Centrifugal multi-stage reaction device and operation method therefor

Abstract

Provided are a centrifugal multi-stage reaction device and an operation method therefor. The centrifugal multi-stage reaction device mainly consists of a reaction vessel (5) and an inlet flow channel (3). The reaction vessel (5) comprises at least one inclined portion (55) and at least one reagent storage area (500), and the at least one reagent storage area (500) is arranged in the at least one inclined portion (55) of the reaction vessel (5). The at least one reagent storage area (500) stores a reagent (4). A plurality of reagent storage areas (500) of the reaction vessel (5) can independently store different lyophilized reagent beads. By means of adjusting and controlling the centrifugal speed and direction, different stages of reactions of a sample with a plurality of reagents can be achieved in the same reaction vessel (5).

Description

Centrifugal multi-stage reaction device and operation method thereof

[0001] This application claims priority to Chinese patent application No. 202311422470.2, filed on October 30, 2023, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present disclosure relates to the field of centrifugal microfluidics for biomedical testing, and more particularly to a centrifugal multi-stage reaction device and an operating method thereof. Background Art

[0003] In modern society, the fast pace of life leads to high stress and poor eating habits, increasing the risk of developing diseases and chronic illnesses. This trend is driving a surge in demand for biomedical testing for health checks and disease diagnosis. Improving the complex procedures of traditional testing and reducing the time required for data processing have become significant issues. Advances in testing technology have led to the integration of microelectromechanical systems (MEMS) and biomedical testing, gradually replacing the inconveniences of traditional testing with novel technologies.

[0004] Microfluidics is a recently emerging biomedical testing technology. It primarily involves microfluidic channels placed on a centrifugal disk. Because microfluidics are micrometer-scale channels within the laminar flow range, centrifugation allows for precise control of fluid direction and volume. Advantages of this technology include requiring only a small sample volume, which reduces reagent usage. Its low energy consumption and ease of quantification allow for reduced power consumption during testing and the ability to obtain large amounts of data in a short period of time.

[0005] The most common application of microfluidics is lab-on-a-disk (LOAD), which generally refers to a technology that can integrate multiple chemical and biological analysis functions on a single disk to process very small amounts of liquid. It is also called a micro total analysis system (μTAS). Through this technology, micron-level channels can be created to manipulate micron-scale substances. The main advantages of micro total analysis systems are: (1) low cost; (2) ability to perform multiple tests in parallel; (3) reduction in human error; (4) ease of use and fast analysis speed; (5) testing can be performed using a small amount of test sample.

[0006] Prior art microfluidic disks utilizing microfluidics can be found in Chinese Patent Publication No. CN107206334B. FIG12 of this patent discloses a device comprising a fluid module 10 in the form of a rotating body (see FIG11 of the present invention for details). The device primarily comprises a base 12 and a top cover 14, which are circular in shape and have a central opening. The rotating body 10 can be attached to a rotating member 18 of a drive device 20 via a conventional fixing mechanism 16. The rotating member 18 is supported for rotation on a stationary member 22 of the drive device 20. The drive device 20 can be a conventional centrifuge with an adjustable speed, or a CD or DVD drive. Furthermore, a control mechanism 24 can be provided, configured to control the drive device 20 so that the rotating body 10 rotates once or multiple times at varying rotational frequencies. While the art generally employs the aforementioned method to control the speed and frequency of the microfluidic disk, the chamber type or number of the microchannels provided on the microfluidic disk can be modified or altered to meet specific usage requirements.

[0007] Even though microfluidics technology has the above-mentioned excellent characteristics, it still faces certain difficulties in different usage requirements and needs to be improved. Existing technologies already include a variety of miniature full analysis systems designed for specific purposes, for example, separating specific targets from blood samples, testing drug effects, and other tests. In addition, there are also designs that integrate multiple reagents into the same microfluidic system. In detail, multiple tanks for storing reagents are connected to the main tank in a serial manner through capillaries. Although this can effectively reduce the time-consuming and complicated process of injecting multiple reagents in the past, the system is designed with a fixed flow channel and cannot be changed arbitrarily. In practice, common problems include the reagent flowing to the next reaction reagent tank before it is completely dissolved or the inability to freely adjust the reaction order of the reagents, resulting in a lack of flexibility in use. Therefore, the above-mentioned problems need to be further improved to solve them.

[0008] Summary of the Invention

[0009] In view of the problems faced by the related art, the present disclosure provides a centrifugal multi-stage reaction device and its operating method, which are mainly applied to centrifugal microfluidics technology in biomedical testing. The design of integrating multiple reagent storage locations into a reaction tank can effectively improve the efficiency and integrity of multiple reactions and effectively enhance the flexibility of adjusting the reagent processing sequence during the reaction.

