Micro-fluidic chip and application thereof in nucleic acid detection

Abstract

The embodiment of the invention discloses a micro-fluidic chip and application thereof in nucleic acid detection.The micro-fluidic chip comprises a chip part, the chip part comprises a substrate, a plurality of cavities and a plurality of micro-fluidic channels are formed in the substrate, at least part of the cavities form cavity openings in the upper surface of the substrate, and the micro-fluidic channels are communicated with the cavity openings; at least part of the microfluidic channel forms a channel opening in the upper surface of the substrate; the connector assembly is arranged on the upper surface of the base plate in a protruding mode, a through hole is formed in the connector assembly, and the connector assembly is fixed to the cavity opening, so that the through hole of the connector assembly is communicated with the cavity corresponding to the cavity opening to form a cavity; and / or the connector assembly is fixed at the channel opening, so that the through hole of the connector assembly is communicated with the microfluidic channel corresponding to the channel opening. The substrate is provided with the opening, and the connector assembly is connected into the opening, so that at least one of the functions of enlarging the reaction system, adjusting the volume of the reaction cavity, flexibly feeding, filtering products, pressing solid reactants pre-buried in the reaction cavity and the like can be realized.

Description

[0001] This application claims priority to Chinese Patent Application No. 202411132058.1, filed on August 19, 2024, with the invention title “Microfluidic Chip, Microfluidic Chip System and Its Application in DNA Testing”, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This application relates to the field of biological nucleic acid detection equipment technology, and in particular to a microfluidic chip. Background Technology

[0003] Microfluidic chips, as a representative of micrototal analysis systems, have extremely broad application value in the field of nucleic acid extraction and detection. Their related application technologies and equipment manufacturing have become a cutting-edge hot topic in academia and industry. Microfluidic chip technology is used to control fluids at the micrometer scale, enabling the automation and integration of biochemical reactions. Applying microfluidic chips to the field of biomedical detection allows complex detection reactions that were originally performed by professionals in specialized laboratories to be automated and rapidly completed on the chip, showing broad application prospects in in vitro diagnostics, point-of-care testing, and liquid biopsy.

[0004] In existing technologies, when reactions are carried out in microfluidic chips, the limited size of the microfluidic chip often prevents it from providing the reaction system required for large-scale reactions. Furthermore, higher-order functions such as adjusting the size of the reaction chamber, filtering products, and pressing pre-embedded solid reactants within the reaction chamber are difficult to achieve in current microfluidic chips. Summary of the Invention

[0005] A first aspect of this application provides a microfluidic chip, which includes a chip portion, the chip portion comprising:

[0006] The substrate has multiple chambers and multiple microfluidic channels, with at least some of the chambers forming chamber openings on the upper surface of the substrate, and at least some of the microfluidic channels forming channel openings on the upper surface of the substrate.

[0007] A connector assembly protrudes from the upper surface of the substrate and has through holes.

[0008] At least a portion of the connector assembly is fixed to the chamber opening so that the through hole of the connector assembly communicates with the chamber corresponding to the chamber opening and forms a cavity; and / or, at least a portion of the connector assembly is fixed to the channel opening so that the through hole of the connector assembly communicates with the microfluidic channel corresponding to the channel opening.

[0009] Optionally, the connector assembly includes at least one of the following:

[0010] A single-hole connector assembly, wherein the single-hole connector assembly has a through hole;

[0011] A multi-hole connector assembly has multiple through holes. When the connector assembly protrudes from the upper surface of the substrate, at least one of the multiple through holes is connected to a cavity corresponding to the cavity opening to form a cavity. At least one of the multiple through holes is connected to a microfluidic channel corresponding to the channel opening. The cavity connected by at least one through hole and the microfluidic channel connected by at least one through hole are connected through the microfluidic channel.

[0012] A laminated connector assembly, comprising an upper connector assembly and a lower connector assembly, wherein the lower connector assembly has a through hole and is fixed to the opening of the chamber, and the upper connector assembly is detachably fixed in the through hole of the lower connector assembly;

[0013] A filter connector assembly; the filter connector assembly has a connector filter section inside the through hole and a chamber filter section inside the corresponding cavity. When the filter connector assembly is fixed to the open cavity, the connector filter section and the chamber filter section form a filter section. The cavity formed by the through hole and the cavity of the filter connector assembly has a second subspace isolated within the cavity. The second subspace is connected to other spaces in the cavity only through the second subspace entrance formed by the top of the filter section. The second subspace entrance is located at a preset height inside the through hole of the filter connector assembly. The discharge port for discharging materials from the cavity is located within the second subspace.

[0014] Optionally, the connector assembly includes a porous connector assembly, with at least one through-hole forming a cavity for carrying out the reaction, and at least one through-hole communicating a microfluidic channel for feeding into the cavity.

[0015] Optionally, the connector assembly includes a filter connector assembly;

[0016] The filter section is generally a portal-shaped column or a semi-cylindrical column, abutting against the inner wall of the cavity to define a second subspace together with the inner wall of the cavity; or,

[0017] The filter section is a cylindrical column, which defines a second subspace. The filter section is fixed to the inner wall of the cavity.

[0018] Optionally, the connector assembly includes a laminated connector assembly;

[0019] The upper connector assembly has a through hole, and the through hole and the chamber of the upper connector assembly form a cavity; multiple upper connector assemblies with different through hole diameters are detachably fixed in the through holes of the lower connector assembly to form multiple cavities with different volumes.

[0020] Optionally, the connector assembly includes a laminated connector assembly;

[0021] The solid reactant is pre-embedded in the bottom pressing chamber of the upper connector assembly.

[0022] Optionally, each connector assembly protrudes from the upper surface of the substrate at the same height, and the diameter of the through holes in at least some connector assemblies is determined according to the required reaction system.

[0023] Optionally, the substrate includes a top layer, a middle layer, and a bottom layer; the substrate is provided with microfluidic channels;

[0024] The upper and lower surfaces of the intermediate layer have three-dimensional patterns. The pattern on the upper surface of the intermediate layer and the top layer form the upper microfluidic channel within the microfluidic channel; the pattern on the lower surface of the intermediate layer and the bottom layer form the lower microfluidic channel within the microfluidic channel; the extension direction of the upper and lower microfluidic channels is parallel to the plane of the substrate.

[0025] The intermediate layer includes through-holes for forming vertical microfluidic channels, the extension direction of which is perpendicular to the plane of the substrate, and at least a portion of the vertical microfluidic channels are used to connect the upper microfluidic channels and the lower microfluidic channels.

[0026] The intermediate layer also includes through-holes for forming chambers for the reaction to take place;

[0027] The top layer has through holes at positions corresponding to at least a portion of the chambers to form chamber openings, and the top layer has through holes at positions corresponding to at least a portion of the upper microfluidic channels or vertical microfluidic channels to form channel openings.

[0028] Optionally, the microfluidic chip includes a clamping part, which includes an upper clamping plate and a lower clamping plate. The upper clamping plate and the lower clamping plate form a receiving cavity. When the chip part is located in the receiving cavity, the lower surface of the upper clamping plate contacts the upper end face of the connector assembly to seal the connector assembly.

[0029] Optionally, the upper clamping plate also includes a sealing gasket, and the lower surface of the upper clamping plate contacts the upper end face of the connector assembly, including: the sealing gasket of the upper clamping plate contacts the upper end face of the connector assembly, and the sealing gasket of the upper clamping plate is located between the upper clamping plate body and the upper end face of the connector assembly.

[0030] Optionally, the upper clamping plate has a fourth through hole, the sealing gasket has a through hole, and the fourth through hole, the through hole of the sealing gasket, and the through hole of the connector assembly are aligned and connected.

[0031] Optionally, the fixture part also includes a fixture assembly, which has a through hole and is fixed to the fourth through hole. When the chip part is located in the receiving cavity, the through hole of the fixture assembly, the fourth through hole of the upper clamping plate, the through hole of the sealing gasket, the through hole of the connector assembly, and the intermediate connectable component corresponding to the connector assembly are aligned. The intermediate connectable component includes a chamber and / or a microfluidic channel.

[0032] A second aspect of this application provides the application of the microfluidic chip of the first aspect in nucleic acid detection, wherein the substrate includes multiple functional regions, the multiple functional regions including a cell lysis region, a nucleic acid extraction region, a nucleic acid amplification region and a product export region; each functional region includes a cavity composed of a connector assembly for performing the reaction required by that functional region.

[0033] The embodiments of this application, by opening an opening in the substrate and connecting a connector assembly to the opening, can achieve at least one of the following functions: expanding the reaction system, making the reaction chamber volume adjustable, flexibly feeding materials, filtering products, and pressing the solid reactants pre-embedded in the reaction chamber. Attached Figure Description

[0034] Figure 1A A schematic diagram of the structure of a microfluidic chip according to an embodiment of this application is shown, wherein the shaded part represents a virtual body;

[0035] Figure 1B A schematic diagram of the structure of a microfluidic chip according to an embodiment of this application is shown, wherein the shaded part represents a virtual body;

[0036] Figure 1C A schematic diagram of the structure of a microfluidic chip according to an embodiment of this application is shown, wherein the shaded part represents a virtual body;

[0037] Figure 1D A schematic diagram of the structure of a microfluidic chip according to an embodiment of this application is shown, wherein the shaded part represents a virtual body;

[0038] Figure 1E A schematic diagram of the structure of a microfluidic chip according to an embodiment of this application is shown, wherein the shaded part represents a virtual body;

[0039] Figure 1F A schematic diagram of the chamber structure of a microfluidic chip according to an embodiment of this application is shown;

[0040] Figure 1G A top view showing a schematic diagram of the chamber structure of a microfluidic chip according to an embodiment of this application;

[0041] Figure 2 A schematic diagram of the structure of a microfluidic chip according to an embodiment of this application is shown, wherein the shaded part represents a virtual body;

[0042] Figure 3 shows a schematic diagram of the structure of a microfluidic chip according to an embodiment of this application, where the shaded part represents a virtual body;

[0043] Figure 4A and Figure 4B A schematic diagram of the structure of a stacked connector assembly according to an embodiment of this application is shown, wherein the shaded portion represents a virtual body;

[0044] Figure 5A schematic diagram of the structure of a filter connector assembly and its corresponding chamber according to an embodiment of this application is shown, wherein the shaded portion represents the solid part;

[0045] Figure 6 shows a perspective view of the structure of a filter connector assembly and its corresponding chamber according to an embodiment of this application;

[0046] Figure 7 A schematic diagram of the structure of a product derived component according to an embodiment of this application is shown, wherein the shaded portion represents a solid object;

[0047] Figure 8 A schematic diagram of the structure of a displacement component according to an embodiment of this application is shown;

[0048] Figure 9 shows a schematic diagram of the structure of a displacement component according to an embodiment of this application;

[0049] Figure 10A A schematic diagram of the structure of the clamping part according to an embodiment of this application is shown;

[0050] Figure 10B This invention illustrates a schematic diagram of the structure of the first clamping part in a clamping part according to an embodiment of the present application;

[0051] Figure 11 A schematic diagram of the chip section structure of a microfluidic chip according to an embodiment of this application is shown;

[0052] Figure 12A This illustration shows a schematic diagram of the top layer structure of a chip substrate according to an embodiment of this application;

[0053] Figure 12B A schematic diagram showing a three-dimensional pattern on the upper surface of the intermediate layer of a chip substrate according to an embodiment of this application;

[0054] Figure 12C A schematic diagram showing a three-dimensional pattern on the lower surface of the intermediate layer of a chip substrate according to an embodiment of this application;

[0055] Figure 12D This illustration shows a schematic diagram of the underlying structure of a chip substrate according to an embodiment of this application;

[0056] Figure 12E A three-dimensional structural diagram of the bottom layer of a chip substrate according to an embodiment of this application is shown;

[0057] Figure 13A A schematic diagram of the upper surface of the upper clamping plate according to an embodiment of this application is shown;

[0058] Figure 13B A schematic diagram of the structure of the lower surface of the upper clamping plate according to an embodiment of this application is shown;

[0059] Figure 14This application illustrates a schematic diagram showing the structural relationship between the upper clamping plate and the chip section according to an embodiment of the present application.

[0060] Figure 15 A schematic diagram of the structure of the lower surface of the upper clamping plate according to an embodiment of this application is shown;

[0061] Figure 16 A schematic diagram of the structure of a sealing gasket according to one embodiment of this application is shown;

[0062] Figure 17 A schematic diagram of the lower clamping plate according to one embodiment of this application is shown;

[0063] Figure 18 This invention illustrates a structural diagram showing the cooperative relationship between a processing component and a lower clamping plate according to an embodiment of the present application.

[0064] Figure 19 A schematic diagram of the upper and lower clamping plates according to an embodiment of this application is shown;

[0065] Figure 20 A schematic diagram of the upper and lower clamping plates according to an embodiment of this application is shown;

[0066] Figure 21A A schematic diagram illustrating the connection relationship between a microfluidic chip and a feed valve assembly according to an embodiment of this application is provided.

[0067] Figure 21B A schematic diagram illustrating the connection relationship between a microfluidic chip, an exhaust valve assembly, and a drain valve assembly according to an embodiment of this application is provided.

[0068] Figure 22 A schematic diagram illustrating the connection relationship between a microfluidic chip and a valve assembly according to an embodiment of this application is shown.

[0069] Figure 23 A schematic diagram illustrating the connection relationship between a microfluidic chip and a displacement component control component according to an embodiment of this application is provided.

[0070] Figure 24 A schematic diagram of the structure of a microfluidic chip system according to an embodiment of this application is shown.

[0071] Explanation of reference numerals in the attached figures

[0072] 100. Chip section; 110. Substrate; 111. Top layer; 112. Intermediate layer; 113. Bottom layer; 114. Opening; 1141. Chamber opening; 1142. Channel opening; 115. Through-hole of the channel opening as a flow path control channel; 116. First countersunk hole; 130. Chamber; 131. Chamber filter section; 132. First subspace; 133. Interface between the chamber and the microfluidic channel; 134. First subspace inlet; 135. Second subspace; 136. Second subspace inlet; 14 0. Microfluidic channel; 141. Upper microfluidic channel; 142. Lower microfluidic channel; 143. Vertical microfluidic channel; 144. Flow path control channel; 1441. Straight pipe section; 1442. Hemispherical section; 150. Connector assembly; 1501. Single-hole connector assembly; 1502. Multi-hole connector assembly; 1503. Stacked connector assembly; 1503A. Upper connector assembly; 1503B. Lower connector assembly; 1503C. Vent hole; 1503D. Second countersunk hole; 1504. Filter connector assembly Components; 1504A, Connector Filter Section; 160, Product Outlet Section; 180, Elastic Membrane; 101, Lysis Zone; 102, Extraction Zone; 103, Amplification Zone; 104, Product Outlet Zone; 300, Displacement Assembly; 310, Output End; 320, Stationary Section; 330, Moving Section; 340, Elastic Component Fixing Section; 350, Elastic Component; 400, Pressing Section; 410, First Pressing Section; 411, Ninth Through Hole; 412, Bottom of Cylinder; 413, Annular Protrusion; 414, Cylinder Wall; 415. 420. Card slot; 421. Second clamping part; 430. Tenth through hole; 431. Third clamping part; 432. Eleventh through hole; 433. Lug; 500. External pipeline; 600. Valve assembly; 700. Processing assembly; 710. Heating assembly; 720. Magnetic suction assembly; 730. Product collection container; 800. Control circuit; 900. Support; 910. Power circuit layer support; 920. Valve assembly layer support; 930. Gas-liquid layer support; 940. Clamping plate support; 950. Processing assembly support. Detailed Implementation

[0073] 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 a part of the embodiments of this application, and not all of the 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.

[0074] The terms "first," "second," and "third" in this application are for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Therefore, a feature defined as "first," "second," or "third" may explicitly or implicitly include at least one of that feature. In the description of this application, "multiple" means at least two, such as two, three, etc., unless otherwise explicitly specified. All directional indications (such as up, down, left, right, front, back, etc.) in the embodiments of this application are only used to explain the relative positional relationships and movements between components in a specific orientation (as shown in the figures). If the specific orientation changes, the directional indications also change accordingly. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion. For example, a process, method, system, product, or device that includes a series of steps or units is not limited to the listed steps or units, but may optionally include steps or units not listed, or may optionally include other steps or units inherent to these processes, methods, products, or devices.

