CN114308144A

CN114308144A - Micro-fluidic chip

Info

Publication number: CN114308144A
Application number: CN202011042956.XA
Authority: CN
Inventors: 胡立教; 张玙璠; 袁春根
Original assignee: BOE Technology Group Co Ltd; Beijing BOE Health Technology Co Ld
Current assignee: BOE Technology Group Co Ltd; Beijing BOE Health Technology Co Ld
Priority date: 2020-09-28
Filing date: 2020-09-28
Publication date: 2022-04-12
Anticipated expiration: 2040-09-28
Also published as: CN114308144B

Abstract

The invention discloses a micro-fluidic chip, comprising: chip substrate, chip cover board and liquid storage bag, chip substrate has and predetermines the liquid pool, the top surface of predetermineeing the liquid pool is opened, the diapire of predetermineeing the liquid pool is equipped with punctures the structure, chip cover board locates the top surface of chip substrate, chip cover board is including apron body and control valve block, the apron body has the relative open region that sets up with predetermineeing the liquid pool, the control valve block is located the open region and is linked to each other with the local edge of open region, can buckle downwards with relative apron body, liquid storage bag is fixed in the bottom of control valve block, and accomodate in predetermineeing the liquid pool, when control valve block drives liquid storage bag and moves down, liquid storage bag can be impaled by punctures the structure, so that the liquid inflow in the liquid storage bag predetermines the liquid pool. According to the micro-fluidic chip disclosed by the invention, a reagent can be injected by adopting the puncture mechanism consisting of the control valve plate, the liquid storage bag and the puncture structure, the puncture mechanism is simple and easy to realize, and the manual operation time and the manual operation errors can be reduced.

Description

Micro-fluidic chip

Technical Field

The invention relates to the technical field of microfluidic chips, in particular to a microfluidic chip.

Background

The microfluidic chip is also called a lab-on-a-chip, and means that basic operation units related to the fields of biology, chemistry, medicine and the like, such as sample preparation, reaction, separation, detection and the like, are integrated on a chip with a micro-channel with micron scale, and the whole process of reaction and analysis is automatically completed. The analysis and detection device based on the microfluidic chip has the advantages that: the sample dosage is few, and analysis speed is fast, is convenient for make portable instrument, is applicable to instant, on-the-spot analysis very much, and the micro-fluidic chip is mostly disposable product moreover, can save liquid way systems such as complicated washing and waste liquid treatment like this. To realize automation and integration of the microfluidic chip analysis and detection device, each function of reaction analysis should be integrated on the chip as much as possible to reduce dependence on off-chip operation, however, reagent storage of the microfluidic chip in the related art is complicated in structure or process, so that the cost of the microfluidic chip as a consumable material is too high, and reagent delivery cannot be accurately and precisely controlled.

Disclosure of Invention

The present invention is directed to solving at least one of the problems of the prior art. Therefore, the invention provides a microfluidic chip which can reduce the manual operation time and errors easily caused by manual operation.

According to the embodiment of the invention, the microfluidic chip comprises: the chip substrate is provided with a preset liquid pool, the top surface of the preset liquid pool is open, and the bottom wall of the preset liquid pool is provided with a puncture structure; the chip cover plate is arranged on the top surface of the chip substrate and comprises a cover plate body and a control valve plate, the cover plate body is provided with an opening area opposite to the preset liquid pool, and the control valve plate is arranged in the opening area and is connected with the local edge of the opening area so as to be capable of being bent downwards relative to the cover plate body; the liquid storage bag is fixed at the bottom of the control valve plate and is contained in the preset liquid pool, and when the control valve plate drives the liquid storage bag to move downwards, the liquid storage bag can be pierced by the piercing structure, so that liquid in the liquid storage bag flows into the preset liquid pool.

According to the micro-fluidic chip provided by the embodiment of the invention, the puncture mechanism consisting of the control valve plate, the liquid storage bag and the puncture structure can be used for injecting the reagent, the puncture mechanism is simple and easy to realize, the processing is easy, the material consumption cost is low, the operation of injecting the reagent through the puncture mechanism can be accurately controlled, the manual operation time is reduced, the error easily caused by manual operation is reduced, and the use reliability, the effectiveness and the convenience of the micro-fluidic chip are improved.

In some embodiments, the edge of the control flap has a connection portion, and the control flap is connected to the partial edge of the opening region through the connection portion.

In some embodiments, the bottom wall of the predetermined liquid pool has a liquid through hole, and the puncturing structure is arranged at an edge position of the liquid through hole.

In some embodiments, the puncture structure includes a plurality of puncture bodies, the plurality of puncture bodies are arranged at intervals along the circumferential direction of the liquid through hole, and each puncture body is a multi-pyramid with a sharp upper end.

In some embodiments, the preset liquid pool has a support table, the liquid storage bag has a stop ledge, and when the stop ledge is supported on the support table, the control valve plate drives the liquid storage bag to move down to a lower limit position.

In some embodiments, the liquid-storing pouch comprises: the upper die is of a plane structure and is fixed at the bottom of the control valve plate, the lower die is fixed at the bottom of the upper die, and the middle of the lower die is concave downward so as to limit a liquid containing cavity between the upper die and the lower die.

