (e.g., whole blood) are used in the description of the workflow, it should be noted that the present disclosure is not limited to them. Other samples, such as those disclosed herein, may be used. Furthermore, the workflow may be automated.

[0095] With reference to FIG. 5A, it is a schematic diagram illustrating a device (e.g., a disc) according to some exemplary embodiments of the present disclosure. For clarity, only a portion of the device is shown. The device 500 is rotatable about a rotational axis, as is the vertical rotational axis 503.In some implementations, the device 500 can be rotated, during one or more processes, at a speed of at least about 1000 rpm, at least about 1200 rpm, at least about 1400 rpm, at least about 1600 rpm, at least about 1800 rpm, at least about 2000 rpm, at least about 2200 rpm, at least about 2400 rpm, at least about 2600 rpm, at least about 2800 rpm, at least about 2900 rpm, at least about 3000 rpm, at least about 3500 rpm, at least about 4000 rpm, at least about 4500 rpm, at least about 5000 rpm, at least about 5500 rpm, at least about 6000 rpm, at least about 6500 rpm, or at least about 7000 rpm. rpm. In some implementations, the 500 device can be rotated, during one or more processes, by one. Petition 870250101629, dated 06 / 11 / 2025, p. 33 / 50 30 / 45 speeds of no maximum near 500 rpm, no maximum near 600 rpm, no maximum near 700 rpm, no maximum near 800 rpm, no maximum near 900 rpm, no maximum near 1000 rpm, no maximum near 1200 rpm, no maximum near 1400 rpm, no maximum near 1600 rpm, no maximum near 1800 rpm, no maximum near 2000 rpm, no maximum near 2200 rpm, no maximum near 2400 rpm, no maximum near 2600 rpm, no maximum near 2800 rpm, no maximum near 2900 rpm, no maximum near 3000 rpm, no maximum near 3500 rpm, no maximum near 4000 rpm, no maximum near 4500 rpm or no maximum near 5000 rpm.

[0096] Device 500 includes one or more cameras, one or more channels and / or other features / components. For example, in the embodiments illustrated, the device 500 includes a ventilation channel 501, a sample chamber 502, a buffer chamber 504, a blood separation chamber 506, a sample overflow chamber 508, an additional plasma overflow chamber 509, a sample dosing chamber 510, a mixing chamber 512, a diluted sample overflow chamber 514, a pneumatic chamber 515, a pneumatic and dilution buffer overflow chamber 516, one or more 1R / detection chambers 518 (e.g., the first well 121 of the reaction chamber 110 shown in FIG. 1C), one or more 2R R1 / detection chambers 520 (e.g., the first well 121 of the reaction chamber 110 shown in FIG. 1A), one or more 2R R2 chambers 522 (for example, the second well 122 of the reaction chamber 110 shown in FIG.1A), one or more aliquot chambers 524 and a mixing / dosing chamber 526.

[0097] Although the 500 device is illustrated with specific components (e.g., specific cameras and channels), it should be noted that this is only an example and is not exhaustive. In some implementations, the 500 device may not include one or more of these specific components. In some implementations, the 500 device may include additional or alternative components, such as those disclosed herein. Furthermore, the Petition 870250101629, dated 06 / 11 / 2025, page 34 / 50 The 31 / 45 500 device can be used in various applications, including but not limited to clinical chemistry.

[0098] With reference to FIG. 5B, a loading process is illustrated according to some exemplary embodiments of the present disclosure. In some embodiments, the buffer and the sample (e.g., whole blood) are loaded into the device 500. For example, in some implementations, a buffer is loaded into the buffer chamber 504, and whole blood is loaded into the sample chamber 502.

[0099] With reference to FIG. 5C, a separation and dosing process is illustrated according to some exemplary embodiments of the present disclosure. In some embodiments, in this process, whole blood is separated and buffer is dosed. For example, in some implementations, the device 500 is rotated about the vertical axis of rotation 503. Whole blood flows from the sample chamber 502 to the blood separation chamber 506 and separates into plasma (radially inward) and cell fractions (radially outward). Buffer flows from the buffer chamber 504, through the mixing chamber 512, to the mixing / dosing chamber 526, where it is measured. If there is excess buffer, the excess flows to the pneumatic overflow and dilution buffer chamber 516. In this process, back pressure is created by air trapped in the pneumatic overflow and dilution buffer chamber 516.The back pressure in the pneumatic overflow chamber and dilution buffer 516 is a function of the rotational speed of the device 500.

