Microfluidic chip and immunoassay instrument

Abstract

The present application relates to a microfluidic chip and an immunoassay instrument. The microfluidic chip comprises a body having a center of rotation, wherein a microfluidic chamber is provided in the body; and in the microfluidic chamber and in order of increasing distance from the center of rotation, a first liquid cavity, a first valve control structure, a first reaction cavity, a fourth valve control structure and a detection reaction cavity are sequentially in communication, and a second liquid cavity, a second valve control structure, the detection reaction cavity, a third valve control structure and a waste liquid cavity are sequentially in communication.

Description

Microfluidic chips and immunoassay instruments

[0001] Related applications

[0002] This application claims priority to Chinese patent application filed on January 14, 2025, with application number 202510052573.7, entitled “Microfluidic Chip and Immunoassay Instrument”, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This application relates to the field of microfluidic chips, and in particular to microfluidic chips and immunoassay instruments. Background Technology

[0004] Microfluidic chips are microanalytical systems that integrate sample pretreatment, mixing, reaction, separation, and detection into one or more chips. Analytical instruments using microfluidic chips can replace a significant amount of traditional laboratory work.

[0005] Furthermore, microfluidic chips offer advantages such as small sample volume, simple operation, and the ability to accurately complete the entire process from sample preparation to result display in a short time, effectively overcoming experimental errors caused by manual operation in traditional laboratory work. Therefore, microfluidic chips are finding increasing applications in fields such as chemical analysis, DNA sequencing, protein analysis, single-cell analysis, single-molecule analysis, food safety, environmental monitoring, and drug screening.

[0006] However, most traditional microfluidic disk chips are designed to achieve biochemical reactions. The few microfluidic disk chips that are adapted to immune reactions are only suitable for one-step processes. There is a lack of microfluidic chips, especially disk chips, that are adapted to delayed one-step processes. Summary of the Invention

[0007] Therefore, it is necessary to provide a microfluidic chip and an immunoassay instrument.

[0008] In one embodiment, a microfluidic chip includes a body having a center of rotation and a microfluidic chamber formed therein;

[0009] The microfluidic chamber includes a first valve-controlled structure, a second valve-controlled structure, a third valve-controlled structure, a fourth valve-controlled structure, a first liquid chamber, a second liquid chamber, a first reaction chamber, a detection reaction chamber, and a waste liquid chamber;

[0010] The first liquid chamber, the first valve-controlled structure, the first reaction chamber, the fourth valve-controlled structure, and the detection reaction chamber are sequentially connected and arranged in order of distance from the rotation center from near to far; the second liquid chamber, the second valve-controlled structure, the detection reaction chamber, the third valve-controlled structure, and the waste liquid chamber are sequentially connected and arranged in order.

[0011] The first valve control structure is configured to close at speeds below a preset first rotational speed;

[0012] The second valve control structure is configured to close at speeds below a preset second rotational speed;

[0013] The third valve control structure is set to close at a speed lower than the preset third rotation speed to control the flow of liquid in the detection reaction chamber to the waste liquid chamber;

[0014] The preset first speed is not higher than the preset second speed, and the preset first speed is lower than the preset third speed.

[0015] The aforementioned microfluidic chip, through the design of a first liquid chamber, a second liquid chamber, a first reaction chamber, and a detection reaction chamber that work together, is suitable for time-delayed one-step and two-step detection schemes that require precise quantification of two reactions. This enables time-delayed one-step or two-step microfluidic detection, allowing the microfluidic chip to be matched with time-delayed one-step and two-step immune reactions, further broadening the versatility of microfluidic chips, especially disk-type microfluidic chips.

[0016] In some embodiments, the first liquid chamber and the first valve control structure are located on one side of the body, while the second liquid chamber, the second valve control structure, the first reaction chamber, the fourth valve control structure, the detection reaction chamber, the third valve control structure, and the waste liquid chamber are located on the other side of the body.

[0017] In some embodiments, the first liquid chamber, the first valve-controlled structure, the first reaction chamber, and the fourth valve-controlled structure are located on one side of the body, while the second liquid chamber, the second valve-controlled structure, the detection reaction chamber, the third valve-controlled structure, and the waste liquid chamber are located on the other side of the body.

[0018] In some embodiments, the first liquid chamber, the first valve-controlled structure, the first reaction chamber, the fourth valve-controlled structure, the detection reaction chamber, the third valve-controlled structure, and the waste liquid chamber are located on one side of the body, while the second liquid chamber and the second valve-controlled structure are located on the other side of the body.

[0019] In some embodiments, the first valve control structure, the second valve control structure, the third valve control structure, the fourth valve control structure, the first liquid chamber, the second liquid chamber, the first reaction chamber, the detection reaction chamber, and the waste liquid chamber are located on the same side of the body.

[0020] In some embodiments, in order of distance from the rotation center from near to far, the inlet of the first valve-controlled structure is connected to the bottom of the first liquid chamber, the inlet of the second valve-controlled structure is connected to the bottom of the second liquid chamber, the inlet of the third valve-controlled structure is connected to the bottom of the detection reaction chamber, the inlet of the fourth valve-controlled structure is connected to the bottom of the first reaction chamber, and the outlet of the fourth valve-controlled structure is connected to the top of the detection reaction chamber, with the top of the fourth valve-controlled structure being higher than the top of the first reaction chamber.

[0021] In some embodiments, the first valve-controlled structure, the second valve-controlled structure, and the third valve-controlled structure are steam traps or capillary valves.

[0022] In some embodiments, the fourth valve-controlled structure is a siphon valve, and the surface of the fourth valve-controlled structure is a hydrophilic layer.

[0023] In some embodiments, the microfluidic chip includes at least one of the following:

[0024] The first liquid chamber and the second liquid chamber are metering chambers used to meter the liquid.

[0025] The first reaction chamber and the detection reaction chamber are each pre-filled with reaction reagents;

[0026] The volume of the detection reaction chamber is greater than or equal to the sum of the volumes of the first liquid chamber and the second liquid chamber;

[0027] The volume of the waste liquid chamber is greater than or equal to the sum of the volumes of the first liquid chamber and the second liquid chamber;

[0028] The first liquid chamber and the second liquid chamber are quantitative chambers, and the volumes of the first liquid chamber and the second liquid chamber are the same or different.

[0029] The shape of the first reaction chamber is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, pear-shaped, rectangular, and regular polygonal.

