A microfluidic chip, a sample processing culture device and a molecular detection system

Abstract

The application relates to the technical field of molecular detection, in particular to a microfluidic chip, a sample processing culture device and a molecular detection system. The microfluidic chip is used for connecting the sample processing device. The sample processing device has a sample outlet channel. The sample outlet channel can output any one of positive pressure gas, oil and a to-be-dispersed liquid. The microfluidic chip comprises a chip main body and a first switching valve. The chip main body has an oil storage cavity, a sample storage cavity and a reaction cavity which are arranged at intervals, and a sample mixing flow channel. One end of the sample mixing flow channel is communicated with the sample storage cavity and the oil storage cavity. The other end of the sample mixing flow channel is communicated with the reaction cavity. The first switching valve is connected to the chip main body and can communicate any one of the oil storage cavity and the sample storage cavity with the sample outlet channel. The first switching valve can also simultaneously communicate the sample storage cavity and the oil storage cavity with the sample outlet channel. The microfluidic chip can use the positive pressure gas provided by the sample processing device as a power source to drive the oil and the to-be-dispersed liquid to focus in the sample mixing flow channel to form microdroplets, so that droplet dispersion PCR detection is realized.

Description

Technical Field

[0001] This application relates to the field of molecular detection technology, and in particular to a microfluidic chip, a sample processing and culture device, and a molecular detection system. Background Technology

[0002] Polymerase chain reaction (PCR) is a core technology for rapidly amplifying specific DNA fragments in vitro. Droplet dispersion PCR is the mainstream form of third-generation PCR technology. Its core principle is to divide the sample into independent droplets, amplify single molecules independently, and achieve high testing accuracy.

[0003] Chinese invention patent CN115895869B discloses a disc-type microfluidic chip for molecular diagnostic detection, comprising a disc substrate, a piston cavity at the center of the disc substrate, sample chambers arranged around the piston cavity, and a rotary valve at the bottom of the piston cavity. Rotating the rotary valve connects each sample chamber to the piston cavity, enabling sample processing. Multiple sample amplification chambers are arranged around the outer periphery of the disc substrate. The centrifugal force generated by the rotation of the disc substrate discharges the processed sample into the sample amplification chambers. This achieves automated sample processing and automatic injection into the sample amplification chambers. However, it cannot perform droplet-dispersive PCR testing, and its detection accuracy is lower than that of droplet-dispersive PCR. Summary of the Invention

[0004] Based on this, the purpose of this application is to provide a microfluidic chip capable of realizing droplet-dispersed PCR detection. The microfluidic chip is used to connect to a sample processing device, which has a sample outlet channel capable of outputting any one of positive pressure gas, oil, and the liquid to be dispersed. The microfluidic chip includes:

[0005] The chip body has an oil storage chamber, a sample storage chamber and a reaction chamber arranged at intervals. The chip body also has a mixing channel, one end of which is connected to the sample storage chamber and the oil storage chamber, and the other end is connected to the reaction chamber.

[0006] A first switching valve is connected to the chip body. The first switching valve is configured to connect either the oil storage chamber or the sample storage chamber to the sample outlet channel. The first switching valve is also configured to connect the sample storage chamber and the oil storage chamber to the sample outlet channel simultaneously, so that the positive pressure gas in the sample outlet channel drives the oil in the oil storage chamber and the liquid to be dispersed in the sample storage chamber to the mixing channel and form microdroplets.

[0007] As an optional solution, the chip body includes a substrate, the substrate having a first groove and a second groove;

[0008] The first switching valve includes a pressure block connected to the substrate. The pressure block and the groove wall of the first groove form a first branch channel, and the substrate and the groove wall of the second groove form a second branch channel. When the microfluidic chip is connected to the sample processing device, the first branch channel connects the sample outlet channel and the sample storage chamber, and the second branch channel connects the sample outlet channel and the oil storage chamber.

[0009] A first valve core assembly is connected to the pressure block and / or the base plate and is configured to open or close the first branch channel;

[0010] The second valve core assembly is connected to the pressure block and / or the base plate and is configured to open or close the second branch channel.

[0011] As an optional embodiment, the first valve core assembly includes a first partition and a first flexible sheet. The first partition is fixed within the first groove, and the first flexible sheet is disposed between the pressure block and the substrate, with a portion of the first flexible sheet opposite to the first partition. The pressure block has a first vent at the position opposite to the first partition. Applying negative pressure to the first vent causes a portion of the first flexible sheet to protrude into the first vent, thereby forming a first gap between the first flexible sheet and the first partition, thus opening the first branch channel. Applying positive pressure to the first vent causes a portion of the first flexible sheet to adhere to the first partition, thereby closing the first branch channel.

[0012] And / or, the second valve core assembly includes a second partition and a second flexible sheet, the second partition being fixed within the second groove, the second flexible sheet being disposed between the pressure block and the substrate, with a portion of the second flexible sheet opposite to the second partition; the pressure block has a second vent at the position opposite to the second partition; applying negative pressure to the second vent causes a portion of the second flexible sheet to protrude into the second vent, thereby forming a second gap between the second flexible sheet and the second partition, thus opening the second branch channel; applying positive pressure to the second vent causes a portion of the second flexible sheet to adhere to the second partition, thereby closing the second branch channel.

[0013] As an optional embodiment, the mixing channel has an inlet and an outlet, the outlet being connected to the reaction chamber, and the chip body further comprises:

[0014] A first flow resistance channel is located between the sample storage chamber and the reaction chamber, connecting the sample storage chamber and the liquid inlet. The first flow resistance channel is used to adjust the flow rate of the liquid to be dispersed at the liquid inlet.

[0015] The second flow resistance channel is located between the oil storage chamber and the reaction chamber, connecting the oil storage chamber and the liquid inlet. The second flow resistance channel is used to adjust the flow rate of the oil at the liquid inlet.

[0016] This application also provides a sample processing and culture device, including a sample processing device and the above-mentioned microfluidic chip, wherein the sample processing device includes:

[0017] The main body includes a sample loading tank, a transfer tank, a sample dispensing channel, an oil storage tank, and multiple processing tanks.

[0018] A second switching valve is connected to the main body and is configured to connect any one of the sample loading tank, the sample discharging channel, the oil storage tank and the plurality of processing tanks to the transfer tank.

[0019] The microfluidic chip is detachably connected to the main body.

[0020] As an optional solution, the sample processing device further includes:

[0021] A sealing membrane is attached to the main body to seal the oil storage tank and the plurality of processing tanks;

[0022] The pressure cap includes a connecting part, a mounting part, and multiple puncture parts. The connecting part is slidably connected to the main body, and the mounting part is connected to the connecting part. The mounting part is spaced apart from the main body and has a third air hole. All of the multiple puncture parts are connected to the mounting part.

[0023] Moving the pressure cap allows each of the puncture parts to pierce the sealing membrane, thereby connecting the oil storage tank to the outside and the multiple processing tanks to the outside. When each of the puncture parts pierces the sealing membrane, the mounting part at the edge of the third air hole seals against the opening of the transfer groove, thereby connecting the third air hole with the transfer groove.

[0024] As an optional solution, the sample processing device further includes:

[0025] An elastic element, one end of which abuts against the main body and the other end of which abuts against the mounting portion.

[0026] As an optional option, one of the plurality of processing tanks is a pyrolysis tank;

[0027] The sample processing device further includes a first processing component and / or a second processing component; the first processing component is detachably connected to the main body and includes a heating element, wherein when the first processing component is connected to the main body, the heating element abuts against the wall of the pyrolysis tank; the second processing component is detachably connected to the main body and includes a magnetic element, wherein when the second processing component is connected to the main body, the magnetic element abuts against the wall of the pyrolysis tank.

[0028] This application also provides a molecular detection system, which includes the above-mentioned sample processing and culture device, and a second switching valve rotatably connected to the main body; the second switching valve has a first channel, and a first port and a second port communicating with the first channel; the first port is communicating with a transfer tank, and rotating the second switching valve can enable the second port to communicate with any one of a sample loading tank, a sample discharging channel, an oil storage tank and a plurality of processing tanks;

[0029] The molecular detection system further includes an operating device, which includes a mounting platform and a first driving component. The first driving component is connected to the mounting platform and is throttle-connected to the second switching valve to drive the second switching valve to rotate.

