CN111111798A - Micro-fluidic detection chip - Google Patents
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- CN111111798A CN111111798A CN201910700750.2A CN201910700750A CN111111798A CN 111111798 A CN111111798 A CN 111111798A CN 201910700750 A CN201910700750 A CN 201910700750A CN 111111798 A CN111111798 A CN 111111798A
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Abstract
本发明涉及一种微流控检测芯片，其包括：储存模块，包括多个储存仓，各储存仓内均用于储存试剂，在检测前，各储存仓的顶部和底部均被构造为封闭状；以及反应模块，设于储存模块的下方，反应模块包括：连接部，包括多个连接头，各连接头内均设有第一通道，多个连接头与多个储存仓一一对应；连接头用于破坏储存仓的底部的密封，以使储存仓内的试剂通过连接头内的第一通道流出；以及反应仓，设于连接部的下方，用于接收通过连接头内的第一通道流出的试剂。本发明将储存模块上的各个储存仓的底部与反应模块上的各个连接头一一对齐，之后将二者用力压紧，使连接头穿透储存仓的底部的密封，芯片组装完成，操作简单、方便，效率高。
The invention relates to a microfluidic detection chip, which comprises: a storage module, including a plurality of storage bins, each storage bin is used for storing reagents, and before detection, the top and bottom of each storage bin are configured to be closed and a reaction module, which is arranged below the storage module, the reaction module includes: a connection part, including a plurality of connection heads, each connection head is provided with a first channel, and the plurality of connection heads are in one-to-one correspondence with a plurality of storage bins; The head is used to break the seal of the bottom of the storage chamber, so that the reagent in the storage chamber flows out through the first channel in the connecting head; and the reaction chamber is arranged below the connecting part and is used for receiving through the first channel in the connecting head effluent reagent. The invention aligns the bottom of each storage bin on the storage module with each connector on the reaction module one by one, and then presses the two together with force, so that the connector penetrates the seal of the bottom of the storage bin, the chip assembly is completed, and the operation is simple , convenient and efficient.
Description
The present application is based on and claims priority from application No. 201910478598.8, filed 2019, 6, month 4, the disclosure of which is incorporated herein by reference in its entirety.
Technical Field
The invention relates to the field of microfluidic detection, in particular to a microfluidic detection chip.
Background
Due to its high integration and strong automation, the microfluidic chip technology is increasingly applied to point-of-care testing (POCT) in clinical testing projects. However, in order to transplant the existing reagent system to the microfluidic platform, the problems of large reaction system, complex reaction steps and high requirement on reaction efficiency need to be overcome. On the other hand, the pre-filling, long-term preservation and release of the reaction reagent, and the orientation and sequential flow of the released reagent in the chip also put high design and processing requirements on the connection and matching of the chip reagent preservation module and the reaction chip.
Therefore, how to store and seal the reagent, how to connect the opened reagent with the reaction chip, how to sequentially conduct the flow of the reagent in a directional manner, how to efficiently complete the reaction between the reagent and the sample, and become a main barrier for limiting the clinical application of the microfluidic technology.
Disclosure of Invention
One of the objectives of the present invention is to provide a microfluidic chip for alleviating the problems of complicated operation and low efficiency of the reaction between the reagent and the sample.
Some embodiments of the invention provide a microfluidic detection chip comprising:
the storage module comprises a plurality of storage bins, each storage bin is used for storing a reagent, and the top and the bottom of each storage bin are closed before detection; and
the reaction module is arranged below the storage module and comprises:
the connecting part comprises a plurality of connectors, a first channel is arranged in each connector, and the connectors correspond to the storage bins one by one; the connector is used for breaking the seal at the bottom of the storage bin so that the reagent in the storage bin flows out through a first channel in the connector; and
the reaction bin is arranged below the connecting part and used for receiving the reagent flowing out through the first channel in the connector.
In some embodiments, the top of the storage bin is larger in size than the bottom of the storage bin.
In some embodiments, the storage module includes a sealing member, the bottom of the storage bin is a throat shape and is provided with a bottom outlet, the sealing member fills the bottom of the storage bin and closes the bottom outlet of the storage bin, and the sealing member is configured to be separated from the bottom of the storage bin under the action of the connector and move toward the middle of the storage bin to open the bottom outlet of the storage bin.