[0030] The volume of the first reaction chamber is greater than or equal to the volume of the first liquid chamber;

[0031] The shape of the detection reaction chamber is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, pear-shaped, rectangular, and regular polygonal.

[0032] The shape of the detection reaction chamber is set according to the photodetector;

[0033] The volume of the detection reaction chamber is greater than the volume of the first liquid chamber, the volume of the first reaction chamber, and the volume of the second liquid chamber.

[0034] The shape of the waste liquid chamber is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, rectangular, and regular polygonal.

[0035] The volume of the waste liquid chamber is greater than or equal to the sum of the volumes of the first liquid chamber, the second liquid chamber, the first reaction chamber, and the detection reaction chamber.

[0036] In some embodiments, the microfluidic chip is a disk-type microfluidic chip.

[0037] In some embodiments, an immunoassay instrument includes a centrifuge and a microfluidic chip, as described in any embodiment, disposed on the rotating shaft of the centrifuge. Attached Figure Description

[0038] To more clearly illustrate the technical solutions in the embodiments of this application or the conventional technology, the drawings used in the description of the embodiments or the conventional technology will be briefly introduced below. Obviously, the drawings described below are only embodiments of this application. For those skilled in the art, other drawings can be obtained based on the disclosed drawings without creative effort.

[0039] Figure 1 is a schematic diagram of the structure of a microfluidic chip according to an embodiment of the present application.

[0040] Figure 2 is a schematic diagram of another embodiment of the microfluidic chip of this application.

[0041] Figure 3 is a partial structural schematic diagram of the embodiment shown in Figure 2.

[0042] Figure 4 is an application diagram of the embodiment shown in Figure 3.

[0043] Reference numerals: microfluidic chip 100, body 110, rotation center 120, microfluidic chamber 130, first valve control structure 131, second valve control structure 132, third valve control structure 133, fourth valve control structure 134, first liquid chamber 135, second liquid chamber 136, first reaction chamber 137, detection reaction chamber 138, waste liquid chamber 139, centrifugation direction 140, liquid 200. Detailed Implementation

[0044] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0045] In one embodiment of this application, a microfluidic chip includes a body with a center of rotation and a microfluidic chamber formed therein. The microfluidic chamber includes a first valve-controlled structure, a second valve-controlled structure, a third valve-controlled structure, a fourth valve-controlled structure, a first liquid chamber, a second liquid chamber, a first reaction chamber, a detection reaction chamber, and a waste liquid chamber. The first liquid chamber, the first valve-controlled structure, the first reaction chamber, the fourth valve-controlled structure, and the waste liquid chamber are sequentially connected in order of distance from the center of rotation, and the second liquid chamber, the second valve-controlled structure, the detection reaction chamber, the third valve-controlled structure, and the waste liquid chamber are also sequentially connected. This microfluidic chip, through the design of a matching first liquid chamber, second liquid chamber, first reaction chamber, and detection reaction chamber, is suitable for time-delayed one-step and two-step detection schemes requiring precise quantification of two reactions. This achieves time-delayed one-step or two-step microfluidic detection, making the microfluidic chip compatible with time-delayed one-step and two-step immune reactions, further broadening the versatility of microfluidic chips, especially disk-type microfluidic chips.

[0046] In some embodiments, a microfluidic chip 100 is shown in FIG1, which includes a body 110 having a rotation center 120 and a microfluidic chamber 130 formed in the body 110; in the illustrated embodiment, the rotation center 120 of the microfluidic chip 100 is located outside the body 110, and the microfluidic chip 100 is a disk-type microfluidic chip, and multiple microfluidic chips 100 can be placed in a rotating system.

[0047] Exemplary examples include a microfluidic chip 100 as shown in FIG2. The microfluidic chip 100 is a single-disk microfluidic chip with a circular body 110. The rotation center 120 is located within the body 110 and is the center of the circle. In this embodiment, the microfluidic chip 100 has only one microfluidic chamber 130 in its body 110. In other embodiments, the microfluidic chip 100 may also have two or more microfluidic chambers 130 in its body 110. The embodiments of this application do not impose additional limitations on this.

[0048] Referring to Figures 2 and 3, in each embodiment, the microfluidic chamber 130 includes a first valve-controlled structure 131, a second valve-controlled structure 132, a third valve-controlled structure 133, a fourth valve-controlled structure 134, a first liquid chamber 135, a second liquid chamber 136, a first reaction chamber 137, a detection reaction chamber 138, and a waste liquid chamber 139. Referring to Figure 4, in this embodiment, the first liquid chamber 135 and the second liquid chamber 136 are used for metering liquid 200; the valve-controlled structures, including the first valve-controlled structure 131, the second valve-controlled structure 132, the third valve-controlled structure 133, and the fourth valve-controlled structure 134, are used for transferring liquid 200; taking the fourth valve-controlled structure 134 as an example, the fourth valve-controlled structure 134 is used to transfer liquid 200 in the first reaction chamber 137 to the detection reaction chamber 138 under specific conditions. The first reaction chamber 137 and the detection reaction chamber 138 are used to carry out reactions such as immune reactions. Reaction reagents for the reaction can be pre-set in the first reaction chamber 137 and the detection reaction chamber 138. The waste liquid chamber 139 is used to receive waste liquid.

[0049] Furthermore, in the microfluidic chamber 130, the first liquid chamber 135 and the second liquid chamber 136 are closest to the rotation center 120 relative to other structures, while the waste liquid chamber 139 is farthest from the rotation center 120 relative to other structures.

[0050] Specifically, the first liquid chamber 135, the first valve control structure 131, the first reaction chamber 137, the fourth valve control structure 134, and the detection reaction chamber 138 are sequentially connected in order of distance from the rotation center 120, and the second liquid chamber 136, the second valve control structure 132, the detection reaction chamber 138, the third valve control structure 133, and the waste liquid chamber 139 are sequentially connected in order of distance from the rotation center 120; that is, in the centrifugal direction 140, the first liquid chamber 135, the first valve control structure 131, the first reaction chamber 137, the fourth valve control structure 134, and the detection reaction chamber 138 are sequentially connected in order of distance from the rotation center 120, and the second liquid chamber 136, the second valve control structure 132, the detection reaction chamber 138, the third valve control structure 133, and the waste liquid chamber 139 are sequentially connected in order of distance from the rotation center 120.