[0010] As described above, the present invention provides a centrifugal multi-stage reaction device, comprising: a reaction tank body, the reaction tank body comprising at least one inclined portion and at least one reagent storage position; and an inlet flow channel connected to the reaction tank body; wherein the at least one reagent storage position is arranged on the at least one inclined portion of the reaction tank body; the at least one reagent storage position stores a reagent.

[0011] In one preferred embodiment, when the reaction tank comprises two reagent storage locations, a first reagent storage location is located at the bottom of the at least one inclined portion, and a second reagent storage location is located at the other end of the at least one inclined portion.

[0012] The present invention further provides an operating method for a centrifugal multi-stage reaction device, comprising the following steps: (A) providing a centrifugal multi-stage reaction device as described in the preceding paragraph; (B) centrifuging a sample through an inlet flow channel into a reaction tank body by a power module; (C) after the sample enters a first reagent storage position, the sample reacts with a first reagent in the first reagent storage position; (D) rapidly changing the rotation direction of the power module so that the sample is forced to flow upward along at least one inclined portion until it enters a second reagent storage position; (E) after the sample enters the second reagent storage position, the sample reacts with a second reagent in the second reagent storage position; and (F) continuing centrifugation of the power module to return the sample to the first reagent storage position.

[0013] In another embodiment, when the reaction tank comprises three reagent storage locations, a first reagent storage is located at the bottom of the inclined portion, a third reagent storage is located at the other end of the inclined portion, and a second reagent storage is located in the center of the inclined portion.

[0014] The present invention further provides an operating method of a centrifugal multi-stage reaction device, comprising the following steps: (a) providing a centrifugal multi-stage reaction device as described in the preceding paragraph; (b) centrifugally rotating a power module to a certain speed, causing a sample to enter a reaction tank through an inlet flow channel; (c) after the sample enters a first reagent storage position, the sample reacts with a first reagent in the first reagent storage position; (d) rapidly changing the rotation direction of the power module at a first acceleration, causing the sample to be forced to flow upward along at least one inclined portion until it enters a second reagent storage position; (e) the sample enters the After entering the second reagent storage position, the sample reacts with a second reagent in the second reagent storage position; (f) the power module continues to centrifuge to return the sample to the first reagent storage position and then stops rotating; (g) after the power module rotates to the rotation speed, the rotation direction is rapidly changed at a second acceleration, so that the sample is forced to flow upward along the at least one inclined portion until it enters a third reagent storage position; (h) after the sample enters the third reagent storage position, the sample reacts with a third reagent in the third reagent storage position; and (i) the power module continues to centrifuge to return the sample to the first reagent storage position.

[0015] In yet another embodiment, when the at least one inclined portion is provided as two, the at least one inclined portion comprises a first inclined portion and a second inclined portion connected at one end thereof, and the plurality of storage positions are arranged on the two inclined portions at a predetermined interval. In addition, the plurality of storage positions comprises five reagent storage positions, a first reagent storage position provided at the bottom end where the first and second inclined portions are connected, a second reagent storage position provided at the center of the first inclined portion, a third reagent storage position provided at the center of the second inclined portion, a fourth reagent storage position provided at the other end of the first inclined portion, and a fifth reagent storage position provided at the other end of the second inclined portion.

[0016] In other possible embodiments, the present invention further provides an operating method of a centrifugal multi-stage reaction device, comprising the following steps: (S01) providing a centrifugal multi-stage reaction device as described in the previous paragraph; (S02) centrifugally rotating a power module to a rotation speed, causing a sample to enter a reaction tank from an inlet flow channel; (S03) after the sample enters a first reagent storage position, the sample reacts with a first reagent in the first reagent storage position; (S04) the power module rapidly changes the rotation direction at a first acceleration, causing the sample to be forced to flow upward along a first inclined portion until it enters a second reagent storage position; (S05) after the sample enters the second reagent storage position, the sample reacts with a second reagent in the second reagent storage position; (S06) the power module continues to centrifuge to return the sample to the first reagent storage position and then stops rotating; (S07) the power module rapidly changes the rotation direction at a first acceleration, causing the sample to be forced to flow upward along a second inclined portion until it enters a third reagent storage position; (S08) the sample After entering the third reagent storage position, the sample reacts with a third reagent in the third reagent storage position; (S09) the power module continues to centrifuge to return the sample to the first reagent storage position and then stops rotating; (S10) after the power module rotates to the rotation speed, the rotation direction is quickly changed with a second acceleration, so that the sample is forced to flow upward along the first inclined portion until it enters a fourth reagent storage position; (S11) after the sample enters the fourth reagent storage position, the sample reacts with a fourth reagent in the fourth reagent storage position. (S12) the power module continues to centrifuge to return the sample to the first reagent storage position and then stops rotating; (S13) after the power module rotates to the rotation speed, the rotation direction is quickly changed with a second acceleration, so that the sample is forced to flow upward along the second inclined portion until it enters a fifth reagent storage position; (S14) after the sample enters the fifth reagent storage position, the sample reacts with a fifth reagent in the fifth reagent storage position; and (S15) the power module continues to centrifuge to return the sample to the first reagent storage position.