[0075] In this document, the term "embodiment" means that a particular feature, structure, or characteristic described in connection with an embodiment may be included in at least one embodiment of this application. The appearance of this phrase in various places throughout the specification does not necessarily refer to the same embodiment, nor is it a separate or alternative embodiment mutually exclusive with other embodiments. It will be explicitly and implicitly understood by those skilled in the art that the embodiments described herein can be combined with other embodiments.

[0076] The term "correspondence" as used in this article can be interpreted broadly. It can refer to components having a corresponding relationship in terms of position (e.g., the projection centers of components coincide in the thickness direction of the substrate), or one component being located upstream or downstream of another component.

[0077] A first aspect of this application provides a microfluidic chip, including a chip portion 100. Exemplarily, it may also include at least one of the following: a clamp portion 200 for accommodating the chip portion 100; a displacement component 300 for applying mechanical action to the chip portion 100 to control the opening and closing of the microfluidic channels of the chip portion 100; and a clamping portion 400 for pressing a local area of ​​the chip portion 100 to prevent leakage of the chip portion 100 when the displacement component 300 applies mechanical action to the chip portion 100.

[0078] The chip section 100 includes a substrate 110, and, by way of example, may also include at least one of a connector assembly 150, a product outlet 160, and an elastic membrane 180. The specific structure and function of each component will be described below. By way of example, the substrate may be a rigid material.

[0079] For example, the substrate 110 includes a chamber 130 and a microfluidic channel 140. The chamber 130 is mainly used to contain materials and carry out reactions, while the microfluidic channel 140 is mainly used for fluid communication between chambers and between chambers and the outside. By rationally arranging the chambers 130 and the microfluidic channel 140 on the substrate 110, the functions of the chip section 100 can be realized, such as carrying out biochemical reactions in the chip section, especially nucleic acid detection reactions, such as DNA testing.

[0080] For example, substrate 110 includes multiple substrate layers to form chambers 130 and microfluidic channels 140 in the substrate layers. For example, a three-dimensional pattern (understood to be a pattern that is not located on a two-dimensional plane but has a certain thickness) is formed on the surface of at least one of two adjacent substrate layers, and then combined with the other, for example by bonding, lamination or other means, so that chambers 130 or microfluidic channels 140 can be formed from the two adjacent substrate layers.

[0081] like Figure 1A As shown, the substrate includes a top layer 111, an intermediate layer 112, and a bottom layer 113. The upper and lower surfaces of the intermediate layer 112 have three-dimensional patterns. The intermediate layer 112 and the top layer 111 are bonded together, forming a microfluidic channel (hereinafter referred to as the upper microfluidic channel 141) between the top layer 111 and the intermediate layer 112. The intermediate layer 112 and the bottom layer 113 are bonded together, forming a microfluidic channel (hereinafter referred to as the lower microfluidic channel 142) between the intermediate layer 112 and the bottom layer 113. The height of the horizontal axis of the microfluidic channel is referred to as the "height of the microfluidic channel." Obviously, the height of the upper microfluidic channel 141 is different from the height of the lower microfluidic channel 142. For example, when the substrate includes three or more substrate layers, a larger through-hole can be formed in the intermediate layer 112 to form a cavity 130, or a smaller through-hole can be formed in the intermediate layer 112 to form a microfluidic channel (hereinafter referred to as the vertical microfluidic channel 143), as shown. Figure 1B As shown. It is understood that the upper microfluidic channel 141 and the lower microfluidic channel 142 are oriented parallel to the plane of the substrate 110, and the vertical microfluidic channel 143 is oriented substantially perpendicular to the plane of the substrate 110. That is, the intermediate layer 112 can form the chamber 130, the upper microfluidic channel 141, the lower microfluidic channel 142, and the vertical microfluidic channel 143, collectively referred to as the intermediate connectable component. The intermediate layer 112 can be thicker than the top layer 111 and the bottom layer 113 to form a larger chamber; for example, the thicknesses of the top layer 111, the intermediate layer 112, and the bottom layer 113 are 0.5 mm, 4 mm, and 0.5 mm, respectively.

[0082] For example, through holes corresponding to the intermediate connectable component can be formed in the top layer 111 or the bottom layer 113, creating an opening 114 that connects the outside to the intermediate connectable component. For example, it can be formed as follows: Figure 1CThe diagram shows the chamber opening 1141 of the connecting chamber 130 and the channel opening 1142 of the connecting microfluidic channel. It is understood that the upper microfluidic channel 141, the lower microfluidic channel 142, and the vertical microfluidic channel 143 may or may not have a channel opening 1142. For example, the vertical microfluidic channel, which is only used to connect the upper microfluidic channel 141 and the lower microfluidic channel 142, usually does not have a channel opening 1142.

[0083] Figure 1D As shown, openings, microfluidic channels, and chambers can be flexibly designed to connect with each other, forming various fluid pathways on the substrate 110. Examples include: outside - channel opening 1142 of top layer 111 - vertical microfluidic channel 143 - lower microfluidic channel 142 - channel opening 1142 of bottom layer 113 - outside; outside - chamber opening 1141 of top layer 111 - chamber 130 - lower microfluidic channel 142 - vertical microfluidic channel 143 - upper microfluidic channel 141, etc. This allows for flexible control of material flow within the substrate 110, adapting to various complex reactions.

[0084] It is understood that the upper microfluidic channel, lower microfluidic channel, vertical microfluidic channel and chamber in the embodiments of this application can not only be formed by preparing and combining multiple substrate layers separately, but also can be integrally formed by 3D printing and other methods. The openings on the upper surface and lower surface of the substrate connect to the outside world and the intermediate connecting components.

[0085] This application embodiment forms microfluidic channels at different heights and with different orientations in the substrate, creating an "overpass" type of microfluidic channel network. This achieves at least one of the following effects: First, by fully utilizing the substrate height dimension and forming microfluidic channels at multiple heights, the distribution density of microfluidic channels on the substrate can be increased, reducing the difficulty of arranging microfluidic channels on the substrate during high-throughput reactions. Second, the height of the substrate can be used to construct a more flexible connection between the external environment, microfluidic channels, and chambers, thereby forming multiple fluid pathways on the substrate to adapt to various complex reactions. Third, it can simultaneously meet the needs of flow path control (requiring the flow path control channel to be located on the upper layer), operational needs (e.g., for blowing operations, air needs to be introduced from the bottom), and reaction conditions (e.g., for reactions in some chambers that need to be heated, the bottom of the chamber needs to be located near the bottom of the entire substrate to heat the chamber from the bottom). Furthermore, by processing three-dimensional patterns on the upper and lower surfaces of the intermediate layer and then combining them with the top and bottom layers, the processing difficulty of complex chips can be greatly reduced.

[0086] The chamber 130 and the chamber opening 1141 will be further described below.

[0087] The chamber 130 is used to contain materials, and the reaction can take place in the chamber. When the substrate is composed of multiple substrate layers, the chamber 130 is surrounded by the through-hole of the intermediate layer 112 and the bottom layer 114.

[0088] For example, to meet different reaction requirements and consider processing difficulty, the chamber 130 and chamber opening 1141 can be one-to-one (i.e., one chamber 130 corresponds to one chamber opening 1141), one-to-many (i.e., one chamber 130 forms multiple chamber openings 1141), or many-to-one (i.e., multiple chambers 130 share one chamber opening 1141). The chamber 130 can also share an opening with a microfluidic channel. For example... Figure 1D As shown, the chamber 130 and the vertical microfluidic channel 143 are formed by two through holes opened in the intermediate layer 114. They share an opening 114, and the projection of the opening 114 onto the plane of the substrate intersects with the projections of the two through holes onto the same plane, allowing the chamber 130 and the vertical microfluidic channel 143 to communicate with the outside through the opening 114. For example, the projection of the opening 114 onto the plane of the substrate and the projections of the two through holes onto the same plane are inclusive; that is, the projection P of the opening 114 onto the plane of the substrate includes the projection P1 of the through hole 1 corresponding to the chamber 130 onto the same plane and the projection P2 of the through hole 2 corresponding to the vertical microfluidic channel 143 onto the same plane. For ease of processing, the opening 114 can be a closed convex hull that includes the projections of the through holes 1 and 2, and its shape can be circular, elliptical, teardrop-shaped, etc.

[0089] The chamber opening 1141 can serve multiple functions, such as acting as an inlet / outlet for the chamber 130. A connector assembly 150 can be externally connected to the chamber opening 1141, allowing the connector assembly 150 and the chamber 130 to form a larger reaction system or achieve additional functions. The connector assembly 150 will be described later.

[0090] The chamber 130 can communicate with the upper microfluidic channel 141 and / or the lower microfluidic channel 142. For example, the chamber 130 is a cylindrical structure, and the interface 133 connecting the chamber 130 and the microfluidic channel is located on the sidewall of the cylindrical structure, such as... Figure 1D As shown. Understandably, the interface 133 connecting the chamber 130 and the microfluidic channel can be located at different heights on the sidewall of the cylindrical structure.

[0091] Just as materials can be filtered in a macroscopic system, a filtration function can also be achieved in chamber 130. For example, the chamber may also include a chamber filter unit 131 to filter the materials within the chamber. The following describes... Figure 1E , Figure 1F and Figure 1G The chamber filter section 131 will be described below. For example... Figure 1E and Figure 1FAs shown, a chamber filter section 131 is provided on the inner wall of the chamber 130. The chamber filter section 131 isolates a first subspace 132 from the chamber 130. The discharge port 1331 of the chamber 130 (i.e., the interface between the chamber 130 and the microfluidic channel for discharge) is located in the first subspace 132. The first subspace 132 can only be connected to other spaces of the chamber 130 through the first subspace inlet 134. The first subspace inlet 134 can be formed at the top of the chamber filter section 131 and has a preset height. The preset height can be flush with the top of the chamber or located at other heights in the chamber. Fluid in the space of chamber 130 other than the first subspace 132 can only enter the first subspace 132 through the first subspace inlet 134 and exit through the outlet in the first subspace. Thus, only the supernatant above a preset height in chamber 130 can enter the first subspace 132 through the first subspace inlet 134 and exit through the outlet 133. Impurities in chamber 130 will settle and remain in the space outside the first subspace 132, thereby achieving filtration in chamber 130. The chamber filter section 131 can be integrally formed with the inner wall of chamber 130, or it can be formed separately from the chamber and then fixed to the inner wall of chamber 130. The chamber filter section 131 can have various structures, such as... Figure 1G The chamber filtration section 131 can be a cylindrical structure (without top or bottom) that is approximately tangent to the inner wall of the chamber 130, or it can be an approximately "door-shaped" structure or a semi-cylindrical structure. Understandably, the feed inlet 1332 of the chamber 130 (i.e., the interface between the chamber 130 and the microfluidic channel for feed) is located in the space of the chamber 130 other than the first subspace 132.

[0092] The structure and function of the microfluidic channel 140 are described below.

[0093] As mentioned above, based on the location and orientation of the microfluidic channels, the microfluidic channels 140 of the substrate 110 are divided into upper microfluidic channels 141, lower microfluidic channels 142, and vertical microfluidic channels 143. Based on whether the sealing and opening of the microfluidic channels are controllable, the microfluidic channels 140 of the substrate 110 can be divided into flow path control channels 144 and non-flow path control channels. Based on the role of the microfluidic channels in the reaction, the microfluidic channels 140 of the substrate 110 can be divided into microfluidic channels for connecting chambers, product outlet channels for exporting products, and waste outlet channels for exporting waste, etc. Based on whether channel openings are formed on the surface of the substrate 110, they can be divided into microfluidic channels with channel openings and microfluidic channels without channel openings.

[0094] Understandably, an opening for an upper microfluidic channel 141 or a vertical microfluidic channel 143 can be formed on the upper surface of the substrate. For a substrate 110 comprising a top layer 111, an intermediate layer 112, and a bottom layer 113, the opening 114 on the upper surface of the substrate can be formed by providing a through-hole in the top layer 111. An opening for a lower microfluidic channel 142 or a vertical microfluidic channel 143 can be formed on the lower surface of the substrate. For a substrate 110 comprising a top layer 111, an intermediate layer 112, and a bottom layer 113, the opening on the lower surface of the substrate can be formed by providing a through-hole in the bottom layer 113.

[0095] The microfluidic channel 140 may include a flow path control channel 144, which is a microfluidic channel that can be blocked and opened.

[0096] For example, a variable component can be used to block and open the flow path control channel. For instance, a variable component can be placed at a suitable location in the flow path control channel. When the variable component deforms, it blocks a portion of the flow path control channel; when it returns to its original position, it opens the corresponding area. For example, the deformation and return of the variable component can be controlled by external force; applying external force causes deformation, and removing the external force causes it to return to its original position.

[0097] In existing technologies, flow control components are typically integrated into microfluidic chips. For example, a microfluidic chip includes a base layer and a chip layer, with the chip layer made of an elastic material. The microfluidic chip contains flow control channels and deformable channels located above and intersecting the flow control channels. In the intersection region, the top surface of the flow control channel is the bottom surface of the deformable channel. When fluid is introduced into the deformable channel, the bottom surface deforms downwards, blocking the portion of the flow control channel located in the intersection region; when no fluid is introduced, the bottom surface of the deformable channel returns to its original position, and the flow control channel is opened. However, integrating the deformable channels into the microfluidic chip makes the fabrication process extremely complex. Furthermore, controlling the deformable channels involves manipulating numerous fluid lines and control valves, making operation difficult. Additionally, when the microfluidic chip is made of a rigid material, integrating the deformable channels within the microfluidic chip becomes challenging.

[0098] For example, to solve at least one of the above problems, embodiments of this application provide an elastic membrane 180 located on the upper surface of the substrate 110 and a displacement component 300 located outside the substrate 110 to control the on / off state of the flow path control channel 144 in the substrate 110. Compared to variable components integrated into the substrate and components for external force control, by providing an elastic membrane 180 on the upper surface of the substrate 110 and a displacement component 300 outside the substrate, and by applying a mechanical pressing action to the elastic membrane through switching the displacement component at different heights, thereby controlling the on / off state of the flow path control channel within the chip section, the processing difficulty of the chip section 100 and the operational difficulty of flow path control can be greatly reduced.

[0099] For example, in this embodiment, an opening of a flow control channel 144 is provided on the upper surface of the substrate 110, and an elastic membrane 180 is covered on the opening. Thus, after deformation, the elastic membrane 180 can pass through the opening of the flow control channel 144 and directly act on the flow control channel. Even if the substrate 110 is made of a non-elastic material, the flow control channel 144 can be blocked and opened. Simultaneously, in this embodiment, a displacement component 300 disposed outside the chip portion 100 is used to apply external force to deform the elastic membrane 180. For example, the displacement component 300 disposed above the substrate 110 presses the elastic membrane 180 downwards, causing the elastic membrane 180 to deform in the pressing direction and block the flow control channel 144 corresponding to the elastic membrane 180.

[0100] The flow control channel is an upper microfluidic channel 141 that provides an opening for the flow control channel 144 on the upper surface of the substrate 110. The flow control channel 144 can be formed between the top layer 111 and the intermediate layer 112, rather than between the intermediate layer 112 and the bottom layer 113.

[0101] For example, to facilitate processing, a through-hole (which penetrates the top layer when the substrate comprises multiple substrate layers) can be formed at the location where the flow control channel 144 is to be blocked, serving as an opening for the flow control channel 144. For example, the through-hole 115 serving as the opening for the flow control channel 144 is circular, and its diameter can be 1.2-4 times the width of the flow control channel. This ensures that a sufficiently large area of ​​elastic membrane can deform and pass through the through-hole to completely block the flow control channel 144.