[0030] The second switching valve is connected to a magnet, the magnetic field direction of which is perpendicular to the rotation axis of the second switching valve. The first drive unit and / or the mounting platform are connected to a detection unit for detecting the magnetic field direction of the magnet.

[0031] As an optional solution, the operating device further includes a lifting device, a first docking member, a second docking member, and a third docking member, wherein the first docking member has a first vent, the second docking member has a second vent, and the third docking member has a third vent.

[0032] The lifting device includes a drive mechanism and a lifting component. The drive mechanism connects the lifting component and the mounting platform. The lifting component is located above the pressure cover. The first docking component, the second docking component, and the third docking component are all connected to the lifting component.

[0033] The driving mechanism drives the lifting member to descend, which enables the first docking member to press the cover downward, causing each puncture part to pierce the sealing film. It also enables the first vent hole to communicate with the first air hole, the second vent hole to communicate with the second air hole, and the third vent hole to communicate with the third air hole.

[0034] Compared with the prior art, the beneficial effects of this application are as follows:

[0035] The microfluidic chip of this application is used to connect to a sample processing device. The sample processing device has a sample outlet channel capable of outputting any one of positive pressure gas, oil, and a liquid to be dispersed. The microfluidic chip includes a chip body and a first switching valve. The chip body has an oil storage chamber, a sample storage chamber, and a reaction chamber arranged at intervals. The chip body also has a mixing channel, one end of which connects to the sample storage chamber and the oil storage chamber, and the other end of which connects to the reaction chamber. The first switching valve is connected to the chip body and can connect either the oil storage chamber or the sample storage chamber to the sample outlet channel. When the first switching valve connects the oil storage chamber to the sample outlet channel, the sample processing device can output the oil. When the sample is fed into the oil storage chamber through the sample outlet channel, and the first switching valve connects the sample storage chamber to the sample outlet channel, the sample processing device can input the liquid to be dispersed into the sample storage chamber through the sample outlet channel. Subsequently, the first switching valve simultaneously connects the sample storage chamber and the oil storage chamber to the sample outlet channel. The sample processing device then introduces positive pressure gas from the sample outlet channel into the sample outlet channel. This positive pressure gas drives the oil in the oil storage chamber and the liquid to be dispersed in the sample storage chamber to the mixing channel, forming microdroplets. Therefore, the microfluidic chip of this application can utilize the positive pressure gas provided by the sample processing device as a power source to drive the oil and the liquid to be dispersed to focus and form microdroplets in the mixing channel, thereby achieving droplet-dispersed PCR detection. Therefore, the microfluidic chip of this application does not require an additional micropump or complex driving mechanism to achieve droplet dispersion, reducing the chip's manufacturing cost. Attached Figure Description

[0036] Figure 1 This is a schematic diagram of the top structure of the microfluidic chip of this application;

[0037] Figure 2 A schematic diagram of the bottom structure of the microfluidic chip in this application, hidden behind the sealing sheet;

[0038] Figure 3 This is an exploded view of the microfluidic chip of this application;

[0039] Figure 4 This is an exploded view of the first switching valve;

[0040] Figure 5 This is a schematic diagram of the mounting structure of the flexible valve plate on the substrate.

[0041] Figure 6 This is an isometric view of the sample processing and culture apparatus of this application;

[0042] Figure 7 This is a schematic diagram showing the installation method of the first processing component on the sample processing device;

[0043] Figure 8 This is a schematic diagram showing the installation method of the second processing component on the sample processing device;

[0044] Figure 9A schematic diagram of the installation structure of the elastic element in the main body to conceal the barrel-shaped shell, the gland, and the sealing membrane;

[0045] Figure 10 An exploded view of the sample processing and culture apparatus of this application, concealing the barrel-shaped shell;

[0046] Figure 11 This is an exploded structural diagram of the second switching valve, the second component, and the main body component;

[0047] Figure 12 This is a cross-sectional structural schematic diagram of the sample processing and culture device of this application;

[0048] Figure 13 This is a top view of the second switching valve;

[0049] Figure 14 for Figure 13 Sectional view along line AA;

[0050] Figure 15 A isometric drawing of the operating equipment;

[0051] Figure 16 This is a schematic diagram of the installation structure of the sample processing and culture device of this application on the operating equipment;

[0052] In the figure, 1 is the microfluidic chip, 11 is the chip body, 111 is the main board, 1110 is the oil storage chamber, 1111 is the sample storage chamber, 1112 is the reaction chamber, 1113 is the mixing channel, 1114 is the first flow resistance channel, 1115 is the second flow resistance channel, 1116 is the waste liquid channel, 11171 is the waste liquid chamber, 11172 is the vent, 11181 is the first positioning hole, 11182 is the second recess, 11183 is the third recess, 11184 is the second through hole, 11185 is the third through hole, 1119 is the positioning protrusion, 112 is the substrate, 1121 is the first branch channel, 1122 is the second branch channel, 1123 is the first through hole, 1 124. First receiving hole; 1125. Second receiving hole; 113. Sealing piece; 12. First switching valve; 121. Pressure block; 1211. First vent; 1212. Second vent; 122. First valve core assembly; 1221. First partition; 123. Second valve core assembly; 1231. Second partition; 124. Flexible valve piece; 1241. First flexible piece; 1242. Second flexible piece; 1243. Third flexible piece; 13. Breathable membrane; 2. Sample processing device; 21. Main body; 211. First piece; 2111. Base; 21111. Sample outlet channel; 21112. First docking part; 21113. Second positioning part. 2112, Hole, First tubular section, 21121, Transfer trough, 2113, Second tubular section, 21131, Sample loading trough, 21132, Tube cap, 2114, Third tubular section, 21141, Oil storage trough, 2115, Fourth tubular section, 21151, Processing trough, 21152, Contact area, 2116, Connecting rib, 212, Barrel-shaped shell, 2121, Mounting hole, 213, Second component, 2131, Clearance hole, 22, Second switching valve, 221, Large diameter section, 2211, First channel, 222, Small diameter section, 2221, Slot, 23, Sealing membrane, 24, Pressure cap, 241, Connecting part, 242, Mounting part, 242 1. Third vent; 243. Puncture section; 244. First sealing ring; 25. Elastic element; 26. First processing component; 261. Heating element; 27. Second processing component; 271. Magnetic element; 28. Sealing sheet; 29. ​​Magnet; 3. Operating equipment; 31. Mounting platform; 311. Clip protrusion; 32. First driving component; 33. Lifting device; 331. Drive mechanism; 3311. Support column; 3312. Guide column; 3313. Lead screw; 3314. Slider; 3315. Drive motor; 332. Lifting component; 34. First docking component; 35. Second docking component; 36. Third docking component; 361. Small diameter pipe section; 362. Large diameter pipe section. Detailed Implementation

[0053] The specific embodiments of this application will be described in further detail below with reference to the accompanying drawings and examples. The following examples are used to illustrate this application, but are not intended to limit the scope of this application.

[0054] In the description of this application, it should be understood that the terms "upper," "lower," "left," "right," "top," and "bottom," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this application. It should be understood that the terms "first," "second," etc., are used in this application to describe various information, but this information should not be limited to these terms, and these terms are only used to distinguish information of the same type from each other. For example, without departing from the scope of this application, "first" information can also be referred to as "second" information, and similarly, "second" information can also be referred to as "first" information.

[0055] like Figure 16 As shown, this application proposes a molecular detection system, including a sample processing and culture device and an operating device 3. The sample processing and culture device includes a sample processing device 2 and a microfluidic chip 1. The sample processing device 2 is used to complete functions such as sample lysis and nucleic acid extraction. The microfluidic chip 1 is connected to the sample processing device 2 and can realize single-molecule droplet dispersion, reaction and detection.