In some embodiments, the microfluidic chip further includes a membrane pump sheet, a first membrane valve sheet, and a second membrane valve sheet disposed on the bottom surface of the chip substrate, wherein a membrane pump cavity is defined between the membrane pump sheet and the bottom surface of the chip substrate, the chip substrate has a sample cell, an extraction cell, a first flow channel, and a second flow channel, the first flow channel communicates the sample cell with the membrane pump cavity, the second flow channel communicates the membrane pump cavity with the extraction cell, the first membrane valve sheet acts on the first flow channel, and the second membrane valve sheet acts on the second flow channel.

In some embodiments, the microfluidic chip further includes a membrane pump sheet, a second membrane valve sheet, a third membrane valve sheet, and a fourth membrane valve sheet disposed on the bottom surface of the chip substrate, wherein a membrane pump cavity is defined between the membrane pump sheet and the bottom surface of the chip substrate, the chip substrate has an extraction pool, an amplification detection pool, a second flow channel, a third flow channel, and a fourth flow channel, the second flow channel communicates with the membrane pump cavity and the extraction pool, the third flow channel and the fourth flow channel are independent of each other, the third flow channel and the fourth flow channel both communicate with the membrane pump cavity and the amplification detection pool, the second membrane valve sheet acts on the second flow channel, the third membrane valve sheet acts on the third flow channel, and the fourth membrane valve sheet acts on the fourth flow channel.

In some embodiments, the chip substrate has a plurality of liquid pools, each of which has an open top surface, and the plurality of liquid pools includes a lysis pool, a sample pool, an extraction pool, an eluent pool, a first washing pool, a second washing pool, a waste liquid pool, and an amplification detection pool, wherein the lysis pool, the extraction pool, the eluent pool, the first washing pool, and the second washing pool are the preset liquid pools.

In some embodiments, the chip substrate has a plurality of flow channels at the bottom and/or top, and the microfluidic chip further comprises: the chip bottom film is arranged on the bottom surface of the chip substrate, and a film pump area and a plurality of film valve areas acting on the flow channel are arranged on the chip bottom film.

Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.

Drawings

Fig. 1 is an exploded view of a microfluidic chip according to one embodiment of the present invention;

FIG. 2 is a cross-sectional view of the microfluidic chip shown in FIG. 1;

FIG. 3 is a cross-sectional view of the reservoir bladder shown in FIG. 2;

FIG. 4 is a top view of the chip cover plate shown in FIG. 1;

FIG. 5 is a top view of the chip substrate shown in FIG. 1;

FIG. 6 is a bottom view of the chip substrate shown in FIG. 1;

fig. 7 is a bottom view of the chip carrier film shown in fig. 1;

FIG. 8 is a bottom view of the microfluidic chip shown in FIG. 1 after assembly;

fig. 9 is a cross-sectional view of a microfluidic chip according to another embodiment of the present invention.

Reference numerals:

a microfluidic chip 100;

a chip substrate 1; a liquid bath 101; presetting a liquid pool 11;

sample cell 1a 1; extraction tank 1a 2; amplification detection cell 1a 3; lysis cell 1a 4;

eluent pool 1a 5; the first washing liquid tank 1a 6; the second washing liquid tank 1a 7; waste liquid pool 1a 8;

a flow channel 102;

the first flow passage 1b 1; the second flow passage 1b 2; the third flow passage 1b 3; the fourth flow passage 1b 4;

the fifth flow passage 1b 5; the sixth flow passage 1b 6; the seventh flow passage 1b 7; the eighth flow passage 1b 8;

the ninth flow passage 1b 9;

a piercing structure 12; a piercing body 121; a liquid through hole 13; a support table 14;

a chip cover plate 2; a cover plate body 21; an open region 210; a control valve plate 22; a connecting portion 220; a cover sheet 23;

a liquid storage bag 3; a stop ledge 30; an upper die 31; a lower die 32; a liquid containing chamber 33;

a chip base film 4; a membrane pump region 41; a membrane valve region 42; a membrane pump sheet 401; a membrane pump chamber 402;

the first membrane valve sheet 1c 1; the second membrane valve sheet 1c 2; the third membrane valve sheet 1c 3; the fourth membrane valve sheet 1c 4;

the fifth membrane valve sheet 1c 5; the sixth membrane valve sheet 1c 6; the seventh diaphragm valve sheet 1c 7; the eighth membrane valve sheet 1c 8.

Detailed Description

Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the drawings are illustrative and intended to be illustrative of the invention and are not to be construed as limiting the invention.

The following disclosure provides many different embodiments, or examples, for implementing different features of the invention. To simplify the disclosure of the present invention, the components and arrangements of specific examples are described below. Of course, they are merely examples and are not intended to limit the present invention. Furthermore, the present invention may repeat reference numerals and/or letters in the various examples. This repetition is for the purpose of simplicity and clarity and does not in itself dictate a relationship between the various embodiments and/or configurations discussed. In addition, the present invention provides examples of various specific processes and materials, but one of ordinary skill in the art may recognize the applicability of other processes and/or the use of other materials.

Next, a microfluidic chip 100 according to an embodiment of the present invention is described with reference to the drawings. The microfluidic chip 100 according to the embodiment of the present invention may be used for molecular detection of biological samples.