[0051] In this structural design, under rotating conditions, the first liquid chamber 135, the first valve control structure 131, and the first reaction chamber 137 form a pre-reaction system. The resulting preliminary reactants enter the detection reaction chamber 138 via the fourth valve control structure 134. At this time, the second valve control structure 132 can be opened in conjunction with this, or the second valve control structure 132 can be opened in advance. The substances in the second liquid chamber 136 enter the detection reaction chamber 138 via the second valve control structure 132, react with the preliminary reactants, and obtain the final reactants. After the detection reaction chamber 138 completes the detection, the final reactants enter the waste liquid chamber 139 via the third valve control structure 133.

[0052] As can be seen from the above description, although the above embodiments are essentially two-step methods, they employ a microfluidic chip and, within a single system, can achieve two reactions requiring precise quantification through a single centrifugation operation. This improves detection efficiency and is compatible with three immunological methodologies: one-step, delayed one-step, and two-step. It can adapt to a wider range of projects. Some embodiments may also be referred to as delayed one-step detection or two-step detection, providing both a delayed one-step detection scheme and a two-step detection scheme. This achieves microfluidic detection of delayed one-step and two-step immunological methods, enabling the microfluidic chip 100 to be matched with delayed one-step and two-step immunological reactions, further broadening the versatility of the microfluidic chip 100, especially the disc-type microfluidic chip.

[0053] Exemplary examples, in some embodiments, as shown in FIG4, the second liquid chamber 136 contains liquid 200, which can be functionally categorized as a reaction solution, diluent, and cleaning solution, etc.; in other embodiments, the first liquid chamber 135 may also contain liquid 200. For stringent reaction conditions, the liquid 200 contained in the first liquid chamber 135 and the second liquid chamber 136 is precisely quantified. In some embodiments, the first reaction chamber 137 and / or the detection reaction chamber 138 may also contain liquid 200 to perform functions such as participating in the reaction, diluting the solute, or cleaning. This structural design facilitates the implementation of two separately controlled reactions in the same centrifugation operation without replacing the microfluidic chip 100. Therefore, it provides a microfluidic detection scheme compatible with both two-step and delayed one-step methods, which not only improves the efficiency of immunoassay detection but also enhances the precision of the detection results and reduces the amount of microfluidic chip 100 used.

[0054] In some embodiments, the first valve control structure 131 is configured to close at a speed lower than a preset first rotational speed; the second valve control structure 132 is configured to close at a speed lower than a preset second rotational speed; and the third valve control structure 133 is configured to close at a speed lower than a preset third rotational speed, thereby controlling the flow of liquid 200 in the detection reaction chamber 138 or the first reaction chamber 137 to the waste liquid chamber 139; wherein the preset first rotational speed is not higher than the preset second rotational speed, and both the preset first rotational speed and the preset second rotational speed are lower than the preset third rotational speed. For example, the preset first rotational speed is the same as the preset second rotational speed, and both the preset first rotational speed and the preset second rotational speed are lower than the preset third rotational speed. Exemplarily, in some embodiments, the preset first rotational speed is 500 rpm, i.e., 500 revolutions per minute; the preset second rotational speed is 800 rpm; and the preset third rotational speed is 1000 rpm; that is, the preset first rotational speed is lower than the preset second rotational speed, and the preset second rotational speed is lower than the preset third rotational speed. Alternatively, the first preset rotational speed is 800 rpm, the second preset rotational speed is 800 rpm, and the third preset rotational speed is 1000 rpm; that is, the first preset rotational speed is the same as the second preset rotational speed, and both the first and second preset rotational speeds are lower than the third preset rotational speed. Alternatively, the first preset rotational speed is 1000 rpm, the second preset rotational speed is 2000 rpm, and the third preset rotational speed is 3000 rpm. Other embodiments follow the same principle and will not be elaborated further. The specific settings or adjustments of the first, second, and third preset rotational speeds can be made according to actual conditions, and no additional limitations are imposed on these settings in the embodiments of this application. This design, through simple centrifugation speed control, can realize the on / off control of the first valve-controlled structure 131, the second valve-controlled structure 132, and the third valve-controlled structure 133, thereby completing the time-delayed one-step and two-step detection schemes requiring precise quantitative measurement of two reactions, offering advantages of convenience, practicality, and speed.

[0055] For example, the liquid transfer process achieved by the microfluidic chip 100 through rotational speed control is described as follows: The second liquid enters the second liquid chamber 136 from the back channel and enters the first reaction chamber 137 under the action of the second rotational speed; and under the action of the second rotational speed, the second liquid will not pass through the fourth valve control structure 134, at which time the fourth valve control structure 134 is in a closed state. When the microfluidic chip 100 stops rotating, the second liquid passes through the fourth valve control structure 134 under capillary action, at which time the fourth valve control structure 134 is in a closed state, and the second liquid enters the detection reaction chamber 138 from the first reaction chamber 137 under the action of the second rotational speed; after the second liquid has been incubated in the detection reaction chamber 138, it enters the waste liquid chamber 139 under the action of the third rotational speed; similarly, the first liquid enters the first liquid chamber 135 from the front channel and enters the detection reaction chamber 138 under the action of the first rotational speed; after the first liquid has been incubated in the detection reaction chamber 138, it enters the waste liquid chamber 139 under the action of the third rotational speed.

[0056] In some embodiments, in order of increasing distance from the rotation center 120, the inlet of the first valve-controlled structure 131 connects to the bottommost end of the first liquid chamber 135, and the inlet of the second valve-controlled structure 132 connects to the bottommost end of the second liquid chamber 136. That is, the inlet of the first valve-controlled structure 131 connects to the lowest point of the first liquid chamber 135, which is also the position of the first liquid chamber 135 furthest from the rotation center 120. Other embodiments follow the same principle and will not be elaborated further. In some embodiments, in order of increasing distance from the rotation center 120, the inlet of the third valve-controlled structure 133 connects to the bottommost end of the detection reaction chamber 138. In some embodiments, in order of increasing distance from the rotation center 120, the inlet of the fourth valve-controlled structure 134 connects to the bottommost end of the first reaction chamber 137. In some embodiments, in order of increasing distance from the rotation center 120, the bottommost end of the first reaction chamber 137 is higher than the topmost end of the detection reaction chamber 138. For example, in some embodiments, the bottom of the first reaction chamber 137 is higher than the top of the detection reaction chamber 138 in order of distance from the rotation center 120. Traditional methodologies for disc-type microfluidic chips are mostly biochemical reactions, which are difficult to adapt to delayed one-step or two-step immunoreaction processes. This is mainly due to the following limitations: immunoreaction requires precise quantification, multiple rounds of washing, repeated reuse of valve-controlled structures, and two incubation cycles required for delayed one-step or two-step processes. Therefore, two reaction chambers for incubation need to be designed, and the reaction proceeds in a specific order. Furthermore, the process of dispensing liquid from the disc-type microfluidic chip to the reaction chambers is typically controlled by a hydrophobic valve or capillary valve, but neither hydrophobic valves nor capillary valves can effectively control the liquid in both reaction chambers. This application provides a microfluidic chamber design that enables quantitative mixing and reaction by designing a first liquid chamber 135, a second liquid chamber 136, a first reaction chamber 137, and a detection reaction chamber 138 that work together. This design enables the application of a disc-type microfluidic chip with delayed one-step or two-step immunoreaction methods, allowing the microfluidic chip 100 to be matched with more methodologies. This solves the problem that traditional disc-type microfluidic chips are difficult to adapt to delayed one-step and two-step immunoreaction methods, and further broadens the versatility of the microfluidic chip 100, especially the disc-type microfluidic chip.