[0017] In other possible embodiments, the order of subsequent reagent reactions after the second reagent reaction can be adjusted as needed, as long as centrifugal balance is maintained. The user can select the reagent storage location to be reacted by adjusting the rotation direction and acceleration during centrifugation. Furthermore, the number of reagent storage locations (reagents) and the number of inclined portions can also be adjusted as needed and are not limited to the above.

[0018] The above brief description of the present invention is intended to provide a basic overview of several aspects and technical features of the present invention. This summary is not a detailed description of the present invention, and therefore does not specifically enumerate the key or important elements of the present invention, nor does it serve to define the scope of the present invention. It simply presents several concepts of the present invention in a concise manner. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] FIG1 is a schematic diagram of a centrifugal multi-stage reaction apparatus according to embodiment 1 of the present invention.

[0020] FIG2 is a schematic diagram of a centrifugal multi-stage reaction device according to embodiment 2 of the present invention.

[0021] FIG3 is a schematic diagram of a centrifugal multi-stage reaction device according to embodiment 3 of the present invention.

[0022] FIG4 is a schematic diagram of a centrifugal multi-stage reaction apparatus according to embodiment 4 of the present invention.

[0023] FIG5 is a schematic diagram showing the steps of the operation method of the centrifugal multi-stage reaction device according to Example 1 of the present invention.

[0024] FIG6 is a schematic diagram showing the steps of the operation method of the centrifugal multi-stage reaction device according to the third embodiment of the present invention.

[0025] FIG. 7 is a schematic diagram showing the steps of the operation method of the centrifugal multi-stage reaction device according to Example 4 of the present invention.

[0026] FIG8 is a flow chart showing the steps of the operation method of the centrifugal multi-stage reaction device according to the present invention.

[0027] FIG9 is a flow chart showing the steps of the operation method of the centrifugal multi-stage reaction device according to the third embodiment of the present invention.

[0028] FIG10 is a flow chart showing the steps of the operation method of the centrifugal multi-stage reaction apparatus according to the fourth embodiment of the present invention.

[0029] FIG. 11 is a schematic diagram of a microfluidic disk and its power module using microfluidic technology according to the prior art of the present invention. DETAILED DESCRIPTION

[0030] In order to understand the technical features and practical effects of the present disclosure and to implement them according to the contents of the specification, the embodiments shown in the accompanying drawings are further described in detail below.

[0031] In order to solve the above technical problems, some embodiments of the present disclosure provide a preferred embodiment, please refer to Figure 1, Figure 1 is a centrifugal multi-stage reaction device 1, which mainly includes a reaction tank body 5 and an inlet flow channel 3 connected to the reaction tank body 5. Among them, the reaction tank body 5 further includes an inclined portion 55 and a reagent storage position 500. The reagent storage position 500 is arranged on the inclined portion 55 included in the reaction tank body 5, and each reagent storage position 500 stores a reagent 4. Further, the types of the reaction tank body 5 include triangular chambers, elongated channels, U-shaped channels or combinations thereof. The embodiment of the reaction tank body 5 shown in Figure 1 is a triangular chamber; the embodiment of the reaction tank body 5 shown in Figure 2 is an elongated channel or a U-shaped channel.

[0032] For detailed structures in the reaction tank body 5, please refer to Figures 1, 2, 3 and 4, which will be described in detail in the following paragraphs. Looking at Figures 1 to 4, a plurality of reagent storage positions 500 are provided in the reaction tank body 5, and the plurality of reagent storage positions 500 can be arranged on the inclined portion 55 at a certain interval. For example, Figure 1 shows embodiment 1 of the present invention, which is a triangular reaction tank body 5 with two reagent storage positions 500. The two reagent storage positions 500 are each provided with a reagent 4, wherein the first reagent storage position 500a is provided at the bottom end of the inclined portion 55. The second reagent storage position 500b is provided at the other end relative to the inclined portion 55. In addition, the inlet flow channel 3 is connected to one short side of the triangular tank body so that the sample can flow directly into the first reagent storage position 500a located at the bottom of the inclined portion 55 after flowing into the inlet flow channel 3.