[0100] With reference to FIG. 5D, a dosing process is illustrated according to some exemplary embodiments of the present disclosure. In some embodiments, in this process, plasma is dosed. For example, in some implementations, the rotation speed of disc 500 is decreased. As the rotation speed of disc 500 decreases, the measured buffer flows from mixing / dosing chamber 526 to mixing chamber 512, due to the decrease in pressure in the buffer overflow chamber. Petition 870250101629, dated 06 / 11 / 2025, p. 35 / 50 32 / 45 pneumatic and dilution 516. Furthermore, as the speed of the disc 500 decreases, the plasma in the blood separation chamber 506 flows through a connecting siphon towards the sample dosing chamber 510. In some implementations, the plasma completely fills the sample dosing chamber 510, with any excess plasma overflowing into the additional plasma overflow chamber 509. At this point in the process, both the plasma and the buffer have been measured.

[0101] With reference to FIGS. 5E-1 and 5E-2, a dilution process is illustrated according to some exemplary embodiments of the present disclosure. In some embodiments, in this process, the dosed plasma is diluted and / or mixed with the dosed buffer. For example, in some implementations, the rotation speed of device 500 is increased. By increasing the rotation speed of device 500, the dosed plasma flows from the sample dosing chamber 510 to the mixing chamber 512. Then, the rotation speed of device 500 is decreased, causing the air trapped in the pneumatic and dilution buffer overflow chamber 516 to expand, thus pushing the dosed buffer from the mixing / dosing chamber 526 to the mixing chamber 512.To mix the metered plasma and the metered buffer, the rotation speed of device 500 is increased and decreased, causing the air trapped in the pneumatic overflow chamber and the dilution buffer 516 to expand and contract, forcing the mixture of the metered buffer and the metered plasma back and forth between the mixing / metering chamber 526 and the mixing chamber 512. Increasing / decreasing the rotation speed of device 500 can be repeated as desired, programmed, or until the solution is adequately / completely mixed.

[0102] With reference to FIGS. 5F-1 and 5F-2, a transfer and dissolution process is illustrated according to some exemplary embodiments of the present disclosure. In some embodiments, in this process, the diluted plasma buffer mixture is transferred to the chambers of Petition 870250101629, dated 06 / 11 / 2025, page 36 / 50 33 / 45 outermost detection chambers, dissolving lyophilized reagents in these chambers. For example, in some implementations, the rotation speed of device 500 is initially decreased, preparing the connecting channel(s) between mixing chamber 512 and aliquot chamber(s) 524. Then, the rotation speed of device 500 is increased to transfer the diluted plasma buffer mixture to aliquot chamber(s) 524, to 2R R1 / detection chamber(s) 520 and to 1R / detection chamber(s) 518, with the excess (if any) flowing to the diluted sample overflow chamber 514. FIG. 5F-1 shows this step before completion, while FIG. 5F-2 shows the complete transfer. The diluted plasma buffer mixture does not reach the 2R R2 522 chambers at this stage due to back pressure in the pneumatic chamber 515.

[0103] Referring now to FIG. 5G, a mixing process is illustrated according to some exemplary embodiments of the present disclosure. In some embodiments, in this process, the dissolved lyophilized reagents and the diluted plasma buffer mixture are adequately or completely mixed. For example, in some implementations, the rotational speed of device 500 is increased and decreased, causing the air trapped in pneumatic chamber 515 (and / or other pneumatic chambers connected to 2R R1 / detection chamber(s) 520 and 1R / detection chamber(s) 518) to expand and contract, forcing the mixture back and forth between 2R R1 / detection chamber 520, 1R / detection chamber 518, and aliquot chamber 524. The increase / decrease in the rotational speed of device 500 can be repeated as desired, programmed, or until a proper / complete mixture is achieved.Once the mixture is complete (for example, once proper mixing is achieved), the reaction mixture can be transferred back to the 2R R1 / detection chamber(s) 520 and to the 1R / detection chamber(s) 518, where optical measurements can be made, if desired.