In some embodiments, the storage module comprises a stepped bore provided below the storage bin; the stepped hole comprises a first hole and a second hole, the aperture of the first hole is smaller than that of the second hole, the top of the first hole is connected with the bottom of the stored bin, and the bottom of the first hole is connected with the top of the second hole.
In some embodiments, the connector comprises:
the first connecting section is positioned in the first hole in a detection state, and the size of the first connecting section is matched with that of the first hole so as to realize sealing; and
and the second connecting section is positioned in the second hole in a detection state, and the size of the second connecting section is matched with that of the second hole so as to realize sealing.
In some embodiments, the storage module comprises a sealing member, the bottom of the storage bin is of a necking type and is provided with a bottom outlet, and the sealing member is filled in the bottom of the storage bin and closes the bottom outlet of the storage bin;
the bottom of the second hole of the stepped hole is sealed;
the first connecting section of the connector is configured to break the bottom seal of the second hole, and push the sealing member to separate from the bottom of the storage bin, and move towards the middle of the storage bin to open the bottom outlet of the storage bin.
In some embodiments, the connecting portion is provided with a groove, and the bottom of the connector is arranged in the groove.
In some embodiments, the storage module includes a sample chamber and a filter element disposed in the sample chamber, wherein a bottom of the sample chamber extends toward a top of the sample chamber to form a pointed structure, and the pointed structure is inserted into the filter element.
In some embodiments, the storage compartment comprises a lyophilized reagent compartment for storing a lyophilized reagent, the lyophilized reagent compartment having a top that is at a lower height than the tops of the other storage compartments.
In some embodiments, the microfluidic detection chip comprises a desiccant module, the desiccant module comprises a desiccant cabin for placing a desiccant, and the desiccant module is used for being inserted into the top of the freeze-dried reagent cabin; and under the state that the desiccant module is inserted into the top of the freeze-drying reagent bin, the top of the desiccant bin and the top of the storage bin are located at the same height.
In some embodiments, the storage compartment comprises a freeze-dried reagent compartment for storing freeze-dried reagents, and the upper part of the freeze-dried reagent compartment is provided with a limiting structure for limiting the falling of the drying agent so as to isolate the drying agent from the freeze-dried reagents in the freeze-dried reagent compartment.
In some embodiments, the reaction module includes a plurality of second channels and a valve area, the second channels are connected with the plurality of connectors in a one-to-one correspondence, and the second channels are used for guiding the reagent flowing in through the first channels of the connectors to the valve area, so that the reagent is selectively guided to the reaction chamber through a valve arranged in the valve area.
Based on the technical scheme, the invention at least has the following beneficial effects:
in some embodiments, the storage module and the reaction module are independent modules, when detecting reaction, the storage module is placed above the reaction module, the bottoms of the storage bins on the storage module are aligned with the connectors on the reaction module one by one, and then the bottoms of the storage bins and the connectors are pressed tightly by force to enable the connectors to penetrate through the sealing of the bottoms of the storage bins, the first channel inside the connectors is communicated with the inside of the storage bins, and therefore the assembly of the detection chip is completed.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the invention and together with the description serve to explain the invention without limiting the invention. In the drawings:
fig. 1 is a schematic view of an assembly direction of a microfluidic detection chip provided in some embodiments of the present invention in preparation for detection;
fig. 2 is an assembled schematic view of a microfluidic detection chip according to some embodiments of the present invention;
fig. 3 is a schematic view illustrating a flow direction of liquid inside an assembled microfluidic detection chip according to some embodiments of the present invention;
FIG. 4 is a schematic layout of the connections, reaction chambers and channels of the reaction module according to some embodiments of the present invention;
FIG. 5 is a schematic diagram of a storage module according to some embodiments of the present invention;
FIG. 6 is a schematic cross-sectional view of a storage module according to some embodiments of the present invention;
FIG. 7 is a schematic view of an assembly direction of a storage module and a desiccant module according to some embodiments of the present invention;
FIG. 8 is a schematic cross-sectional view of the partial structure A shown in FIG. 7;
fig. 9 is a schematic view of a storage compartment of a storage module provided with a seal according to some embodiments of the present invention.
Reference numerals in the drawings indicate:
1-a storage module; 11-a storage bin; 111-freeze drying the reagent bin; 12-a sample bin; 13-a seal;
2-a reaction module; 21-a connecting part; 211-a connecting head; 2111-first connection segment; 2112-second connection segment; 212-a fabrication hole; 22-a reaction bin; 23-a second channel; 24-a valve zone; 25-waste liquid bin; 26-an amplification reaction bin; 27-sample quantification bin;
3-a desiccant module; 31-a desiccant bin;
4-valve.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments. It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without any inventive step, are within the scope of the present invention.