As shown in fig. 1 to 3, the microfluidic chip 100 includes: chip substrate 1, chip cover plate 2 and liquid storage bag 3, chip cover plate 2 locates the top surface of chip substrate 1, chip substrate 1 has predetermined liquid pool 11, the top surface of predetermined liquid pool 11 is opened, the diapire of predetermined liquid pool 11 is equipped with punctures structure 12, combine figure 4, chip cover plate 2 includes cover plate body 21 and control valve plate 22, cover plate body 21 has the open region 210 with predetermineeing liquid pool 11 relative setting, control valve plate 22 locates open region 210 and links to each other with the local edge of open region 210, with relative cover plate body 21 can buckle downwards, get back to figure 2, liquid storage bag 3 is fixed in the bottom of control valve plate 22, and accomodate in predetermined liquid pool 11, when control valve plate 22 drives liquid storage bag 3 and moves down, liquid storage bag can be punctured by punctures structure 12, so that the liquid in the liquid storage bag 3 flows into predetermined liquid pool 11.

Therefore, liquid (for example, a reagent) to be used can be pre-contained in the liquid storage bag 3, when the liquid in the liquid storage bag 3 needs to be injected into the preset liquid pool 11, the control valve plate 22 can be pressed down in a mode of an instrument ejector rod and the like to drive the liquid storage bag 3 to move down, so that the liquid storage bag 3 can be punctured by the puncturing structure 12 at the bottom wall of the preset liquid pool 11, the liquid in the liquid storage bag 3 can flow out and be injected into the preset liquid pool 11, and when the liquid in the liquid storage bag 3 does not need to be injected into the preset liquid pool 11, the control valve plate 22 is not touched, the control valve plate 22 is maintained at an initial position, the liquid storage bag 3 is suspended above the puncturing structure 12 and is not punctured by the puncturing structure 12, and the liquid can be maintained in the liquid storage bag 3.

Therefore, according to the micro-fluidic chip 100 provided by the embodiment of the invention, the puncture mechanism composed of the control valve plate 22, the liquid storage bag 3 and the puncture structure 12 is used for injecting the reagent, the puncture mechanism is simple and easy to implement, the processing is easy, the consumable cost is low, the operation of injecting the reagent through the puncture mechanism can be accurately controlled, the manual operation time is reduced, the error easily caused by manual operation is reduced, and the use reliability, effectiveness and convenience of the micro-fluidic chip 100 are improved.

In some embodiments of the present invention, as shown in fig. 4, the edge of the control valve plate 22 has a connection portion 220, and the control valve plate 22 is connected to a partial edge of the opening region 210 through the connection portion 220. Therefore, the control valve plate 22 can be bent downward by the deformation of the connecting portion 220, and since the control valve plate 22 is connected to the edge of the opening region 210 only by one connecting portion 220, the size of the connecting portion 220 can be reduced to improve the deformation easiness of the connecting portion 220, that is, the connecting portion 220 is more easily deformed, and it is ensured that the puncturing operation can be performed as desired.

Moreover, the connection between the opening region 210 and the control valve plate 22 is realized by providing one connection portion 220, and the chip cover plate 2 can be manufactured conveniently, for example, the chip cover plate 2 can be an integrated piece, an opening annular through hole is reserved on the chip cover plate 2, or the chip cover plate 2 is subsequently processed with an opening annular through hole, so that the control valve plate 22 can be obtained. Of course, the invention is not limited to this, and the control valve plate 22 and the cover plate body 21 may also be a separate piece and fixedly connected through a subsequent process. In addition, in other embodiments of the present invention, the control valve plate 22 may be connected to a plurality of local edges of the opening region 210 through a plurality of connection portions 220 arranged at intervals, so as to improve the reliability of maintaining the control valve plate 22 at the initial position.

In some embodiments of the present invention, as shown in fig. 2 and 5, the bottom wall of the predetermined liquid pool 11 has a liquid through hole 13, so that after the liquid storage pouch 3 is punctured by the puncturing structure 12, the liquid injected into the predetermined liquid pool 11 can simply, effectively, quickly and reliably flow out from the liquid through hole 13, thereby simplifying the structure of the microfluidic chip 100, wherein the puncturing structure 12 can be disposed at an edge position of the liquid through hole 13, that is, the puncturing structure 12 is attached to or spaced from an edge of the liquid through hole 13 at a small distance, thereby greatly shortening the distance between the puncturing position and the liquid through hole 13, so that the liquid in the liquid storage pouch 3 can flow into the liquid through hole 13 more directly and quickly after the puncturing structure 12 punctures the liquid storage pouch 3. Of course, in the embodiment of the present invention, the bottom of the predetermined liquid pool 11 may also be set to be uneven, for example, the terrain at the liquid through hole 13 is low, so that the liquid can flow to the liquid through hole 13 more conveniently.

In some embodiments of the present invention, as shown in fig. 2 and 5, the puncturing structure 12 comprises a plurality of puncturing bodies 121, each puncturing body 121 is a polygonal pyramid with a sharp upper end, that is, the cross section of the puncturing body 121 is polygonal, and the cross section area of the puncturing body 121 is gradually reduced from bottom to top, so that the puncture body 121 can easily puncture the liquid storage bag 3, and since the puncture body 121 is a polygonal pyramid, thereby making the puncture opening of the liquid storage bag 3 irregular, avoiding the puncture opening being blocked by the puncture body 121, further, when the liquid storage bag 3 is punctured, the liquid in the liquid storage bag 3 can be ensured to flow out more reliably, and the plurality of puncturing bodies 121 are arranged at intervals along the circumferential direction of the liquid through hole 13, therefore, the liquid injected into the preset liquid pool 11 can also flow into the liquid through hole 13 through the gap between two adjacent puncture bodies 121, and the reliable work of the microfluidic chip 100 is ensured.