[0056] like Figures 1 to 5As shown, the microfluidic chip 1 of this application is used to connect to a sample processing device 2. The sample processing device 2 has a sample outlet channel 21111, which can output any one of positive pressure gas, oil, and liquid to be dispersed. The microfluidic chip 1 of this application includes a chip body 11 and a first switching valve 12. The chip body 11 has an oil storage chamber 1110, a sample storage chamber 1111, and a reaction chamber 1112 arranged at intervals. The chip body 11 also has a mixing channel 1113, one end of which is connected to the sample storage chamber 1111 and the oil storage chamber 111. 0, the other end is connected to the reaction chamber 1112; the first switching valve 12 is connected to the chip body 11, and the first switching valve 12 is configured to connect either the oil storage chamber 1110 or the sample storage chamber 1111 to the sample outlet channel 21111; the first switching valve 12 is also configured to connect the sample storage chamber 1111 and the oil storage chamber 1110 to the sample outlet channel 21111 at the same time, so that the positive pressure gas in the sample outlet channel 21111 drives the oil in the oil storage chamber 1110 and the liquid to be dispersed in the sample storage chamber 1111 to the mixing channel 1113 to focus and form microdroplets. Specifically, when the first switching valve 12 connects the oil storage chamber 1110 to the sample outlet channel 21111, the sample processing device 2 can input the oil into the oil storage chamber 1110 through the sample outlet channel 21111. When the first switching valve 12 connects the sample storage chamber 1111 to the sample outlet channel 21111, the sample processing device 2 can input the liquid to be dispersed into the sample storage chamber 1111 through the sample outlet channel 21111. After that, the first switching valve 12 connects the sample storage chamber 1111 and the oil storage chamber 1110 to the sample outlet channel 21111 at the same time. The sample processing device 2 introduces the positive pressure gas in the sample outlet channel 21111 into the sample outlet channel 21111. The positive pressure gas drives the oil in the oil storage chamber 1110 and the liquid to be dispersed in the sample storage chamber 1111 to the mixing channel 1113 to focus and form microdroplets. Therefore, the microfluidic chip 1 of this application can utilize the positive pressure gas provided by the sample processing device 2 as a power source to drive the oil and the liquid to be dispersed to focus and form microdroplets in the mixing channel 1113, thereby realizing droplet dispersion PCR detection. Thus, the microfluidic chip 1 of this application does not require an additional micropump or complex drive mechanism 331 to achieve droplet dispersion, reducing the chip manufacturing cost.

[0057] In some embodiments, the chip body 11 includes a substrate 112, which has a first groove and a second groove. The first switching valve 12 includes a pressure block 121, a first valve core assembly 122, and a second valve core assembly 123. The pressure block 121 is connected to the substrate 112. The pressure block 121 and the groove wall of the first groove form a first branch channel 1121, and the substrate 112 and the groove wall of the second groove form a second branch channel 1122. This arrangement facilitates the processing of the first branch channel 1121 and the second branch channel 1122. When the microfluidic chip 1 is connected to the sample processing device 2, the first branch channel 1121 connects the sample outlet channel 21111 and the sample storage chamber 1111. The second branch channel 1122 connects the sample outlet channel 21111 and the oil storage chamber 1110; the first valve core assembly 122 is connected to the pressure block 121 and / or the base plate 112, and the first valve core assembly 122 is configured to open or close the first branch channel 1121 to achieve individual control of the first branch channel 1121; the second valve core assembly 123 is connected to the pressure block 121 and / or the base plate 112, and the second valve core assembly 123 is configured to open or close the second branch channel 1122 to achieve individual control of the second branch channel 1122. During use, when it is necessary to introduce the liquid to be dispersed into the sample storage chamber 1111, the first valve core assembly 122 is used to open the first branch channel 1121, and the second valve core assembly 123 is used to close the second branch channel 1122, so that the first branch channel 1121 connects the sample outlet channel 21111 and the sample storage chamber 1111, and the liquid to be dispersed in the sample outlet channel 21111 can flow from the first branch channel 1121 to the sample storage chamber 1111. When it is necessary to introduce the oil to be dispersed into the sample storage chamber 1111, the first valve core assembly 122 is used to close the first branch channel 1121, and the second valve core assembly 123 is used to open the second branch channel 1122, so that the second branch channel 1122 connects the sample outlet channel 21111 and the sample storage chamber 1111, and the oil in the sample outlet channel 21111 can flow from the first branch channel 1121 to the oil storage chamber 1110.

[0058] In some embodiments, the first valve core assembly 122 includes a first partition 1221 and a first flexible sheet 1241. The first partition 1221 is fixed in a first groove, and the first flexible sheet 1241 is arranged between the pressure block 121 and the substrate 112. A portion of the first flexible sheet 1241 is clamped by the pressure block 121 and the substrate 112, and another portion is opposite to the first partition 1221. The pressure block 121 has a first vent 1211 at the position opposite to the first partition 1221, allowing air to pass through to the first... Applying negative pressure to the vent 1211 can draw the portion of the first flexible sheet 1241 opposite to the first partition 1221 into the vent 1211, thereby forming a first gap between the first flexible sheet 1241 and the first partition 1221, thus opening the first branch channel 1121; applying positive pressure to the vent 1211 can cause the portion of the first flexible sheet 1241 opposite to the first partition 1221 to adhere to the first partition 1221, thereby closing the first branch channel 1121.

[0059] Specifically, such as Figure 4 As shown, the first partition 1221 is disposed in the middle of the first groove. The first partition 1221 has a plate-like structure. The side of the first partition 1221 opposite to the first flexible sheet 1241 is the first surface. The first surface is flat and flush with the groove end face of the first groove. The first vent 1211 is arranged opposite to the first partition 1221. Applying positive pressure to the first vent 1211 can make a part of the first flexible sheet 1241 tightly fit the first surface and the groove end face of the first groove, ensuring the sealing effect of the first flexible sheet 1241 with the first surface and the groove end face of the first groove. The first partition 1221 and the substrate 112 are integrally formed.

[0060] In some embodiments of this application, such as Figure 3 As shown, the second valve core assembly 123 includes a second partition 1231 and a second flexible sheet 1242. The second partition 1231 is fixed in the second groove, and the second flexible sheet 1242 is arranged between the pressure block 121 and the substrate 112. A portion of the second flexible sheet 1242 is clamped by the pressure block 121 and the substrate 112, while another portion is opposite to the second partition 1231. The pressure block 121 has a second vent 1212 at the position opposite to the second partition 1231. Applying negative pressure to the second vent 1212 causes the portion of the second flexible sheet 1242 opposite to the second partition 1231 to protrude into the second vent 1212, thereby forming a second gap between the second flexible sheet 1242 and the second partition 1231, thus opening the second branch channel 1122; applying positive pressure to the second vent 1212 causes the portion opposite to the second partition 1231 to adhere to the second partition 1231, thereby closing the second branch channel 1122.

[0061] Specifically, such as Figure 4As shown, the second partition 1231 is disposed in the middle of the second groove. The second partition 1231 has a plate-like structure. The side of the second partition 1231 opposite to the second flexible sheet 1242 is the second surface, which is flat and flush with the groove end face of the second groove. The second vent 1212 is arranged opposite to the second partition 1231. Applying positive pressure to the second vent 1212 can make the second flexible sheet 1242 tightly fit the second surface and the groove end face of the second groove, ensuring the sealing effect between the second flexible sheet 1242 and the second surface and the groove end face of the second groove. The second partition 1231 and the substrate 112 are integrally formed, and the second surface, the first surface, the groove end face of the second groove, and the groove end face of the first groove are all flush.

[0062] In some embodiments of this application, such as Figure 3 As shown, both the first and second grooves are disposed on the upper surface of the substrate 112, and the pressure block 121 is stacked on the substrate 112. The substrate 112 has a first through hole 1123, which extends vertically through the substrate 112. The upper end of the first through hole 1123 is connected to the end of the first branch channel 1121 away from the sample storage chamber 1111, and the upper end of the first through hole 1123 is also connected to the end of the second branch channel 1122 away from the oil storage chamber 1110. After the microfluidic chip 1 is connected to the sample processing device 2, the lower end of the first through hole 1123 is connected to the sample outlet channel 21111. The upper end of the first through hole 1123 is covered with a third flexible sheet 1243, which is located between the substrate 112 and the pressure block 121.