In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicate orientations or positional relationships based on those shown in the drawings, and are used merely for convenience in describing the present invention and for simplifying the description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore, should not be taken as limiting the scope of the present invention.
Fig. 1 and 2 are schematic diagrams of a microfluidic detection chip according to some embodiments.
In some embodiments, the microfluidic detection chip includes a reservoir module 1 and a reaction module 2. The storage module 1 can realize the sealed storage and release of the reagent. The reaction module 2 can realize the directional flow guide and the reaction execution of the liquid.
The storage module 1 includes a plurality of storage compartments 11, each of the storage compartments 11 is used for storing a reagent therein, and a top and a bottom of each of the storage compartments 11 are configured to be closed before testing. During detection, the sealing of the top and the bottom of the storage bin 11 is broken, the reagent flows out from the bottom of the storage bin 11, and the sealing of the top of the storage bin 11 is broken to ensure the atmospheric pressure balance in the storage bin 11.
Optionally, the bottom of the storage compartment 11 is sealed by a sealing membrane.
Optionally, the bottom of the storage compartment 11 is sealed by a filling seal 13.
The reaction module 2 is arranged below the storage module 1, and the reaction module 2 comprises a connecting part 21 and a reaction bin 22. The connecting part 21 of the reaction module 2 can be integrally formed with the position where the reaction bin 22 is located, or the connecting part 21 of the reaction module and the position where the reaction bin 22 is located are of a split structure.
The connection portion 21 includes a plurality of connection heads 211. The respective connection heads 211 may be arranged side by side in sequence. Each connector 211 has a first channel therein. The first channel is used for communicating the storage bin 11 and guiding the reagent in the storage bin 11 to the reaction bin 22.
The plurality of connectors 211 correspond to the plurality of storage bins 11 one by one; the connectors 211 are used to break the seal of the bottom of the storage chamber 11, that is, each connector 211 breaks the seal of the bottom of one storage chamber 11, so that the reagent in the storage chamber 11 flows out through the first channel in the connector 211, as shown in fig. 3.
The reaction chamber 22 is disposed below the connection portion 21, and is used for receiving the reagent flowing out through the first channel in the connection head 211.
In some embodiments, the assembly method of the microfluidic detection chip is as follows: the storage module 1 is placed above the reaction module 2, the bottoms of the storage bins 11 on the storage module 1 are aligned with the connectors 211 on the reaction module 2 one by one, and then the two connectors are pressed tightly by force to enable the connectors 211 to penetrate through the seal of the bottoms of the storage bins 11, a first channel in the connectors 211 is communicated with the interior of the storage bins 11, and therefore the assembly of the detection chip is completed, the operation is simple and convenient, and the efficiency is high; and because connector 211 locates the below of storing storehouse 11, do benefit to reagent and flow to connector 211 through the action of gravity.
The seal at the top of the storage compartment 11 may be broken by means on the detection instrument to equalise the gas pressure within the storage compartment 11.
As shown in fig. 5, the storage module 1 includes more than two storage compartments 11 arranged side by side. Each storage bin 11 is provided with a top outlet at the top and a bottom outlet at the bottom, which are the air pressure balance channel and the liquid flow channel of the storage bin 11, respectively.
In some embodiments, in the initial state, the top outlet and the bottom outlet are each sealed by a sealing membrane.
The microfluidic detection chip provided by the embodiment of the disclosure is simple and convenient to operate, can realize the assembly of the chip and the operation of reaction only by manual or simple external instruments, and is convenient to adjust. And the adaptation to different detection systems can be realized simply by overlapping the number of storage bins 11 in the storage module 1 without redesigning the chip.
In some embodiments, the top and bottom of the storage compartment 11 may be sealed with a polymeric film.
The high polymer film can be made of organic high polymer materials, and the thickness range can be adjusted according to needs. The polymer film can be closely attached to the top and bottom of the storage chamber 11 by hot pressing, ultrasonic or adhesive.
Before the detection starts, the storage module 1 and the reaction module 2 are separately stored, and reagents required by the detection experiment are hermetically stored in the storage bin 11 of the storage module 1.