In some embodiments of the present invention, as shown in fig. 2, the predetermined liquid pool 11 has a support platform 14, the liquid storage bag 3 has a stop ledge 30, and the control valve plate 22 drives the liquid storage bag 3 to move down to the lower limit position when the stop ledge 30 is supported on the support platform 14. That is to say, when stock solution bag 3 moved down to the lower extreme position, by the supporting role of brace table 14, stock solution bag 3 can not continue to move down again to can guarantee to keep a determining deviation between stock solution bag 3 and the predetermined liquid pool 11 diapire, in order to avoid stock solution bag 3 to plug up logical liquid hole 13, guarantee that logical liquid hole 13 can carry liquid smoothly, guarantee that micro-fluidic chip 100 can reliably work.

In some embodiments of the invention, as shown in fig. 2 and 3, the reservoir pouch 3 comprises: the liquid valve comprises an upper die 31 and a lower die 32, wherein the upper die 31 is of a plane structure and is fixed at the bottom of the control valve plate 22, the lower die 32 is fixed at the bottom of the upper die 31, and the middle part of the lower die 32 is concave to define a liquid containing cavity 33 with the upper die 31. Therefore, the liquid storage bag 3 is simple in structure, convenient to process and inject liquid, and convenient to connect with the control valve plate 22. It should be noted that the connection manner of the liquid storage bag 3 and the control valve plate 22 is not limited, and for example, the liquid storage bag and the control valve plate may be connected by gluing. In addition, it should be noted that the material of the liquid storage pouch 3 is not limited, for example, the liquid storage pouch may be made of an aluminum foil material, for example, in a specific example, the upper die 31 and the lower die 32 are both formed by hot-pressing an aluminum foil film, the center of the lower die 32 is a concave shape, the shape can be formed by hot-pressing a hot-pressing plate, the lower die 32 is placed into a lower substrate of a hot press after being hot-pressed into the concave shape, then, corresponding reagent liquid is injected through a peristaltic pump, and then, the reagent liquid enters the next station through a flow line to perform hot-pressing connection of the upper die 31 and the lower die 32.

In some embodiments of the present invention, as shown in fig. 8, the microfluidic chip 100 further includes a membrane pump plate 401, a first membrane valve plate 1c1, and a second membrane valve plate 1c2, which are disposed on the bottom surface of the chip substrate 1, a membrane pump cavity 402 is defined between the membrane pump plate 401 and the bottom surface of the chip substrate 1, the chip substrate 1 has a sample cell 1a1, an extraction cell 1a2, a first flow channel 1b1, and a second flow channel 1b2, the first flow channel 1b1 communicates the sample cell 1a1 with the membrane pump cavity 402, the second flow channel 1b2 communicates the membrane pump cavity 402 with the extraction cell 1a2, the first membrane valve plate 1c1 acts on the first flow channel 1b1, and the second membrane valve plate 1c2 acts on the second flow channel 1b 2.

Therefore, in the process of nucleic acid extraction, the liquid can reciprocate between the sample cell 1a1, the membrane pump cavity 402 and the extraction cell 1a2 through the first flow channel 1b1 and the second flow channel 1b2 by controlling the movement of the first membrane valve plate 1c1, the second membrane valve plate 1c2 and the membrane pump plate 401, so that the mixing efficiency can be effectively improved, and the mixing efficiency is further improved through the vibration of the first membrane valve plate 1c1, the second membrane valve plate 1c2 and the membrane pump plate 401, thereby being beneficial to nucleic acid extraction.

In some embodiments of the present invention, as shown in fig. 1 and 7, a chip base film 4 may be disposed at a bottom of a chip substrate 1, the chip base film 4 has a film pump region 41 and a plurality of film valve regions 42, portions of the chip base film 4 other than the film pump region 41 and the film valve region 42 are fixed (e.g., welded or adhered) to a bottom surface of the chip substrate 1, the film pump region 41 and the film valve region 42 are not adhered to be capable of vibrating up and down relative to the chip substrate 1, the film pump region 41 may serve as the film pump sheet 401, and the plurality of film valve regions 42 may include the first film valve sheet 1c1 and the second film valve sheet 1c 2. Thus, the film pump sheet 401, the first film valve sheet 1c1, and the second film valve sheet 1c2 can be conveniently disposed. Of course, the invention is not limited thereto, and in other embodiments of the invention, the membrane pump plate 401, the first membrane valve plate 1c1 and the second membrane valve plate 1c2 may also be elastic membrane plates which are independent from each other and are adhered to the bottom surface of the core substrate 1 through edge rings separately, so as to meet different practical requirements.

In some embodiments of the present invention, as shown in fig. 8, the microfluidic chip 100 further includes a membrane pump sheet 401, a second membrane valve sheet 1c2, a third membrane valve sheet 1c3, and a fourth membrane valve sheet 1c4 disposed on the bottom surface of the chip substrate 1, the membrane pump sheet 401 and the bottom surface of the chip substrate 1 define a membrane pump chamber 402 therebetween, the chip substrate 1 has an extraction cell 1a2, an amplification detection cell 1a3, a second flow channel 1b2, a third flow channel 1b3, and a fourth flow channel 1b4, the second flow channel 1b2 communicates the membrane pump chamber 402 with the extraction cell 1a2, the third flow channel 1b3 and the fourth flow channel 1b4 are independent of each other, the third flow channel 1b 638 and the fourth flow channel 1b4 both communicate the membrane pump chamber 402 with the amplification detection cell 1a3, the second membrane valve sheet 1c2 acts on the second flow channel 1b2, the third membrane valve sheet 6861 c3 acts on the fourth flow channel 3, and the fourth membrane 461 c4 acts on the fourth flow channel 1b 4.