[0033] In Example 2, as shown in FIG2 , the reaction vessel 5 having two reagent storage locations 500 may also be a U-shaped flow channel, with the first reagent storage location 500a located in the concave portion of the U-shape, and the second reagent storage location 500b located at the end of the U-shape. Similar to Example 1 shown in FIG1 , the inlet flow channel 3 is connected to the other end of the U-shaped flow channel opposite the end where the second reagent storage location 500b is located, so that the sample flowing through the inlet flow channel 3 can flow directly into the first reagent storage location 500a located in the concave portion of the U-shape.

[0034] In Example 3, as shown in FIG3 , a triangular reaction cell 5 is shown with three reagent storage locations 500. The three reagent storage locations 500 are spaced apart on an inclined portion 55 at regular intervals. The first reagent storage location 500a is located at the bottom of the triangle, the second reagent storage location 500b is located in the center of the inclined portion 55, and the third reagent storage location 500c is located at the top of the inclined portion 55. The three reagent storage locations 500 can each accommodate three reagents 4. The positions of the reagent storage locations 500 can be adjusted as needed based on the reaction sequence between the sample and each reagent 4. Similar to the previous two embodiments, an inlet channel 3 is connected to one short side of the triangular cell, allowing the sample to flow directly into the first reagent storage location 500a at the bottom of the inclined portion 55 after entering the inlet channel 3.

[0035] Embodiment 4 of the present invention, as shown in FIG4 , illustrates an isosceles triangle-shaped reaction cell 5 comprising five reagent storage locations 500. In Embodiment 4, the reaction cell 5 comprises two inclined portions 55 , a first inclined portion 55 a and a second inclined portion 55 b connected at one end thereof and symmetrically disposed on either side of the isosceles triangle-shaped reaction cell 5 . The five reagent storage locations 500 are symmetrically arranged within the cell and spaced a certain distance apart. Specifically, the five reagent storage locations 500 are arranged within the reaction vessel 5 as follows: a first reagent storage location 500a is located at the intersection of the first inclined portion 55a and the second inclined portion 55b (i.e., the bottom end of the isosceles triangle-shaped reaction vessel 5); a second reagent storage location 500b is located at the center of the first inclined portion 55a; a third reagent storage location 500c is located at the center of the second inclined portion 55b; a fourth reagent storage location 500d is located at the other end of the first inclined portion 55a; and a fifth reagent storage location 500e is located at the other end of the second inclined portion 55b. Furthermore, the inlet channel 3 of this embodiment is positioned so that the sample flows directly into the first reagent storage location 500a.

[0036] In the aforementioned embodiment, the multiple reagent storage locations 500 can store different reagents 4, and the reaction reagents 4 can be in the form of volatile reagents, freeze-dried reagents, freeze-dried reagent microspheres, or a combination thereof. As long as the reagents 4 are stored in a manner that does not alter the enzyme activity, all are included in this embodiment. Specifically, volatile reagents are solid or colloidal reagents obtained through natural volatilization or other methods. Furthermore, freeze-dried reagent microsphere technology can convert chemical reagents that are unstable at room temperature into a high-quality, stable, and quantitative form for long-term storage at room temperature. The solid spheres after freeze-drying have a loose network structure and are quickly redissolved, thereby improving the stability of the reaction reagents 4 stored in the multiple reagent storage locations 500 of the centrifugal multi-stage reaction device 1 of the present invention in microfluidic reactions.

[0037] In summary, the internal designs of the reaction tank 5 described in the four embodiments of the centrifugal multi-stage reaction apparatus 1 are all mounted on the rotating body 10 shown in Figure 11 , and the rotational frequency and direction of the power module 20' (i.e., the drive device 20 of the related art) are adjusted by the control mechanism 24. Whether it is the number or position of the reagent storage positions 500 or the number or position of the inclined portions 55, they can be adjusted accordingly based on the reaction objectives or requirements of the sample. The arrangement is not limited to the above embodiments; it only needs to ensure that the centrifugation process can maintain the balance of the entire apparatus and maintain smooth flow during different rotational directions.

[0038] Unlike prior art techniques in which multiple reagent storage locations 500 are often fixed or serially connected to the sample tank or the external tank body of the reaction tank, the order and position of the sample and each reagent 4 reaction stage must be preset in advance and the flow direction is single. As a result, the reagents 4 in each reaction stage must be injected one by one in advance and the order cannot be freely adjusted, or there is a possibility that the reagents 4 will not react completely. To solve the above problems, the centrifugal multi-stage reaction device 1 of the present invention is based on the problems of the prior art and improves it. The sample can be simultaneously placed in a single reaction tank body 5 with multiple reagents 4, and the reaction order of these reagents 4 can be adjusted in time. If one of the reagents 4 does not react completely, it can be adjusted in time through multiple centrifugation, rotation or speed change.