[0057] In some embodiments, the first valve control structure 131, the second valve control structure 132, and the third valve control structure 133 are hydrophobic valves or capillary valves. In some embodiments, the opening speed of the first valve control structure 131 and the second valve control structure 132 is less than the opening speed of the third valve control structure 133. At low speeds, the first valve control structure 131 and the second valve control structure 132 prevent the liquid 200 in the first liquid chamber 135 and the second liquid chamber 136 (e.g., the metering chamber) from flowing to the reaction chamber, thus achieving the metering function of the metering chamber. The opening speed of the third valve control structure 133 is higher than that of the first valve control structure 131 and the second valve control structure 132, preventing the liquid 200 in the reaction chamber from flowing to the waste liquid chamber 139 when the first valve control structure 131 is opened. The inlet of the first valve control structure 131 should be located at the bottom of the first liquid chamber 135, and the inlet of the second valve control structure 132 should be located at the bottom of the second liquid chamber 136. This design facilitates accurate control of the flow direction of the liquid 200 in the microfluidic chamber 130, ensuring that the reactions in the first reaction chamber 137 and the detection reaction chamber 138 can be precisely quantified according to the design objectives.

[0058] In some embodiments, the fourth valve-controlled structure 134 is a siphon valve. In some embodiments, the fourth valve-controlled structure 134 is a siphon valve, and its surface is a hydrophilic layer. In order of increasing distance from the rotation center 120, the outlet of the fourth valve-controlled structure 134 connects to the uppermost end of the detection reaction chamber 138, and the uppermost end of the fourth valve-controlled structure 134 is higher than the uppermost end of the first reaction chamber 137. That is, the minimum distance between the fourth valve-controlled structure 134 and the rotation center 120 is less than the minimum distance between the first reaction chamber 137 and the rotation center 120. For example, the uppermost end of the fourth valve-controlled structure 134 is higher than the uppermost end of the detection reaction chamber 138, meaning the minimum distance between the fourth valve-controlled structure 134 and the rotation center 120 is less than the minimum distance between the detection reaction chamber 138 and the rotation center 120. The uppermost end is the position closest to the rotation center 120, i.e., the position with the smallest distance from the rotation center 120. In some embodiments, the surface of the fourth valve-controlled structure 134 is made of a hydrophilic material, allowing the liquid 200 to fill the fourth valve-controlled structure 134 under capillary action. The inlet of the fourth valve-controlled structure 134 is located at the bottom of the first reaction chamber 137, and the outlet of the fourth valve-controlled structure 134 is located at the top of the detection reaction chamber 138. The highest point of the fourth valve-controlled structure 134 should be higher than the highest point of the first reaction chamber 137. This design, compared to traditional hydrophobic valves and capillary valves, allows for precise control of the opening and closing of the fourth valve-controlled structure 134 by controlling the centrifugation speed. This effectively meets the technical requirements of both one-step and two-step delayed-time immunoassays, ensuring that the microfluidic chip 100 is compatible with both one-step and two-step delayed-time immunoassays.

[0059] In some embodiments, as shown in Figures 1 to 4, the microfluidic chip 100 includes a body 110 having a rotation center 120 and a microfluidic chamber 130 formed therein; the microfluidic chamber 130 includes a first valve control structure 131, a second valve control structure 132, a third valve control structure 133, a fourth valve control structure 134, a first liquid chamber 135, a second liquid chamber 136, a first reaction chamber 137, a detection reaction chamber 138, and a waste liquid chamber 139; the distance from the rotation center 120 is... From near to far, the first liquid chamber 135, the first valve-controlled structure 131, the first reaction chamber 137, the fourth valve-controlled structure 134, and the detection reaction chamber 138 are sequentially connected, and the second liquid chamber 136, the second valve-controlled structure 132, the detection reaction chamber 138, the third valve-controlled structure 133, and the waste liquid chamber 139 are sequentially connected. The first valve-controlled structure 131 is configured to close at a speed lower than a preset first rotational speed; the second valve-controlled structure 132 is configured to close at a speed lower than a preset second rotational speed; and the third valve-controlled structure 133 is configured to close at a speed lower than a preset third rotational speed, so as to control the flow of liquid in the detection reaction chamber 138 to the waste liquid chamber 139. The preset first rotational speed is not higher than the preset second rotational speed, and both the preset first rotational speed and the preset second rotational speed are lower than the preset third rotational speed. From near to far from the rotation center 120, the inlet of the first valve-controlled structure 131 is connected to the bottom of the first liquid chamber 135, the inlet of the second valve-controlled structure 132 is connected to the bottom of the second liquid chamber 136, and the inlet of the third valve-controlled structure 133 is connected to the bottom of the second liquid chamber 136. The bottom of the detection reaction chamber 138 is connected to the bottom of the first reaction chamber 137, and the outlet of the fourth valve control structure 134 is connected to the top of the detection reaction chamber 138, with the top of the fourth valve control structure 134 being higher than the top of the first reaction chamber 137. The first valve control structure 131, the second valve control structure 132, and the third valve control structure 133 are hydrophobic valves or capillary valves; the fourth valve control structure 134 is a siphon valve, and its surface is a hydrophilic layer. This design, using a microfluidic chip, allows for precise quantification of two reactions in a single system through a single centrifugation operation combined with centrifugation speed control, thus improving detection efficiency. It is compatible with three immunoassay methodologies: one-step, delayed one-step, and two-step, making it suitable for a wider range of projects. Furthermore, it provides detection schemes adapted to delayed one-step and two-step methods, thereby realizing microfluidic detection of delayed one-step and two-step immune methods. This allows the microfluidic chip 100 to be matched with one-step, delayed one-step, and two-step immune reactions, respectively, thus broadening the versatility of the microfluidic chip 100, especially the disc-type microfluidic chip.