Therefore, in the process of conveying nucleic acid to the amplification detection cell, the movement of the second membrane valve plate 1c2 and the membrane pump plate 401 is controlled, so that the nucleic acid in the extraction cell 1a2 is conveyed to the membrane pump cavity 402 through the second flow channel 1b2, the movement of the third membrane valve plate 1c3 and the membrane pump plate 401 is controlled, so that the nucleic acid in the membrane pump cavity 402 is conveyed to the amplification detection cell 1a3 through the third flow channel 1b3, and the movement of the fourth membrane valve plate 1c4 and the membrane pump plate 401 is controlled, so that the gas in the amplification detection cell 1a3 flows back to the membrane pump cavity 402, thereby ensuring that the nucleic acid can be reliably and smoothly conveyed to the amplification detection cell 1a 3.

In short, by arranging the third flow channel 1b3 and the fourth flow channel 1b4, a closed-loop flow channel loop is formed between the amplification detection cell 1a3 and the membrane pump cavity 402 through the liquid inlet flow channel (i.e., the third flow channel 1b3) and the gas outlet flow channel (i.e., the fourth flow channel 1b4), so that a gas outlet structure is omitted, the flow channel loop is fully utilized, and the nucleic acid can smoothly flow into the amplification detection cell 1a 3.

In some embodiments of the present invention, as shown in fig. 1 and 7, a chip bottom film 4 may be disposed at the bottom of a chip substrate 1, the chip bottom film 4 has a film pump region 41 and a plurality of film valve regions 42, portions of the chip bottom film 4 except the film pump region 41 and the film valve region 42 are fixed (e.g., welded or adhered) to the bottom surface of the chip substrate 1, the film pump region 41 and the film valve region 42 are not adhered to be capable of vibrating up and down relative to the chip substrate 1, the film pump region 41 may serve as the film pump sheet 401, and the plurality of film valve regions 42 may include the second film valve sheet 1c2, the third film valve sheet 1c3, and the fourth film valve sheet 1c 4. Thus, the film pump sheet 401, the second film valve sheet 1c2, the third film valve sheet 1c3, and the fourth film valve sheet 1c4 can be conveniently provided. Of course, the invention is not limited thereto, and in other embodiments of the invention, the membrane pump plate 401, the second membrane valve plate 1c2, the third membrane valve plate 1c3, and the fourth membrane valve plate 1c4 may also be elastic membrane plates which are independent from each other and are adhered to the bottom surface of the core substrate 1 through edge rings separately, so as to meet different practical requirements.

In some embodiments of the present invention, as shown in fig. 1 and 5, the chip substrate 1 has a plurality of liquid pools 101, a top surface of each liquid pool 101 is open, and at least one liquid pool 101 is a predetermined liquid pool 11. For example, in the example shown in fig. 5 and 8, the plurality of liquid cells 101 includes a lysis cell 1a4, a sample cell 1a1, an extraction cell 1a2, an eluent cell 1a5, a first eluent cell 1a6, a second eluent cell 1a7, a waste liquid cell 1a8, and an amplification detection cell 1a3, wherein the lysis cell 1a4, the extraction cell 1a2, the eluent cell 1a5, the first eluent cell 1a6, and the second eluent cell 1a7 are all preset liquid cells 11, so that reagents can be injected into the preset liquid cells 11, and all the reagents can be injected through the puncturing mechanism, so that the manual operation time can be reduced, the error easily caused by the manual operation can be avoided, and the use reliability, effectiveness, and convenience of the microfluidic chip 100 can be improved.

Referring to fig. 6 and 8, the chip substrate 1 has a plurality of flow channels 102, and referring to fig. 1 and 7, the microfluidic chip 100 further includes a chip base film 4, the chip base film 4 is disposed on the bottom surface of the chip substrate 1, and the chip base film 4 has a film pump region 41 and a plurality of film valve regions 42 acting on the flow channels 102, that is, the portions of the chip base film 4 other than the film pump region 41 and the film valve regions 42 are fixed (e.g., welded or adhered) on the bottom surface of the chip substrate 1, and the film pump region 41 and the film valve regions 42 are not fixed, so as to be capable of vibrating up and down relative to the chip substrate 1, thereby playing a role of pumping liquid.

For example, the plurality of flow channels 102 may include the first flow channel 1b1, the second flow channel 1b2, and the third flow channel 1b3, the membrane pump region 41 may serve as the membrane pump plate 401, the plurality of membrane valve regions 42 may include the first membrane valve plate 1c1, the second membrane valve plate 1c2, the third membrane valve plate 1c3, and the first membrane valve plate 1c1, the second membrane valve plate 1c2, and the membrane pump plate 401 cooperate to allow the first flow channel 1b1 and the second flow channel 1b2 to flow liquid, and the third membrane valve plate 1c3 cooperates with the membrane pump plate 401 to allow the third flow channel 1b3 to flow liquid. In addition, it should be noted that the flow channel 102 is used to connect two liquid reservoirs 101 designed to communicate with each other, for example, when the plurality of liquid reservoirs 101 may include the above-mentioned sample reservoir 1a1, extraction reservoir 1a2, and amplification detection reservoir 1a3, the first flow channel 1b1 and the second flow channel 1b2 are used to communicate the sample reservoir 1a1 with the extraction reservoir 1a2, and the second flow channel 1b2 and the third flow channel 1b3 are used to communicate the extraction reservoir 1a2 with the amplification detection reservoir 1a 3.