[0039] The present invention further provides an operating method for implementing the centrifugal multi-stage reaction device 1. A preferred embodiment thereof is shown in FIG5 and FIG8. The operating method for the centrifugal multi-stage reaction device 1 comprises the following steps:

[0040] In step (A), a centrifugal multi-stage reaction apparatus 1 as described in Example 1 or Example 2 is provided, as shown in Figures 1, 2, and 5(I). In step (B), the centrifugal multi-stage reaction apparatus 1 is centrifuged by the power module 20, allowing the sample to enter the reaction tank 5 through the inlet flow channel 3 (see Figure 5(II)). The centrifugal speed in step (B) is sufficient to allow the sample to flow into the reaction tank 5. After the sample enters the first reagent storage position 500a in step (C), the sample will flow directly into the first reagent storage position 500a and react with the first reagent 4a (see the grid pattern in Figure 5(II)). In step (D), the power module 20 rapidly changes its rotational direction, causing the sample to flow upward along the inclined portion 55 under the Euler force until it enters the second reagent storage position 500b (see the triangular patterns in Figures 5 (III) and (IV)). Then, according to step (E), after the sample enters the second reagent storage position 500b, it reacts with the second reagent 4b in the second reagent storage position 500b (see Figures 5 (III) and (IV)). After the reaction, in step (F), the power module 20 continues centrifugation to return the sample to the first reagent storage position 500a. At this stage, as shown in Figure 5 (IV), the liquid at the bottom of the reaction tank 5 is a mixed solution 4a' / 4b' formed after the sample reacts with the first reagent 4a and the second reagent 4b.

[0041] The present invention further provides an operating method for implementing the centrifugal multi-stage reaction device 1. A preferred embodiment thereof is shown in FIG6 and FIG9. The operating method for the centrifugal multi-stage reaction device 1 comprises the following steps:

[0042] In step (a), a centrifugal multi-stage reaction device 1 as described in the aforementioned embodiment 3 is provided, as shown in FIG3 and FIG6(I) for details. In step (b), the power module 20 is centrifugally rotated to a certain speed, and the sample is allowed to enter the reaction tank body 5 from the inlet flow channel 3. The speed only needs to enable the sample to overcome the viscosity or friction force and enter the reaction tank body 5, and the present invention is not limited thereto. In step (c), the sample flows into the reaction tank body 5 and enters the first reagent storage position 500a at the bottom of the reaction tank body 5 by inertia, so that the sample reacts with the first reagent 4a in the first reagent storage position 500a. The liquid at the bottom of the tank body as shown in FIG6(II) is a mixture 4a' of the sample and the first reagent 4a.

[0043] In step (d), the power module 20 is used to quickly change the direction of rotation at a first acceleration, so that the sample is subjected to the Euler force and flows upward along the inclined portion 55 until it enters a second reagent storage position 500b. The liquid at the bottom of the tank body as shown in Figure 6 (III) moves upward along the inclined portion 55 to the second reagent storage position 500b. In step (e), after the sample enters the second reagent storage position 500b, the sample reacts with the second reagent 4b in the second reagent storage position 500b. In step (f), the power module 20 continues centrifugation to return the sample to the first reagent storage position 500a and then stops rotating. As shown in Figure 6 (IV), the liquid at the bottom of the tank body is the mixed liquid 4a' / 4b' after the sample reacts with the first reagent 4a and the second reagent.

[0044] In step (g), after the power module 20 rotates to the specified rotation speed, the rotation direction is rapidly changed at a second acceleration, causing the sample to flow upward along the inclined portion 55 under the Euler force until it enters the third reagent storage position 500c. As shown in Figure 6(V), the aforementioned mixed liquid flows upward along the inclined portion 55 at a second acceleration greater than the first acceleration, so the overall path length of the fluid is greater than that shown in Figure 6(III). In step (h), after the sample enters the third reagent storage position 500c, the sample reacts with the third reagent 4c in the third reagent storage position 500c, and then step (i) is continued. The power module 20 is centrifuged continuously to return the sample to the first reagent storage position 500a. As shown in Figure 6(VI), the liquid at the bottom of the tank at this stage is a mixed liquid 4a' / 4b' / 4c' formed after the sample reacts with the first reagent 4a, the second reagent 4b, and the third reagent 4c.

[0045] The present invention further provides an operating method for implementing the centrifugal multi-stage reaction device 1. A preferred embodiment thereof is shown in FIG7 and FIG10. The operating method for the centrifugal multi-stage reaction device 1 comprises the following steps:

[0046] In step (S01), a centrifugal multi-stage reaction device 1 as described in Example 4 is provided, as shown in Figures 4 and 7(I) for details. In step (S02), the power module 20 is centrifugally rotated to a certain speed, and the sample is allowed to enter the reaction tank body 5 from the inlet flow channel 3. The speed only needs to enable the sample to overcome viscosity or friction and enter the reaction tank body 5, and the present invention is not limited thereto. In step (S03), the sample flows into the reaction tank body 5 and enters the first reagent storage position 500a located at the bottom of the reaction tank body 5 by inertia, so that the sample reacts with the first reagent 4a in the first reagent storage position 500a. The liquid at the bottom of the tank body, as shown in Figure 7(II), is the mixed liquid 4a' after the sample and the first reagent 4a react.