[0060] In some embodiments, the first liquid chamber 135 and the second liquid chamber 136 are metering chambers for metering liquid 200; in some embodiments, the first liquid chamber 135 and the second liquid chamber 136 are metering chambers, and their volumes are the same or different. This structural design facilitates the accurate addition of liquid 200 to the first liquid chamber 135 and / or the second liquid chamber 136, controlling the two reactions requiring precise metering from the source, thereby improving the testing precision of the microfluidic chip 100.

[0061] In some embodiments, the first reaction chamber 137 and the detection reaction chamber 138 are respectively pre-filled with reaction reagents. The reaction chamber, including the first reaction chamber 137 and the detection reaction chamber 138, contains the reagent to be reacted, i.e., the reaction reagent. The reaction reagent can be liquid, solid, or a solid-liquid mixture, etc. Exemplarily, in some embodiments, the reaction reagent is one or more of solid reagents, lyophilized reagents, liquid reagents, and solidified reagents adsorbed onto the chamber. Exemplarily, in some embodiments, the reaction reagent is encapsulated in the first reaction chamber 137 and / or the detection reaction chamber 138 for release under preset conditions, including at least one of centrifugal speed and ambient temperature. This structural design allows the reaction reagent to have a relatively long shelf life and prevents it from being transferred to other locations before the valve-controlled structure is opened, thus avoiding affecting the accuracy of the two reactions.

[0062] In some embodiments, the shape of the first reaction cavity 137 is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, pear-shaped, rectangular, and regular polygonal; or, in some embodiments, the volume of the first reaction cavity 137 is greater than or equal to the volume of the first liquid cavity 135. Exemplarily, in some embodiments, the volume of the first reaction cavity 137 is 1.5 to 2 times the volume of the first liquid cavity 135. In some embodiments, the shape of the first reaction cavity 137 can be circular, square, pear-shaped, rectangular, etc., and its volume should be greater than the volume of the first liquid cavity 135, more preferably 1.5 to 2 times the volume of the first liquid cavity 135. The first reaction cavity 137 is closer to the center of the microfluidic disk-type chip than the detection reaction cavity 138, and more preferably, the bottom of the first reaction cavity 137 is higher than the top of the detection reaction cavity 138. This structural design ensures that the reaction in the first reaction chamber 137 starts first, completes first, and obtains preliminary reactants. Then, after adjusting the centrifugation speed, the fourth valve control structure 134 is opened, and the preliminary reactants enter the detection reaction chamber 138. At this time, the substances in the second liquid chamber 136 can be simultaneously introduced into the detection reaction chamber 138 through the second valve control structure 132. Alternatively, the substances in the second liquid chamber 136 can be introduced into the detection reaction chamber 138 in advance through the second valve control structure 132, and then the reaction and detection are completed in the detection reaction chamber 138.

[0063] In some embodiments, the shape of the detection reaction cavity 138 is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, pear-shaped, rectangular, and regular polygonal; or, in some embodiments, the shape of the detection reaction cavity 138 is set according to the photodetector. In some embodiments, the shape of the detection reaction cavity 138 can be various shapes such as circular, square, pear-shaped, and rectangular, and its shape matches the photodetector. In some embodiments, the volume of the detection reaction cavity 138 is greater than the volume of the first liquid cavity 135, the volume of the first reaction cavity 137, and the volume of the second liquid cavity 136, that is, the volume of the detection reaction cavity 138 is greater than the volume of the second liquid cavity 136, the volume of the detection reaction cavity 138 is greater than the volume of the first liquid cavity 135, and the volume of the detection reaction cavity 138 is greater than the volume of the first reaction cavity 137. More preferably, the volume of the detection reaction cavity 138 is 1.5 to 2 times the larger of the volumes of the second liquid cavity 136 and the first liquid cavity 135. In some embodiments, the volume of the detection reaction chamber 138 is greater than or equal to the sum of the volumes of the first liquid chamber 135 and the second liquid chamber 136; in some embodiments, the volume of the detection reaction chamber 138 is greater than or equal to the sum of the volumes of the first liquid chamber 135, the first reaction chamber 137, and the second liquid chamber 136. In some embodiments, the volume of the detection reaction chamber 138 is greater than or equal to 1.5 times the volume of the first liquid chamber 135, the volume of the detection reaction chamber 138 is greater than or equal to 1.5 times the volume of the second liquid chamber 136, and the volume of the detection reaction chamber 138 is greater than or equal to 1.5 times the volume of the first reaction chamber 137. Exemplarily, in some embodiments, the volume of the detection reaction chamber 138 is greater than or equal to 1.5 to 2 times the sum of the volumes of the first liquid chamber 135 and the second liquid chamber 136; or, the volume of the detection reaction chamber 138 is greater than or equal to 1.5 to 2 times the larger of the volumes of the first liquid chamber 135 and the second liquid chamber 136. This structural design is suitable for two reactions that require precise quantification, so that the second reaction can be accurately controlled in the detection reaction chamber 138, and all the liquid will not overflow, and finally be controlled to enter the waste liquid chamber 139.

[0064] In some embodiments, the waste liquid chamber 139 is used to receive the liquid 200 discharged from the detection reaction chamber 138, and its shape can be circular, elliptical, square, rectangular, or other shapes. In some embodiments, the shape of the waste liquid chamber 139 is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, rectangular, and regular polygonal. In some embodiments, the volume of the waste liquid chamber 139 is greater than or equal to the sum of the volumes of the first liquid chamber 135 and the second liquid chamber 136. In some embodiments, the volume of the waste liquid chamber 139 is greater than or equal to the sum of the volumes of the first liquid chamber 135, the second liquid chamber 136, the first reaction chamber 137, and the detection reaction chamber 138. This structural design facilitates the concentration of all the reacted liquid in the waste liquid chamber 139, making cleaning easier after the detection is completed.