When the sealing of the storage chamber 11 needs to be opened, the sealing film at the bottom of the storage chamber 11 is pierced and fixed by the connector 211 of the connecting part 21 at the upper part of the reaction module 2, so that the directional release of the liquid can be realized. Meanwhile, the punctured sealing membrane seals the joint, so that sealing in the liquid flowing process is guaranteed.
Through the interaction between connector 211, seal membrane and passageway three sealed, sealing performance is good, and the reaction process is secure, and the principle is simple, and processing is convenient, need not complicated design and can realize, greatly reduced the design processing cost of chip.
In other embodiments, as shown in fig. 9, the storage module 1 includes a sealing member 13, the bottom of the storage compartment 11 is a throat type and is provided with a bottom outlet, the sealing member 13 is filled in the bottom of the storage compartment 11 and closes the bottom outlet of the storage compartment 11, and the sealing member 13 is configured to be separated from the bottom of the storage compartment 11 under the force of the connecting head 211 and move toward the middle of the storage compartment 11 to open the bottom outlet of the storage compartment 11.
In some embodiments, as shown in fig. 6, the top of the storage compartment 11 is larger in size than the bottom of the storage compartment 11. The top size of storing storehouse 11 is big, is convenient for pour into reagent into through the top of storing storehouse 11 into, and the bottom size of storing storehouse 11 is little, does benefit to and realizes sealing connection with connector 211.
In some embodiments, the cross-section of the storage compartment 11 between the top and bottom is circular, and the bottom of the storage compartment 11 is of a reduced-diameter shape.
As shown in fig. 6, in some embodiments, the storage module 1 comprises a stepped hole provided below the storage compartment 11; the stepped hole comprises a first hole and a second hole, the aperture of the first hole is smaller than that of the second hole, the top of the first hole is connected with the bottom of the stored bin 11, and the bottom of the first hole is connected with the top of the second hole.
Optionally, the bottom of the second hole is sealed by a sealing membrane.
In some embodiments, joining head 211 includes a first joining segment 2111 and a second joining segment 2112. The width of the first connection section 2111 is smaller than the width of the second connection section 2112.
The first connection section 2111 is located in the first hole in the detection state, and the size of the first connection section 2111 is adapted to the size of the first hole. For example: the first connection section 2111 has a size corresponding to the size of the first hole, or the first connection section 2111 is in interference fit with the first hole for sealing.
The second connecting section 2112 is located in the second hole in the testing state, and the size of the second connecting section 2112 is adapted to the size of the second hole. For example: the second connection section 2112 has a size corresponding to the size of the second hole, or the second connection section 2112 is in interference fit with the second hole for sealing.
As shown in fig. 6, the stepped hole at the lower part of the storage chamber 11 is matched with the first connection section 2111 and the second connection section 2112 of the connection head 211, so as to realize the sealing of the connection contact part of the connection head 211 and the storage chamber 11 after the sealing of the bottom of the storage chamber 11 is broken, prevent the leakage of the reagent led out from the storage chamber 11 through the first channel of the connection head 211, and ensure that the reagent is led to the reaction chamber 22 completely.
In other embodiments, as shown in fig. 9, the storage module 1 comprises a sealing member 13, the bottom of the storage bin 11 is a necking type and is provided with a bottom outlet, and the sealing member 13 is filled in the bottom of the storage bin 11 and closes the bottom outlet of the storage bin 11. The bottom of the second hole of the stepped hole is sealed.
The first connection section 2111 of the connection head 211 is configured to break the bottom seal of the second hole and push the sealing member 13 away from the bottom of the storage compartment 11, moving towards the middle of the storage compartment 11 to open the bottom outlet of the storage compartment 11.
Since the second hole of the stepped hole is sealed in some embodiments, if the bottom opening of the storage chamber 11 connected with the first hole of the stepped hole is not sealed, the reagent stored in the storage chamber 11 may flow into the stepped hole at the lower part of the storage chamber 11 under the influence of physical factors such as bumping and vibration, thereby causing leakage of the reagent liquid during the process of piercing the reaction module 2 and connecting the storage module 1.
Therefore, in another embodiment, to prevent the leakage, the sealing member 13 is filled in the bottom of the storage compartment 11 to close the bottom outlet of the storage compartment 11.
Optionally, the seal 13 comprises a sealing ball or block.
Optionally, the material of the sealing member 13 includes polymer materials such as paraformaldehyde, polycarbonate, polytetrafluoroethylene, and the like.