[0104] Referring now to FIG. 5H, a process is illustrated of Petition 870250101629, dated 06 / 11 / 2025, p. 37 / 50 34 / 45 Dissolution according to some exemplary embodiments of the present disclosure. In some embodiments, in this process, the lyophilized reagents in the 2R R2 522 chamber(s) are dissolved. For example, in some implementations, the rotation speed of the device 500 is increased, causing the compressed air in the pneumatic chamber 515 (and / or other pneumatic chambers connected to the 2R R1 / detection chamber(s) 520 and to the 1 R / detection chamber(s) 518) to contract, forcing the mixture into the 2R R2 522 chamber(s), dissolving the lyophilized reagents in the 2R R2 522 chamber(s).

[0105] Referring now to FIG. 5I, a mixing and detection process is illustrated according to some exemplary embodiments of the present disclosure. In some embodiments, in this process, the mixture of lyophilized reagents and diluted buffer is mixed and / or optical measurements are made. For example, in some implementations, the rotational speed of device 500 is increased and decreased, causing the air trapped in pneumatic chamber 515 (and / or other pneumatic chambers connected to 2R R1 / detection chamber(s) 520 and 1R / detection chamber(s) 518) to expand and contract, forcing the mixture back and forth between chamber 2R R2 522, 2R R1 / detection chamber 520, 1R / detection chamber 518, and aliquot chamber 524. The increase / decrease in the rotational speed of device 500 can be repeated as desired, programmed, or until a proper / complete mixture is achieved.Once the mixture is complete (for example, once the proper mixing is achieved), the reaction mixture can be transferred back to the 2R R2 522 chamber, the 2R R1 / detection chamber 520 and the 1R / detection chamber 518, where optical measurements can be made, if desired.

[0106] In some embodiments, the workflow is completed at this point. The rotation of device 500 can be stopped and device 100 can be discarded.

[0107] Referring to FIG. 6, a workflow is illustrated. Petition 870250101629, dated 06 / 11 / 2025, p. 38 / 50 35 / 45 exemplary 600 (e.g., workflow) according to some exemplary embodiments of this disclosure. Workflow 600 can be run on any device disclosed herein (e.g., device 500). Workflow 600 can also be automated.

[0108] In some embodiments, workflow 600 includes a process 602 that loads a buffer (e.g., water) into the device and a process 606 that loads a sample (e.g., whole blood) into the device. In some embodiments, process 602 and process 606 are the same or similar to those disclosed here with respect to FIG. 5B.

[0109] In some embodiments, workflow 600 includes a process 604 that doses the buffer and a process 608 that separates the sample (e.g., separating whole blood into plasma and cell fractions). In some embodiments, process 604 and process 608 are the same or similar to those disclosed here with respect to FIG. 5C.

[0110] In some embodiments, workflow 600 includes a process 610 that measures the sample or a component of the sample. For example, in embodiments where the sample is whole blood that has been separated into plasma and cell fractions, process 610 measures the plasma. In some embodiments, process 610 is the same as or similar to that disclosed here with respect to FIG. 5D.

[0111] In some embodiments, workflow 600 includes a process 612 that mixes the dosed buffer and the dosed sample (e.g., dosed plasma). In some embodiments, process 612 is the same as or similar to that disclosed herein with respect to FIGS. 5E-1 and 5E-2.

[0112] In some embodiments, workflow 600 includes a process 614 that divides the dosed buffer mixture and the dosed sample (e.g., dosed plasma) into aliquots. In some embodiments, process 614 is the same as or similar to that disclosed herein with respect to FIGS. 5F-1 and 5F-2.

[0113] In some modes, workflow 600 includes a Petition 870250101629, dated 06 / 11 / 2025, p. 39 / 50 36 / 45 process 616 which resuspends the dry R1 granule(s), for example, by dissolving lyophilized reagents in the 2R R1 / detection chamber(s) 520 and in the 1 R / detection chamber(s) 518. In some embodiments, process 616 is the same or similar to that disclosed herein with respect to FIGS. 5F-1 and 5F-2.

[0114] In some embodiments, the workflow includes a process 618 that incubates the mixture of the measured buffer, dosed plasma, and dissolved lyophilized reagents. In some embodiments, process 618 may be carried out in a controlled environment (e.g., with a controlled temperature and / or during a predetermined period of time).

[0115] In some embodiments, workflow 600 includes a process 620 that resuspends the dry R2 granule(s), for example, by dissolving lyophilized reagents in chamber(s) 2R R2 522. In some embodiments, process 620 is the same or similar to that disclosed here with respect to FIG. 5H.