[0063] In some embodiments of this application, such as Figure 4 , Figure 5As shown, the second flexible sheet 1242, the first flexible sheet 1241, and the third flexible sheet 1243 are integrally formed, constituting a flexible valve plate 124. In the vertical direction, the third flexible sheet 1243 is completely offset from the first vent 1211 and the second vent 1212. When a negative pressure is applied to the first vent 1211 and the second vent 1212, the first flexible sheet 1241 and the second flexible sheet 1242 deform, while the third flexible sheet 1243 does not deform. The third flexible sheet 1243 acts as a constraint force on the first flexible sheet 1241 and the second flexible sheet 1242, thereby preventing the first flexible sheet 1241 from being completely sucked into the first vent 1211 and preventing the second flexible sheet 1242 from being completely sucked into the second vent 1212. This ensures that when positive pressure is applied to the first vent 1211 and the second vent 1212, the first flexible sheet 1241 and the second flexible sheet 1242 can be smoothly reset, achieving the cutoff of the first branch channel 1121 and the second branch channel 1122. This configuration, compared to setting the first flexible sheet 1241 and the second flexible sheet 1242 as separate structures, eliminates the need to additionally connect one end of the first flexible sheet 1241 to the substrate 112 or the pressure block 121. The assembly of the first flexible sheet 1241 and the second flexible sheet 1242 can be achieved using the holding force of the pressure block 121 on the third flexible sheet 1243. The first flexible sheet 1241, the second flexible sheet 1242, and the third flexible sheet 1243 are made of polymer materials with high elasticity and toughness, such as thermoplastic elastomers, silicone rubber, and low-density polyethylene. In this embodiment, the gas pressure applied to the first vent 1211 and the second vent 1212 can be maintained between -60 kPa and 60 kPa.

[0064] In some embodiments of this application, such as Figure 3As shown, the chip body 11 includes a main plate 111 and a sealing piece 113. The lower end face of the main plate 111 is provided with a third groove and a first pit. The sealing piece 113 is connected to the lower end of the main plate 111. The sealing piece 113 and the groove wall of the third groove form a mixing channel 1113. The sealing piece 113 and the pit wall of the first pit form a reaction chamber 1112. The substrate 112 is connected to the upper end of the main plate 111. The upper end of the main plate 111 has a second pit 11182 and a third pit 11183. The substrate 112 has a first receiving hole 1124 and a second receiving hole 1125 that are connected vertically. The first receiving hole 1124 communicates with the second pit 11182 to form a sample storage chamber 1111. The second receiving hole 1125 communicates with the third pit 11183 to form an oil storage chamber 1110. The bottom wall of the second recess 11182 has a second through hole 11184, the upper end of which is connected to the sample storage chamber 1111 and the lower end of which is connected to the third groove. The bottom wall of the third recess 11183 has a third through hole 11185, the upper end of which is connected to the oil storage chamber 1110 and the lower end of which is connected to the third groove.

[0065] In some embodiments of this application, such as Figure 2 As shown, the mixing channel 1113 has an inlet and an outlet, with the outlet connected to the reaction chamber 1112. The chip body 11 also has a first flow resistance channel 1114 and a second flow resistance channel 1115. The first flow resistance channel 1114 is located between the sample storage chamber 1111 and the reaction chamber 1112, connecting the sample storage chamber 1111 to the inlet. The first flow resistance channel 1114 is used to adjust the flow rate of the liquid to be dispersed at the inlet. The second flow resistance channel 1115 is located between the oil storage chamber 1110 and the reaction chamber 1112, connecting the oil storage chamber 1110 to the inlet. The first flow resistance channel 1114 is used to adjust the flow rate of the oil at the inlet. Specifically, in this embodiment, the driving force for droplet dispersion comes from the positive pressure gas of the sample processing device 2, and the oil and the liquid to be dispersed are subjected to the same gas pressure. By designing the geometric parameters, such as length and degree of bending, and the surface conditions, such as hydrophobic or hydrophilic surfaces, of the first flow resistance channel 1114 and the second flow resistance channel 1115, the flow resistance of the oil and the liquid to be dispersed can be preset during the chip manufacturing stage. This allows the liquid to be dispersed and the oil to flow towards the inlet at the required flow rate ratio under the same driving force, which is beneficial for obtaining microdroplets with high dispersibility and uniform size.

[0066] In some embodiments of this application, such as Figure 2As shown, there are two second flow resistance channels 1115, which are symmetrically arranged on the left and right sides of the mixing channel 1113. The liquid outlet of the first flow resistance channel 1114, the liquid outlet of the two second flow resistance channels 1115 and the liquid inlet form a cross-shaped flow channel structure. This arrangement can ensure that the liquid to be dispersed is surrounded by oil, further improving the quality of droplet dispersion.

[0067] In some embodiments of this application, the first flow resistance channel 1114 and the second flow resistance channel 1115 have the same depth and width, which are set to 60 micrometers to 120 micrometers. The length ratio of the first flow resistance channel 1114 and the second flow resistance channel 1115 is set to 1:2, and the droplet generation driving force is 20 kPa. Under these parameters, high-speed generation of droplets with a diameter of 90 μm can be achieved.

[0068] In some embodiments of this application, such as Figure 2 As shown, the reaction chamber 1112 has a waste liquid outlet at the end furthest from the mixing channel 1113. The main body plate 111 also has a waste liquid channel 1116 and a waste liquid chamber 11171. The waste liquid channel 1116 is located between the waste liquid outlet and the waste liquid chamber 11171, connecting the waste liquid outlet and the waste liquid chamber 11171. The waste liquid channel 1116 and the waste liquid chamber 11171 are used to guide and store excess waste liquid.

[0069] Furthermore, such as Figure 2 As shown, the waste liquid flow channel 1116 is designed with multiple curved sections. These curved sections increase flow resistance, thereby reducing reagent flow during isothermal or thermal cycling reactions and preventing useless reagents in the waste liquid chamber 11171 from flowing back into the reaction chamber 1112. Depending on the size of the waste liquid chamber 11171, reinforcing ribs can be provided inside. These ribs support the sealing plate 113, preventing channel collapse and adhesion during bonding. The tail end of the waste liquid chamber 11171 has an exhaust port 11172, which extends upwards through the main body plate 111 to its upper surface. The exhaust port 11172 is used for air discharge and pressure balance within the reaction chamber 1112 and the waste liquid chamber 11171. Simultaneously, a breathable membrane 13 is provided at the upper end of the main body plate 111. This breathable membrane 13 allows air to pass through but not liquid, preventing leakage of waste liquid and aerosol contamination.

[0070] In some embodiments of this application, such as Figure 10As shown, the sample processing device 2 has a first docking part 21112, which is tubular and vertically arranged, and its inner cavity is connected to the sample outlet channel 21111. The main body plate 111 has a first positioning hole 11181, which extends vertically through the main body plate 111. After the microfluidic chip 1 is connected to the sample processing device 2, the first docking part 21112 passes through the first positioning hole 11181, and the first through hole 1123 is vertically aligned with the inner cavity of the first docking part 21112, thereby connecting the inner cavity of the first docking part 21112 with the first through hole 1123. The oil or liquid to be dispersed in the sample outlet channel 21111 first flows into the first through hole 1123 through the inner cavity of the first docking part 21112, and then enters the first branch channel 1121 or the second branch channel 1122 through the first through hole 1123.

[0071] In some embodiments of this application, such as Figure 2 As shown, a first positioning protrusion 1119 is also provided at the lower end of the main body plate 111, and the first positioning protrusion 1119 is arranged parallel to and spaced apart from the first positioning hole 11181; as shown Figure 10 As shown, the sample processing device 2 is provided with a second positioning hole 21113, which is arranged parallel to and spaced apart from the first docking part 21112. After the microfluidic chip 1 is connected to the sample processing device 2, the first positioning protrusion 1119 is inserted into the second positioning hole 21113.

[0072] In some embodiments of this application, the main body plate 111 and the sealing plate 113 are manufactured using light-transmitting materials such as PC and COC. This arrangement allows the fluorescence imaging component of an external detection instrument to perform fluorescence droplet imaging through the main body plate 111 or the sealing plate 113. To enhance the reaction efficiency within the reaction chamber 1112, in some embodiments of this application, the thickness of the sealing plate 113 ranges from 0.1 mm to 0.5 mm, ensuring both sealing and thermal conductivity.