[0102] For example, the pattern on the upper surface of the intermediate layer includes a straight pipe pattern 1441 and a matching pattern 1442 that matches the shape of the output end. The position of the matching pattern corresponds to the position of the channel opening of the flow control channel, and it cuts the straight pipe pattern into two segments. The widest width of the matching pattern is greater than the width of the straight pipe pattern and / or the deepest depth of the matching pattern is greater than the depth of the straight pipe pattern. For example, if the output end is hemispherical, the matching pattern is also a hemispherical pattern. The position of the hemispherical pattern 1442 corresponds to the position of the through-hole 115, and the hemispherical pattern 1442 cuts the straight pipe pattern 1441 into two segments. The diameter of the hemispherical pattern 1442 is greater than the width of the straight pipe pattern 1441 and / or the depth of the hemispherical pattern 1442 is greater than the depth of the straight pipe pattern 1441. Accordingly, the flow control channel includes a straight pipe portion and a matching portion. For example, the highest point of the hemispherical pattern 1442 is the same as the highest point of the flow control channel 144, that is, the hemispherical pattern starts from the bottom surface of the top layer 111. Thus, after the elastic membrane 180 deforms, it passes through the through-hole and adheres to the surface of the hemispherical pattern 1442, completely sealing the flow control channel 144. For example, when the matching pattern is a hemispherical pattern, the diameter of the through-hole 115, which serves as the opening of the flow control channel 144, is equal to the diameter of the hemispherical pattern 1442. For example, the diameter of the hemispherical pattern is approximately 0.5 mm.

[0103] For example, to ensure the flatness of the upper surface of the substrate 110 and the stability of the elastic membrane 180 fixed at the channel opening 1142, and to facilitate the positioning of the elastic membrane 180, a first countersunk hole 116 for placing the elastic membrane 180 can be formed on the upper surface of the substrate 110 (e.g., if the substrate includes multiple substrate layers, i.e., on the upper surface of the top layer 111). For example, the thickness of the first countersunk hole 116 and the elastic membrane 180 are matched. When the elastic membrane 180 is placed in the first countersunk hole 116, the upper surface of the elastic membrane 180 is flush with the upper surface of the top layer 111, which facilitates the assembly of the elastic membrane 180 and the substrate 110 as a whole with other components. It is understood that the diameter of the first countersunk hole 116 is larger than the diameter of the through hole 115, and the center of the projection of the through hole 115 on the substrate coincides with the center of the projection of the first countersunk hole 116 on the substrate 110. For example, the diameter of the first countersunk hole 116 is 9.5-12 mm, the depth of the first countersunk hole 116 is 0.3 mm, and the thickness of the elastic membrane 180 is 0.2-0.3 mm. (The text repeats itself here.) Figure 2 As shown.

[0104] As mentioned above, channels can have channel openings 1142, and chambers can have chamber openings 1141. When the substrate includes multiple substrate layers, through holes can be formed on the top layer 111 to form different types of openings 114, such as: first, only serving as chamber openings 1141; second, only serving as channel openings 1142 for flow path control channels 144; third, only serving as channel openings 1142 for non-flow path control channels (including vertical microfluidic channels 143 and upper microfluidic channels 141); and fourth, serving as both chamber openings 1141 and vertical microfluidic channels 143. Of course, if needed, there can also be openings that simultaneously serve as multiple chamber openings 1141 and multiple channel openings 1142. The bottom layer 113 may not have through holes and only serve as a base, or it may have through holes to form the channel opening 1142 of the vertical microfluidic channel 143 and / or the lower microfluidic channel 142.

[0105] Of these, only the channel opening 1142, which serves as the flow path control channel 144, can be used for flow path on / off control, while the remaining openings can be used for material feeding / discharging or as interfaces with other components. This allows for flexible control of the material's path within the substrate 110 and the communication between the material in the substrate and the external environment, adapting to various complex reactions.

[0106] For example, the chip section 100 may also include a connector assembly 150, which has a through hole and may be a bottomless cylindrical component. The connector assembly 150 may be made of the same material as the substrate 110.

[0107] Existing chip substrates suffer from at least one of the following problems: the substrate has a limited volume, making it difficult to form high-throughput, large-capacity reaction chambers on the substrate, thus failing to meet the needs of the reaction system; if the reaction system is contained within the chip, the chip becomes too large, making chip processing and manipulation very difficult; in some cases, the volume of the reaction chamber needs to be adjustable, which is difficult to meet by forming the reaction chamber using a substrate; in some cases, the reaction chamber needs to perform additional functions, such as filtering products, pressing down solid reactants pre-embedded in the reaction chamber, forming more inlet and outlet ports, etc., which requires a more complex structure, and integrating a complex structure onto a substrate is quite difficult.

[0108] To solve at least one of the above problems, a connector assembly 150 with a through hole can be introduced. The connector can be installed at the opening on the substrate surface of the intermediate connectable assembly, so that the through hole of the connector and the intermediate connectable assembly are connected. When the intermediate connectable assembly is a chamber, when the connector assembly 150 is installed on the substrate 110, the through hole of the connector assembly 150 and the chamber 130 of the substrate 110 together form a cavity for carrying out the reaction.

[0109] When the connector assembly 150 has a more complex structure, it can work in conjunction with the chamber 130 of the substrate 110 to achieve additional functions. Depending on the structure of the connector assembly 150, its functions may include expanding the reaction system, filtering products, pressing down solid reactants embedded in the reaction chamber, and serving as an extended inlet / outlet for intermediate connected components.

[0110] Furthermore, the through holes of the connector assembly 150 can also be of various sizes. When the connector assembly 150 is detachably connected to the substrate 110, a reaction chamber with adjustable volume can be obtained by replacing the connector assembly 150 with different through hole sizes.

[0111] Understandably, the connector assembly 150 has a certain height and protrudes from the surface of the substrate 110. Understandably, for intermediate connectable components with the connector assembly 150 installed, material can be fed into and out of the intermediate connectable component through the through-hole of the connector assembly 150; for intermediate connectable components without the connector assembly 150 installed, material can be fed into and out directly through the open end of the intermediate connectable component. To ensure that the inlet and outlet heights of intermediate connectable components requiring material feeding and discharging on the substrate 110 are the same for sealing and other operations, the connector assembly 150 can be installed on all intermediate connectable components requiring material feeding and discharging, and the connector assembly 150 can be protruding from the upper surface of the substrate 110 at the same height. This reduces the difficulty of operation and packaging.

[0112] All connector assemblies 150 protrude from the upper surface of the substrate 110 at the same height. That is, the height of the connector assemblies 150 protruding from the upper surface of the substrate 110 is consistent, and the height of the through holes of the connector assemblies 150 is consistent. The volume of the cavity formed by the connector assembly 150 and the cavity 130 can be adjusted by adjusting the diameter of the through holes of the connector assembly 150.

[0113] Considering the ease of processing, installation, and component arrangement, one connector assembly 150 can correspond to one or more intermediate connectable components. For example... Figure 3A As shown, one connector assembly 150 can correspond to one chamber 130, vertical microfluidic channel 143, or upper microfluidic channel 141; as Figure 3B As shown, one connector assembly 150 can correspond to one chamber 130 and one vertical microfluidic channel 143. The connector assembly 150 has through holes, and the number of through holes on a connector assembly 150 matches the number of intermediate connectable components corresponding to that connector assembly 150. For example... Figure 3B As shown, connector assembly 150A corresponds to chamber 130 and vertical microfluidic channel 143, and has two through holes; connector assembly 150B corresponds to upper microfluidic channel 141, and has one through hole.

[0114] The connector assembly 150 can be detachably fixed to the substrate 110, or it can be non-detachably fixed to the substrate 110 by means of adhesive bonding, chemical bonding, etc.

[0115] As previously mentioned, when the substrate comprises multiple substrate layers, the openings of the chamber 130, upper microfluidic channel 141, and vertical microfluidic channel 143 in the intermediate connectable components are typically formed by through-holes formed in the top layer 111. One opening may correspond to one or more intermediate connectable components, and the projection of the opening onto the substrate 110 includes the union of the projections of its corresponding intermediate connectable component onto the substrate 110. Figure 3C As shown, chamber 130 corresponds to a chamber opening, the size of which matches the size of the through-hole forming the chamber. Vertical microfluidic channel 143 corresponds to a channel opening, the size of which matches the size of the through-hole forming the vertical microfluidic channel 143. That is, chamber 130 and vertical microfluidic channel 143 each correspond to an opening. Connector assembly 150 has two through-holes, corresponding to both chamber 130 and vertical microfluidic channel 143. In this case, connector assembly 150 can be fixed to the upper surface of top layer 111, and the bottom surface of connector assembly 150 contacts the upper surface of top layer 111. The two through-holes of connector assembly 150 communicate with the chamber opening and channel opening of top layer 111, respectively, and further communicate with the corresponding intermediate connectable component. At this point, the through-holes, openings, and intermediate connectable component of connector assembly 150 correspond one-to-one. Figure 3B As shown, the size of the opening can be basically matched with the size of the outer contour of the bottom surface of the connector assembly 150. Thus, when the connector assembly 150 is fixed to the substrate 110, it can be inserted into the opening of the top layer 111. The bottom surface of the connector assembly 150 contacts the upper surface of the intermediate layer 112, and the through-hole of the connector assembly 150 directly communicates with the corresponding intermediate connectable component. This makes it easier to position the connector assembly 150 on the substrate 110 and makes the connection between the connector assembly 150 and the substrate 110 more stable. For example, the correspondence between the connector assembly, the opening, and the intermediate connectable component can be such that the through-hole of the connector assembly 150 corresponds one-to-one with the intermediate connectable component, and the connector assembly 150 corresponds one-to-one with the opening. One connector assembly includes multiple through-holes, and one opening can correspond to multiple intermediate connectable components. Figure 3B The connector assembly 150 corresponds to the chamber 130 and the vertical microfluidic channel 143, and the opening also corresponds to the chamber 130 and the vertical microfluidic channel 143. The connector assembly 150 has through holes that correspond to the chamber 130 and the vertical microfluidic channel 143 respectively. The size of the opening matches the size of the outer contour of the bottom surface of the connector assembly 150, so that the connector assembly 150 can be inserted into the opening.

[0116] For example, the connector assembly 150 may be of various types to meet different needs, and the types of the connector assembly 150 may include:

[0117] A single-hole connector assembly 1501, i.e., a connector assembly 150, has one through hole. For example, the size of the through hole of the connector assembly matches the size of its corresponding intermediate connectable component. If the intermediate connectable component corresponding to the through hole of the connector assembly is a chamber 130, then the through hole size matches the size of the through hole forming the chamber 130; if the intermediate connectable component corresponding to the through hole of the connector assembly is a vertical microfluidic channel 143, then the through hole size matches the size of the through hole forming the vertical microfluidic channel 143; if the intermediate connectable component corresponding to the through hole of the connector assembly is an upper microfluidic channel 141, then the through hole size can match the width of the upper microfluidic channel 141, for example, 1.2 times the width of the upper microfluidic channel 141. For the cavity formed by the single-hole connector assembly 1501 and the chamber 130, its inlet and outlet may include: the through hole of the single-hole connector assembly 1501, and inlet and outlet ports (if any) opened on the side wall of the chamber and communicating with the microfluidic channels. Understandably, the material may include gaseous and / or liquid phase materials. The material can enter or exit through the inlet / outlet connecting the chamber to the microfluidic channel, or through the through-hole of the connector assembly 150. This applies to all types of connector assemblies, and will not be elaborated further below. Understandably, for the single-hole connector assembly 1501, the material from the inlet / outlet connecting the chamber 130 to the microfluidic channel often originates from upstream or downstream chambers, while the material from the through-hole of the connector assembly 150 often originates from the outside environment. Figure 3B The 150B is a single-hole connector assembly.

[0118] The multi-hole connector assembly 1502 is a connector assembly 150 with multiple through holes, each corresponding to a different intermediate connectable component. Similar to a regular connector assembly 150, the dimensions of the through holes in the multi-hole connector assembly match the dimensions of their corresponding intermediate connectable components, which will not be elaborated further. Figure 3B The 150A is a multi-hole connector assembly.

[0119] For example, the porous connector assembly 1502 has two through holes, corresponding to the chamber 130 and the vertical microfluidic channel 143 respectively. The chamber 130 and the vertical microfluidic channel 143 are interconnected through the upper microfluidic channel 141 or the lower microfluidic channel 142. This effectively adds an extra opening to the chamber 130 for material feeding and discharging, expanding the inlet and outlet of the chamber 130. This allows the chamber 130 to simultaneously feed two materials, simultaneously discharge two materials, or simultaneously feed one material and discharge another, thus providing greater flexibility and convenience for the reaction. Figure 3BAs shown, the inlet and outlet ports of the cavity formed by the large through-hole and the chamber 130 in the porous connector assembly 150A include: two through-holes of the porous connector assembly 150A, and an interface 133A opened on the side wall of the chamber 130, which communicates with the lower microfluidic channel 142 on the left, for a total of three inlet and outlet ports. The interface 133B opened on the side wall of the chamber 130, which communicates with the lower microfluidic channel 142 on the right, belongs to the same flow path as the small through-hole in the porous connector assembly 150A and will not be calculated again. In practical applications, when the cavity is filled with liquid, air can be introduced into the vertical microfluidic channel 143. The gas enters the cavity connected by the vertical microfluidic channel 143 through the lower microfluidic channel 142. In this way, gas can enter the cavity from the bottom and create bubbles in the cavity, better simulating the blowing and stirring actions in a conventional laboratory reaction system. In addition, compared to the cavity and vertical microfluidic channel 143 being interconnected through the upper microfluidic channel 141, when the cavity and vertical microfluidic channel 143 are interconnected through the lower microfluidic channel 142, air and liquid can be introduced into the bottom corner of the cavity, causing the liquid level in the cavity to rise slowly and allowing the reactants to be fully soaked.

[0120] Stacked connector assembly 1503, such as Figure 4A and Figure 4B As shown, the laminated connector assembly 1503 may include an upper connector assembly 1503A and a lower connector assembly 1503B, with the lower connector assembly 1503B fixed at the opening 1142. The lower connector assembly 1503B has a through hole, and the upper connector assembly 1503A is detachably fixed in the through hole of the lower connector assembly 1503B, for example, the upper connector assembly 1503A and the lower connector assembly 1503B are threaded together.

[0121] The upper connector assembly 1503A can press and position the solid reactants in the cavity, or it can expand the reaction system. Furthermore, by replacing the upper connector assembly 1503A with different through-hole diameters, cavities with different reaction volumes can be obtained.

[0122] For example, such as Figure 4A As shown, the upper connector assembly 1503A can be cylindrical. For cases requiring pre-embedding of reactants, the upper connector assembly 1503A can be disassembled, the reactant to be pre-embedded can be placed into the chamber 130, and then the upper connector assembly 1503A can be reinstalled. The bottom end of the upper connector assembly 1503A can press and position the pre-embedded reactant. For example, when the upper connector assembly 1503A is fixed in the through hole of the lower connector assembly 1503B, the upper connector assembly 1503A can be tightened to press and position the pre-embedded reactant. For example, for reactions requiring venting, a vent hole 1503C can be provided on the upper connector assembly 1503A. For example, a second countersunk hole 1503D can also be provided on the top of the upper connector assembly 1503A as a buffer for overflow from the chamber.

[0123] For example, such as Figure 4B As shown, the upper connector assembly 1503A can be cylindrical. The through hole of the upper connector assembly 1503A and the corresponding chamber 130 of the stacked connector assembly 1503 form a cavity. Furthermore, by replacing the upper connector assembly 1503A with different through hole diameters, the volume of the cavity can be adjusted.

[0124] For the cavity formed by the laminated connector assembly 1503 and the chamber 130, its inlet and outlet may include: a through hole (if any) in the connector assembly 150, and an inlet and outlet opened on the side wall of the chamber that communicates with the microfluidic channel 140.

[0125] The filter connector assembly 1504 has a through hole corresponding to the chamber 130. This through hole and the chamber 130 form a cavity. A filter section is disposed within the cavity. The filter section isolates a second subspace 132 within the cavity formed by the through hole of the filter connector assembly 1504 and the chamber 130. The second subspace 132 can only communicate with other spaces in the cavity through the second subspace inlet 136. The second subspace inlet 136 is formed at the top of the filter section and is located at a preset height within the through hole of the filter connector assembly 1504 corresponding to the chamber 130. The outlet 1331 of the chamber 130, which communicates with the microfluidic channel 142, is located within the second subspace 132. Only the supernatant above the preset height in the cavity can enter the second subspace 135 through the second subspace inlet 136 and be discharged through the cavity outlet 1331. Impurities will settle and remain in the cavity, thus achieving filtration within the cavity. The discharge port 1331, which connects the cavity and the microfluidic channel 140, is usually the discharge port that connects the cavity and the lower microfluidic channel 142. It is located at the bottom of the middle layer 112. In order to make the discharge port 1331 located in the second subspace 135, the bottom end of the filter section can contact the bottom layer.