[0116] In some embodiments, workflow 600 includes a process 622 that incubates the mixture of metered buffer, dosed plasma, and dissolved lyophilized reagents. Like process 618, in some embodiments, process 622 can be carried out in a controlled environment (e.g., with a controlled temperature and / or during a predetermined period of time).

[0117] In some embodiments, workflow 600 may include one or more measurements, which may be performed at any suitable stage when desired. For example, a measurement may be performed to measure absorbance before, during, or after the lyophilized reagents are dissolved. A measurement may be performed in the same or similar manner to those disclosed herein with respect to FIGS. 5G and 5I. In some embodiments, workflow 600 may include a measurement process 624 after resuspension of the dried granule(s) R1, a measurement process 626 after the first incubation but before resuspension of the dried granule(s) R2, a measurement process 628 after resuspension of the Petition 870250101629, dated 06 / 11 / 2025, pages 40 / 50 37 / 45 dry R2 granules, but before the second incubation, a measurement process 630 after the second incubation, or any combination thereof.

[0118] The devices and methods of the present disclosure have a number of advantages. For example, the devices of the present disclosure are more compact, without active valves. They are independent of coatings / surface tension and the total number of reaction structures and reactions. They are also insensitive to sample / biological variability and are easily prototyped (not dependent on materials, surface roughness, etc.). The devices and methods of the present disclosure allow for a well-controlled mixing strategy, reading during mixing cycles, and execution of multiple reaction methodologies simultaneously.

[0119] The devices and methods disclosed herein may be used in a variety of applications, including but not limited to clinical chemistry, immunoassays, and hematology. Examples of clinical chemistry, immunoassays, and / or hematology are disclosed in WO2018 / 119437, WO2018 / 140719, WO2022 / 029731, and WO2022 / 029732, the contents of each application being incorporated herein by reference in their entirety. The devices and methods disclosed herein may be operated or performed by a system similar to those disclosed in U.S. Patent Application No. 17 / 371,746, the contents of which are incorporated herein by reference in their entirety.

[0120] Illustration of Matter Technology as Clauses.

[0121] Several examples of aspects of the disclosure are described as numbered clauses (1, 2, 3, etc.) for convenience. These are provided as examples and are not limited to the technology in question.

[0122] Clause 1. A device comprising: a rotating shaft; and a reaction chamber comprising an inlet for receiving a fluid and a first well for serving as a reagent and reading well, wherein (i) a first reagent is disposed in the first well when the device is made, (ii) a radial position of a fluid meniscus in the reaction chamber depends, at least in part, on the rotational speed of the device, and (iii) the Petition 870250101629, dated 06 / 11 / 2025, pp. 41 / 50 38 / 45 first well has a substantially flat wall perpendicular to the rotational axis to allow for reproducible light transmission. In some embodiments, the connection between the inlet and the first well is designed to ensure that a portion of the liquid within the first well can return to the inlet.

[0123] Clause 2. The device in Clause 1, where the first reagent is lyophilized.

[0124] Clause 3. The device of any previous Clause, in which the first reagent is in the form of granules.

[0125] Clause 4. The device of any previous Clause, in which the reaction chamber is not ventilated to promote homogeneous mixing of the first reagent with the fluid.

[0126] Clause 5. The device of Clause 4, in which the reaction chamber comprises an outlet connected to an air chamber.

[0127] Clause 6. The device of any preceding Clause, further comprising: an upstream chamber positioned radially into the reaction chamber and connected to the inlet of the reaction chamber.

[0128] Clause 7. The device of any previous Clause, in which the first well is connected to an air chamber.

[0129] Clause 8. The device of any of Clauses 1 to 4, wherein the reaction chamber further comprises a second well and an intermediate chamber between the first and second wells, in which a second reagent is disposed in the second well when the device is made. In some embodiments, the structure or reaction chamber is not vented, so that the meniscus level in the aliquot chamber is variable during operation or similar.

[0130] Clause 9. The provision of Clause 8, in which the second reagent is different from the first reagent.

[0131] Clause 10. The device of any of Clauses 8 to 9, where the inlet is located in the first well and the second well is Petition 870250101629, dated 06 / 11 / 2025, pp. 42 / 50 39 / 45 positioned radially into the first well.

[0132] Clause 11. The device of Clause 10, where the outlet is located in the second well.