The diameter of the seal 13 should be slightly larger than the diameter of the bottom of the storage chamber 11 to form an interference fit to seal the bottom outlet of the storage chamber 11.
In operation, the first connecting section 2111 of the connecting head 211 first pierces the sealing membrane at the bottom of the second hole of the stepped hole. At this time, the reagent is prevented from flowing down due to the presence of the seal 13, and thus, leakage of the reagent does not occur.
As the first connection section 2111 goes deeper, the first connection section 2111 contacts the liquid-resistant seal 13 when the connection head 211 is about to be completely fitted into the stepped hole. At this time, as the first connection section 2111 continues to go deep, the sealing member 13 is pushed open, and complete embedding and sealing between the connector 211 and the stepped hole are realized, at this time, the reagent starts to flow along the pipeline, so that the problem of leakage of the reagent during module connection can be effectively avoided, and loss of the reagent and pollution to a detection instrument and the like are avoided.
In some embodiments, the storage module 1 is provided with a plurality of storage compartments 11 for storing the reaction reagents, the bottom of each storage compartment 11 is provided with a channel which is downward connected with the reaction module 2 and is used for guiding the flow of the reagents, and the top of each storage compartment 11 is opened upward for connecting with the external atmosphere, so as to balance the air pressure in the storage compartment 11.
In some embodiments, first connection segment 2111 and second connection segment 2112 of connection head 211 may be integrally formed from plastic. Alternatively, the first connection section 2111 is assembled with the second connection section 2112 using a metal needle.
In some embodiments, as shown in fig. 6, a process hole 212 is further disposed between two adjacent stepped holes on the storage module 1 to reduce the weight of the chip and save materials. As shown in fig. 3, in some embodiments, connecting portion 21 is provided with a recess, and the bottom of connecting head 211 is disposed in the recess.
Optionally, the connecting joint 211 further includes a third connecting section, and the third connecting section is disposed in the groove. The width of the third connecting section is smaller than that of the second connecting section, the width of the third connecting section is consistent with that of the groove, or the third connecting section is in interference fit with the groove.
A third channel is arranged on the connecting part 21, the third channel connects the first channel in the connecting joint 211 and the second channel 23 on the reaction module 2, and the opening of the third channel is positioned in the groove.
Of course, the third channel and the second channel 23 may be an integral channel, the second channel 23 is communicated with the first channel of the connector 211, and the opening of the second channel 23 is located in the groove.
As shown in fig. 6, in some embodiments, the storage module 1 includes a sample chamber 12 and a filter cartridge disposed within the sample chamber 12.
The bottom of the sample chamber 12 extends towards the top of the sample chamber 12 to form a pointed structure, and the pointed structure is inserted into the filter element.
Disposed within the sample compartment 12 is typically a whole blood sample. The filter element is used to adsorb most of the red blood cells in a whole blood sample. The sharp structure penetrating into the filter element can efficiently and quickly lead out the plasma primarily separated from the whole blood sample from the filter element. The plasma initially separated is plasma containing white blood cells, platelets and a small fraction of red blood cells.
In some embodiments, the storage compartments 11 include lyophilized reagent compartments 111 for storing lyophilized reagents, wherein one of the storage compartments 11 contains a reconstitution reagent. When the freeze-dried reagent is redissolved, the corresponding reagent for redissolution is pumped into the reaction bin 22 through the matching of the liquid flow driving pump and the valve 4, then the reagent is reversely pumped to the freeze-dried reagent bin 111 from the reaction bin 22, and the redissolved freeze-dried reagent can enter the reaction bin 22 through the matching of the liquid flow driving pump and the valve 4.
As shown in fig. 7, 8, in some embodiments, the height of the top of the lyophilized reagent compartment 111 is lower than the height of the tops of the other storage compartments 11.
The microfluidic detection chip further comprises a desiccant module 3, the desiccant module 3 comprises a desiccant bin 31 for placing a desiccant, and the desiccant module 3 is used for being inserted into the top of the freeze-drying reagent bin 111; and the top of the desiccant storage 31 is located at the same height as the top of the storage 11 in a state where the desiccant module 3 is inserted into the top of the lyophilized reagent storage 111.
The seal at the bottom of the desiccant cartridge 31 is broken and the bottom of the desiccant cartridge 31 is smaller in size than the desiccant to prevent the desiccant from escaping the desiccant cartridge 31 and falling into the lyophilized reagent cartridge 111. Due to the desiccant, the dryness of the freeze-dried reagent can be better maintained, and the long-term storage of the reagent is facilitated.