[0126] If a filter connector assembly is installed in chamber 130, the product in the cavity formed by the chamber and the through hole of the connector assembly can be filtered through the filter section formed by the abutment of the connector filter section 1504A and the chamber filter section 131. It is understood that when the connector filter section 1504A and the chamber filter section 131 form the filter section, their shapes and dimensions, except for height, are the same, so that when the filter connector assembly 1504 is fixed to the corresponding opening of chamber 130, the top end of the chamber filter section 131 and the bottom end of the connector filter section 1504A can abut. It is understood that at this time, the first subspace inlet 134 no longer exists, the second subspace inlet 136 becomes the only inlet of the second subspace 132, and the outlet 1331 (which is also the outlet of the first subspace when the filter connector assembly is not installed) is the only outlet of the second subspace. The first subspace is a part of the second subspace.

[0127] It is worth noting that the outlet 1331, which connects the cavity and the microfluidic channel 140, is located inside the second subspace 135, while the inlet 1332, which connects the cavity and the microfluidic channel 140, is located outside the second subspace 135. For example, the inlet 1332 and the second subspace 135 may be located on opposite sides of the cavity.

[0128] For example, the connector filter portion 1504A can extend from the filter connector assembly 1504 within the through hole corresponding to the chamber 130 to the bottom end of the filter connector assembly at a preset height. The height of the chamber filter portion 131 can be the same as the height of the chamber 130, and the two can form a filter portion extending from the preset height to the bottom layer. For example, the connector filter portion 1504A can extend from the filter connector assembly 1504 within the through hole corresponding to the chamber 130 to below the bottom end of the filter connector assembly, and the height of the chamber filter portion 131 is correspondingly shortened, so that the two can still form a filter portion extending from the preset height to the bottom layer. For example, as... Figure 6A As shown, the filter connector assembly 1504 has a connector filter section 1504A in the through hole corresponding to the chamber, and a chamber filter section 131 is provided in the chamber. When the filter connector assembly 1504 is fixed to the substrate 110, the connector filter section 1504A and the chamber filter section 131 abut against each other to form a filter section. For example, as shown... Figure 6B As shown, a connector filter section 1504A extending to the bottom of the chamber may be provided only in the filter connector assembly 1504, and the connector filter section 1504A may serve as the filter section alone. Alternatively, a chamber filter section 131 extending to a preset height may be provided only in the chamber 130, and the chamber filter section 131 may serve as the filter section alone.

[0129] Understandably, as previously described, chamber 130 and chamber filter 131 can be used to filter products in chamber 130 if the filter connector assembly is not installed in chamber 130.

[0130] The filter section can have various shapes, such as Figure 6A As shown, the filter section can be a cylindrical structure that is approximately tangent to the cavity, and can independently form a second subspace; as Figure 6B and Figure 1G As shown, the filter section can also be a gate-shaped column or a semi-cylindrical structure, forming a second subspace together with the inner wall of the cavity.

[0131] It is understood that the connector assembly 150 can belong to multiple types of connector assemblies mentioned above. For example, the connector assembly 150 can be both a porous connector assembly 1502 and a filter connector assembly 1504, with a filter section provided in the through hole of the connector assembly 150 corresponding to the chamber 130. Figure 5As shown in Figure 6, exemplarily, in a connector assembly 150 that belongs to both the porous connector assembly 1502 and the filter connector assembly 1504, the connector assembly 150 includes a through hole corresponding to the chamber 130 and a through hole corresponding to the vertical microfluidic channel 143, which are connected by a lower microfluidic channel 142. The interface between the lower microfluidic channel 142 and the chamber 130 is an inlet 1332. The filter is disposed in the corresponding through hole of the chamber 130. A second subspace 135 is located on the opposite side of the inlet, and an outlet 1331 is located within the second subspace 135. Exemplarily, the inlet 136 of the second subspace is located at approximately 5 / 9 of the height of the through hole of the connector assembly. Exemplarily, the outer contour of the connector assembly 150 is generally teardrop-shaped.

[0132] For example, a product exporter 160 protruding from the lower surface of the substrate 110 can be provided on the lower surface of the substrate 110 bottom layer 113 to export the product. The substrate 110 bottom layer 113 has a channel opening 1142 corresponding to the vertical microfluidic channel 143 or the lower microfluidic channel 142. The product exporter 160 has a through hole. When the product exporter 160 is fixed to the lower surface of the substrate 110 bottom layer 113, the through hole of the product exporter 160 and the channel opening 1142 for exporting the product are aligned, thereby exporting the product to a product collection container (e.g., a perforated plate) disposed below the substrate 110. For example, the product exporter 160 has an inverted frustum structure, and various types of needles can be inserted into its through hole. When the product exporter 160 is inserted into the product collection container, the needle punctures the film covering the product collection container and introduces the product into the product collection container. Understandably, the lower surface of the bottom layer 113 may have a countersunk hole to position the product guide 160 on the lower surface, and the size of the countersunk hole matches the outer contour size of the product guide 160.

[0133] The chip section 100 may further include a displacement component 300, which has an output terminal 310. The displacement component 300 is disposed above the substrate 110, its position corresponding to the position of the elastic membrane 180. Its output terminal 310 can switch between a first height and a second height. When the output terminal 310 is at the first height, it presses against the corresponding elastic membrane 180, causing the elastic membrane 180 to deform in the pressing direction and block the flow control channel 144 corresponding to the elastic membrane 180. When the output terminal 310 is at the second height, it no longer presses against the corresponding elastic membrane 180, the deformation of the elastic membrane 180 recovers, and it no longer blocks the flow control channel 144 corresponding to the elastic membrane 180. For example, the output terminal 310 is spherical or hemispherical in shape, adapted to the matching pattern of the flow control channel at the through-hole 113. For example, the displacement assembly 300 includes an electromagnet and an output end 310, with the output end 310 fixedly connected to the moving iron core of the electromagnet; or, the displacement assembly 300 includes a lead screw nut and an output end 310, with the output end 310 fixedly connected to the nut.

[0134] For example, such as Figure 8 As shown, besides the output end 310, the displacement assembly 300 includes a stationary part 320 and a moving part 330. The stationary part 320 is used to fix the displacement assembly 300 (for example, the displacement assembly 300 is fixed to the clamp part 200 by the thread 3201 of the stationary part). The output end 310 is fixedly connected to the lower end of the moving part 330. The moving part 330 can move in the height direction relative to the stationary part 320 under the control of a control signal, thereby driving the output end 310 to switch between a first height and a second height. For example, the displacement assembly 300 includes a coil, a stationary iron core, and a moving iron core. The stationary iron core is the stationary part 320, and the moving iron core is the moving part 330. The output end 310 is fixedly connected to the lower end of the moving iron core. It should be noted that, for ease of demonstration, Figure 8 The top layer 111 is not shown in the chip section 100.

[0135] For example, the fixed connection between the output terminal 310 and the lower end of the moving part 330 includes: a detachable connection between the output terminal 310 and the lower end of the moving part 330, for example, via a threaded connection, which facilitates the replacement of different output terminals 310 according to the needs of the chip part 100. The threaded connection between the output terminal 310 and the lower end of the moving part 330 can be such that the output terminal 310 has an internal thread and the moving part 330 has an external thread, such as... Figure 9A As shown, or the output end 310 has an external thread and the moving part 330 has an internal thread, such as Figure 9B As shown. Compared to Figure 9A The proposed scheme Figure 9B The proposed solution reduces the size and weight of the output end 310, thus reducing the load. Furthermore, it minimizes the possibility of inconsistent screw-in depths when the internal thread is screwed into the external thread, making it easier to ensure the uniformity of the height of the output ends 310 across multiple displacement components 300. Additionally, the output end 310 can be made of stainless steel, which offers higher strength at a microscale, is easier to process, and has lower costs.

[0136] like Figure 8 , Figure 9A and Figure 9B As shown, the stationary portion 320 is sleeved on the outer periphery of the moving portion 330, and the moving portion 330 extends out of the stationary portion 320 from below. The displacement assembly 300 also includes an elastic element fixing portion 340 and an elastic element 350. The elastic element fixing portion 340 is circumferentially disposed on the portion of the moving portion 330 that extends out of the stationary portion 320; for example, the elastic element fixing portion 340 is a retaining ring. The elastic element 350 is sleeved on the outer periphery of the moving portion 330, and both ends of the elastic element 350 are located between the elastic element fixing portion 340 and the lower surface of the stationary portion 320; for example, the elastic element 350 is a spring.

[0137] When the output end 310 is at the first height, the elastic element 350 is in a first compressed state; when the output end 310 is at the second height, the elastic element 350 is in a second compressed state; wherein the compression amount in the first compressed state is less than that in the second compressed state. For example, when the coil is de-energized, the stationary iron core has no attraction to the moving iron core, and the output end 310 is at a third height lower than the first height. At this time, the chip part 100 is placed below the output end 310, and the output end 310 is lifted to the first height by the chip part 100. Since the height of the stationary part 320 remains unchanged, the distance between the lower surface of the elastic element fixing part 340 and the stationary part 320 decreases, and the elastic element 350 is compressed, generating elastic force. Under the action of the elastic force, the output end 310 presses its corresponding elastic membrane 180, causing the elastic membrane 180 to deform in the pressing direction and block the flow path control channel 144 corresponding to the elastic membrane 180. When the coil is energized, the moving iron core moves upward under the attraction, causing the elastic element fixing part 340 to move upward and the output end 310 to rise to the second height. Since the height of the stationary part 320 remains unchanged, the distance between the lower surfaces of the elastic element fixing part 340 and the stationary part 320 is further reduced, and the elastic element 350 is further compressed. The output end 310 no longer presses its corresponding elastic membrane 180, and the deformation of the elastic membrane 180 is restored, no longer blocking the flow path control channel 144 corresponding to the elastic membrane 180. When the coil is de-energized again, the attraction between the moving iron core and the stationary iron core disappears, and the moving iron core will separate from the stationary iron core under the elastic force of the elastic element 350, causing the output end 310 to return to the first height. The output end 310 presses its corresponding elastic membrane 180, causing the elastic membrane 180 to deform in the pressing direction and block the flow path control channel 144 corresponding to the elastic membrane 180.

[0138] For example, the microfluidic chip also includes a clamping part 400 for clamping the elastic membrane 180 in the circumferential direction outside the through hole 115 (i.e., the through hole opened on the top layer as the channel opening of the flow path control channel 144), and the clamping part 400 has a through hole at the center for the output end 310 to pass through.

[0139] As mentioned earlier, the elastic membrane 180 can be a circular silicone sheet, fixedly connected to the first countersunk hole 116 on the top layer of the substrate 110 by means of adhesive bonding, bonding, etc. Understandably, when the elastic membrane 180 and the substrate 110 are made of different materials, they may not be able to connect securely. If the connection between the elastic membrane 180 and the substrate 110 is not secure enough, when the output end 310 of the displacement component 300 presses the elastic membrane 180, the elastic membrane 180 may easily detach from the substrate 110, causing leakage from the substrate. Therefore, a pressing part 400 can be provided, fixed to the upper surface of the elastic membrane 180, to press the elastic membrane 180 in the circumferential direction outside the through hole 115, especially at the connection point between the elastic membrane 180 and the substrate 110. This prevents the elastic membrane 180 from detaching from the substrate 110 and causing leakage when the output end 310 presses the elastic membrane 180 through the through hole of the pressing part 400. When leakage occurs at a single elastic membrane 180, the clamping part 400 can be used to clamp it, allowing the chip portion 100 of the microfluidic chip to continue to be used, rather than being scrapped due to the damage of one elastic membrane 180. Understandably, the clamping part 400 can be made of the same material as the substrate 110, so that the fixed connection between the clamping part 400 and the substrate 110 is sufficiently stable.

[0140] For example, refer to Figure 10A and Figure 10B The clamping part 400 includes:

[0141] The first pressing part 410 has a ninth through hole 411. The first pressing part 410 is fixed to the upper surface of the elastic membrane 180, so that the position of the ninth through hole 411 corresponds to the position of the through hole 115. The first pressing part 410 includes at least two annular protrusions 413 of different diameters arranged circumferentially outside the through hole 115, so that at least two layers of circumferential protection can be formed in the circumferential direction outside the through hole 115.

[0142] The second pressing part 420 has a tenth through hole 421. The second pressing part 420 is fixedly connected to the first pressing part 410 and applies downward pressure to the first pressing part 410. When the second pressing part 420 is fixedly connected to the first pressing part 410, the position of the tenth through hole 421 is aligned with the ninth through hole 411. It can be understood that the ninth through hole of the first pressing part 410 and the tenth through hole of the second pressing part 420 are aligned so that the output end of the displacement component 300 can pass through the ninth through hole 411 and the tenth through hole 421 to press the center of the elastic membrane 180. The second pressing part 420 is fixed above the first pressing part 410 and applies downward pressure to the first pressing part 410 to ensure that the first pressing part 410 presses the elastic membrane 180. In this way, when the center of the elastic membrane 180 is pressed by the output end 310 of the displacement component, the entire elastic membrane 180 will not detach from the first countersunk hole 116. For example, the second clamping part 420 is threadedly connected to the first clamping part 410.

[0143] For example, the first pressing part 410 is a cylindrical part, which includes a cylindrical wall 414 and a cylindrical bottom 412, and the cylindrical wall 414 and the cylindrical bottom 412 define the inner cavity of the first pressing part 410; the cylindrical bottom 412 of the first pressing part 410 is provided with a ninth through hole 411, and the cylindrical bottom 412 of the first pressing part 410 includes at least two annular protrusions 413 of different diameters that are circumferentially disposed outside the ninth through hole 411 and protrude toward the upper surface of the elastic membrane 180; the second pressing part 420 is threadedly connected to the first pressing part 410, and the second pressing part 420 is fixed in the inner cavity of the first pressing part 410.

[0144] For example, the first pressing part 410 has a groove 415 on its cylindrical wall 414, and the second pressing part 420 is threadedly connected to the first pressing part 410. The pressing part 400 also includes a third pressing part 430, which is engaged in the groove 415 and located between the first pressing part 410 and the second pressing part 420. The third pressing part 430 has an eleventh through hole 431, and when the third pressing part 430 is engaged in the groove 415, the position of the eleventh through hole 431 corresponds to the position of the ninth through hole 411. When the second pressing part 420 and the first pressing part 410 are threadedly connected, and downward pressure is applied to the first pressing part 410 in a rotating manner, in order to prevent the rotational force from being transmitted to the first pressing part 410, the third pressing part 430 can be provided between the first pressing part 410 and the second pressing part 420, so that the third pressing part 430 is fixed above the first pressing part 410 without rotation. For example, a groove 415 is provided in the cylindrical wall 414 of the first pressing part 410; the lug 432 of the third pressing part 430 is engaged in the groove 415, and the lower surface of the bottom of the third pressing part 430 contacts the upper surface of the cylindrical bottom 412 of the first pressing part 410. When the first pressing part 410 and the second pressing part 420 are screwed into place, the bottom of the second pressing part 420 contacts the top of the third pressing part 430. In this way, the downward pressure of the second pressing part 420 can be transmitted to the first pressing part 410, while the rotational force is not transferred.

[0145] Understandably, leakage from the substrate when the elastic membrane 180 is pressed can also be avoided by selecting a relatively low fluid pressure, selecting a more robust connection between the elastic element 350 and the substrate 110, or a combination of other means.

[0146] Based on the various possible combinations of substrate 110, connector assembly 150, elastic membrane 180, product outlet 160, displacement assembly 300, etc., as described above, a reaction that meets at least one of the following characteristics can be realized in the chip section 100: high throughput, controllable fluid flow direction, large reaction system, adjustable reaction system, need for filtration, need for pre-embedded reactants, and need for blowing operation.