[0133] Clause 12. The device of any of Clauses 8 to 11, in which the reaction chamber, the upstream chamber, the air chamber or a combination thereof are configured to allow the initial filling of the first well with fluid without wetting the second well.

[0134] Clause 13. A device comprising: a rotational axis; a plurality of reaction structures arranged circumferentially on at least a portion of the device, wherein each reaction structure in the plurality of reaction structures comprises an aliquot chamber with an outlet and a reaction chamber with an inlet connected to the outlet of the aliquot chamber, wherein the reaction chamber is positioned radially outward from the aliquot chamber; and one or more connecting siphons radially leveled to each other and radially inward to the outlets of the aliquot chambers of the plurality of reaction structures, wherein each respective connecting siphon in the one or more connecting siphons connects the corresponding adjacent reaction structure aliquot chambers in the plurality of reaction structures.

[0135] Clause 14. The device of Clause 13, wherein for each reaction structure in the plurality of reaction structures, the reaction chamber comprises a first well.

[0136] Clause 15. The device of Clause 14, wherein for each reaction structure in the plurality of reaction structures, a first reagent is disposed in the first well when the device is made.

[0137] Clause 16. The device of any of Clauses 14 to 15, wherein for each reaction structure in the plurality of reaction structures, the first corresponding well serves as both a reagent and a readout well.

[0138] Clause 17. The device of any of the Clauses Petition 870250101629, dated 06 / 11 / 2025, page 43 / 50 40 / 45 to 16, where the first wells of the plurality of reaction structures are radially leveled relative to each other.

[0139] Clause 18. The device of any of Clauses 13 to 17, wherein for each reaction structure in at least one subset of the plurality of reaction structures, the reaction chamber comprises a second well and an intermediate chamber between the first and second wells.

[0140] Clause 19. The device of Clause 18, wherein for each reaction structure at least in the subset of the plurality of reaction structures, a second reagent is disposed in the second well when the device is made.

[0141] Clause 20. The device of any of Clauses 18 to 19, in which the second wells of at least the subset of the plurality of reaction structures are radially leveled to each other.

[0142] Clause 21. The device of any of Clauses 18 to 20, wherein the inlet of the reaction chamber is located in the first well and the second well is positioned radially into the first well.

[0143] Clause 22. The device of any of Clauses 13 to 21, wherein each respective reaction structure in the plurality of reaction structures comprises an air chamber, wherein the reaction chamber of the respective reaction structure has an outlet connected to the air chamber.

[0144] Clause 23. The device of any of Clauses 13 to 22, further comprising: an overflow chamber; and an overflow channel connecting the aliquot chamber of a last reaction structure in the plurality of reaction structures to the overflow chamber.

[0145] Clause 24. The device of Clause 23, wherein the overflow channel comprises a siphon portion and the siphon portion comprises (optionally or additionally): a radially level inlet with an inlet and an outlet from each connecting siphon in one or more siphons; and a level crest with or radially outward from a crest from each connecting siphon in one or more siphons. Petition 870250101629, dated 06 / 11 / 2025, pp. 44 / 50 41 / 45

[0146] Clause 25. The device of Clause 24, wherein the overflow channel further comprises a U-shaped channel having an outlet connected to the overflow chamber. In some embodiments, the outlet of the U-shaped channel is radially outside the siphon inlet. In some embodiments, an outer crest of the U-shaped channel is outside the channel outlet.

[0147] Clause 26. A method for using the device of any preceding Clause to mix one or more fluids, resuspend reagents, detect mixing or reaction, or a combination thereof.

[0148] Clause 27. A method comprising: (A) obtaining a device comprising: a rotational axis; and a plurality of reaction structures arranged circumferentially on at least a portion of the device, wherein each reaction structure in the plurality of reaction structures comprises an aliquot chamber having an outlet and a reaction chamber having an inlet connected to the outlet of the aliquot chamber, wherein the reaction chamber is positioned radially outward from the aliquot chamber; and one or more connecting siphons radially leveled to each other and radially inward to the outlets of the aliquot chambers of the plurality of reaction structures, wherein each respective connecting siphon in the one or more connecting siphons connects the aliquot chambers of the corresponding adjacent reaction structures in the plurality of reaction structures;(B) rotate the device at a speed to fill a first portion of the reaction chamber of each reaction structure in the plurality of reaction structures with a fluid; (C) increase the speed to empty the fluid from each corresponding connecting siphon in one or more connecting siphons; and (D) decrease the speed to create a volume of trapped gas within each corresponding connecting siphon in one or more connecting siphons, thus interrupting the fluidic connection between the plurality of reaction structures. In some embodiments, the reaction chamber is not vented;