In some embodiments, the upper portion of the lyophilized reagent compartment 111 is provided with a limiting structure for limiting the falling of the desiccant so that the desiccant is isolated from the lyophilized reagent in the lyophilized reagent compartment 111.
In some embodiments, the limiting structure may include an upper limiting platform disposed inside the lyophilized reagent compartment 111, and the desiccant is placed on the upper limiting platform of the lyophilized reagent compartment 111, and is limited by the limiting platform, so as to avoid the contact between the desiccant and the lyophilized reagent.
In some embodiments, the upper portion of the lyophilized reagent storage container 111 is provided as a necking section, a flaring section is arranged above the necking section, a desiccant is placed in the flaring section, the size of the desiccant is larger than that of the necking section, and the desiccant is prevented from contacting the lyophilized reagent by limiting through the necking section.
As shown in fig. 4, in some embodiments, the reaction module 2 includes a plurality of second channels 23 and a valve area 24. The second channels 23 are connected to the plurality of connection heads 211 in a one-to-one correspondence. Namely: each connecting head 211 is correspondingly connected to a second channel 23.
The second channel 23 is used to guide the reagent introduced through the first channel of the connection head 211 to the valve section 24, so that the reagent is selectively introduced to the reaction chamber 22 through the valve 3 provided at the valve section 24.
The reaction module 2 comprises a planar valve area 24 which cooperates with the valve 4.
The reaction chamber 22 of the reaction module 2 may be any shape suitable for accommodating the reaction, and the reaction chamber 22 has at least two channels connected to serve as a liquid flow and air pressure balance channel of the reaction chamber 22.
The reaction module 2 comprises a waste liquid bin 25. The waste liquid bin 25 is connected with the reaction bin 22 through a channel, and a hole is drilled at one side of the liquid inlet channel far away from the waste liquid bin 25, or another channel is formed for balancing the air pressure in the waste liquid bin 25.
Further, the waste liquid bin 25 may have any shape and capacity suitable for the application.
Further, the reaction module 2 comprises any number of waste liquid bins 25 for containing waste liquid generated in different reaction steps, so as to achieve better biological pollution prevention effect.
Optionally, a sufficient amount of filter paper, absorbent paper or other absorbent material with liquid fixing ability is placed in the waste liquid bin 25 to fix the waste liquid and prevent the waste liquid from overflowing.
The reaction module 2 further comprises an amplification reaction chamber 26 and a sample quantification chamber 27.
The reaction module 2 is provided with a plurality of channels, for example: and a plurality of second channels for guiding the flow of the liquid flow. The connection between the channels of the reaction module 2 is then realized by means of the valves 4 arranged in the valve area 24.
In some embodiments, the microfluidic chip comprises a valve 4 to enable directional conduction of the liquid flow. Storage module 1 and reaction module 2 are connected through connecting portion 21, and reagent flows out the back from storage module 1, gets into reaction module 2 through connector 211, thereby the valve 4 on the rethread reaction module 2 carries out the water conservancy diversion and realizes the sequential release and the reaction of reagent.
The reaction module 2 integrates a reaction bin 22, a waste liquid bin 25 and a plurality of channels. The reaction chamber 22 is connected to the valve 4 through a separate access passage and can be connected to the remaining passages of the valve area 24 by rotation of the valve 4.
The microfluidic chip provided by the embodiment of the disclosure is made of common materials, is low in price, has a millimeter-scale pipeline design scale, can realize large-scale mold opening injection molding, and is easy to realize large-batch production and manufacturing.
The microfluidic chip provided by the embodiment of the disclosure integrates reagent storage, release, sequential and directional flow, mixing reaction and final signal detection.
The micro-fluidic chip provided by the embodiment of the disclosure can simply adjust the size and the number of the storage bins 11 as required, can meet the requirements of different detection items, can seamlessly butt joint the existing detection reagent, can greatly reduce the production, research, development and processing costs of the micro-fluidic chip, and can improve the clinical application value of the micro-fluidic chip.
Some specific embodiments of microfluidic chips are listed below.
The main body of the storage module 1 is made of PC material and has dimensions 78mm × 23mm × 8 mm. Wherein, distributed have 11 diameter 6mm, the storage storehouse 11 of the cup-shaped structure of the depth 20 mm.