[0065] As an example, the body 110 has two opposing sides. In some embodiments, the body 110 is cylindrical with two opposing sides. In some embodiments, the first liquid chamber 135 and the first valve control structure 131 are located on one side of the body 110, and the second liquid chamber 136, the second valve control structure 132, the first reaction chamber 137, the fourth valve control structure 134, the detection reaction chamber 138, the third valve control structure 133, and the waste liquid chamber 139 are located on the other side of the body 110; as an example, the first liquid chamber 135 and the first valve control structure 131 are both located on the back side of the chip, and the second liquid chamber 136, the second valve control structure 132, the third valve control structure 133, the first reaction chamber 137, the fourth valve control structure 134, the detection reaction chamber 138, and the waste liquid chamber 139 are all located on the front side of the chip. It is understood that the front side and the back side of the chip are opposite each other, and either side can be used as the front side. Since the first liquid chamber 135 and the second liquid chamber 136 need to contain liquid 200, after the first liquid chamber 135 and / or the second liquid chamber 136 contain liquid 200, they are sealed with a thin film or sheet to prevent liquid 200 from splashing out after inversion or high-speed centrifugation. This design makes full use of the thickness of the body 110, making it particularly suitable for detection applications with small reaction liquid volumes. The rational distribution of the first liquid chamber 135, the second liquid chamber 136, the first reaction chamber 137, and the detection reaction chamber 138 significantly reduces the area occupied by the microfluidic chambers 130 on the body 110. Applied to a single-disk microfluidic chip, this allows for a greater number of microfluidic chambers 130 to be placed on a single microfluidic chip 100. Compared to the traditional single-sided distribution design, this embodiment, through a double-sided distribution design, maintains the original detection throughput without increasing the number of microfluidic chambers 130, resulting in a smaller microfluidic chip 100 and a reduction in the weight of the body 110 by approximately 13% to 19%. This not only helps to reduce the product's size and production costs but also reduces the motor load.

[0066] In some embodiments, the first liquid chamber 135, the first valve-controlled structure 131, the first reaction chamber 137, and the fourth valve-controlled structure 134 are located on one side of the body 110, while the second liquid chamber 136, the second valve-controlled structure 132, the detection reaction chamber 138, the third valve-controlled structure 133, and the waste liquid chamber 139 are located on the other side of the body 110; or, in some embodiments, the first liquid chamber 135, the first valve-controlled structure 131, the first reaction chamber 137, the fourth valve-controlled structure 134, and the detection reaction chamber 139 are located on the other side of the body 110; The first liquid chamber 138, the third valve control structure 133, and the waste liquid chamber 139 are located on one side of the main body 110, while the second liquid chamber 136 and the second valve control structure 132 are located on the other side of the main body 110. Alternatively, in some embodiments, the first valve control structure 131, the second valve control structure 132, the third valve control structure 133, the fourth valve control structure 134, the first liquid chamber 135, the second liquid chamber 136, the first reaction chamber 137, the detection reaction chamber 138, and the waste liquid chamber 139 are located on the same side of the main body 110. Other embodiments follow the same principle and will not be described in detail.

[0067] In some embodiments, an immunoassay analyzer includes a centrifuge and a microfluidic chip 100 of any embodiment, the microfluidic chip 100 being disposed on the rotating shaft of the centrifuge. Since the immunoassay analyzer employs the microfluidic chip 100 of any embodiment, it also possesses the beneficial technical effects brought about by the microfluidic chip 100 of the relevant embodiments, which will not be elaborated upon here.

[0068] As an example, an immunoassay analyzer includes a centrifuge and a microfluidic chip 100. The microfluidic chip 100 includes a body 110 with a rotation center 120 and a microfluidic chamber 130 formed therein. The microfluidic chamber 130 includes a first valve control structure 131, a second valve control structure 132, a third valve control structure 133, a fourth valve control structure 134, a first liquid chamber 135, a second liquid chamber 136, a first reaction chamber 137, a detection reaction chamber 138, and a waste liquid chamber 139. The first liquid chamber 135, the first valve control structure 131, the first reaction chamber 137, and the first reaction chamber 136 are arranged in order of increasing distance from the rotation center 120. The fourth valve control structure 134 and the detection reaction chamber 138 are sequentially connected, and the second liquid chamber 136, the second valve control structure 132, the detection reaction chamber 138, the third valve control structure 133, and the waste liquid chamber 139 are sequentially connected. The first valve control structure 131 is configured to close at a speed lower than a preset first speed; the second valve control structure 132 is configured to close at a speed lower than a preset second speed; and the third valve control structure 133 is configured to close at a speed lower than a preset third speed, so as to control the flow of liquid in the detection reaction chamber 138 to the waste liquid chamber 139. The preset first speed is not higher than the preset second speed, and the preset first speed is lower than the preset third speed. Other embodiments follow the same principle and will not be described in detail here. In this embodiment, the body 110 is disposed on the rotating shaft of the centrifuge, and the rotation center 120 is correspondingly disposed with respect to the rotating shaft. For example, the body 110 can be directly mounted on the rotating shaft or indirectly mounted on the rotating shaft through an intermediate component. The embodiments of this application do not impose additional restrictions on this. A centrifuge can be used to drive the body 110 to rotate through the rotating shaft. In this way, the various valve control structures of the microfluidic chamber 130 can be controlled by rotation.

[0069] In some embodiments, in the immunoassay analyzer, the first liquid chamber 135 and the first valve control structure 131 are located on one side of the body 110, while the second liquid chamber 136, the second valve control structure 132, the first reaction chamber 137, the fourth valve control structure 134, the detection reaction chamber 138, the third valve control structure 133, and the waste liquid chamber 139 are located on the other side of the body 110.

[0070] In some embodiments, in the immunoassay analyzer, the first liquid chamber 135, the first valve control structure 131, the first reaction chamber 137 and the fourth valve control structure 134 are located on one side of the body 110, and the second liquid chamber 136, the second valve control structure 132, the detection reaction chamber 138, the third valve control structure 133 and the waste liquid chamber 139 are located on the other side of the body 110.

[0071] In some embodiments, in the immunoassay analyzer, the first liquid chamber 135, the first valve control structure 131, the first reaction chamber 137, the fourth valve control structure 134, the detection reaction chamber 138, the third valve control structure 133 and the waste liquid chamber 139 are located on one side of the body 110, and the second liquid chamber 136 and the second valve control structure 132 are located on the other side of the body 110.

[0072] In some embodiments, in the immunoassay instrument, the first valve control structure 131, the second valve control structure 132, the third valve control structure 133, the fourth valve control structure 134, the first liquid chamber 135, the second liquid chamber 136, the first reaction chamber 137, the detection reaction chamber 138, and the waste liquid chamber 139 are located on the same side of the body 110.