[0149] Clause 28. The method of Clause 27, wherein the increase (C) moves at least a portion of the fluid out of the aliquot chamber of Petition 870250101629, dated 06 / 11 / 2025, pages 45 / 50 42 / 45 each respective reaction structure in the plurality of reaction structures, so that the fluid contained in the aliquot chamber of each respective reaction structure in the plurality of reaction structures is positioned out of either the inlet or outlet of each corresponding connection siphon in the one or more connection siphons, thereby emptying the fluid from each corresponding connection siphon in the one or more connection siphons.

[0150] Clause 29. The method of any of Clauses 27 to 28, wherein the decrease (D) moves at least a portion of the fluid contained in the reaction chamber of each respective reaction structure in the plurality of reaction structures to the aliquot chamber of each respective reaction structure in the plurality of reaction structures, so that the fluid contained in the aliquot chamber of each respective reaction structure in the plurality of reaction structures is positioned into the inlet and outlet of each corresponding connection siphon in the one or more connection siphons, thereby wetting the inlet and outlet of each corresponding connection siphon in the one or more connection siphons with the volume of gas trapped within each corresponding connection siphon.

[0151] Clause 30. The method of any of Clauses 27 to 29, wherein the first portion of the reaction chamber of each reaction structure in the plurality of reaction structures comprises a first well with a first reagent, the method further comprising: (E) repeating the increase (C) and decrease (D) one or more times to promote mixing of the fluid with the first reagent.

[0152] Clause 31. The method of any of Clauses 27 to 30, further comprising: (F) detecting, for each reaction structure in the plurality of reaction structures, light transmitted through the first well or a wall of the first well of the reaction chamber.

[0153] Clause 32. The method of any of Clauses 27 to 31, further comprising: (G) increasing the rate to fill a second portion of the reaction chamber of each reaction structure by at least Petition 870250101629, dated 06 / 11 / 2025, pp. 46 / 50 43 / 45 minus a subset of the plurality of reaction structures, and (H) perform the decrease (D).

[0154] Clause 33. The method of Clause 32, wherein the second portion of the reaction chamber of each reaction structure in at least the subset of the plurality of reaction structures comprises a second well with a second reagent, the method further comprising: (I) repeating the increase (G) and performing (H) one or more times to promote mixing of the fluid with the second reagent.

[0155] Clause 34. The method of any of Clauses 27 to 33, further comprising: (J) detecting, for each reaction structure at least in the set of plurality of reaction structures, light transmitted through the first well or a wall of the first well of the reaction chamber.

[0156] Clause 35. A system for operating the device or executing the method of any preceding Clause. TERMINOLOGIES AND REFERENCES CITED

[0157] The terminology used in this document is intended to describe particular implementations only and is not intended to limit the claims. As used in the description of the implementations and in the appended claims, the singular forms “a / an”, “a / an”, and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be understood that the terms “left” or “right”, “top” or “bottom”, “lower” or “upper”, “inside” or “outside”, “inside” or “outside”, etc. are used to describe features of the exemplary embodiments with reference to the positions of such features as shown in the figures. It will also be understood that, although terms first, second, etc. may be used in this document to describe various elements, these elements should not be limited by these terms. These terms are used only to distinguish one element from another.For example, a first element could be called a second element, and so on. Petition 870250101629, dated 06 / 11 / 2025, pp. 47 / 50 44 / 45 In the same way, a second element could be called a first element, without altering the meaning of the description, as long as the “first element” and the “second element” were renamed consistently.

[0158] Also used in this document, the term “and / or”, as used herein, refers to and encompasses any and all possible combinations of one or more of the associated listed items. It will be further understood that the terms include, includes, including, comprise, comprises and / or comprising, when used in this descriptive report, specify the presence of declared features, integers, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integers, steps, operations, elements, components and / or groups thereof.