[0147] The following explanation uses a microfluidic chip 100, which can be used for nucleic acid detection, as an example. Specific applications of nucleic acid detection include, for example, DNA testing.

[0148] refer to Figure 11The microfluidic chip's chip section 100 includes multiple functional regions: a lysis region 101, an extraction region 102, an amplification region 103, and a product export region 104. The lysis region 101 is used for cell lysis reactions, the extraction region 102 is used for extracting nucleic acids from the cell lysis products, the amplification region 103 is used for amplifying the nucleic acids, and the product export region 104 is used for exporting the amplified products to a product collection container, such as a well plate. It should be noted that, to clearly illustrate the distribution of components such as chambers and microfluidic channels, Figure 11 The image shows components that may actually be obscured. Figure 11 The upper connector assembly in the multilayer connector assembly of the amplification region is not shown.

[0149] The lysis region 101 may include N (e.g., N ≥ 24) independent cavities. Correspondingly, the extraction region 102 and the amplification region 103 each include N independent cavities. Each cavity consists of a substrate chamber and a connector assembly. The product export region 104 includes a channel opening for a product export channel communicating with the cavity of the amplification region 103. A product export component 160 communicating with the channel opening of the product export region 104 is provided on the lower surface of the substrate. The cavities of each functional region correspond one-to-one, and the upstream and downstream cavities are connected through a flow path control channel 144. The cavities of the lysis region, extraction region, and amplification region are composed of substrate chambers and connector assemblies. The connector assemblies of the lysis region, extraction region, and amplification region are respectively called the lysis connector assembly, extraction connector assembly, and amplification connector assembly.

[0150] For the chambers of pyrolysis zone 101, extraction zone 102, and amplification zone 103, the operating sequence includes: feeding according to the reaction flow, controlling reaction conditions, etc. After the reaction is completed, the flow path control channel 144 between the current chamber and the downstream chamber is opened, allowing the reaction product of the current chamber to enter the downstream chamber. For example, multiple chambers in the same reaction zone can be fed sequentially and react synchronously. The pyrolysis zone 101, extraction zone 102, and amplification zone 103 need to be set relatively far apart to ensure that the temperature of each functional zone is maintained at its required reaction temperature without affecting each other.

[0151] In the substrate of the chip section, an "overpass"-style microfluidic channel, flow path control channel, and intermediate connectable components that can be connected to the connector assembly are constructed to meet the needs of nucleic acid detection, such as high throughput, large system reaction, fluid control, heating and magnetic attraction of reactants, filtration, blowing and agitation, and pre-embedding of reactants.

[0152] For example, substrate 110 includes a top layer 111, an intermediate layer 112, and a bottom layer 113.

[0153] The components of each functional area are explained below with reference to Figure 12.

[0154] Figure 12AThe structure of the top layer 111 is shown. The top layer 111 has through-holes in the pyrolysis region 101, extraction region 102, and amplification region 103, serving as openings for interconnected components. Each opening in the pyrolysis region 101 and extraction region 102 corresponds to a chamber and a vertical microfluidic channel, while each opening in the amplification region 103 corresponds to a chamber. The size of the openings matches the outer contour of the connector assembly, allowing the bottom of the connector assembly to be inserted into the opening. Through-holes 115 are formed between the pyrolysis region 101 and extraction region 102, between the extraction region 102 and amplification region 103, and between the amplification region 103 and product export region 104, serving as channel openings for the flow path control channel 144. A first countersunk hole 116, with an area larger than the through-hole 115 and concentric with it, is also formed at the through-hole 115 on the top surface. When the elastic membrane 180 is placed in the first countersunk hole 116, the elastic membrane 180 is substantially flush with the upper surface of the substrate 110. The number of openings in each functional area is the same, and the number of through holes 115 and the number of first countersunk holes between each functional area are also the same.

[0155] Figure 12B and Figure 12C The three-dimensional patterns on the upper surface and lower surface of the intermediate layer 112 are shown respectively (the chamber filter section is not shown). Reference Figure 12B and Figure 12C In the pyrolysis zone 101 and the extraction zone 102, the intermediate layer has N larger through holes and N smaller through holes. The larger through holes form chambers 130, and the smaller through holes form vertical microfluidic channels 143. A combination of a larger through hole and a smaller through hole corresponds to the same opening and also to the same two-hole connector assembly. The chambers 130 and the vertical microfluidic channels 143 are connected through a lower microfluidic channel 142A formed on the lower surface of the intermediate layer. Figure 11 and Figure 5As shown in Figure 6, the connector assembly 150 of the pyrolysis zone is the two-hole connector assembly described above and is also a filter connector assembly. One through hole of the pyrolysis zone connector assembly 150 forms a cavity with the chamber 130, and the other through hole communicates with the vertical microfluidic channel 143. The vertical microfluidic channel 143 communicates with the chamber 130 through the lower microfluidic channel 142. Air can be vented into the vertical microfluidic channel 143 and enter the cavity 130 through the lower microfluidic channel 142, which can create a bubbling and blowing effect on the liquid sample in the cavity 130. The second subspace 136 enclosed by the filter section in the cavity also includes an outlet 1331 communicating with the lower microfluidic channel 142. The lower microfluidic channel 142 communicates with the flow path control channel 144 between the pyrolysis zone and the extraction zone through the outlet 1331, so as to export the pyrolysis products to the extraction zone at an appropriate time. The filter section in the chamber prevents solid waste or scum generated after the pyrolysis reaction in the pyrolysis zone from entering the downstream chamber, thus achieving a filtering effect. The pyrolysis zone chamber needs to be large enough to facilitate the placement of the sample to be tested, such as breaking a swab containing a biological sample before placing it in, and to form a reaction system that meets the requirements of the pyrolysis reaction. The chamber 130 is connected to another lower microfluidic channel on the opposite side of the lower microfluidic channel 142A, which is used to export the products in the chamber. This lower microfluidic channel is connected to the flow path control channel 144 between the pyrolysis zone 101 and the extraction zone 102. After the pyrolysis reaction is completed, the reaction products enter the chamber of the extraction zone 102 through the flow path control channel 144 between the pyrolysis zone 101 and the extraction zone 102.

[0156] The chamber 130 of the extraction zone 102 is also connected to the corresponding vertical microfluidic channel 143 through the lower microfluidic channel 142A to create a bubbling and blowing effect on the sample in the chamber. The chamber 130 of the extraction zone 102 is also connected to the lower microfluidic channel for introducing the product of the lysis zone 101 into the extraction zone chamber (understandably, this lower microfluidic channel connects the flow path control channel 144 between the lysis zone 101 and the extraction zone 102), and optionally connected to the lower microfluidic channel for exporting waste liquid in the extraction zone chamber, which is further connected to the vertical microfluidic channel and the connector assembly (a single-hole connector assembly) for exporting waste liquid to discharge waste liquid from the chip section 100. Figure 12C The diagram illustrates a one-to-one correspondence between the lower microfluidic channels and the chambers used to drain waste liquid from the extraction zone. This prevents waste liquid from flowing into other chambers and causing contamination via a shared waste liquid draining channel. Alternatively, multiple chambers can share a single microfluidic channel for draining waste liquid from the extraction zone, thus reducing space requirements. (Reference) Figure 11As shown in Figure 12, the connector assembly for waste liquid discharge can be located near the pyrolysis zone 101. It is understood that if the pyrolysis reaction process does not require waste liquid discharge, then a lower microfluidic channel for discharging waste liquid from the extraction zone cavity is unnecessary. After the extraction reaction is completed, the reaction products enter the cavity of the amplification zone 103 through the flow path control channel 144 between the extraction zone 102 and the amplification zone 103.

[0157] The cavity 130 of the amplification region 103 is connected to an upper microfluidic channel for introducing the product from the extraction region 102 into the extraction region cavity, and a lower microfluidic channel for exporting the product from the cavity 130. The connector assembly 150 of the amplification region is a multilayer connector assembly 1503. Figure 11 The upper connector assembly in the stacked connector assembly is not shown. As shown in Figure 4, before the reaction on the chip begins, the upper connector assembly 1503A can be removed, solid reactants can be placed into the cavity, and then the upper connector assembly 1503A can be fixed onto the lower connector assembly 1503B to complete the pre-embedding of the solid reactants. After the reaction product is extracted and enters the cavity of the amplification region 103, it can undergo an amplification reaction with the pre-embedded solid reactants if the reaction conditions are met. For example, the pre-embedded amplification solid reactants can be lyophilized pellets. The detachable connection between the upper connector assembly 1503A and the lower connector assembly 1503B allows the reactant pre-embedding to be performed after the chip fabrication is completed, avoiding the stringent requirement of completing the pre-embedding step simultaneously during chip fabrication. Furthermore, the pre-embedding operation can be performed under conditions that meet the requirements of the pre-embedded amplification solid reactants, such as in a sterile environment. After the amplification reaction is completed, the reaction product enters the product export region 104 through the flow path control channel 144 between the amplification region 103 and the product export region 104. It is understandable that the adapter assembly of the amplification region and the chamber constitute the amplification chamber used for amplification reactions. This can be interpreted broadly, as amplification reactants or products can enter the chamber, while adapter assemblies that do not enter the amplification region can enter.

[0158] The product export area 104 has multiple vertical microfluidic channels connecting to the upper microfluidic channels to export the product through the channel openings in the lower layer, such as... Figure 12D As shown in the top view of the bottom layer, the channel openings of the bottom layer are designed according to the specifications of the product collection container. Some vertical microfluidic channels have openings on the lower surface of the intermediate layer directly aligned with the channel openings; others have openings on the lower surface of the intermediate layer that are not directly aligned with the channel openings and require further guidance through the lower layer microfluidic channels to reach the channel openings. The product enters the product collection container from the channel openings via the product outlet 160. Figure 12E This is a three-dimensional image of the bottom layer.

[0159] By designing the pipe shape, multiple waste liquid outlet channels can have similar flow resistance, and appropriate microfluidic pipe sizes can be selected to reduce flow resistance. For example, widening the size of the flow path control channel 144 from 0.4mm*0.4mm to 0.6mm*0.6mm can reduce flow resistance by 80%. Flow resistance can also be further reduced by reducing straight-flow channels and adding curved channels.

[0160] The lysis, extraction, and amplification reactions all require heating. Considering that it's easier to operate the components for feeding and discharging materials above the chip section, and the displacement component 300 is already positioned above the chip section, the heating components for heating the cavities in the lysis, extraction, and amplification zones are positioned below the chip section, heating the cavities from below. In other words, the cavities and the microfluidic channels for feeding materials into the cavities to achieve bubbling and blowing effects need to be positioned on the lower layer of the substrate, while the flow control channel 144 needs to be positioned on the upper layer. Therefore, a "flyover" type chip section is designed to meet the requirement of forming microfluidic channels at different heights within the chip for nucleic acid detection reactions.

[0161] In some lysis reaction processes, a magnetic chuck is required to adsorb the magnetic beads used in the reaction. The magnetic chuck can be positioned below the chip section. For example, the magnetic chuck is used to adsorb tiny magnetic beads encapsulating sample DNA during the rinsing step of the lysis reaction process, preventing them from being washed into the waste liquid microfluidic channel by the rinsing solution. The effects of the heating and magnetic chuck components on the materials within the chamber can be controlled by switching the heating and magnetic chuck components on and / or their distance from the chamber.

[0162] To achieve precise alignment between the auxiliary components and the chip section and reduce the user's operational difficulty, the microfluidic chip in this embodiment of the application further includes a clamping section 200. The clamping section 200 can be used to perform at least one of the following functions: connecting external pipelines, supporting displacement components, providing a seal for the chip section, providing an interaction path between a heating component, a magnetic suction component, a product discharge container, and the chip section, thereby forming automatic alignment between the chip section 100 and other auxiliary components. The structure of the clamping section 200 will be described below.

[0163] The clamping part 200 includes an upper clamping plate 210, a lower clamping plate 220, and a clamping assembly 230 for connecting external pipelines. When the upper clamping plate 210 and the lower clamping plate 220 are closed, they form a receiving cavity, in which the chip part 100 is located.

[0164] like Figure 13AAs shown, the upper clamping plate 210 has a third through hole 211 penetrating through it. The stationary part 320 of the displacement assembly 300 is fixed in the third through hole 211, and the two can be connected by threads. When the displacement assembly 300 is fixed in the third through hole and the chip part 100 is located in the receiving cavity, the output end 310 of the displacement assembly 300 is directly opposite the center of the elastic membrane 180. When the output end 310 of the displacement assembly 300 is at the first height, it can press the elastic membrane 180 to deform it and completely block the corresponding flow path control channel 144. In this way, the automatic alignment of the displacement assembly 300 and the elastic membrane 180 when the chip part 100 is placed in the receiving cavity facilitates subsequent control of the flow path control channel 144 and greatly reduces the difficulty of operation.

[0165] For example, such as Figure 14 As shown, the upper clamping plate 210 also has a fourth through hole 213 penetrating through the upper clamping plate, and the clamping assembly 230 is fixed in the fourth through hole 213. For example, the lower half of the clamping assembly 230 is fixed in the fourth through hole 213, and the upper half protrudes from the upper surface of the upper clamping plate for communication with the external pipeline. The clamp assembly 230 has a through hole 231, which corresponds one-to-one with the through hole 151 of the connector assembly 150. When one end of the external pipeline is inserted into the through hole 231 of the clamp assembly 230 and the chip part 100 is located in the receiving cavity, the through hole 231 of the clamp assembly 230, the fourth through hole 213 of the upper clamping plate, the through hole 151 of the connector assembly 150, and the corresponding intermediate connectable component of the connector assembly 150 are automatically aligned. The external pipeline connects to the through hole of the connector assembly 150 through the through hole 231 of the clamp assembly 230 to feed and discharge the corresponding intermediate connectable component of the connector assembly 150. After replacing the chip part, simply placing the chip part into the receiving cavity will re-align the chip part and the components on the feed and discharge flow path and automatically connect the external pipeline fixed on the upper clamping plate, greatly reducing the user's operating difficulty. For example, the clamp assembly 230 is fixed to the upper clamping plate. This can mean that the clamp assembly 230 is integrally formed with the upper clamping plate, or it can mean that the clamp assembly 230 is separately formed with and fixedly connected to the upper clamping plate. When the clamp assembly 230 is fixed in the upper fourth through hole, the through hole 231 of the clamp assembly 230 is connected to the fourth through hole 213. For example, the fourth through hole 213 is a stepped hole. After the clamp assembly 230 is fixed in the large radius portion of the stepped hole 213, the through hole 231 of the clamp assembly 230 and the small radius portion of the stepped hole 213 are connected.

[0166] To reserve space for the displacement assembly 300 and the connector assembly 150 between the upper clamping plate 210 and the base plate 110, a first recess 212 can be formed on the lower surface of the upper clamping plate 210, such as... Figure 13B , Figure 14 and Figure 15 As shown, the projection of the third through hole onto the plane of the substrate is located within the projection of the first recess 212 onto the plane of the substrate.