In the embodiment of the present invention, all the flow channels 102 may be disposed at the bottom and/or the top of the chip substrate 1, that is, all the flow channels 102 may be disposed at the top of the chip substrate 1 and have an open top (as shown in fig. 1), or all the flow channels 102 may be disposed at the bottom of the chip substrate 1 and have an open bottom, or a plurality of the flow channels 102 may be disposed at the top of the chip substrate 1 and have an open top, and the rest of the flow channels 102 are disposed at the bottom of the chip substrate 1 and have an open bottom, or some of the flow channels 102 may be disposed at the top of the chip substrate 1 and have an open top, and the rest of the flow channels 102 may be disposed at the bottom of the chip substrate 1 and have an open bottom (as shown in fig. 9). Therefore, different practical requirements can be met, and the design is more flexible. The open bottom surface of the flow channel 102 (or a part of the flow channel 102) may be closed by the bottom chip carrier film 4, and the open top surface of the flow channel 102 (or a part of the flow channel 102) may be closed by the top chip cover plate 2, with respect to the flow channel 102 (or a part of the flow channel 102) which is provided at the bottom of the chip substrate 1 and has an open bottom.

Next, a microfluidic chip 100 according to an embodiment of the present invention is described.

As shown in fig. 1 to 3, the microfluidic chip 100 may include: chip base plate 1, chip apron 2, stock solution bag 3 and chip basement membrane 4, chip apron 2 locates the top of chip base plate 1, and chip basement membrane 4 locates the below of chip base plate 1. As shown in fig. 4, the material of the chip cover plate 2 may be ABS, as shown in fig. 5-6, the material of the chip substrate 1 may be PP, and the chip substrate 1 is processed by an injection molding process, as shown in fig. 5, the chip substrate 1 has a plurality of liquid pools 101 with open tops, as shown in fig. 6, the bottom of the chip substrate 1 has a plurality of flow channels 102 with open bottoms, and all reagents flow and react in the liquid pools 101 and the flow channels 102.

As shown in fig. 7, the chip base film 4 may be an elastic plastic composite film, and may be a composite film of a PP film and a PET film, wherein the PET film has certain elasticity and toughness and is located at a lower layer, the PP film is located at an upper layer and is the same as the chip substrate 1, and the compatibility is better, the PP film of the upper layer is welded to the lower surface of the chip substrate 1 by a laser welding method, during the laser welding, the film pump region 41 and the film valve region 42 on the chip base film 4 are hidden by a mask plate and are not welded (for example, the film valve region 42 is a small circular non-welded region shown in fig. 7, and the film pump region 41 is a large circular non-welded region shown in fig. 7), the closing and opening actions of the film valve region 42 and the film pump region 41 are realized by driving the film valve region 42 and the film pump region 41 of the chip base film 4 to be pushed up and pulled down after a positive negative pressure is given to the outside, and a single push up or pull down action is realized, the high frequency push-pull up and down effects the mixing of the reagents in the space above the chip bottom film 4, wherein the membrane pump area 41 is the membrane pump plate 401 and defines the membrane pump cavity 402 with the bottom surface of the chip substrate 1.

As shown in fig. 5, the chip substrate 1 has a plurality of liquid pools 101, at least one liquid pool 101 is a preset liquid pool 11, and the plurality of liquid pools 101 are a sample pool 1a1, a lysis pool 1a4, a first wash pool 1a6, a second wash pool 1a7, an eluent pool 1a5, an extraction pool 1a2, an amplification detection pool 1a3, and a waste liquid pool 1a8, respectively. Referring to fig. 6, the bottom of the chip substrate 1 has a first flow channel 1b1 (for communicating the membrane pump chamber 402 with the sample cell 1a1), a second flow channel 1b2 (for communicating the membrane pump chamber 402 with the extraction cell 1a2), a third flow channel 1b3 (for communicating the membrane pump chamber 402 with the amplification detection cell 1a3), a fourth flow channel 1b4 (for communicating the membrane pump chamber 402 with the amplification detection cell 1a3), a fifth flow channel 1b5 (for communicating the membrane pump chamber 402 with the eluent cell 1a5), a sixth flow channel 1b6 (for communicating the membrane pump chamber 402 with the first eluent cell 1a6), a seventh flow channel 1b7 (for communicating the membrane pump chamber 402 with the second eluent cell 1a7), an eighth flow channel 1b8 (for communicating the membrane pump chamber 402 with the waste liquid cell 1a8), and a ninth flow channel 1b9 (for communicating the lysis cell 4 with the sample cell 1a 1).