[0047] In step (S04), the power module 20 is used to quickly change the direction of rotation at a first acceleration, so that the sample is subjected to the Euler force and flows upward along the first inclined portion 55a until it enters a second reagent storage position 500b. The liquid at the bottom of the tank body as shown in Figure 7 (III) moves upward along the first inclined portion 55a to the second reagent storage position 500b. After the sample enters the second reagent storage position 500b, the sample reacts with the second reagent 4b in the second reagent storage position 500b. In step (S06), the power module 20 continues to centrifuge to return the sample to the first reagent storage position 500a and then stops rotating. As shown in Figure 7 (IV), the liquid at the bottom of the tank body is the mixed liquid 4a' / 4b' after the sample reacts with the first reagent 4a and the second reagent 4b.

[0048] Step (S07) rotates the power module 20 to the rotation speed set forth in step (S02) and then rapidly changes the direction of rotation with a first acceleration, causing the sample to flow upward along the second inclined portion 55b under the Euler force until it enters the third reagent storage position 500c. As shown in FIG7(V), the third reagent storage position 500c of the second inclined portion 55b corresponds to the second reagent storage position 500b of the first inclined portion 55a. Therefore, the acceleration implemented in this step is the same as that in step (S04), with centrifugation being performed with the same first acceleration. After the sample enters the third reagent storage position 500c, the sample reacts with the third reagent 4c in the third reagent storage position 500c, and step (S09) is subsequently performed. The power module 20 continues centrifugation to return the sample to the first reagent storage position 500a and then stops rotating. As shown in FIG. 7( VI ), the liquid at the bottom of the tank at this stage is a mixed solution 4 a ′ / 4 b ′ / 4 c ′ formed by the reaction of the sample with the first reagent 4 a , the second reagent 4 b , and the third reagent 4 c .

[0049] In step (S10), after the power module 20 rotates to the rotation speed set in the aforementioned step (S02), the rotation direction is rapidly changed at a second acceleration, so that the sample is forced to flow upward along the first inclined portion 55a until it enters the fourth reagent storage position 500d. As shown in Figure 7 (VII), the aforementioned mixed liquid flows upward along the first inclined portion 55a at a second acceleration greater than the first acceleration, so that the overall path length of the fluid is greater than that shown in Figures 7 (III) and 7 (V). In step (S11), after the sample enters the fourth reagent storage position 500d, the sample reacts with the fourth reagent 4d in the fourth reagent storage position 500d, and then the power module 20 continues to centrifuge in step (S12), so that the sample returns to the first reagent storage position 500a and stops rotating. As shown in FIG. 7 (VIII), the liquid at the bottom of the tank at this stage is a mixed solution 4a' / 4b' / 4c' / 4d' formed after the sample reacts with the first reagent 4a, the second reagent 4b, the third reagent 4c and the fourth reagent 4d.

[0050] In step (S13), after the power module 20 rotates to the rotation speed described in the aforementioned step (S02), the rotation direction is quickly changed with a second acceleration, so that the sample is forced to flow upward along the second inclined portion 55b until it enters the fifth reagent storage position 500e. As shown in Figure 7 (IX), the aforementioned mixed liquid flows upward along the second inclined portion 55b with a second acceleration greater than the first acceleration, so that the overall path length of the fluid is greater than that shown in Figures 7 (III) and 7 (V). After the sample enters the fifth reagent storage position 500e in step (S14), the sample reacts with the fifth reagent 4e in the fifth reagent storage position 500e. Finally, in step (S15), the power module 20 is continuously centrifuged to return the sample to the first reagent storage position 500a. At this stage, as shown in FIG. 7(X), the liquid at the bottom of the reaction tank 5 is a mixture 4a / 4b / 4c / 4d / 4e formed by the reaction of the sample with the first reagent 4a, the second reagent 4b, the third reagent 4c, the fourth reagent 4d and the fifth reagent 4e.