[0167] For example, in order to make the seal between the upper clamping plate 210 and the connector assembly 150 tighter so as to control the fluid flow direction by the pressure within the system, the upper clamping plate 210 may also include a sealing gasket 214, which is disposed between the upper clamping plate 210 body and the connector assembly 150. The sealing gasket 214 has a through hole 2141 corresponding to the through hole 151 of the connector assembly 150. The through hole 2141 connects the through hole 151 of the connector assembly 150 and the fourth through hole 213. When one end of the external pipeline is inserted into the through hole 231 of the clamp assembly 230 and the chip part 100 is located in the receiving cavity, the through hole 231 of the clamp assembly 230, the fourth through hole 213 of the upper clamping plate, the through hole 2141 of the sealing gasket 214, the through hole 151 of the connector assembly 150, and the corresponding intermediate connectable component of the connector assembly 150 are automatically aligned. The external pipeline connects to the through hole of the connector assembly 150 through the through hole 231 of the clamp assembly 230 and the through hole 2414 of the sealing gasket 214 to feed and discharge the corresponding intermediate connectable component of the connector assembly 150. Typically, the sealing gasket 214 and the connector assembly 150 can be configured as needed; for example, three connector assemblies 150 can share two sealing gaskets 214. Since each cavity is independent, crosstalk will not occur even if the connector assemblies 150 share a single sealing gasket. The sealing gasket can have various shapes, such as... Figure 15 and Figure 16 As shown, the shape of the sealing gasket 214 can be the shape of the convex bulge formed by the outer contour of its corresponding connector assembly 150, and the shape of the sealing gasket 214 can be an irregular structure. For example, one connector assembly 150 of the pyrolysis zone 101 and an adjacent connector assembly 150 for discharging waste liquid share a common shape. Figure 16 The sealing gasket shown in the left figure; the two adjacent connector assemblies 150 in the extraction area 102 share a common one, as shown in the left figure. Figure 16 The sealing gasket shown in the middle figure; one connector assembly 150 of the amplification region 103 corresponds to one such as Figure 16 The sealing gasket is shown in the figure. The sealing gasket can be made of materials such as nitrile rubber, fluororubber, or silicone rubber, with a Shore hardness between 10 and 90. For example, silicone rubber with a Shore hardness of 30 is used. The thickness of the sealing gasket 214 can be selected according to the design of the chip section and the clamping plate section, as well as the elastic deformation of the sealing gasket 214, for example, 1.5 mm.

[0168] For example, in order to facilitate the positioning and installation of the sealing gasket 214, a second countersunk hole 215 matching the outer contour of the sealing gasket 214 can be opened on the lower surface of the upper clamping plate 210. When the sealing gasket 214 is located in the second countersunk hole 215 and the chip part 100 is placed in the receiving cavity, the sealing gasket 214 and its corresponding connector assembly 150 and the upper clamping plate body are in contact to seal the connector assembly 150.

[0169] For example, such as Figure 17 and Figure 18 As shown, the lower clamping plate 220 has a through slot, allowing the heating assembly 710 to approach or contact the lower surface of the chip section 100 to heat the bottom of the cavity that needs to be heated, and / or allowing the magnetic suction assembly 720 to approach or contact the lower surface of the chip section 100 to magnetically attract the material, and / or allowing the product outlet 160 to extend from the through slot into the perforated plate for collecting the product. Exemplarily, whether the heating assembly 710 and / or the magnetic suction assembly 720 act on the material in the cavity can be controlled by switching the heating assembly 710 and the magnetic suction assembly 720 on and / or by the distance between the heating assembly 710 and the magnetic suction assembly 720 and the cavity; the greater the distance, the smaller the effect. For example, the lower clamping plate 220 has a first through groove 221, a second through groove 222 and a third through groove 223. The heating components 710 of the pyrolysis zone 101 and the extraction zone 102 are fixed in the first through groove 221 by the fixing plate 240. The fixing plate is fixed below the first through groove 221. The magnetic suction component 720 is located below the fixing plate and on the lifting platform. The distance between the magnetic suction component 720 and the bottom of the cavity can be adjusted by the lifting platform. The heating component 710 of the amplification zone 103 is fixed in the second through groove 222. The product outlet 160 extends out of the third through groove 223 and into the product collection container located below the third through groove 223.

[0170] For example, such as Figure 19 As shown, the upper clamping plate 210 and the lower clamping plate 220 are pivotally connected so that the chip portion 100 can be placed into or removed from the receiving cavity.

[0171] For example, such as Figure 18 and Figure 20 As shown, the distance between the upper clamping plate 210 and the lower clamping plate 220 can be controlled by the lifting platform supporting the lower clamping plate, so that the chip part 100 can be placed into or removed from the receiving cavity. When the lower clamping plate 220 is at the third height, the distance between the upper and lower clamping plates is large enough that the chip part 100 can be placed on or removed from the lower clamping plate 220; when the lower clamping plate 220 is at the fourth height, the bottom surface of the upper clamping plate 210 and the top surface of the lower clamping plate 220 are in contact, and the two are closed to form the receiving cavity. For example, the upper clamping plate 210 and / or the lower clamping plate 220 are provided with locking devices for locking the two when the lower clamping plate 220 is at the fourth height. The upper clamping plate is equipped with a displacement component 300, external pipelines, etc. Frequent movement of the upper clamping plate is not conducive to system stability. In this embodiment, the upper and lower clamping plates are opened or closed by moving the lower clamping plate, which is beneficial to improving system stability.

[0172] In another aspect of the embodiments of this application, a microfluidic chip system is also provided, including the microfluidic chip described above, and further including:

[0173] An external pipeline 500 is connected at one end to a clamp assembly 230, and at the other end is directly or indirectly connected to a material container.

[0174] Valve assembly 600 is used to control whether the material container and the chamber in the chip section used for reaction are connected, for example, to control whether a certain one of a plurality of material containers and a certain one of a plurality of chambers are connected. According to its control function, the valve assembly may include at least one of a feed control valve, an exhaust control valve, and a drain control valve. According to its controlled functional area, the valve assembly may include a control valve for a pyrolysis zone, a control valve for an extraction zone, and a control valve for an amplification zone.

[0175] For example, the working mechanism of the valve assembly 600 is illustrated using the lysis region 101 of a microfluidic chip for nucleic acid detection as an example. During the lysis reaction, materials need to be fed from M lysis material containers into N chambers of the lysis region 101. For example, materials are first fed from the first material container into the first to N chambers sequentially, then from the second material container into the first to N chambers sequentially, until materials are fed from the last material container into the first to N chambers sequentially. Of course, materials can also be fed from the first to M material containers into the first chamber, then into the second chamber into the first to M material containers, and so on, until materials are fed from the first to M material containers into the Nth chamber. The former will be used as an example below. M pyrolysis material containers (including gaseous and liquid materials) correspond to M feed lines. A primary feed control valve is installed between the M feed lines and the intermediate line to control which of the M feed lines is connected to the intermediate line. For example, the primary feed control valve can be a multi-port valve or a combination of a multi-port connector and multiple solenoid valves. N external pipelines connect to the clamping assemblies corresponding to the vertical microfluidic channels connected to the N cavities (in the pyrolysis zone, the external pipelines connected to the clamping assemblies corresponding to the cavities are used for venting, and the external pipelines connected to the clamping assemblies corresponding to the vertical microfluidic channels connected to the cavities are used for feeding). A secondary feed control valve is installed between the intermediate line and the N external pipelines to control which of the intermediate line and the N external pipelines is connected. When the primary feed control valve connects the m-th feed line to the intermediate line, and the secondary feed control valve connects the intermediate line to the n-th feed line, the M-th feed container feeds into the N-th cavity.

[0176] Understandably, a primary feed control valve and a secondary feed control valve can be set for the pyrolysis zone 101, and a primary feed control valve and a secondary feed control valve can be set for the extraction zone 102. Alternatively, the pyrolysis zone 101 and the extraction zone 102 can share a primary feed control valve, while each can have its own secondary feed control valve. Separate valves avoid cross-contamination, while sharing valves reduces costs and saves space. For example, the primary feed control valve could be a 10-way valve, and the secondary feed valve could be a 24-way valve.

[0177] Understandably, if a cavity in a functional area only needs to be fed from a single feed container, then there is no need to set up a primary feed control valve or the primary control valve is only used for on / off control, and only the secondary feed control valve is retained.

[0178] During the pyrolysis reaction, exhaust gas also needs to be released from the N chambers of pyrolysis zone 101. For example, an exhaust control valve can be installed between the N external pipelines connected to the clamp assemblies corresponding to the N chambers and the exhaust terminal (such as a gas filter bottle or the atmosphere) to control which of the N external pipelines is connected to the exhaust terminal. The exhaust control valve can be, for example, a 24-way valve. Understandably, each functional area can have its own exhaust control valve.

[0179] During the extraction reaction, in addition to feeding and venting, waste liquid may also need to be discharged. For example, a discharge control valve can be installed between the N external pipelines connected to the clamp assemblies corresponding to the N waste liquid discharge connector assemblies and the discharge terminal (such as a gas washing bottle). This valve controls which of the N external pipelines is connected to the discharge terminal. The discharge control valve can be, for example, a 24-way valve.

[0180] Understandably, if the same external pipeline is used for at least two of the functions of feeding, venting, and draining, the corresponding functions can share a single control valve. For example, if the same external pipeline is used for both air intake and exhaust, its secondary feed control valve and exhaust control valve can share a single 24-way valve.

[0181] Understandably, for each multi-way control valve, a corresponding master valve can be set upstream or downstream to simultaneously control the on / off state of multiple channels.

[0182] For example, Figure 21 and Figure 22 Two different valve assembly setup schemes are provided, among which Figure 21A and Figure 21B The images show the valve assembly setup for feeding and the valve assembly setup for venting, respectively. Figure 21A In the elliptical section, a single line represents 24 parallel connections.

[0183] The processing assembly 700 includes a heating assembly 710 for heating a cavity in at least a portion of the functional area, a magnetic adsorption assembly 720 for adsorbing magnetic particles in a cavity in at least a portion of the functional area, and / or a product collection container 730 for collecting products generated by the chip section 100.

[0184] The controller 800 is used to control the displacement assembly 300, the valve assembly 600, and / or the processing assembly 700.

[0185] For example, controller 800 includes a displacement component controller to control the displacement component, thereby switching its output between a first height and a second height. It is understood that if N flow path control channels between two functional areas only need to be opened and closed simultaneously without individual control, then synchronous control of the displacement components controlling these N flow path control channels is sufficient, see reference... Figure 23 The illustrated displacement assembly includes a displacement assembly controller for an electromagnet. Controller 800 may include a valve controller for controlling a valve assembly. Controller 800 may include a heating assembly controller for controlling the switching and / or temperature of the heating assembly. Controller 800 may include a magnetic attraction assembly controller for controlling the switching and / or lifting of the magnetic attraction assembly. Controller 800 may include a product collection container for controlling the lifting of the product collection container.

[0186] For example, such as Figure 24 As shown, the microfluidic chip system also includes a support 900, which, from top to bottom, includes: a power circuit layer support 910 for supporting the power supply and control circuit 800; a valve assembly layer support 920 for supporting the valve assembly 600; a gas-liquid layer support 930 for supporting the external pipeline 500; a clamping plate support 940 for supporting the upper clamping plate 210 and the lower clamping plate 220; and a processing component support 950 for supporting at least part of the processing components 700. The support 900 may also include a housing. For example, some processing components, such as heating components, are fixedly connected to the lower clamping plate and move with the movement of the lower clamping plate. Some processing components, such as magnetic components, are supported on the processing component support 950. For example, the clamping plate support 940 for supporting the upper clamping plate 210 and the lower clamping plate 220 can drive the lower clamping plate 220 to rise and fall. This layered design can fully isolate the circuitry and the gas-liquid path, avoiding the risk of short circuits caused by liquid leakage.

[0187] The following describes some of the operating procedures. When feeding into the extraction zone chamber, the vent is open. Theoretically, the waste liquid microfluidic channel should not be open, but in practice, to prevent cross-contamination between samples within the extraction zone chamber, the drain valve assembly can be opened, and a slight positive pressure is introduced into the waste liquid microfluidic channel. When pushing the reaction product from the pyrolysis zone chamber into the extraction zone chamber, the vent valve assembly of the pyrolysis zone chamber is closed, and the drain valve assembly of the extraction zone is opened, with a slight positive pressure introduced. When pushing the reaction product from the extraction zone chamber into the amplification zone chamber, the vent valve assembly of the extraction zone is closed, and the drain valve assembly is open to prevent cross-contamination between different samples. When pushing liquid from the amplification zone chamber into the product export zone, the amplification zone chamber is vented to expel the liquid. During the pyrolysis reaction, the vent valve assembly of the pyrolysis zone chamber is open. During most of the reaction within the extraction zone chamber, the vent valve assembly of the extraction zone chamber is open, the drain valve assembly is open, and a slight positive pressure is introduced to prevent cross-contamination between samples. During the drainage operation in the extraction zone chamber, the vent valve assembly is closed, while the drain valve assembly is open and a slight negative pressure is applied to facilitate waste liquid drainage and prevent cross-contamination of waste liquid from different samples. When venting from the extraction zone chamber, the drain valve assembly is open and a slight positive pressure is applied to prevent sample cross-contamination. Conversely, the vent valve assembly must be closed when draining from the extraction zone chamber.

[0188] Furthermore, embodiments of this application also provide an application of the microfluidic chip of the first aspect of this application or the microfluidic chip system of another aspect in nucleic acid detection.

[0189] Combined with the valve assembly in Figure 21 and Figure 23The control components are described below. Specifically, during the pyrolysis reaction, after the sample to be tested is placed into the cavity of the pyrolysis zone, firstly, the first valve inlet of the first gate valve is controlled to connect the container of reactants required for the pyrolysis reaction to the first valve outlet. The first valve outlet is connected to the second valve inlet of the second gate valve corresponding to the pyrolysis reaction. The second valve outlet of the second gate valve corresponding to the pyrolysis reaction sequentially connects to cavity 1, cavity 2, ..., cavity N of the pyrolysis zone until reactants are added to each cavity of the pyrolysis zone. Then, the first valve inlet of the first gate valve is controlled to connect to the air pump and the first valve outlet, thereby sequentially generating air bubbles into cavity 1, cavity 2, ..., cavity N of the pyrolysis zone. At the same time, the third valve outlet of the third gate valve corresponding to the cavity of the pyrolysis zone is connected to the third valve inlet of the cavity of the pyrolysis zone that is generating air bubbles, so as to facilitate the exhaust of the air bubbles and the gas generated by the reaction. Finally, after the pyrolysis reaction is completed, the third valve outlet of the third selector valve corresponding to the cavity of the pyrolysis zone stops selecting any third valve inlet. The first control component controls the displacement component corresponding to the first flow path control channel between the cavity of the pyrolysis zone and the cavity of the extraction zone to work, so that the output end of the displacement component is at the second height and no longer blocks the first flow path control channel. The first selector valve continues to select the gas pump and the first valve outlet, and the second selector valve sequentially selects the second valve inlet and the second valve outlet. The introduced gas is used to force the reactants in the cavity of the pyrolysis zone into the cavity of the corresponding extraction zone through the first flow path control channel. Each cavity of the pyrolysis zone corresponds to a cavity of the extraction zone. After the reactants are forced into the cavity of the extraction zone, the first control component controls the displacement component corresponding to the first flow path control channel between the cavity of the pyrolysis zone and the cavity of the extraction zone to work, so that the output end of the displacement component is at the first preset height to block the first flow path control channel.

[0190] During the extraction reaction, after the reactants in the pyrolysis zone are introduced into the extraction zone, firstly, the first valve inlet of the first selector valve is controlled to connect the container of reactants required for the extraction reaction to the first valve outlet. The first valve outlet is connected to the second valve inlet of the second selector valve corresponding to the extraction reaction. The second valve outlet of the second selector valve corresponding to the extraction reaction sequentially connects to extraction zone cavity 1, extraction zone cavity 2, ..., extraction zone cavity N, until reactants are added to each extraction zone cavity. Then, the first valve inlet of the first selector valve is controlled to connect the air pump and the first valve outlet, thereby sequentially generating air bubbles into extraction zone cavity 1, extraction zone cavity 2, ..., extraction zone cavity N. At the same time, the third valve outlet of the third selector valve corresponding to the extraction zone cavity is connected to the third valve inlet of the extraction zone cavity that is generating air bubbles, facilitating the extraction of the generated gas and the gas produced by the reaction. Finally, after the extraction reaction is completed, the third valve outlet of the third gate valve corresponding to the cavity of the extraction zone stops selecting any third valve inlet. The first control component controls the displacement component corresponding to the second flow path control channel between the cavity of the extraction zone and the cavity of the amplification zone to work, so that the output end of the displacement component is at the second height and no longer blocks the second flow path control channel. The first gate valve continues to select the gas pump and the first valve outlet, and the second gate valve sequentially selects the second valve inlet and the second valve outlet. The introduced gas is used to force the reactants in the cavity of the extraction zone into the cavity of the corresponding amplification zone through the second flow path control channel.