[0073] In practical use, the microfluidic chip 1 is used as a disposable consumable. In this application, it is not necessary to add a driving mechanism 331 to the microfluidic chip 1 to drive the flow channels of the liquid to be dispersed and the oil to form a droplet dispersion effect, which can reduce the manufacturing cost of the microfluidic chip 1. Moreover, the microfluidic chip 1 adopts a multi-layer structure, consisting of a first switching valve 12 and a chip body 11 from top to bottom. The first switching valve 12 includes a pressure block 121, a flexible valve plate 124, and a substrate 112 arranged from top to bottom. The pressure block 121, the flexible valve plate 124, and the substrate 112 can be connected by processes such as bonding, hot pressing, and laser welding. The chip body 11 consists of a main plate 111 and a sealing plate 113 from top to bottom. The substrate 112 and the main plate 111, and the sealing plate 113 and the main plate 111 can also be connected by processes such as bonding, hot pressing, and laser welding. The overall structure of the microfluidic chip 1 facilitates its fabrication and further reduces its processing cost.

[0074] The technical features of the above embodiments of microfluidic chip 1 can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0075] This application also provides an embodiment of a sample processing and culture apparatus, such as... Figures 6 to 14 As shown, the device includes a sample processing device 2 and the aforementioned microfluidic chip 1. The sample processing device 2 includes a main body 21 and a second switching valve 22. The main body 21 has a sample loading slot 21131, a transfer slot 21121, a sample outlet channel 21111, an oil storage tank 21141, and multiple processing slots 21151. The second switching valve 22 is connected to the main body 21 and is configured to connect any one of the sample loading slot 21131, the sample outlet channel 21111, the oil storage tank 21141, and the multiple processing slots 21151 to the transfer slot 21121. The microfluidic chip 1 is connected to the main body 21.

[0076] Specifically, during use, the transfer tank 21121 is connected to an external air source, and positive or negative pressure is applied to the transfer tank 21121 using the external air source. After adding the sample to the sample loading tank 21131, the second switching valve 22 is used to connect the sample loading tank 21131 and the transfer tank 21121. Then, negative pressure is applied to the transfer tank 21121 using the external air source, thereby drawing the sample in the sample loading tank 21131 into the transfer tank 21121. At this time, the sample in the transfer tank 21121 is the primary sample. Subsequently, the transfer tank 21121 and one of the multiple processing tanks 21151 are connected by the second switching valve 22. Positive pressure is then applied to the transfer tank 21121 by an external air source, which can squeeze the primary sample in the transfer tank 21121 into the corresponding processing tank 21151 for processing. After the primary sample is processed into a dispersion liquid, the dispersion liquid is transferred to the transfer tank 21121. Then, the transfer tank 21121 and the sample outlet channel 21111 are connected by the second switching valve 22, and the first branch channel 1121 is opened by the first switching valve 12. Positive pressure is then applied to the transfer tank 21121, which can squeeze the dispersion liquid in the transfer tank 21121 into the sample storage chamber 1111 through the sample outlet channel 21111 and the first branch channel 1121. After the liquid to be dispersed is squeezed into the sample storage chamber 1111, the second switching valve 22 is used to connect the oil storage tank 21141 and the transfer tank 21121. Then, negative pressure is applied to the transfer tank 21121, so that the oil in the oil storage tank 21141 enters the transfer tank 21121. Then, the first switching valve 12 is used to connect the transfer tank 21121 and the oil storage chamber 1110. Then, positive pressure is applied to the transfer tank 21121, so that the oil in the transfer tank 21121 is squeezed into the oil storage chamber 1110. After the oil storage chamber 1110 contains oil and the sample storage chamber 1111 contains the liquid to be dispersed, the transfer tank 21121 and the oil storage chamber 1110 are connected through the first switching valve 12, and the transfer tank 21121 and the sample storage chamber 1111 are also connected, so that both the oil storage chamber 1110 and the sample storage chamber 1111 are connected to the transfer tank 21121; then, a positive pressure is applied to the transfer tank 21121, which can push the oil in the oil storage chamber 1110 and the liquid to be dispersed in the sample storage chamber 1111 to flow simultaneously to the mixing channel 1113, thereby achieving droplet dispersion.

[0077] The sample processing and culture device of this embodiment integrates the microfluidic chip 1 and the sample processing device 2, combining traditionally dispersed pretreatment steps such as sample lysis, purification, and reagent mixing with the core droplet generation and PCR reaction into a coherent system, thus improving operational efficiency. Furthermore, the drive mechanism 331 required for droplet formation in the microfluidic chip 1 is shared with the drive mechanism 331 required for sample transfer during the sample processing, reducing the manufacturing cost of the sample processing and culture device.

[0078] In some embodiments of this application, such as Figure 10 , Figure 12As shown, the sample processing device 2 also includes a sealing membrane 23 and a pressure cap 24. The sealing membrane 23 is connected to the main body 21 to seal the oil storage tank 21141 and multiple processing tanks 21151. During storage and transportation, the sealing membrane 23 seals the oil storage tank 21141 and all processing tanks 21151, preventing reagent evaporation and contamination. Figure 10 As shown, the pressure cap 24 includes a connecting part 241, a mounting part 242, and multiple puncture parts 243. The connecting part 241 is slidably connected to the main body 21, and the mounting part 242 is connected to the connecting part 241. The mounting part 242 is spaced apart from the main body 21 and has a third air hole 2421. The multiple puncture parts 243 are all connected to the mounting part 242. When the pressure cap 24 is moved, each puncture part 243 can puncture the sealing membrane 23 to connect the oil storage tank 21141 with the outside, and the multiple processing tanks 21151 with the outside. When each puncture part 243 punctures the sealing membrane 23, the mounting part 242 at the edge of the third air hole 2421 seals against the opening of the transfer tank 21121, so that the third air hole 2421 communicates with the transfer tank 21121. In use, by moving the pressure cap 24, multiple piercing parts 243 can pierce the sealing film 23 covering the oil storage tank 21141 and the processing tank 21151, eliminating the need to tear or puncture multiple sealing films 23 one by one. Moreover, when the pressure cap 24 is in place, at the same time as the piercing parts 243 pierce the sealing film 23, the mounting part 242 will press and seal the opening of the transfer tank 21121, so that the third air hole 2421 is connected to the transfer tank 21121. By connecting the transfer tank 21121 with an external air source, the preparatory work for the initial processing can be completed, improving the efficiency of the operation.

[0079] It should be noted that in this embodiment, the sealing film 23 can be set in multiple ways, with multiple sealing films 23 being set one-to-one at the openings of the processing tank 21151 and the oil storage tank 21141. Alternatively, only one sealing film 23 can be set, covering all the openings of the processing tank 21151 and the oil storage tank 21141.

[0080] Among them, such as Figure 9 , Figure 10 , Figure 12 As shown, the sample processing device 2 also includes an elastic element 25, one end of which abuts against the main body 21, and the other end abuts against the mounting part 242. Before the sealing film 23 is punctured, the pressure cap 24 and the puncture part 243 are positioned above the sealing film 23 under the action of the elastic element 25. When it is necessary to puncture the sealing film 23, the pressure cap 24 is pressed down, the elastic element 25 is compressed and shortened, and the puncture part 243 approaches and punctures the sealing film 23.

[0081] Specifically, in some embodiments of this application, such as Figure 6 , Figure 9As shown, the main body 21 includes a first part 211 and a barrel-shaped shell 212. The first part 211 includes a base portion 2111, a first tubular portion 2112, a second tubular portion 2113, a third tubular portion 2114, and a plurality of fourth tubular portions 2115. The first tubular portion 2112 is located at the upper middle part of the base portion 2111. The second tubular portion, the third tubular portion 2114, and the plurality of third tubular portions 2114 are arranged around the first tubular portion 2112 at intervals. Connecting ribs 2116 are provided between the first tubular portion 2112 and the second tubular portion 2113, between the first tubular portion 2112 and the third tubular portion 2114, and between the first tubular portion 2112 and each of the fourth tubular portions 2115. The elastic element 25 is a compression spring, and there are multiple elastic elements 25. An elastic element 25 is provided between two adjacent connecting ribs 2116. The cavity of the first tubular portion 2112 forms a transfer trough 21121, the cavity of the second tubular portion 2113 forms a sample loading trough 21131, the cavity of the third tubular portion 2114 forms an oil storage trough 21141 and a processing trough 21151, and the cavity of the fourth tubular portion 2115 forms a processing trough 21151. The barrel-shaped shell 212 is arranged with its opening facing downwards, and the opening is engaged with the base portion 2111. The first tubular portion 2112, the third tubular portion 2114, and multiple fourth tubular portions 2115 are all disposed in the cavity of the barrel-shaped shell 212. The lower part of the second tubular portion 2113 is located in the cavity of the barrel-shaped shell 212, and the upper end of the second tubular portion 2113 protrudes upwards to the outside of the barrel-shaped shell 212 and is threadedly connected to a cap 21132. The pressure cap 24 can be slidably connected to the upper inner side of the barrel-shaped shell 212, or the pressure cap 24 can be slidably connected to the outer side of the first tubular part 2112, the second tubular part 2113, and the third tubular part 2114. The sample outlet channel 21111, the first docking part 21112, and the second positioning hole 21113 are all provided on the base part 2111.