The material of the storage module 1 can also be selected from high polymer materials such as PP, PET and the like, the thickness of the material has no special requirement, and optimization is needed according to the material characteristics and the bonding technology. The bonding technique can be realized by adopting the techniques of gluing, thermal bonding, ultrasonic bonding, ion bonding and the like.
The specific embodiment of the storage module 1 is as follows: the stepped hole at the bottom of the storage bin 11 is sealed by using a film, then the reagent is filled into the storage bin 11 from the opening at the upper part, and the two freeze-drying reagents are respectively placed into the storage bins 11 corresponding to the chips.
In this embodiment, 100uL of the extract, 200uL × 2 of the first washing solution, 200uL × 2 of the second washing solution, 200uL of the third washing solution, 100uL of the eluent, 100uL of paraffin oil, one each of the lyophilized beads A \ B of the extraction reagent, and one each of the lyophilized beads of the amplification reagent were separately poured. Meanwhile, a whole blood separation filler is placed in the sample chamber 12. The upper end of the chip is then sealed with another film.
The reaction module 2 is made of PC material. The appearance of the plastic is an irregular plastic three-dimensional structure. The reaction module 2 comprises five parts: the waste liquid bin 25, the connecting part 21, the reaction bin 22, the valve area 24 and the amplification reaction bin 26 are respectively positioned right below, right above, one side of the middle part, the other side of the middle part and the external hanging area of the reaction module 2.
The waste liquid bin 25 is located at the lower part of the reaction module 2, and has internal dimensions of 51mm × 15mm × 2mm, a chamfer radius of four sides of 1.5mm, a total volume of 1530uL and a wall thickness of 1 mm. Both the liquid and gas passages are connected to the through-holes of the valve area 24 by means of pipes.
The connecting part 21 is located at the upper part of the reaction module 2 and is a row of needle-like structures with truncated pyramids. The outer diameter of the needle (first connection section 2111) is 1.5mm, the inner diameter is 0.8mm, and the length is 2.0 mm. The diameter of the prism base (second connecting section 2112) is 3.0mm, and the height is 1.5 mm. Which is sized to fit into a stepped hole below the storage bin 11 to effect a seal.
The reaction chamber 22 is positioned at one side of the chip, the main body is circular, the diameter is 38mm, the front surface is raised by about 2mm from the center of a circle and gradually descends to the circumference. The depth of the interior of the chamber was 2 mm. A liquid outlet channel extends tangentially from the position right below the reaction bin 22 and is connected with the central hole of the valve area 24. An air pressure channel extends out of the position right above the reaction chamber 22 in a straight line, is used as a channel for adjusting the air pressure of the chamber, and is matched with an instrument for use to provide power for the flowing of the reagent in the chip.
An amplification reaction chamber 26, i.e., a secondary reaction chamber, is located at the other side of the chip and connected to the valve region 24 through a channel. The inner volume is 100uL, and the depth is 0.5 mm. The main body of the utility model is composed of a square with 8mm multiplied by 8mm and a semicircle with a radius of 4 mm.
The experimental preparation for detecting the chip is as follows: the reaction module 2 and the storage module 1 are taken out, and the liquid flow interface of the storage module 1 is aligned with the liquid flow interface of the reaction module 2 and vertically placed on a frame. While it is determined that the valve 3 is in the closed state. The sample is added through the sample dosing bin 27.
The detection chip starts to work, the connector 211 of the reaction module 2 is aligned with the bottom of the storage bin 11, and is pressed down forcibly, so that the connector 211 of the reaction module 2 pierces the sealing film at the bottom of the storage bin 11, the connection between the reaction module 2 and the storage module 1 is completed, and the detection chip is placed into a matched instrument.
The following steps are included in the working procedures for completing nucleic acid extraction and amplification detection by adopting the microfluidic chip provided by the embodiment:
the valve 4 rotates to the passage of the sample quantitative bin 27, and the air is pumped from the air pressure port of the reaction bin 22 to generate negative pressure, so that the sample is pumped into the reaction bin 22 after being filtered by the filler.
The valve 4 rotates to the storage chamber 11 containing the lysis reagent, and the negative pressure is generated by pumping air from the air pressure port of the reaction chamber 22, so as to pump the lysis reagent into the reaction chamber 22.
The valve 4 is rotated to the freeze-drying reagent bin 111, the pressure is applied from the air pressure port of the reaction bin 22, the lysis reagent and the sample mixed solution are pushed back to flow through the freeze-drying reagent bin 111, and the freeze-drying reagent is re-dissolved and then pumped into the reaction bin 22 under negative pressure.