[0159] The term about or approximately is used here to provide literal support for the exact quantity that precedes it, as well as a number that is close to or approximately the number that the term precedes. To determine whether a number is close to or approximately a specifically cited number, the uncited approximation or near number may be a number that, in the context in which it is presented, provides the substantial equivalent of the specifically cited number. It should be understood that all numerical values ​​and ranges disclosed here are approximate values ​​and ranges, regardless of whether “about” is used in conjunction with them.It should also be understood that the term "approximately," as used herein in conjunction with a numeral, refers to a value that may be ±0.01% (inclusive), ±0.1% (inclusive), ±0.5% (inclusive), ±1% (inclusive) of that numeral, ±2% (inclusive) of that numeral, ±3% (inclusive) of that numeral, ±5% (inclusive) of that numeral, ±10% (inclusive) of that numeral, or ±15% (inclusive) of that numeral. It should further be understood that when a numerical range is disclosed herein, any numerical value within that range is also specifically disclosed.

[0160] The term “if” used here is optionally interpreted Petition 870250101629, dated 06 / 11 / 2025, pp. 48 / 50 45 / 45 as meaning “when” or “after” or “in response to the determination” or “in response to the detection” or “according to a determination that”, depending on the context. Similarly, the phrase “if it is determined” or “if [a stated condition or event] is detected” used here is optionally interpreted as meaning “upon determining” or “in response to the determination” or “upon detecting [the stated condition or event]” or “in response to the detection [of the stated condition or event]” or “according to a determination that [a stated condition or event] is detected”, depending on the context.

[0161] When a reference number is given the denotation “ith”, the reference number refers to a generic component, set, or modality. For example, a “unit i” refers to the i’th unit in a plurality of units.

[0162] All references cited in this document are incorporated herein by reference in their entirety and for all purposes, to the same extent as if each individual publication or patent or patent application had been specifically and individually indicated to be incorporated by reference in its entirety and for all purposes. Petition 870250101629, dated 06 / 11 / 2025, pp. 49 / 50

Claims

1 / 5 CLAIMS 1. Device, characterized by comprising: a rotation axis; and a reaction chamber comprising an inlet for receiving a fluid and a first well to serve as both a reagent and a reading well, wherein (i) a first reagent is disposed in the first well when the device is made, (ii) a radial position of a fluid meniscus in the reaction chamber depends at least in part on a rotation speed of the device, and (iii) the first well has a substantially flat wall perpendicular to the rotation axis to allow reproducible light transmission.

2. Device according to claim 1, characterized in that the reaction chamber is not ventilated to promote homogeneous mixing of the first reagent with the fluid.

3. Device according to claim 2, characterized in that the reaction chamber comprises an outlet connected to an air chamber.

4. Device according to claim 3, characterized in that the first well is connected to the air chamber.

5. Device according to claim 3, characterized in that the reaction chamber further comprises a second well and an intermediate chamber between the first and second wells, in which a second reagent is disposed in the second well when the device is made.

6. Device according to claim 5, characterized in that the inlet is located in the first well and the second well is positioned radially inside the first well.

7. Device according to claim 6, characterized in that the outlet is located in the second well.

8. Device, characterized by comprising: a rotation axis; and a plurality of reaction structures arranged circumferentially on at least one portion of the device, wherein each reaction structure in the plurality of reaction structures comprises an aliquot chamber having an outlet and a reaction chamber having an inlet connected to the outlet of the aliquot chamber, wherein the reaction chamber is positioned radially outward from the aliquot chamber; and one or more connecting siphons radially leveled to each other and radially inward to the outlets of the aliquot chambers of the plurality of reaction structures, wherein each respective connecting siphon in the one or more connecting siphons connects the aliquot chambers of the corresponding adjacent reaction structures in the plurality of reaction structures.

9. Device according to claim 8, characterized in that each reaction structure in the plurality of reaction structures, the reaction chamber comprises a first well, and a first reagent is disposed in the first well.

10. Device according to claim 9, characterized by a corresponding reaction structure in the plurality of reaction structures, the first corresponding well serving as both a reagent and a readout well.

11. Device according to claim 8, characterized in that the first wells of the plurality of reaction structures are radially leveled with respect to each other.

12. Device according to claim 9, characterized in that each reaction structure in at least one subset of the plurality of reaction structures, the reaction chamber comprises a second well and an intermediate chamber between the first and second wells, and a second reagent is disposed in the second well.

13. Device according to claim 12, characterized in that the second wells of at least the subset of the plurality of reaction structures are radially leveled with respect to each other.