[0082] In some embodiments of this application, one of the multiple processing tanks 21151 is a pyrolysis tank, which pre-stores a mixture of pyrolysis fluid and magnetic beads. After connecting the transfer tank 21121 and the pyrolysis tank using the second switching valve 22, a positive pressure is applied to the transfer tank 21121 using an external air source. This forces the primary sample in the transfer tank 21121 into the pyrolysis tank. The primary sample mixes with the pyrolysis fluid and magnetic beads mixture in the pyrolysis tank to form a secondary sample. Subsequently, the secondary sample is repeatedly transferred between the transfer tank 21121 and the pyrolysis tank using an external air source, ensuring thorough mixing of the primary sample with the pyrolysis fluid and the mixture.

[0083] During the pyrolysis and magnetic adsorption processes, the secondary sample needs to be heated. In some embodiments of this application, to facilitate heating of the secondary sample in the pyrolysis tank, such as... Figure 7 , Figure 8As shown, the sample processing device 2 further includes a first processing component 26; the first processing component 26 is detachably connected to the main body 21, and includes a heating element 261. When the first processing component 26 is connected to the main body 21, the heating element 261 abuts against the wall of the pyrolysis tank. Specifically, the barrel wall of the barrel-shaped shell 212 is provided with a mounting hole 2121, the axial direction of which extends radially along the fourth tubular portion 2115 forming the pyrolysis tank. The heating element 261 can enter the barrel cavity of the barrel-shaped shell 212 through the mounting hole 2121 and fit against the fourth tubular portion 2115 forming the pyrolysis tank.

[0084] In some embodiments, such as Figure 7 , Figure 8 As shown, the lower end of the fourth tubular portion 2115 forming the lysis groove is provided with a contact area 21152. The contact area 21152 has a conical surface, and the heating element 261 contacts the conical surface, which can increase the heat exchange area and improve the heating effect. In this embodiment, the heating temperature is 60°C, and the thermal lysis operation is performed for 3 minutes to release nucleic acid molecules in the sample to be further processed, so that the nucleic acid molecules can be adsorbed by the magnetic beads in the magnetic bead mixture.

[0085] After lysis, the magnetic beads in the lysis tank need to be adsorbed. In some embodiments of this application, at least one of the multiple processing tanks 21151 is provided with a washing solution, at least one with an elution solution, and at least one with a PCR reaction solution. The sample processing device 2 also includes a second processing component 27; the second processing component 27 is detachably connected to the main body 21, and the second processing component 27 includes a magnetic component 271. When the second processing component 27 is connected to the main body 21, the magnetic component 271 abuts against the tank wall of the thermal lysis tank. Specifically, the magnetic component 271 can also enter the barrel cavity of the barrel-shaped shell 212 through the mounting hole 2121 and contact the conical surface. After the first processing component 26 is removed from the mounting hole 2121, the second processing component 27 is inserted into the mounting hole 2121, so that the magnetic component 271 contacts the conical surface and adsorbs the magnetic beads in the lysis tank. The magnetic beads are magnetically adsorbed onto the inner wall of the lysis tank and will not be discharged with the secondary sample transfer. After the second processing component 27 is placed, the waste liquid in the pyrolysis tank is transferred to the transfer tank 21121 by applying negative pressure. Then, the sample loading tank 21131 and the transfer tank 21121 are connected by the second drive valve, and positive pressure is applied to the transfer tank 21121 to discharge the waste liquid in the transfer tank 21121 to the sample loading tank 21131. Subsequently, the cleaning solution in other processing tanks 21151 is transferred to the pyrolysis tank by the second switching valve 22 and by applying positive and negative pressure to the transfer tank 21121 to clean the magnetic beads. Multiple cleaning of the magnetic beads can be achieved through multiple processing tanks 21151. At least one of the multiple processing tanks 21151 contains eluent. After the magnetic beads are cleaned, the eluent is introduced into the lysis tank using the positive and negative pressure of the second switching valve 22 and the transfer tank 21121. Then, the magnetic suction device is removed and placed into the first processing component 26. Elution is accelerated by external heating, with the heating time and temperature set to 60°C for 1 minute. This operation adsorbs the magnetic beads and releases nucleic acid molecules into the eluent. After obtaining the eluent containing nucleic acid molecules, the eluent is transferred to the processing tank 21151 containing PCR reaction solution through the transfer tank 21121 to form the dispersion solution. Subsequently, the dispersion solution is transferred to the transfer tank 21121 through the transfer tank 21121 and the second switching valve 22, and then the dispersion solution and oil are transferred to the sample storage chamber 1111 and the oil storage chamber 1110, respectively, using the first switching valve 12.

[0086] It should be noted that in this embodiment, the oil in the oil storage tank 21141 is first transferred to the oil storage chamber 1110, and then the liquid to be dispersed is transferred to the sample storage chamber 1111. This allows the oil to first wet the mixing channel 1113, the first flow resistance channel 1114, the second flow resistance channel 1115 and the reaction chamber 1112 under the guidance of gravity, thus avoiding the generation of bubbles in the subsequent droplet dispersion operation.

[0087] During droplet dispersion, a negative pressure is first applied to the first vent 1211 and the second vent 1212, so that both the first branch channel 1121 and the second branch channel 1122 are in a conductive state. The second switching valve 22 is used to connect the transfer tank 21121 and the sample outlet channel 21111. Then, a positive pressure is applied to the transfer tank 21121 to push the oil and the liquid to be dispersed to achieve droplet generation based on flow focusing at the cross-shaped flow channel. The droplets are spread out in the reaction chamber 1112, and the excess oil is discharged into the waste liquid chamber 11171 through the waste liquid flow channel 1116. The excess oil acts as a seal for the reaction chamber 1112 to prevent aerosol contamination.

[0088] After the droplets are dispersed, the heating device of the external instrument is placed close to the lower end face of the sealing sheet 113 to perform heating and cooling cycles on the droplets in the reaction chamber 1112, completing the PCR amplification reaction process. The heating device can be a semiconductor cooling chip. After amplification, the external instrument performs fluorescence imaging detection through the light-transmitting sealing sheet 113 or the main plate 111.

[0089] In some embodiments of this application, such as Figures 11 to 14 As shown, the second switching valve 22 is rotatably connected to the main body 21; the second switching valve 22 has a first channel 2211, and a first port and a second port connected to the first channel 2211; the first port is connected to the transfer tank 21121, and rotating the second switching valve 22 can connect the second port to any one of the sample loading tank 21131, the sample discharging channel 21111, the oil storage tank 21141 and the multiple processing tanks 21151; in order to facilitate the detection of the angular state of the second switching valve 22, the second switching valve 22 is connected to a magnet 29, and the magnetic field direction of the magnet 29 is perpendicular to the rotation axis of the second switching valve 22. The position state of the second switching valve 22 can be determined by detecting the magnetic field direction of the magnet 29. The second switching valve 22 is rotatably connected to the lower end of the base. The base portion 2111 has corresponding first connecting holes at the vertically opposite positions of the first tubular portion 2112, the second tubular portion 2113, the third tubular portion 2114, and each of the fourth tubular portions 2115. Each first connecting hole communicates with the lumen of the first tubular portion 2112, the second tubular portion 2113, the third tubular portion 2114, and each of the fourth tubular portions 2115. The extension path of the first channel 2211 is a U-shape with the opening facing upwards. A sealing plate 28 is provided between the base and the second switching valve 22. The sealing plate 28 has second connecting holes at the positions opposite to the first connecting holes. The sealing plate 28 is made of soft materials such as silicone, rubber, or TPU. The thickness of the sealing plate 28 is greater than or equal to 0.2 mm and less than or equal to 2 mm. Too thin a thickness is detrimental to sealing and rotational dynamic stability, while too thick a thickness will result in a large uneven area, posing a risk of reagent leakage and crosstalk.