The valve 4 is closed and the liquid in the reaction chamber 22 is reacted using ultrasound.
The magnet adsorbs magnetic particles in the reaction system, the valve 4 rotates to the opening of the waste liquid bin 25, the pressure is applied from the air pressure port of the reaction bin 22, and the waste liquid is discharged into the waste liquid bin 25.
The valve 4 rotates to the first washing liquid storage chamber 11, and the first washing liquid is sucked in by negative pressure caused by air pressure suction from the air pressure port of the reaction chamber 22.
The valve 4 is closed and the liquid in the reaction chamber 22 is reacted using ultrasound.
The magnet adsorbs magnetic particles in the reaction system, the valve 4 rotates to the opening of the waste liquid bin 25, the pressure is applied from the air pressure port of the reaction bin 22, and the waste liquid is discharged into the waste liquid bin 25.
Repeating the steps for 5-7 times.
The valve 4 rotates to the storage bin 11 of the second washing liquid, and the second washing liquid is sucked in by negative pressure caused by air pressure suction from the air pressure port of the reaction bin 22.
The valve 4 is closed and the liquid in the reaction chamber 22 is reacted using ultrasound.
The magnet adsorbs magnetic particles in the reaction system, the valve 4 rotates to the opening of the waste liquid bin 25, the pressure is applied from the air pressure port of the reaction bin 22, and the waste liquid is discharged into the waste liquid bin 25.
Repeating the steps for 9-11 times.
The valve 4 rotates to the storage bin 11 of the third washing liquid, and the third washing liquid is sucked in due to negative pressure caused by air pressure suction from the air pressure port of the reaction bin 22.
The valve 4 is closed, and the magnet is kept in an attracted state and soaked and washed.
The valve 4 is rotated to the opening of the waste liquid bin 25, the air pressure port of the reaction bin 22 is pressurized, and waste liquid is discharged into the waste liquid bin 25.
The valve 4 is rotated to the eluent storage chamber 11, and the eluent is sucked in by sucking air from the air pressure port of the reaction chamber 22 to cause negative pressure.
The valve 4 is closed and the liquid in the reaction chamber 22 is reacted using ultrasound.
The magnet adsorbs magnetic particles in the reaction system, the valve 4 rotates to the freeze-drying reagent bin 111, the pressure is applied from the air pressure port of the reaction bin 22, and the eluted product is pushed into the freeze-drying reagent bin 11 to redissolve the reagent.
The magnet keeps adsorption, air is pumped from the air pressure port of the reaction bin 22, and the re-dissolved reaction system is pumped back to the reaction bin 22.
The valve 4 is rotated to the amplification reaction chamber 26, and the reaction system is slowly pushed into the amplification reaction chamber 26 by pressurizing the reaction chamber 22 through the air pressure port.
The valve 4 is rotated to the storage chamber 11 for paraffin oil, which is pumped to the reaction chamber 22. Then, the sample is rotated to the opening of the amplification reaction chamber 26, and the paraffin oil is pushed into the amplification reaction chamber 26 to seal the opening.
And closing the valve 4 and detecting amplification.
After the reaction is finished, the chip is taken out of the instrument and discarded as a whole.
Some embodiments of the present disclosure can achieve reagent flow control simply by the cooperation of the valve 4 with a variable air pressure source, greatly reducing the technical requirements of the detection instrument. On the other hand, adopt this technical scheme, only need replace the storage module 1 of the different reagents of splendid attire, can satisfy different testing item needs, improved the clinical suitability of this design greatly. Meanwhile, due to the universality, the complexity and difficulty of batch production are reduced, and convenience is provided for the mass production and popularization of the product.
The microfluidic detection chip provided by the embodiment can be applied to the field of clinical detection.
In the description of the present invention, it should be understood that the terms "first", "second", "third", etc. are used to define the components, and are used only for the convenience of distinguishing the components, and if not otherwise stated, the terms have no special meaning, and thus, should not be construed as limiting the scope of the present invention.
Furthermore, the technical features of one embodiment may be combined with one or more other embodiments advantageously without explicit negatives.
Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present invention and not to limit the same; although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art will understand that: modifications to the specific embodiments of the invention or equivalent substitutions for parts of the technical features may be made; without departing from the spirit of the present invention, it is intended to cover all aspects of the invention as defined by the appended claims.
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