[0051] In summary, the embodiment of the operation method of the above-mentioned centrifugal multi-stage reaction device 1 can be classified into five major processes: the first reagent 4a reaction process from step (S02) to step (S03), the second reagent reaction process from step (S04) to step (S06), the third reagent reaction process from step (S07) to step (S09), the fourth reagent reaction process from step (S10) to step (S12), and the fifth reagent reaction process from step (S13) to step (S15). Overall, each reagent reaction process includes three main actions: "acceleration", "reaction of the liquid flowing to the reagent storage position at the destination", and "rotation to return the liquid to the first reagent storage position 500a". The final action will return the liquid to its original position. Given the characteristics of the present invention, different reagent 4 reactions are carried out in the same reaction tank body 5, and the entire process can be classified into different stages of reagent 4 reaction processes based on needs, such as the number or position of the reagent storage positions 500 set in the reaction tank body 5 and the number or position of the inclined portion 55. In this way, since each reagent 4 reaction process ends by returning to the first reagent 4a reaction storage position, the reaction order of the reagent 4 reaction process at different stages can be adjusted according to the purpose of use. Moreover, if the previous reaction is incomplete or the reagent 4 remains in the reagent storage position 500, the user can freely adjust the centrifugal direction and acceleration to control the back and forth vibration of the microfluidic plate so that the incompletely reacted reagent 4 can eventually react completely.

[0052] Therefore, based on the reaction principle described above, the embodiment of the centrifugal multi-stage reaction apparatus 1, steps (S01) to (S15), can, after the second reagent reaction process (step (S06)) is completed, select to first perform the fourth reagent reaction process (steps (S10) to (S12)) and then continue with the third reagent reaction process (steps (S07) to (S09)), and finally complete the fifth reagent reaction process (steps (S13) to (S15)). Alternatively, after the third reagent reaction process (step (S09)) is completed, select to first perform the fifth reagent reaction process (steps (S13) to (S15)) and then continue with the fourth reagent reaction process (steps (S10) to (S12)).

[0053] In summary, the centrifugal multi-stage reaction device 1 of the present invention is different from the prior art, which usually has multiple reaction reagent tanks connected or connected in series to the main flow channel in the form of independent tank bodies, so that the sample flows through the main flow channel and flows into each independent reaction reagent tank body in sequence in a fixed reaction order. Since the overall reaction process sequence cannot be freely adjusted, the reagents in each stage of the reaction must be injected one by one in advance according to the reaction sequence with the sample, or there is a possibility that the reaction reagent 4 will not be completely reacted and cannot be refluxed for further reaction. Such difficulties as the above are improved based on the problems of the prior art by the centrifugal multi-stage reaction device 1 of the present invention, so that the sample can be placed in multiple reagent storage positions 500 in a single reaction tank body 5 at the same time, and the reaction order of the reagents 4 can be adjusted in time. If one of the reagents 4 does not react completely, it can be adjusted in time through multiple centrifugation, steering or speed change.

[0054] However, the above is only a preferred embodiment of the present invention and should not be used to limit the scope of implementation of the present invention. Simple changes and modifications made according to the scope of the patent application and the description of the present invention are still within the scope of the present invention.

Claims

1. A centrifugal multi-stage reaction device, comprising: A reaction tank body, the reaction tank body comprising: at least one inclined portion and at least one reagent storage position; and An inlet flow channel connected to the reaction tank body; Wherein, the at least one reagent storage position is disposed on the at least one inclined portion of the reaction tank body; The at least one reagent storage position stores a reagent.

2. The centrifugal multi-stage reaction device according to claim 1, wherein: The reaction tank body comprises a triangular chamber, a long strip channel, a U-shaped flow channel or a combination thereof.

3. The centrifugal multi-stage reaction device according to claim 1, wherein: When the reaction tank body comprises a plurality of reagent storage positions, the reagent storage positions are arranged on the at least one inclined portion at a certain interval.

4. The centrifugal multi-stage reaction device according to claim 3, wherein: When the reaction tank body includes two reagent storage positions, a first reagent storage is arranged at the bottom of the at least one inclined portion, and a second reagent storage is arranged at the other end opposite to the at least one inclined portion.

5. The centrifugal multi-stage reaction device according to claim 3, wherein: When the reaction tank body includes three reagent storage positions, a first reagent storage is arranged at the bottom end of the inclined portion, a third reagent storage is arranged at the other end of the inclined portion, and a second reagent storage is arranged at the center of the inclined portion.

6. The centrifugal multi-stage reaction device according to claim 1, wherein: When the at least one inclined portion is provided in two, the at least one inclined portion comprises a first inclined portion and a second inclined portion connected at one end thereof, and the plurality of storage locations are arranged on the two inclined portions at a certain interval.

7. The centrifugal multi-stage reaction device according to claim 6, wherein: The at least one reagent storage position includes five reagent storage positions, a first reagent storage position, which is arranged at the bottom end where the first inclined portion and the second inclined portion are connected; a second reagent storage position, which is arranged at the center of the first inclined portion; a third reagent storage position, which is arranged at the center of the second inclined portion; a fourth reagent storage position, which is arranged at the other end of the first inclined portion; and a fifth reagent storage position, which is arranged at the other end of the second inclined portion.