[0090] In some embodiments of this application, the second switching valve 22 has a stepped columnar structure. The second switching valve 22 includes a large-diameter section 221 and a small-diameter section 222. The large-diameter section 221 is connected to the upper end of the small-diameter section 222. A sealing plate 28 is disposed between the upper end surface of the large-diameter section 221 and the base portion 2111. The first channel 2211, the first port, and the second port are all disposed in the large-diameter section 221, and the first port and the second port are located at the upper end surface of the large-diameter section 221. The lower end surface of the small-diameter section 222 has a groove 2221. A protrusion 311 is provided on the external driving component. The protrusion 311 can drive the second switching valve 22 to rotate by inserting into the groove 2221. The main body 21 also includes a second part 213, which is detachably connected to the lower end of the base portion 2111. The second part 213 has a clearance hole 2131 for the small diameter section 222 to pass through. After the second part 213 is connected to the base portion 2111, the upper end face of the second part 213 abuts against the lower end face of the large diameter section 221, thereby pressing the large diameter section 221 upward, so that the sealing sheet 28 can be clamped between the large diameter section 221 and the base portion 2111.

[0091] The technical features of the above-described sample processing and culture apparatus embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0092] This application also provides a preferred embodiment of a molecular detection system, such as... Figure 15 , Figure 16 As shown, the molecular detection system includes the sample processing and culture device described above. The molecular detection system also includes an operating device 3, which includes a mounting platform 31 and a first driving member 32. The first driving member 32 is connected to the mounting platform 31 and is driven by a second switching valve 22 to drive the second switching valve 22 to rotate. The second switching valve 22 is connected to a magnet 29, and the magnetic field direction of the magnet 29 is perpendicular to the rotation axis of the second switching valve 22. The first driving member 32 and / or the mounting platform 31 are connected to a detection element for detecting the magnetic field direction of the magnet 29.

[0093] In this embodiment, the cam 311 is connected to the first driving component 32, and the first driving component 32 is connected to the mounting stage 31. The microfluidic chip 1 and the sample processing device 2 in the sample processing and culture device are both disposable items. In this embodiment, the magnet 29 has a lower cost, while the detection device has a higher cost. The magnet 29 is connected to the second switching valve 22, which cannot be reused, and the detection device is connected to the operating device 3, which can be reused, so that the detection device can be reused and the operating cost of the molecular detection system is reduced.

[0094] In some embodiments of this application, the operating device 3 further includes a lifting device 33, a first docking member 34, a second docking member 35, and a third docking member 36. The first docking member 34 has a first vent hole, the second docking member 35 has a second vent hole, and the third docking member 36 has a third vent hole. The lifting device 33 includes a driving mechanism 331 and a lifting member 332. The driving mechanism 331 connects the lifting member 332 and the mounting platform 31, and the lifting member 332 is located above the pressure cover 24. The first docking member 34, the second docking member 35, and the third docking member 36 are all connected to the lifting member 332. The driving mechanism 331 drives the lifting member 332 to descend, which enables the third docking member 36 to press down on the pressure cover 24, so that each puncture part 243 punctures the sealing film 23. It also enables the first vent hole to communicate with the first air hole 1211, the second vent hole to communicate with the second air hole 1212, and the third vent hole to communicate with the third air hole 2421. The lifting device 33 can drive the first docking part 34, the second docking part 35 and the third docking part 36 to move synchronously, thereby pressing down the pressure cap 24, puncturing the sealing film 23, and allowing ventilation through the first air hole 1211, the second air hole 1212 and the third air hole 2421, thus improving work efficiency.

[0095] Specifically, the third docking member 36 has a stepped shaft structure, comprising a small-diameter pipe section 361 and a large-diameter pipe arranged coaxially. The outer diameter of the small-diameter pipe section 361 is smaller than the inner diameter of the third air hole 2421, and the diameter of the large-diameter pipe section is larger than the inner diameter of the third air hole 2421. The lifting member 332 moves the third docking member 36 downward, first inserting the small-diameter pipe section 361 into the third air hole 2421, thus connecting the cavity of the small-diameter pipe section 361 with the third air hole 2421, thereby connecting the small-diameter pipe section 361 with the transfer groove 21121. Further downward movement of the third docking member 36 allows the small-diameter section 222 to push the pressure cap 24 downward, thereby causing the pressure cap 24 to compress the elastic member 25 downward and causing the piercing part 243 to pierce the sealing membrane 23. After the puncture part 243 punctures the sealing membrane 23, the lower end face of the pressure cap 24 abuts against the upper end face of the first tubular part 2112. At the same time, the second mating part 35 and the third mating part 36 abut against the upper end face of the pressure block 121, so that the second vent hole is connected to the second air hole 1212 and the first vent hole is connected to the first air hole 1211. The large-diameter pipe section enhances the structural strength of the third mating part 36, ensuring that the third mating part 36 will not deform when it presses the pressure cap 24 downward.

[0096] In some embodiments of this application, the lower end of the mounting portion 242 is connected to a first sealing ring 244. The first sealing ring 244 is coaxially arranged with the third air hole 2421. By pushing the cover 24 downward, the first sealing ring 244 can be sandwiched between the upper end face of the first tubular portion 2112 and the lower end face of the mounting portion 242, thereby improving the sealing performance between the mounting portion 242 and the first tubular portion 2112.

[0097] In some embodiments of this application, two second sealing rings may be provided at the upper end of the pressure block 121. The two second sealing rings are coaxially arranged with the first air hole 1211 and the second air hole 1212 respectively, thereby improving the sealing performance between the first mating member 34 and the pressure block 121, and between the second mating member 35 and the pressure block 121.

[0098] In some embodiments of this application, the drive mechanism 331 includes a support column 3311, a lead screw 3313, a slider 3314, and a drive motor 3315. The support column 3311 is arranged vertically, and the lead screw 3313 is arranged parallel to and spaced apart from the support column 3311. The support column 3311 has a guide post portion 3312 on the side facing the lead screw 3313. The slider 3314 is slidably connected to the guide post portion 3312, and the slider 3314 has a threaded hole arranged vertically. The lead screw 3313 is threadedly inserted into the threaded hole. The lifting member 332 is fixed on the slider 3314. The drive motor 3315 is connected to the lead screw 3313. The motor drives the lead screw 3313 to rotate, which can drive the slider 3314 to move up and down, thereby driving the lifting member 332 to move up and down.

[0099] The molecular detection system of this application enables efficient detection using droplet-dispersive PCR. The technical features of the above molecular detection system can be combined arbitrarily. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described; however, as long as the combinations of these technical features are not contradictory, they should be considered within the scope of this specification.

[0100] The above description is only a preferred embodiment of this application. It should be noted that for those skilled in the art, several improvements and substitutions can be made without departing from the technical principles of this application, and these improvements and substitutions should also be considered within the scope of protection of this application.