[0073] In some embodiments, in the immunoassay instrument, in order of distance from the rotation center 120 from near to far, the inlet of the first valve control structure 131 is connected to the bottom of the first liquid chamber 135, the inlet of the second valve control structure 132 is connected to the bottom of the second liquid chamber 136, the inlet of the third valve control structure 133 is connected to the bottom of the detection reaction chamber 138, the inlet of the fourth valve control structure 134 is connected to the bottom of the first reaction chamber 137, the outlet of the fourth valve control structure 134 is connected to the top of the detection reaction chamber 138, and the top of the fourth valve control structure 134 is higher than the top of the first reaction chamber 137.

[0074] In some embodiments, the first valve control structure 131, the second valve control structure 132, and the third valve control structure 133 in the immunoassay analyzer are hydrophobic valves or capillary valves.

[0075] In some embodiments, the fourth valve-controlled structure 134 in the immunoassay analyzer is a siphon valve, and the surface of the fourth valve-controlled structure 134 is a hydrophilic layer.

[0076] In some embodiments, the microfluidic chip 100 in the immunoassay analyzer is a disk-type microfluidic chip.

[0077] In some embodiments, the microfluidic chip in the immunoassay analyzer includes at least one of the following: a first liquid chamber 135 and a second liquid chamber 136 are quantitative chambers for quantitatively containing liquid 200; a first reaction chamber 137 and a detection reaction chamber 138 are respectively pre-filled with reaction reagents; the volume of the detection reaction chamber 138 is greater than or equal to the sum of the volumes of the first liquid chamber 135 and the second liquid chamber 136; the volume of the waste liquid chamber 139 is greater than or equal to the sum of the volumes of the first liquid chamber 135 and the second liquid chamber 136; the first liquid chamber 135 and the second liquid chamber 136 are quantitative chambers, and the volumes of the first liquid chamber 135 and the second liquid chamber 136 are the same or different; the shape of the first reaction chamber 137 is selected from partially elliptical, partially circular, elliptical, circular, trapezoidal, pear-shaped, and rectangular. The shape of the detection reaction chamber 138 is selected from one of the following: a partial ellipse, a partial circle, an ellipse, a circle, a trapezoid, a pear shape, a rectangle, and a regular polygon; the shape of the detection reaction chamber 138 is set according to the optical detection device; the volume of the detection reaction chamber 138 is greater than the volume of the first liquid chamber 135, the volume of the first reaction chamber 137, and the volume of the second liquid chamber 136; the shape of the waste liquid chamber 139 is selected from one of the following: a partial ellipse, a partial circle, an ellipse, a circle, a trapezoid, a rectangle, and a regular polygon; the volume of the waste liquid chamber 139 is greater than or equal to the sum of the volumes of the first liquid chamber 135, the second liquid chamber 136, the first reaction chamber 137, and the detection reaction chamber 138.

[0078] It should be noted that other embodiments of this application also include microfluidic chips and immunoassay instruments formed by combining the technical features of the above embodiments.

[0079] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0080] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims.

Claims

1. A microfluidic chip, comprising a body having a center of rotation and a microfluidic chamber formed therein; The microfluidic chamber includes a first valve-controlled structure, a second valve-controlled structure, a third valve-controlled structure, a fourth valve-controlled structure, a first liquid chamber, a second liquid chamber, a first reaction chamber, a detection reaction chamber, and a waste liquid chamber; The first liquid chamber, the first valve-controlled structure, the first reaction chamber, the fourth valve-controlled structure, and the detection reaction chamber are sequentially connected in order of distance from the rotation center from near to far, and the second liquid chamber, the second valve-controlled structure, the detection reaction chamber, the third valve-controlled structure, and the waste liquid chamber are sequentially connected in order. The first valve control structure is configured to close at speeds below a preset first rotational speed; The second valve control structure is configured to close at speeds below a preset second rotational speed; The third valve control structure is configured to close at a speed lower than a preset third rotation speed to control the flow of liquid in the detection reaction chamber to the waste liquid chamber; The preset first rotational speed is not higher than the preset second rotational speed, and the preset first rotational speed is lower than the preset third rotational speed.

2. The microfluidic chip according to claim 1, wherein, The first liquid chamber and the first valve control structure are located on one side of the main body, while the second liquid chamber, the second valve control structure, the first reaction chamber, the fourth valve control structure, the detection reaction chamber, the third valve control structure, and the waste liquid chamber are located on the other side of the main body.

3. The microfluidic chip according to claim 1, wherein, The first liquid chamber, the first valve-controlled structure, the first reaction chamber, and the fourth valve-controlled structure are located on one side of the main body, while the second liquid chamber, the second valve-controlled structure, the detection reaction chamber, the third valve-controlled structure, and the waste liquid chamber are located on the other side of the main body.

4. The microfluidic chip according to claim 1, wherein, The first liquid chamber, the first valve-controlled structure, the first reaction chamber, the fourth valve-controlled structure, the detection reaction chamber, the third valve-controlled structure, and the waste liquid chamber are located on one side of the main body, while the second liquid chamber and the second valve-controlled structure are located on the other side of the main body.

5. The microfluidic chip according to claim 1, wherein, The first valve control structure, the second valve control structure, the third valve control structure, the fourth valve control structure, the first liquid chamber, the second liquid chamber, the first reaction chamber, the detection reaction chamber, and the waste liquid chamber are all located on the same side of the main body.

6. The microfluidic chip according to claim 1, wherein, In order of increasing distance from the rotation center, the inlet of the first valve-controlled structure is connected to the bottom of the first liquid chamber, the inlet of the second valve-controlled structure is connected to the bottom of the second liquid chamber, the inlet of the third valve-controlled structure is connected to the bottom of the detection reaction chamber, the inlet of the fourth valve-controlled structure is connected to the bottom of the first reaction chamber, and the outlet of the fourth valve-controlled structure is connected to the top of the detection reaction chamber, with the top of the fourth valve-controlled structure being higher than the top of the first reaction chamber.

7. The microfluidic chip according to claim 1, wherein, The first valve control structure, the second valve control structure, and the third valve control structure are either steam traps or capillary valves.

8. The microfluidic chip according to claim 1, wherein, The fourth valve-controlled structure is a siphon valve, and the surface of the fourth valve-controlled structure is a hydrophilic layer.