8. The centrifugal multi-stage reaction device according to claim 1, wherein: The reagent comprises at least one of a freeze-dried reagent, a volatile reagent, and a freeze-dried reagent ball.

9. A method for operating a centrifugal multi-stage reaction device, comprising the following steps: (A) providing a centrifugal multi-stage reaction device as claimed in claim 4; (B) centrifuging a sample through a power module to enter a reaction tank through an inlet flow channel; (C) after the sample enters a first reagent storage position, the sample reacts with a first reagent in the first reagent storage position; (D) the power module is rapidly changed in rotation direction, so that the sample is forced to flow upward along at least one inclined portion until it enters a second reagent storage position; (E) after the sample enters the second reagent storage position, the sample reacts with a second reagent in the second reagent storage position; and (F) The power module continues to centrifuge to return the sample to the first reagent storage position.

10. A method for operating a centrifugal multi-stage reaction device, comprising the following steps: (a) providing a centrifugal multi-stage reaction device as claimed in claim 5; (b) after centrifugal rotation to a certain speed by a power module, a sample is allowed to enter a reaction tank through an inlet flow channel; (c) After the sample enters a first reagent storage position, the sample reacts with a first reagent in the first reagent storage position. answer; (d) rapidly changing the rotation direction of the power module at a first acceleration, so that the sample is forced to flow upward along at least one inclined portion until it enters a second reagent storage position; (e) after the sample enters the second reagent storage position, the sample reacts with a second reagent in the second reagent storage position; (f) the power module continues to centrifuge to return the sample to the first reagent storage position and then stops rotating; (g) after the power module rotates to the rotation speed, the rotation direction is quickly changed at a second acceleration, so that the sample is forced to flow upward along the at least one inclined portion until it enters a third reagent storage position; (h) after the sample enters the third reagent storage position, the sample reacts with a third reagent in the third reagent storage position; and (i) The power module continues to centrifuge to return the sample to the first reagent storage position.

11. A method for operating a centrifugal multi-stage reaction device, comprising the following steps: (S01) providing a centrifugal multi-stage reaction device as described in claim 7; (S02) After centrifugation by a power module to a certain speed, a sample is allowed to enter a reaction tank through an inlet flow channel; (S03) After the sample enters a first reagent storage position, the sample reacts with a first reagent in the first reagent storage position; (S04) the power module is used to quickly change the rotation direction at a first acceleration, so that the sample is forced to flow upward along a first inclined portion until it enters a second reagent storage position; (S05) After the sample enters the second reagent storage position, the sample reacts with a second reagent in the second reagent storage position; (S06) the power module continues to centrifuge so that the sample returns to the first reagent storage position and then stops rotating; (S07) The power module is used to quickly change the rotation direction at a first acceleration, so that the sample is forced to flow upward along a second inclined portion until it enters a third reagent storage position; (S08) After the sample enters the third reagent storage position, the sample reacts with a third reagent in the third reagent storage position; (S09) the power module continues to centrifuge so that the sample returns to the first reagent storage position and then stops rotating; (S10) After the power module rotates to the rotation speed, the rotation direction is quickly changed at a second acceleration, so that the sample is forced to flow upward along the first inclined portion until it enters a fourth reagent storage position; (S11) After the sample enters the fourth reagent storage position, the sample reacts with a fourth reagent in the fourth reagent storage position; (S12) the power module continues to centrifuge so that the sample returns to the first reagent storage position and then stops rotating; (S13) After the power module rotates to the rotation speed, the rotation direction is quickly changed at a second acceleration, so that the sample is forced to flow upward along the second inclined portion until it enters a fifth reagent storage position; (S14) After the sample enters the fifth reagent storage position, the sample reacts with a fifth reagent in the fifth reagent storage position; and (S15) The power module continues to centrifuge to return the sample to the first reagent storage position.

12. The method for operating a centrifugal multi-stage reaction device according to claim 11, wherein: Step (S02) to step (S03) is a first reagent reaction process, step (S04) to step (S06) is a second reagent reaction process, step (S07) to step (S09) is a third reagent reaction process, step (S10) to step (S12) is a fourth reagent reaction process, and step (S13) to step (S15) is a fifth reagent reaction process.

13. The method for operating a centrifugal multi-stage reaction device according to claim 12, wherein: After the second reagent reaction process is completed, the fourth reagent reaction process is performed first and then the third reagent reaction process is performed, and finally the fifth reagent reaction process is completed.

14. The method for operating a centrifugal multi-stage reaction device according to claim 12, wherein: After the third reagent reaction process is completed, the fifth reagent reaction process is performed first and then the fourth reagent reaction process is performed.

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