[0191] During the amplification reaction, each extraction chamber corresponds to an amplification chamber. When the amplification reaction occurs within the amplification chamber, firstly, the inlet and outlet of the third valve corresponding to each amplification chamber are sequentially connected to expel the gas generated during the reaction. After the amplification reaction is complete, the connection between the third valve inlet and outlet connected to each amplification chamber is stopped. Then, the first control component controls the displacement component corresponding to the third flow path control channel between the amplification chamber and the product outlet chamber, ensuring that the output of the displacement component is at a second preset height, no longer blocking the third flow path control channel. The second valve inlet of the second selection valve corresponding to the extraction chamber remains connected to the gas pump, sequentially pumping gas into the amplification chamber through the second valve outlet, the extraction chamber, and the second flow path control channel. This forces the reaction product within the amplification chamber into the third flow path control channel, and then out through the product outlet chamber to the designated location.

[0192] Furthermore, in order to facilitate the timely discharge of waste liquid from the waste liquid recovery chamber, in other embodiments based on the above embodiments, the microfluidic chip system further includes: a first negative pressure suction device and a gas washing bottle; the external pipeline also includes a waste pipeline, the waste pipeline is connected to the fourth through hole of the clamp assembly for connecting the waste pipeline, the gas washing bottle is connected to the waste pipeline, and the first negative pressure suction device is connected to the gas washing bottle.

[0193] Specifically, the gas in the gas washing bottle is drawn out by the first negative pressure suction device, so that the gas pressure in the gas washing bottle is lower than the gas pressure in the waste liquid outlet chamber. Under the action of gas pressure, the waste liquid in the waste liquid outlet chamber is discharged into the gas washing bottle through the third selector valve 81, thus completing the recovery of the waste liquid in the waste liquid outlet chamber.

[0194] The following example uses a 10-way rotary valve as the first selector valve, 24-way valves as the second and third selector valves, a dual-way solenoid valve as the first and second on / off control valves, an electromagnet as the displacement component, and a single connector assembly for waste liquid discharge shared by the six extraction chambers, in conjunction with Figure 21. Figure 23 This paper describes the specific operating steps of the microfluidic chip and microfluidic chip system in the embodiments of this application when applied to nucleic acid detection.

[0195] The chip consists of four main biochemical functional areas:

[0196] A. Pyrolysis Zone: Samples and their corresponding inlets / outlets are numbered 1–24, denoted by A1, A2, …; B. Extraction Zone: Samples and their corresponding inlets / outlets are numbered 1–24, denoted by B1, B2, …; C. Amplification Zone: Samples and their corresponding inlets / outlets are numbered 1–24, denoted by C1, C2, …; D. Product Zone: Samples and their corresponding inlets / outlets are numbered 1–24, denoted by D1, D2, … The three sets of fluid control channels between the pyrolysis and extraction zones, between the pyrolysis and amplification zones, and between the amplification and product export zones on the chip are represented by the Greek letters α, β, and γ, respectively. The electromagnets and control areas of each region are denoted by α1–24, β1–24, and γ1–24, respectively.

[0197] 1. Preparation: Confirm all electromagnets are de-energized; confirm the pressure controller, power supply, and electromagnet controller are off; confirm all two-way solenoid valves are closed; confirm the 10-way rotary valve outlet is turned to #10 neutral; confirm all 24-way rotary valves are initially de-energized and set to #1 (for sample 1). Confirm all heating plates are not heated and electromagnets are not raised. For the chip amplification area cavity, embed the lyophilized bulb and tighten the mating components in the amplification area cavity. In the following steps, electromagnets, rotary valves, etc., not mentioned in the next step, should remain in the on / off state as in the previous step.

[0198] 2. Pyrolysis:

[0199] Step 1. Take a sample swab, cut it (length 15mm~20mm, calculated as 18mm, diameter 5mm cylinder, volume approximately 353.25ul) and insert it into the pyrolysis reaction chambers A1-A24 in sequence.

[0200] Introduce lysis buffer and protease into A1.

[0201] Step 2. Power on (open) the following valves in sequence: dual-way solenoid valve 3, 24-way rotary valve #3 (initially set to #1), 24-way rotary valve #1 (initially set to #1), dual-way solenoid valve 1, pressure controller, flow meter (or flow meter), and turn the 10-way rotary valve to #2. At this point, the gas flow path for sample #1 is open, and the lysis buffer and protease can be pumped in.

[0202] Step 3. Using a flow meter or velocity meter, pump 500 μL of a mixture of lysis buffer and proteinase K (mixing ratio 40:1) into the A1 inlet.

[0203] Next, lysis buffer + proteinase K is introduced into A2.

[0204] Step 4.10: Turn the rotary valve to #10 (cut off the airflow first), then turn the #3 24-way valve to #2, and the #1 24-way valve to #2. The airflow path for sample #2 is now open, and the lysis buffer and protease can be pumped in.

[0205] Step 5.10 Turn the rotary valve to #2, and pump 500ul of lysis buffer and proteinase K mixture (mixing ratio 40:1) into the A2 inlet by reading the flow meter or timing the flow rate.

[0206] Next, lysis buffer and proteinase K are introduced into A3.

[0207] Step 6.10: Turn the rotary valve to #10 neutral (cut off the airflow first), then turn the #3 24-way valve to #3, and the #1 24-way valve to #3. The airflow path for sample #3 is now open, and the lysis buffer and protease can be pumped in.

[0208] Step 7.10 Turn the rotary valve to #2, and pump 500ul of lysis buffer and proteinase K mixture (mixing ratio 40:1) into the A3 inlet by reading the flow meter or the flow rate meter.

[0209] Next, lysis buffer and proteinase K are introduced into A4-A24.

[0210] Step 8. By switching between valve 3 (24-way), valve 1 (24-way), and rotary valve 10, lysis buffer + proteinase K solution is injected into the cavity of the A4-24 lysis zone.

[0211] Next, heating and bubbles are used to promote the pyrolysis reaction.

[0212] Step 9. Up to the previous step, all the cavities in the lysis zone are filled with reaction solution and proteinase K, but only the gas path of sample A24 is connected.

[0213] First, turn the 10-way rotary valve to the #10 neutral position to cut off the gas supply. Then, turn on the heating plate in the pyrolysis zone and raise the temperature to 25 degrees Celsius, maintaining this temperature.

[0214] Step 10.1: Turn valve #1 (24-way valve) to #1. Simultaneously, turn valve #3 (24-way valve) to #1, and then turn the 10-way rotary valve to the #1 clean air channel. Pump clean air into the A1 inlet, causing bubbles to rise from the bottom of the cavity in the A1 pyrolysis zone, promoting the A1 pyrolysis reaction. Continue for 10-30 seconds.

[0215] Step 11. Immediately afterwards, turn valve #1 (24-way valve) to #2, and simultaneously turn valve #3 (24-way valve) to #2, pumping clean air into the A2 inlet. Bubbles will rise from the bottom of the cavity in the A2 pyrolysis zone, promoting the A2 pyrolysis reaction. Continue for 10-30 seconds.

[0216] Step 12. Immediately afterwards, turn valve 1 (24-way) to #3, and simultaneously turn valve 3 (24-way) to #3, pumping clean air into the A3 inlet. Bubbles will rise from the bottom of the cavity in the A3 pyrolysis zone, promoting the A3 pyrolysis reaction. Continue for 10-30 seconds.

[0217] Step 13. Continue in this manner, creating clean air bubbles for 10-30 seconds for each of A1-A24. The total time is 240-720 seconds, approximately 4-12 minutes.

[0218] Step 14. Next, begin the second round of bubble aeration starting with A1. Simultaneously, turn 24-way valve #1 to #1, and then turn 24-way valve #3 to #1. Next, turn the 10-way rotary valve to the #1 clean air channel, pumping clean air into the A1 inlet. Bubbles will rise from the bottom of the A1 pyrolysis zone, promoting the A1 pyrolysis reaction. Continue for 10-30 seconds. Repeat this process to complete the second round of bubble aeration for A1-A24. This process takes 4-12 minutes.

[0219] Step 15. Repeat Steps 10-14, performing a total of 5 rounds of bubble aeration. This process takes 20-60 minutes in total. Complete the entire pyrolysis reaction.

[0220] Step 16. Turn off the heating plate.

[0221] 3. Transfer the pyrolysis products to the cavity of the extraction zone.

[0222] First, the A1 pyrolysis products are forced into the B1 chamber.

[0223] Step 17.10: Turn the rotary valve to #10 neutral position, close the double-way solenoid valve 3, and close the double-way solenoid valve 1. First, disconnect the gas circuit.

[0224] Step 18. Open the double-way solenoid valve 4. Prepare to open the exhaust path in section B.

[0225] Step 19. Turn valve #4 (24-way) to #1, and turn rotary valve #1 (24-way) to #1. Prepare to inject the pyrolysis products.

[0226] Step 20. Turn on the negative pressure aspirator 1 and adjust the pressure controller 2 to generate appropriate suction. Open the double-way solenoid valve 6 (this valve controls the drainage of samples 1-6). Prepare to assist in drawing the lysis products into chamber B1 of the extraction zone.

[0227] Step 21. The electromagnet α1 is energized, simultaneously opening the double-way solenoid valve 1 and turning the 10-way rotary valve to the #1 clean air setting. The electromagnet is energized for approximately 1.5-2 minutes. The pyrolysis products are then forced into the extraction chamber.

[0228] Step 21.10: Turn the rotary valve to #10 neutral position to de-energize electromagnet α1. This cuts off the gas supply and re-blocks the extraction channel for sample 1. At this time, the negative pressure suction device 1 remains open to maintain suction.

[0229] Next, the A2 cleavage products are forced into chamber B2.

[0230] At the same time as the electromagnet in Step 22.α1 is de-energized, valve #4 (24-way) is turned to #2, and rotary valve #1 (24-way) is also turned to #2. This step opens the gas path from the pyrolysis of sample #2 to the extraction zone, preparing for the injection of pyrolysis products.

[0231] Step 23. Closely following the previous step, energize magnet α2, simultaneously open dual-way solenoid valve 1, and turn 10-way rotary valve to clean air setting #1. Energize the electromagnet for approximately 1.5-2 minutes. Force the pyrolysis products into the cavity of the A2 extraction zone.

[0232] Step 24.10: Turn the rotary valve to #10 neutral position to de-energize electromagnet α2. This cuts off the gas supply and re-blocks the extraction channel for sample 2. At this time, the negative pressure suction device 1 remains open to maintain suction.

[0233] Next, the pyrolysis products of A3-6 are forced into chamber B3-6.

[0234] Step 25. Similarly, repeat steps 22-24 above, sequentially pressing the pyrolysis products of samples 3-6 into the cavities of their corresponding extraction zones.

[0235] Next, the pyrolysis products of A7-12 are forced into chamber B7-12.

[0236] Step 26. The steps are similar, but you need to switch solenoid valve 7 once because it controls the drainage channel of B7-12.

[0237] When the pyrolysis products of sample A6 are forced into chamber B6, the 10-way rotary valve is turned to position #10, the α6 electromagnet is de-energized, and the double-way solenoid valve 7 opens. Subsequently, similarly, throughout the entire extraction process, the path "exhaust 2—double-way solenoid valve 7—multi-port head 2—gas washing bottle—pressure controller 2—negative pressure suction device 1" remains open, maintaining a certain suction force.

[0238] Next, the pyrolysis products of A13-18 are forced into the B13-18 chamber.

[0239] Step 27. The steps are similar, but you need to switch the two-way solenoid valve 8 once because it controls the drainage channel of B13-18.

[0240] When the pyrolysis products of sample A12 are forced into chamber B12, the 10-way rotary valve is turned to position #10, the α12 electromagnet is de-energized, and the double-way solenoid valve 8 opens. Subsequently, similarly, throughout the entire extraction process, the path "exhaust 3—double-way solenoid valve 8—multi-port head 2—gas washing bottle—pressure controller 2—negative pressure suction device 1" remains open, maintaining a certain suction force.

[0241] Next, the pyrolysis products of A19-24 are forced into chamber B19-24.

[0242] Step 28. The steps are similar, but the two-way solenoid valve 9 needs to be switched once because it controls the drainage channel of B19-24.

[0243] When the pyrolysis products of sample A18 are forced into chamber B18, the 10-way rotary valve is turned to position #10, the α18 electromagnet is de-energized, and the double-way solenoid valve 9 opens. Subsequently, similarly, throughout the entire extraction process, the path "drain 4—double-way solenoid valve 9—multi-port head 2—gas washing bottle—pressure controller 2—negative pressure suction device 1" remains open, maintaining a certain suction force.

[0244] Step 29. After the final B24 chamber is filled with pyrolysis products (assuming all pyrolysis products have been filled into the extraction chamber; if not, start the second round of filling from B1), all electromagnets α1-24 are de-energized, and the pyrolysis to extraction flow channel is closed. Electromagnetic valve #24 is now in position #24, and double-pass solenoid valves 6, 7, 8, and 9 are open. Negative pressure suction device 1 maintains a weak suction force to prevent samples in region B from mixing at the bottom.

[0245] Quickly turn the 10-way rotary valve to the #10 neutral position and disconnect the power to the double-way solenoid valve 1.

[0246] 4. Extraction

[0247] Combine.

[0248] Step 30. Quickly turn valve #4 (24-way valve) to #1, then open the double-way solenoid valve 4 to open the drain path of B1.

[0249] Step 31. Simultaneously, quickly turn the No. 2 dual-way solenoid valve to #1, open the No. 2 dual-way solenoid valve, and turn the 10-way rotary valve to the #4 magnetic bead position. Using the flow meter reading, press 20ul of magnetic bead into chamber B1. Then, quickly turn the 10-way rotary valve to the #3 binding fluid position and press 250ul of magnetic bead binding fluid into chamber B1 using the flow meter reading.

[0250] Step 32. Next, quickly turn valve #4 (24-way valve) to #2 to open the drain path of B2. Turn valve #2 (24-way valve) to #2, read the flow meter reading, and repeat Step 31 to inject 20ul of magnetic bead liquid and 250ul of combined liquid into chamber B2.

[0251] Step 33. Repeat Step 31 to inject the magnetic beads and their binding solution into the extraction chambers of the 24 samples.

[0252] Next, air bubbles are blown into the cavities of the 24 extraction zones in turn to promote the binding reaction.

[0253] Step 34. After completing Step 33, each chamber in Zone B is filled with pyrolysis products, binding fluid, and magnetic beads. 24-way valve #4 and 24-way valve #24 are both in position #24. At this point, turn 24-way valve #4 to #1 and 24-way valve #1, then immediately turn 10-way valve to the #1 clean air setting. Inject clean air into the chambers of Zone B1, causing bubbles to rise from the bottom to promote the binding reaction. The air injection time should be controlled between 10-30 seconds.

[0254] Step 35. Next, turn valve #4 (24-way valve) to #2 and valve #2 (24-way valve) to #2. Inject clean air into the cavity of the B2 extraction zone, causing bubbles to rise from the bottom to promote the binding reaction. The air injection time should be controlled between 10-30 seconds.

[0255] Step 36. Repeat Step 35, sequentially injecting clean air into the extraction chambers of zones B3-B24. The injection time for each chamber should be controlled between 10-30 seconds.

[0256] Step 37. Repeat Steps 34-36 for two rounds. If necessary, perform a third round of inflation.

[0257] Step 38. Close the double-way solenoid valve 4, and simultaneously close the double-way solenoid valve 2. Turn the 10-way rotary valve to the #10 neutral position. Next, raise the permanent magnet (located below the chamber in the chip extraction area; raised to below the chamber using the lifting device when needed; used to attract small magnetic beads encapsulating sample DNA, preventing them from being washed into the waste liquid pipe during rinsing). Use the magnet to hold the magnetic beads in each chamber of area B. Hold them in place.

[0258] Step 39: Turn the 24-way rotary valve #4 to #1, turn the 24-way rotary valve #2 to #1, and turn the 10-way rotary valve to the #1 clean air setting. Then simultaneously open the double-way solenoid valve 2 to use clean air to force the liquid in chamber B1 out from the waste liquid pipe.

[0259] Step 40. Next, turn the No. 4 24-way rotary valve to #2 and the No. 2 24-way rotary valve to #2, and use clean air to discharge the waste liquid in the B2 chamber through the waste liquid pipe.