Claims

1. A microfluidic chip (1), characterized in that, The microfluidic chip (1) is used to connect to a sample processing device (2), the sample processing device (2) having a sample outlet channel (21111), the sample outlet channel (21111) being capable of outputting any one of positive pressure gas, oil, and liquid to be dispersed; the microfluidic chip (1) includes: The chip body (11) has an oil storage chamber (1110), a sample storage chamber (1111) and a reaction chamber (1112) arranged at intervals. The chip body (11) also has a mixing channel (1113), one end of which is connected to the sample storage chamber (1111) and the oil storage chamber (1110), and the other end is connected to the reaction chamber (1112). A first switching valve (12) is connected to the chip body (11). The first switching valve (12) is configured to connect either the oil storage chamber (1110) or the sample storage chamber (1111) to the sample outlet channel (21111). The first switching valve (12) is also configured to simultaneously connect the sample storage chamber (1111) and the oil storage chamber (1110) to the sample outlet channel (21111), so that the positive pressure gas in the sample outlet channel (21111) drives the oil in the oil storage chamber (1110) and the liquid to be dispersed in the sample storage chamber (1111) to the mixing channel (1113) and form microdroplets. The chip body (11) includes a substrate (112), which has a first groove and a second groove. The first switching valve (12) includes a pressure block (121), a first valve core assembly (122), and a second valve core assembly (123); the pressure block (121) is connected to the substrate (112), the pressure block (121) and the groove wall of the first groove form a first branch channel (1121), and the substrate (112) and the groove wall of the second groove form a second branch channel (1122); when the microfluidic chip (1) is connected to the sample processing device (2), the first branch channel (1121) connects the sample outlet channel (21111) and the sample storage chamber (1111), and the second branch channel (1122) connects the sample outlet channel (21111) and the oil storage chamber (1110). The first valve core assembly (122) is connected to the pressure block (121) and / or the substrate (112) and is configured to open or close the first branch channel (1121); the second valve core assembly (123) is connected to the pressure block (121) and / or the substrate (112) and is configured to open or close the second branch channel (1122). The first valve core assembly (122) includes a first partition (1221) and a first flexible sheet (1241). The first partition (1221) is fixed in the first groove, and the first flexible sheet (1241) is arranged between the pressure block (121) and the substrate (112), with a portion of the first flexible sheet (1241) opposite to the first partition (1221). The pressure block (121) has a first vent (1211) at the position opposite to the first partition (1221). Applying negative pressure to the first vent (1211) can cause a portion of the first flexible sheet (1241) to protrude into the first vent (1211), thereby forming a first gap between the first flexible sheet (1241) and the first partition (1221), thus opening the first branch channel (1121); applying positive pressure to the first vent (1211) can cause a portion of the first flexible sheet (1241) to adhere to the first partition (1221), thereby closing the first branch channel (1121); And / or, the second valve core assembly (123) includes a second partition (1231) and a second flexible sheet (1242), the second partition (1231) being fixed in the second groove, the second flexible sheet (1242) being disposed between the pressure block (121) and the substrate (112) with a portion of the second flexible sheet (1242) opposite to the second partition (1231); the pressure block (121) having a second vent (1212) at the position opposite to the second partition (1231). Applying negative pressure to the second vent (1212) can cause a portion of the second flexible sheet (1242) to protrude into the second vent (1212), thereby forming a second gap between the second flexible sheet (1242) and the second partition (1231), thus opening the second branch channel (1122); applying positive pressure to the second vent (1212) can cause a portion of the second flexible sheet (1242) to adhere to the second partition (1231), thereby closing the second branch channel (1122).

2. The microfluidic chip (1) according to claim 1, characterized in that, The mixing channel (1113) has an inlet and an outlet, the outlet being connected to the reaction chamber (1112), and the chip body (11) also has a first flow resistance channel (1114) and a second flow resistance channel (1115). The first flow resistance channel (1114) is arranged between the sample storage chamber (1111) and the reaction chamber (1112), connecting the sample storage chamber (1111) and the liquid inlet. The first flow resistance channel (1114) is used to adjust the flow rate of the liquid to be dispersed at the liquid inlet. The second flow resistance channel (1115) is arranged between the oil storage chamber (1110) and the reaction chamber (1112), connecting the oil storage chamber (1110) and the liquid inlet. The second flow resistance channel (1115) is used to adjust the flow rate of the oil at the liquid inlet.

3. A sample processing and culture device, characterized in that, The sample processing device (2) includes a sample processing device (2) and a microfluidic chip (1) according to any one of claims 1 to 2, wherein the sample processing device (2) includes a main body (21) and a second switching valve (22); The main body (21) has a sample loading tank (21131), a transfer tank (21121), a sample dispensing channel (21111), an oil storage tank (21141), and multiple processing tanks (21151). The second switching valve (22) is connected to the main body (21), and the second switching valve (22) is configured to connect any one of the sample addition tank (21131), the sample outlet channel (21111), the oil storage tank (21141) and the plurality of processing tanks (21151) to the transfer tank (21121); The microfluidic chip (1) is detachably connected to the main body (21).

4. The sample processing and culture apparatus according to claim 3, characterized in that, The sample processing device (2) also includes a sealing membrane (23) and a pressure cap (24). The sealing membrane (23) is connected to the main body (21) to seal the oil storage tank (21141) and the plurality of processing tanks (21151). The pressure cap (24) includes a connecting part (241), a mounting part (242), and a plurality of puncture parts (243). The connecting part (241) is slidably connected to the main body (21). The mounting part (242) is connected to the connecting part (241). The mounting part (242) is spaced apart from the main body (21) and has a third air hole (2421). The plurality of puncture parts (243) are all connected to the mounting part (242). Moving the pressure cap (24) allows each of the puncture parts (243) to puncture the sealing membrane (23), thereby connecting the oil storage tank (21141) with the outside world, and the multiple processing tanks (21151) with the outside world; when each of the puncture parts (243) punctures the sealing membrane (23), the mounting part (242) at the edge of the third air hole (2421) seals against the opening of the transfer groove (21121), so that the third air hole (2421) communicates with the transfer groove (21121).

5. The sample processing and culture apparatus according to claim 4, characterized in that, The sample processing device (2) further includes an elastic element (25), one end of which abuts against the main body (21) and the other end of which abuts against the mounting part (242).

6. The sample processing and culture apparatus according to claim 3, characterized in that, One of the plurality of processing tanks (21151) is a pyrolysis tank; The sample processing device (2) further includes a first processing component (26) and / or a second processing component (27); the first processing component (26) is detachably connected to the main body (21), the first processing component (26) includes a heating element (261), and when the first processing component (26) is connected to the main body (21), the heating element (261) abuts against the wall of the pyrolysis tank; the second processing component (27) is detachably connected to the main body (21), the second processing component (27) includes a magnetic element (271), and when the second processing component (27) is connected to the main body (21), the magnetic element (271) abuts against the wall of the pyrolysis tank.

7. A molecular detection system, characterized in that, The molecular detection system includes the sample processing and culture device according to any one of claims 3 to 6, wherein a second switching valve (22) is rotatably connected to the main body (21); the second switching valve (22) has a first channel (2211), and a first port and a second port communicating with the first channel (2211); the first port is communicating with the transfer tank (21121), and rotating the second switching valve (22) enables the second port to communicate with any one of the sample loading tank (21131), the sample discharging channel (21111), the oil storage tank (21141), and a plurality of processing tanks (21151); The molecular detection system further includes an operating device (3), which includes a mounting platform (31) and a first driving member (32). The first driving member (32) is connected to the mounting platform (31) and is connected to the second switching valve (22) to drive the second switching valve (22) to rotate. The second switching valve (22) is connected to a magnet (29), the magnetic field direction of the magnet (29) is perpendicular to the rotation axis of the second switching valve (22), and the first driving member (32) and / or the mounting platform (31) are connected to a detection member for detecting the magnetic field direction of the magnet (29).

8. The molecular detection system according to claim 7, characterized in that, The molecular detection system includes the sample processing and culture device according to any one of claims 4 to 6, and the sample processing and culture device includes the microfluidic chip (1) according to claim 1 or 2. The operating device (3) further includes a lifting device (33), a first docking member (34), a second docking member (35) and a third docking member (36), wherein the first docking member (34) has a first vent hole, the second docking member (35) has a second vent hole, and the third docking member (36) has a third vent hole; The lifting device (33) includes a drive mechanism (331) and a lifting component (332). The drive mechanism (331) connects the lifting component (332) and the mounting platform (31). The lifting component (332) is located above the pressure cover (24). The first docking component (34), the second docking component (35) and the third docking component (36) are all connected to the lifting component (332). The driving mechanism (331) drives the lifting member (332) to descend, which enables the first docking member (34) to press down on the cover (24), so that each puncture part (243) punctures the sealing film (23), and also enables the first vent hole to communicate with the first air hole (1211), the second vent hole to communicate with the second air hole (1212), and the third vent hole to communicate with the third air hole (2421).

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