14. Device according to claim 12, characterized in that the inlet of the reaction chamber is located in the first well and the second well is positioned radially into the first well. Petition 870250080861, dated 09 / 09 / 2025, pp. 81 / 85 3 / 5 15. Device according to claim 8, characterized in that each respective reaction structure in the plurality of reaction structures comprises an air chamber, wherein the reaction chamber of the respective reaction structure has an outlet connected to the air chamber.

16. Device according to claim 8, further characterized by comprising: an overflow chamber; and an overflow channel connecting the aliquot chamber of a final reaction structure in the plurality of reaction structures to the overflow chamber.

17. Device according to claim 16, characterized in that the overflow channel comprises a siphon portion and the siphon portion comprises: a radially level inlet with an inlet and an outlet of each connecting siphon in one or more siphons; and a crest level with or radially outward from a crest of each connecting siphon in one or more siphons.

18. Device according to claim 17, characterized in that the overflow channel further comprises a U-shaped channel having an outlet connected to the overflow chamber.

19. Method, characterized by comprising: (A) obtaining a device comprising: a rotation axis; and a plurality of reaction structures arranged circumferentially on at least a portion of the device, wherein each reaction structure in the plurality of reaction structures comprises an aliquot chamber having an outlet and a reaction chamber having an inlet connected to the outlet of the aliquot chamber, wherein the reaction chamber is positioned radially outward from the aliquot chamber; and one or more connecting siphons radially leveled to each other and Petition 870250080861, dated 09 / 09 / 2025, page.82 / 85 4 / 5 radially into the outlets of the aliquot chambers of the plurality of reaction structures, wherein each respective connecting siphon in the one or more connecting siphons connects the aliquot chambers of the corresponding adjacent reaction structures in the plurality of reaction structures; (B) rotate the device at a speed to fill a first portion of the reaction chamber of each reaction structure in the plurality of reaction structures with a fluid; (C) increase the speed to empty the fluid from each corresponding connecting siphon in the one or more connecting siphons; and (D) decrease the speed to create a volume of trapped gas within each corresponding connecting siphon in the one or more connecting siphons, thereby interrupting the fluid connection between the plurality of reaction structures.

20. Method according to claim 19, characterized in that increase (C) moves at least a portion of the fluid out of the aliquot chamber of each respective reaction structure in the plurality of reaction structures, so that the fluid contained in the aliquot chamber of each respective reaction structure in the plurality of reaction structures is positioned out of either the inlet or outlet of each corresponding connection siphon in the one or more connection siphons, thereby emptying the fluid from each corresponding connection siphon in the one or more connection siphons.

21. Method according to claim 19, characterized by the decrease (D) moving at least a portion of the fluid contained in the reaction chamber of each respective reaction structure in the plurality of reaction structures to the aliquot chamber of each respective reaction structure in the plurality of reaction structures, such that the fluid contained in the aliquot chamber of each respective reaction structure in the plurality of reaction structures is positioned into the inlet and outlet of each corresponding connection siphon in one or more connection siphons, thereby wetting the inlet and outlet of each corresponding connection siphon in one or more connection siphons with the volume of gas trapped within each corresponding connection siphon.

22. Method according to claim 19, characterized in that the first portion of the reaction chamber of each reaction structure in the plurality of reaction structures comprises a first well with a first reagent, the method further comprising: (E) repeating the increase (C) and decrease (D) one or more times to promote mixing of the fluid with the first reagent.

23. Method according to claim 22, characterized by further comprising: (F) detecting, for each reaction structure in the plurality of reaction structures, light transmitted through the first well or a wall of the first well of the reaction chamber.

24. Method according to claim 19, characterized by further comprising: (G) increasing the rate to fill a second portion of the reaction chamber of each reaction structure in at least one subset of the plurality of reaction structures; and (H) performing the decrease (D).

25. Method according to claim 24, characterized in that the second portion of the reaction chamber of each reaction structure at least in the subset of the plurality of reaction structures comprises a second well with a second reagent, the method further comprising: (I) repeating the increase (G) and perform (H) one or more times to promote mixing of the fluid with the second reagent.

26. Method according to claim 25, characterized by further comprising: (J) detecting, for each reaction structure at least in the subset of the plurality of reaction structures, light transmitted through the first well or a wall of the first well of the reaction chamber. Petition 870250080861, dated 09 / 09 / 2025, pp. 84 / 85

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