9. The microfluidic chip according to claim 1, wherein, The microfluidic chip includes at least one of the following: The first liquid chamber and the second liquid chamber are metering chambers used to meterly contain liquid; The first reaction chamber and the detection reaction chamber are respectively pre-filled with reaction reagents; The volume of the detection reaction chamber is greater than or equal to the sum of the volumes of the first liquid chamber and the second liquid chamber; The volume of the waste liquid chamber is greater than or equal to the sum of the volumes of the first liquid chamber and the second liquid chamber; The first liquid chamber and the second liquid chamber are metering chambers, and the first liquid chamber and the second liquid chamber are configured to have the same or different volumes; The shape of the first reaction chamber is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, pear-shaped, rectangular, and regular polygonal. The volume of the first reaction chamber is greater than or equal to the volume of the first liquid chamber; The shape of the detection reaction chamber is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, pear-shaped, rectangular, and regular polygonal. The shape of the detection reaction cavity is set according to the optical detection device; The volume of the detection reaction chamber is greater than the volume of the first liquid chamber, the volume of the first reaction chamber, and the volume of the second liquid chamber. The shape of the waste liquid chamber is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, rectangular, and regular polygonal. The volume of the waste liquid chamber is greater than or equal to the sum of the volumes of the first liquid chamber, the second liquid chamber, the first reaction chamber, and the detection reaction chamber.

10. The microfluidic chip according to claim 1, wherein, The microfluidic chip is a disk-type microfluidic chip.

11. An immunoassay instrument, wherein, The invention includes a centrifuge and a microfluidic chip, wherein the microfluidic chip is disposed on the rotating shaft of the centrifuge; The microfluidic chip includes a body with a center of rotation and a microfluidic chamber formed in the body; The microfluidic chamber includes a first valve-controlled structure, a second valve-controlled structure, a third valve-controlled structure, a fourth valve-controlled structure, a first liquid chamber, a second liquid chamber, a first reaction chamber, a detection reaction chamber, and a waste liquid chamber; The first liquid chamber, the first valve-controlled structure, the first reaction chamber, the fourth valve-controlled structure, and the detection reaction chamber are sequentially connected in order of distance from the rotation center from near to far, and the second liquid chamber, the second valve-controlled structure, the detection reaction chamber, the third valve-controlled structure, and the waste liquid chamber are sequentially connected in order. The first valve control structure is configured to close at speeds below a preset first rotational speed; The second valve control structure is configured to close at speeds below a preset second rotational speed; The third valve control structure is configured to close at a speed lower than a preset third rotation speed to control the flow of liquid in the detection reaction chamber to the waste liquid chamber; The preset first rotational speed is not higher than the preset second rotational speed, and the preset first rotational speed is lower than the preset third rotational speed.

12. The immunoassay instrument according to claim 11, wherein, The first liquid chamber and the first valve control structure are located on one side of the main body, while the second liquid chamber, the second valve control structure, the first reaction chamber, the fourth valve control structure, the detection reaction chamber, the third valve control structure, and the waste liquid chamber are located on the other side of the main body.

13. The immunoassay instrument according to claim 11, wherein, The first liquid chamber, the first valve-controlled structure, the first reaction chamber, and the fourth valve-controlled structure are located on one side of the main body, while the second liquid chamber, the second valve-controlled structure, the detection reaction chamber, the third valve-controlled structure, and the waste liquid chamber are located on the other side of the main body.

14. The immunoassay instrument according to claim 11, wherein, The first liquid chamber, the first valve-controlled structure, the first reaction chamber, the fourth valve-controlled structure, the detection reaction chamber, the third valve-controlled structure, and the waste liquid chamber are located on one side of the main body, while the second liquid chamber and the second valve-controlled structure are located on the other side of the main body.

15. The immunoassay instrument according to claim 11, wherein, The first valve control structure, the second valve control structure, the third valve control structure, the fourth valve control structure, the first liquid chamber, the second liquid chamber, the first reaction chamber, the detection reaction chamber, and the waste liquid chamber are all located on the same side of the main body.

16. The immunoassay instrument according to claim 11, wherein, In order of increasing distance from the rotation center, the inlet of the first valve-controlled structure is connected to the bottom of the first liquid chamber, the inlet of the second valve-controlled structure is connected to the bottom of the second liquid chamber, the inlet of the third valve-controlled structure is connected to the bottom of the detection reaction chamber, the inlet of the fourth valve-controlled structure is connected to the bottom of the first reaction chamber, and the outlet of the fourth valve-controlled structure is connected to the top of the detection reaction chamber, with the top of the fourth valve-controlled structure being higher than the top of the first reaction chamber.

17. The immunoassay instrument according to claim 11, wherein, The first valve control structure, the second valve control structure, and the third valve control structure are either steam traps or capillary valves.

18. The immunoassay instrument according to claim 11, wherein, The fourth valve-controlled structure is a siphon valve, and the surface of the fourth valve-controlled structure is a hydrophilic layer.

19. The immunoassay instrument according to claim 11, wherein, The microfluidic chip includes at least one of the following: The first liquid chamber and the second liquid chamber are metering chambers used to meterly contain liquid; The first reaction chamber and the detection reaction chamber are respectively pre-filled with reaction reagents; The volume of the detection reaction chamber is greater than or equal to the sum of the volumes of the first liquid chamber and the second liquid chamber; The volume of the waste liquid chamber is greater than or equal to the sum of the volumes of the first liquid chamber and the second liquid chamber; The first liquid chamber and the second liquid chamber are metering chambers, and the first liquid chamber and the second liquid chamber are configured to have the same or different volumes; The shape of the first reaction chamber is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, pear-shaped, rectangular, and regular polygonal. The volume of the first reaction chamber is greater than or equal to the volume of the first liquid chamber; The shape of the detection reaction chamber is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, pear-shaped, rectangular, and regular polygonal. The shape of the detection reaction cavity is set according to the optical detection device; The volume of the detection reaction chamber is greater than the volume of the first liquid chamber, the volume of the first reaction chamber, and the volume of the second liquid chamber. The shape of the waste liquid chamber is selected from one of the following: partially elliptical, partially circular, elliptical, circular, trapezoidal, rectangular, and regular polygonal. The volume of the waste liquid chamber is greater than or equal to the sum of the volumes of the first liquid chamber, the second liquid chamber, the first reaction chamber, and the detection reaction chamber.

20. The immunoassay instrument according to claim 11, wherein, The microfluidic chip is a disk-type microfluidic chip.

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