Referring to fig. 7 and 8, the plurality of film valve regions 42 include a first film valve sheet 1c1 acting on the first flow channel 1b1, a second film valve sheet 1c2 acting on the second flow channel 1b2, a third film valve sheet 1c3 acting on the third flow channel 1b3, a fourth film valve sheet 1c4 acting on the fourth flow channel 1b4, a fifth film valve sheet 1c5 acting on the fifth flow channel 1b5, a sixth film valve sheet 1c6 acting on the sixth flow channel 1b6, a seventh film valve sheet 1c7 acting on the seventh flow channel 1b7, and an eighth film valve sheet 1c8 acting on the eighth flow channel 1b 8.

As shown in fig. 2 and 3, a control valve plate 22 capable of being bent up and down is disposed on the chip cover plate 2, a liquid storage bag 3 for storing a reagent is adhered to the bottom surface of the control valve plate 22 through a double-sided adhesive tape, the liquid storage bag 3 is accommodated in the predetermined liquid pool 11, when the control valve plate 22 on the chip cover plate 2 is pressed down by a corresponding pressing rod in the apparatus, the control valve plate 22 drives the liquid storage bag 3 to move down, the liquid storage bag 3 is punctured by the tip of the puncturing structure 12 at the bottom of the predetermined liquid pool 11, and the reagent in the liquid storage bag 3 flows into the corresponding flow channel 102 at the bottom of the chip substrate 1 through the liquid through hole 13 beside the puncturing structure 12, so as to perform specific flow and reaction.

As shown in FIGS. 4 and 8, the sample cell 1a1 is a through hole, and after the sample is added to the sample cell 1a1, the cover plate 2 is closed, and the corresponding cover plate 23 on the cover plate 2 may be closed (or may not be closed). The bottom of the lysis pool 1a4 is pre-embedded with freeze-dried or air-dried protease K, the lysis solution is stored in the liquid storage bag 3 in the lysis pool 1a4, when the liquid storage bag 3 in the lysis pool 1a4 is punctured, the lysis solution flows out of the lysis pool 1a4 and is mixed with the protease K pre-embedded at the bottom of the lysis pool 1a4, the protease K is re-melted, the sample, the lysis solution and the protease K are mixed by pushing and pulling the membrane pump area 41 and the corresponding membrane valve area 42 up and down, the sample, the lysis solution and the protease K are sucked out and pushed back into the sample pool 1a1 for mixing, the sample, the lysis solution and the protease K are pushed back into the sample pool 1a1 for temperature domain lysis after mixing, and the lysis solution is pumped into the extraction pool 1a2 by pushing and pulling the membrane pump area 41 and the corresponding membrane valve area 42 up and down.

As shown in fig. 8, the bottom of the extraction pool 1a2 has pre-embedded freeze-dried magnetic beads, the lysed sample dissolves the magnetic beads, and then the magnetic beads are mixed by the membrane sucking and pumping pump region 41 and the corresponding membrane valve region 42, and the mixing of the magnetic bead extraction is combined by mixing the upstream and downstream flow channels 102 and mixing the up and down vibration of the bottom membrane 4 of the chip, so as to improve the mixing efficiency. After the magnetic beads capture nucleic acids, firstly, the rinse solution obtained by puncturing the first wash solution tank 1a6 is injected into the extraction tank 1a2, foreign proteins are washed away by the up-and-down vibration of the chip bottom film 4, and then, after the magnetic beads are attracted by a magnet in the instrument, reagents in a membrane pump are pumped into the waste solution tank 1a 8. Then, the rinse liquid obtained by puncturing the second eluent pool 1a7 is injected into the extraction pool 1a2, the rinsing process of the previous step is repeated, salt ions and some small molecules are washed away, finally, the eluent obtained by puncturing the eluent pool 1a5 is injected into the extraction pool 1a2, eluted nucleic acid is melted and injected into the amplification detection pool 1a3 through the membrane pump cavity 402 and the third flow channel 1b3, and the amplification detection pool 1a3 is communicated with the membrane pump cavity 402 through the fourth flow channel 1b4 to realize ventilation, so that the nucleic acid template can be smoothly pumped into the amplification detection pool 1a 3.

In the above process, the magnet in the associated instrument is moved upward to keep the state of adsorbing the magnetic beads, and after the nucleic acid is pumped into the amplification detection cell 1a3, the instrument moves the magnet downward to pump the reagent in the membrane pump chamber 402 and the magnetic beads in the extraction cell 1a2 into the waste liquid cell 1a 8. After the nucleic acid is amplified at constant temperature or subjected to polymerase chain reaction (i.e., PCR) in the amplification detection cell 1a3, the amplified product is analyzed and detected by the optical detection part of the instrument.

The above steps can be briefly described as follows: adding the sample, covering the chip cover plate 2, and pressing down the control valve plate 22 opposite to the lysis cell 1a4 to make the liquid storage bag 3 containing the lysis solution punctured. Sucking and beating a membrane pump area 41 and a corresponding membrane valve area 42 on a chip bottom membrane 4, mixing a lysate and a sample, sucking and beating the membrane pump area 41 and the corresponding membrane valve area 42 on the chip bottom membrane 4, pumping the mixed lysis sample to an extraction pool 1a2, pushing and sucking the membrane pump area 41 and the corresponding membrane valve area 42 on the chip bottom membrane 4, enabling liquid to flow back and forth in an upstream and downstream flow channel 102 to be mixed with magnetic beads of an extraction pool 1a2, driving the corresponding area on the chip bottom membrane 4 to vibrate up and down, mixing more fully, finally grabbing nucleic acid, pumping the nucleic acid and reagents extracted from the extraction pool 1a2 to an amplification detection pool 1a3 through a membrane pump chamber 402 and a third flow channel 1b3, simultaneously, discharging air bubbles in the amplification detection pool 1a3 to the membrane pump chamber 402 through a fourth flow channel 1b4, amplifying the nucleic acid and the reagent in the amplification detection pool 1a3, and performing in-situ optical scanning detection by a detection mechanism, and outputs the detection result.