[0260] Step 41. Repeat step 40 to drain the liquid from cavities B3-B24 in sequence.

[0261] 5. Extract

[0262] First, perform the first round of rinsing.

[0263] Step 42. Retract the permanent magnet, then turn the No. 4 24-way valve back to #1, and turn the No. 2 24-way rotary valve to #1. Prepare to introduce rinsing solution 1 into chamber B1.

[0264] Step 43. Then quickly turn the 10-way rotary valve to #5 rinse solution 1, and through the flow meter reading, introduce 500ul of rinse solution 1 into chamber B1.

[0265] Step 44. Then, quickly turn the No. 4 24-way rotary valve to #2, and at the same time turn the No. 2 24-way rotary valve to #2. According to the flow meter reading, 500ul of rinsing solution 1 is introduced into chamber B2.

[0266] Step 45. Repeat Step 44 in this manner, and press 500ul of rinsing solution into chambers B3-B24 in sequence.

[0267] Step 46. After pressing the last chamber B24, quickly turn valve #4 (24-way valve) to #1, valve #2 (24-way valve) to #1, and then turn valve #10 to the #1 clean air setting. Aerate the air bubbles from the bottom of chamber B1 to ensure even mixing and rinsing. Aerate for 10-30 seconds.

[0268] Step 47. Immediately afterwards, quickly turn valve #4 (24-way valve) to #2 and valve #2 (24-way valve) to #2 to expel air bubbles into B2.

[0269] Step 48. Continue in this manner, repeating Step 47 to blow air bubbles into B3-B24 in sequence. The total time is approximately 6-10 minutes.

[0270] Step 49. Extend the permanent magnet while simultaneously closing the double-way solenoid valve 4. First, turn the 10-way rotary valve to the #10 neutral position. Cut off the gas supply and use magnetic force to hold the magnetic bead.

[0271] Step 50.2 The 24-way solenoid valve is quickly turned to #1, and at the same time the 10-way rotary valve is quickly turned to the #1 clean air position, controlling the flow meter to uniformly pump the rinsing liquid in the B1 chamber into the waste liquid channel.

[0272] Step 51.2 The 24-way solenoid valve is quickly turned to #2, and at the same time the 10-way rotary valve is quickly turned to the #1 clean air position. The flow meter is controlled to uniformly pump the rinsing liquid in the B2 chamber into the waste liquid channel.

[0273] Step 52. Similarly, adjust the position of valve 24 to pressurize the rinsing fluid in chamber B1-24 into the waste liquid pipeline.

[0274] Next, a second round of rinsing will be performed. Before the second round of rinsing, the permanent magnet must be retrieved.

[0275] Step 53. First, retract the permanent magnet, then open the double-way solenoid valve 4. At the same time, turn the 10-way rotary valve to the #6 rinse liquid 2 position, and turn the 24-way valve to #1. Based on the flow meter reading, press 500ul of rinse liquid 2 into the cavity of the B1 extraction zone.

[0276] Step 54.2: Turn the 24-way valve to #2 and, based on the flow meter reading, force 500 μL of rinsing solution 2 into the cavity of the B2 extraction zone.

[0277] Step 55. Repeat Steps 53-54, and press 500ul of rinsing solution 2 into chambers B3-B24 in sequence.

[0278] Step 56. Then repeat Steps 46-52 to complete the air bubble removal and waste liquid drainage operations for each cavity.

[0279] 6. Extract

[0280] Washing off.

[0281] After rinsing is complete, the permanent magnet is in the raised state, the magnetic beads are in the attracted state, and the 10-way rotary valve is in the #1 clean air neutral position.

[0282] Step 57. Quickly turn valve #4 (24-way valve) to #1, and then quickly turn valve #2 (24-way valve) to #1 to introduce clean air for 20-40 seconds. This step is to dry the chamber of the extraction zone.

[0283] Step 58. Similarly, adjust valves 4 and 2 to introduce clean air into the cavity of each sample extraction area for 20-40 seconds.

[0284] Step 59. Retract the permanent magnet, and quickly turn the No. 4 24-way valve to #1, then quickly turn the No. 2 24-way valve to #1, and turn the 10-way rotary valve to the #7 eluent setting. Using the flow meter reading, press 40ul of eluent into chamber B1 of the extraction zone.

[0285] Step 60. Following this pattern, adjust valve #4 (24-way valve) to #2-#24 in sequence, and quickly turn valve #2 (24-way valve) to #2-#24. Apply the washing and extracting pressure to valves B2-B24 respectively.

[0286] Step 61. Turn on the heating plate and set the temperature to 65℃ to allow it to heat up quickly.

[0287] Step 62. At the same time, quickly turn the No. 4 24-way valve to #1, quickly turn the No. 2 24-way valve to #1, and turn the 10-way rotary valve to the #1 clean air setting for about 15 seconds to press clean air into the bottom of the extraction chamber B1 to create air bubbles and make the eluent mix evenly.

[0288] Step 63. Following this pattern, turn valve #4 (24-way) to #2-#24, then quickly turn valve #2 (24-way) to #2-#24, and leave the 10-way rotary valve at #1. Pressurize clean air into the bottom of chamber B1 in the extraction zone to create air bubbles. Introduce clean air into each sample extraction zone chamber for 10-30 seconds to ensure the eluent is thoroughly mixed. This process takes approximately 10 minutes. Then close the double-way solenoid valve 4, using the weak negative attraction of the drain channel to retain the eluent mixture in each chamber of zone B.

[0289] Step 63. Continue to maintain the heating plate at 65 degrees Celsius for about 5-10 minutes. Then turn off the heating plate.

[0290] Step 64. Extend the permanent magnet to perform magnetic separation. Simultaneously, open the double-port solenoid valve 4, and quickly turn the 24-port valve #2 sequentially to #1-#24, approximately 1-5 seconds per sample. This step is for magnetic separation and to quickly expel the waste gas generated by the reactions in Steps 61-63 that has expanded within the extraction chamber.

[0291] 7. Transfer the extracted DNA to the cavity of the amplification region.

[0292] As shown in the system schematic, the transfer of DNA products from the extraction chamber in region B to the amplification chamber relies primarily on two forces: one is the forced inflow of clean air from the inlet of the extraction chamber, and the other is the suction force of the negative pressure aspirator 2. Because the flow channels in regions B and C are relatively long and have high flow resistance, the suction force of the negative pressure aspirator 2 is necessary as one of the driving forces. Simultaneously, the negative pressure aspirator 1 needs to maintain a weak negative pressure, and the negative pressure of the negative pressure aspirator 2 must be greater than that of the negative pressure aspirator 1. This ensures that during DNA transfer, the DNA product is transferred to the amplification chamber while preventing cross-contamination of the liquid in region B within the extraction chamber.

[0293] Step 65. After completing Step 64, close the dual-way solenoid valve 4, and simultaneously close the dual-way solenoid valve 2. Turn the 10-way rotary valve back to #10. First, disconnect the intake path. Then, open the dual-way solenoid valve 5, the negative pressure suction device 2, and the pressure controller 3 to open the exhaust path of the expansion zone.

[0294] Step 66.5: Turn the 24-way valve to #1. Simultaneously, turn the 24-way valve to #1, and energize the β1 electromagnet. Then, quickly turn the 10-way rotary valve to the #1 clean air setting, and simultaneously open the double-way solenoid valve 2. Using the flow meter reading, slowly and evenly inject 15 μL of DNA product into the C1 chamber.

[0295] Step 67.5: Turn valve #2 to #2. Simultaneously, turn valve #2 to #2, energizing electromagnet β1. Using the flow meter reading, force 15 μL of DNA product into chamber C2.

[0296] Step 68. Continue in this manner, pressing 15 μL of DNA product from each sample into the cavity of its corresponding amplification region.

[0297] 8. Amplification

[0298] Step 69. After completing Step 68, turn off all β electromagnets. Perform the following operations in sequence: close the double-way solenoid valve 2, turn the 10-way rotary valve to #10 neutral, close the double-way solenoid valve 5, negative pressure suction device 2, and pressure controller 3.

[0299] Step 70. Turn on heating plate 2 and set it to 100℃ to rapidly raise and maintain the temperature, promoting the amplification reaction. At this time, the amplification reaction will occur in all chambers of region C.

[0300] 9. Amplification products were introduced into a 96-well plate.

[0301] After the amplification reaction is complete, clean air needs to be introduced into the cavity of the extraction zone B to drive the product in the amplification zone cavity to flow into the 96-well plate.

[0302] Step 71. The mechanical structure raises the 96-well plate to the designated position, and the needle on the chip punctures the thin film on the 96-well plate.

[0303] Step 72.2: Turn the 24-way valve to #1. Then, simultaneously, perform the following actions: turn the 10-way rotary valve to the #1 clean air setting; energize electromagnet β1; energize electromagnet γ1. Press the product from chamber C1 into orifice D1 of the 96-well plate. Then de-energize electromagnets β1 and γ1.

[0304] Step 73.2: Turn valve #24 to #2. Then, simultaneously, perform the following actions: energize electromagnet β1; energize electromagnet γ1. Press the product from cavity C2 into hole D2 of the 96-well plate. Then de-energize electromagnets β2 and γ2.

[0305] Step 74. Continue in this manner, pushing the product of all samples into region D.

[0306] Step 75. The mechanical structure is removed along with the 96-hole plate.

[0307] Step 76. Turn off all electromagnets, all solenoid valves, all pressure controllers, and all negative pressure suction devices. End.

[0308] After the product is discharged directly into the collection container, it can be automatically or manually transferred from the collection container to a capillary electrophoresis gene sequencing instrument for sequencing (such as Sanger sequencing). The sequenced sequence is then compared with a database to complete DNA testing.

[0309] Furthermore, the functional units in the various embodiments of this application can be integrated into one processing unit, or each unit can exist physically separately, or two or more units can be integrated into one unit. The integrated units can be implemented in hardware or as software functional units. The above are merely embodiments of this application and do not limit the patent scope of this application. Any equivalent structural or procedural transformations made based on the description and drawings of this application, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of this application.

Claims

1. A microfluidic chip, characterized in that, It includes a chip section, the chip section comprising: A substrate having multiple chambers and multiple microfluidic channels therein, wherein at least some of the chambers form chamber openings on the upper surface of the substrate, and at least some of the microfluidic channels form channel openings on the upper surface of the substrate; A connector assembly protrudes from the upper surface of the substrate and has a through hole. At least a portion of the connector assembly is fixed to the chamber opening such that the through hole of the connector assembly communicates with the chamber corresponding to the chamber opening and forms a cavity; and / or, at least a portion of the connector assembly is fixed to the channel opening such that the through hole of the connector assembly communicates with the microfluidic channel corresponding to the channel opening.

2. The microfluidic chip according to claim 1, characterized in that, The connector assembly includes at least one of the following: A single-hole connector assembly, wherein the single-hole connector assembly has a through hole; A multi-hole connector assembly has multiple through holes. When the connector assembly protrudes from the upper surface of the substrate, at least one of the multiple through holes communicates with a cavity corresponding to the cavity opening to form a cavity. At least one of the multiple through holes communicates with a microfluidic channel corresponding to the channel opening. The cavity communicated by the at least one through hole is connected to the microfluidic channel communicated by the at least one through hole through the microfluidic channel. A laminated connector assembly, comprising an upper connector assembly and a lower connector assembly, wherein the lower connector assembly has a through hole and is fixed to the opening of the chamber, and the upper connector assembly is detachably fixed in the through hole of the lower connector assembly; A filter connector assembly; the filter connector assembly has a connector filter section inside its through hole, and a chamber filter section is provided in the corresponding cavity of the filter connector assembly. When the filter connector assembly is fixed to the open cavity, the connector filter section and the chamber filter section form a filter section. The cavity formed by the through hole of the filter connector assembly and the cavity has a second subspace isolated within the cavity. The second subspace is connected to other spaces of the cavity only through the second subspace entrance formed by the top of the filter section. The second subspace entrance is located at a preset height inside the through hole of the filter connector assembly, and the outlet for discharging material from the cavity is located within the second subspace.

3. The microfluidic chip according to claim 2, characterized in that, The connector assembly includes the porous connector assembly, wherein the cavity formed by the at least one through hole is used for reaction, and the microfluidic channel connected by the at least one through hole is used for feeding into the cavity.

4. The microfluidic chip according to claim 2, characterized in that, The connector assembly includes the filter connector assembly; The filter section is generally a portal-shaped column or a semi-cylindrical column, abutting against the inner wall of the cavity to define the second subspace together with the inner wall of the cavity; or... The filter section is a cylindrical column, which defines the second subspace and is fixed to the inner wall of the cavity.

5. The microfluidic chip according to claim 2, characterized in that, The connector assembly includes the laminated connector assembly; The upper connector assembly has a through hole, and the through hole of the upper connector assembly and the chamber form a cavity; multiple upper connector assemblies with different through hole diameters are detachably fixed in the through holes of the lower connector assembly to form multiple cavities with different volumes.

6. The microfluidic chip according to claim 2, characterized in that, The connector assembly includes the laminated connector assembly; The bottom of the upper connector assembly presses against the solid reactant pre-embedded in the chamber.

7. The microfluidic chip according to claim 1, characterized in that, Each of the connector assemblies protrudes from the upper surface of the substrate at the same height, and the diameter of the through holes in at least a portion of the connector assemblies is determined according to the required reaction system.

8. The microfluidic chip according to claim 1, characterized in that, The substrate includes a top layer, a middle layer, and a bottom layer; the substrate is provided with microfluidic channels. The upper and lower surfaces of the intermediate layer have three-dimensional patterns, and the patterns on the upper surface of the intermediate layer and the top layer form the upper microfluidic channel in the microfluidic channel. The pattern on the lower surface of the intermediate layer and the bottom layer form a lower microfluidic channel in the microfluidic channel; the extension directions of the upper microfluidic channel and the lower microfluidic channel are parallel to the plane of the substrate; The intermediate layer includes through holes for forming vertical microfluidic channels, the extension direction of which is perpendicular to the plane of the substrate, and at least a portion of the vertical microfluidic channels are used to connect the upper microfluidic channels and the lower microfluidic channels. The intermediate layer also includes through-holes for forming chambers for carrying out reactions; The top layer has through holes at positions corresponding to at least a portion of the chambers to form the chamber openings, and the top layer has through holes at positions corresponding to at least a portion of the upper microfluidic channels or the vertical microfluidic channels to form the channel openings.

9. The microfluidic chip according to claim 1, characterized in that, The microfluidic chip includes a clamping part, which includes an upper clamping plate and a lower clamping plate. The upper clamping plate and the lower clamping plate form a receiving cavity. When the chip part is located in the receiving cavity, the lower surface of the upper clamping plate contacts the upper end face of the connector assembly to seal the connector assembly.

10. The microfluidic chip according to claim 9, characterized in that, The upper clamping plate also includes a sealing gasket, and the lower surface of the upper clamping plate contacts the upper end face of the connector assembly, including: the sealing gasket of the upper clamping plate contacts the upper end face of the connector assembly, and the sealing gasket of the upper clamping plate is located between the upper clamping plate body and the upper end face of the connector assembly.

11. The microfluidic chip according to claim 10, characterized in that, The upper clamping plate has a fourth through hole, the sealing gasket has a through hole, and the fourth through hole, the through hole of the sealing gasket, and the through hole of the connector assembly are aligned and connected.

12. The microfluidic chip according to claim 11, characterized in that, The clamping part further includes a clamping assembly, which has a through hole and is fixed to the fourth through hole. When the chip part is located in the receiving cavity, the through hole of the clamping assembly, the fourth through hole of the upper clamping plate, the through hole of the sealing gasket, the through hole of the connector assembly, and the intermediate connectable component corresponding to the connector assembly are aligned. The intermediate connectable component includes the chamber and / or the microfluidic channel.

13. The application of the microfluidic chip according to any one of claims 1-12 in nucleic acid detection, wherein, The substrate includes multiple functional regions, including a cell lysis region, a nucleic acid extraction region, a nucleic acid amplification region, and a product export region; each functional region includes a cavity composed of the connector assembly for performing the reaction required for that functional region.

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