Therefore, according to the microfluidic chip 100 of the embodiment of the present invention, on the basis of completing the lysis, the nucleic acid extraction, the nucleic acid amplification and the optical detection, the storage and the quantitative delivery of the reagent can be achieved, the manual operation time and the easily caused errors can be reduced, the storage structure (the liquid storage bag 3) and the puncture mechanism (i.e., the puncture structure 12) of the reagent are simple and easy to implement, and in addition, the liquid inlet channel (i.e., the third channel 1b3) and the gas outlet channel (i.e., the fourth channel 1b4) are combined between the amplification detection cell 1a3 and the membrane pump cavity 402 to form a closed-loop channel circuit, thereby reducing the design of the gas outlet hole and simplifying the structure.

In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.

While embodiments of the invention have been shown and described, it will be understood by those of ordinary skill in the art that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.

Claims

1. A microfluidic chip, comprising:

the chip substrate is provided with a preset liquid pool, the top surface of the preset liquid pool is open, and the bottom wall of the preset liquid pool is provided with a puncture structure;

the chip cover plate is arranged on the top surface of the chip substrate and comprises a cover plate body and a control valve plate, the cover plate body is provided with an opening area opposite to the preset liquid pool, and the control valve plate is arranged in the opening area and is connected with the local edge of the opening area so as to be capable of being bent downwards relative to the cover plate body;

the liquid storage bag is fixed at the bottom of the control valve plate and is contained in the preset liquid pool, and when the control valve plate drives the liquid storage bag to move downwards, the liquid storage bag can be pierced by the piercing structure, so that liquid in the liquid storage bag flows into the preset liquid pool.

2. The microfluidic chip according to claim 1, wherein the edge of the control valve plate has a connection portion, and the control valve plate is connected to the local edge of the opening region through the connection portion.

3. The microfluidic chip according to claim 1, wherein the bottom wall of the predetermined liquid pool has a liquid through hole, and the puncturing structure is disposed at an edge of the liquid through hole.

4. The microfluidic chip according to claim 3, wherein the piercing structure comprises a plurality of piercing bodies, the plurality of piercing bodies are arranged at intervals along the circumferential direction of the liquid through hole, and each piercing body is a multi-pyramid with a sharp upper end.

5. The microfluidic chip according to claim 1, wherein the predetermined liquid pool has a support platform, the liquid storage bag has a stop ledge, and the control valve plate drives the liquid storage bag to move down to a lower limit position when the stop ledge is supported by the support platform.

6. The microfluidic chip according to claim 1, wherein the reservoir pouch comprises: the upper die is of a plane structure and is fixed at the bottom of the control valve plate, the lower die is fixed at the bottom of the upper die, and the middle of the lower die is concave downward so as to limit a liquid containing cavity between the upper die and the lower die.

7. The microfluidic chip according to claim 1, further comprising a membrane pump sheet, a first membrane valve sheet, and a second membrane valve sheet disposed on the bottom surface of the chip substrate, wherein a membrane pump cavity is defined between the membrane pump sheet and the bottom surface of the chip substrate, the chip substrate has a sample cell, an extraction cell, a first flow channel, and a second flow channel, the first flow channel communicates with the sample cell and the membrane pump cavity, the second flow channel communicates with the membrane pump cavity and the extraction cell, the first membrane valve sheet acts on the first flow channel, and the second membrane valve sheet acts on the second flow channel.

8. The microfluidic chip according to claim 1, further comprising a membrane pump sheet, a second membrane valve sheet, a third membrane valve sheet, and a fourth membrane valve sheet disposed on a bottom surface of the chip substrate, wherein a membrane pump chamber is defined between the membrane pump sheet and the bottom surface of the chip substrate, the chip substrate has an extraction cell, an amplification detection cell, a second flow channel, a third flow channel, and a fourth flow channel, the second flow channel communicates with the membrane pump chamber and the extraction cell, the third flow channel and the fourth flow channel are independent of each other, the third flow channel and the fourth flow channel both communicate with the membrane pump chamber and the amplification detection cell, the second membrane valve sheet acts on the second flow channel, the third membrane valve sheet acts on the third flow channel, and the fourth membrane valve sheet acts on the fourth flow channel.

9. The microfluidic chip according to claim 1, wherein the chip substrate has a plurality of liquid pools, each of which has an open top surface, and the plurality of liquid pools includes a lysis pool, a sample pool, an extraction pool, an eluent pool, a first washing pool, a second washing pool, a waste pool, and an amplification detection pool, wherein the lysis pool, the extraction pool, the eluent pool, the first washing pool, and the second washing pool are the predetermined liquid pools.

10. The microfluidic chip according to any of claims 1 to 9, wherein the chip substrate has a plurality of flow channels at the bottom and/or top, the microfluidic chip further comprising:

the chip bottom film is arranged on the bottom surface of the chip substrate, and a film pump area and a plurality of film valve areas acting on the flow channel are arranged on the chip bottom film.