CN108816301B - Microfluidic chip, packaging method thereof and packaging accessory for packaging microfluidic chip - Google Patents
Microfluidic chip, packaging method thereof and packaging accessory for packaging microfluidic chip Download PDFInfo
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- CN108816301B CN108816301B CN201810901054.3A CN201810901054A CN108816301B CN 108816301 B CN108816301 B CN 108816301B CN 201810901054 A CN201810901054 A CN 201810901054A CN 108816301 B CN108816301 B CN 108816301B
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Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/04—Closures and closing means
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0809—Geometry, shape and general structure rectangular shaped
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- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
Abstract
The invention discloses a microfluidic chip, a packaging method thereof and a packaging accessory for packaging the microfluidic chip, wherein the microfluidic chip is used for high-throughput analysis and comprises the following components: the chip body and set up in the strengthening rib at chip body back. The reinforcing ribs comprise edge reinforcing ribs and/or inner reinforcing ribs, wherein the edge reinforcing ribs are annular and are arranged along the edge of the chip body, and the edge of the chip body surrounds the inner reinforcing ribs. According to the microfluidic chip, the reinforcing ribs are arranged on the back of the chip body, so that the strength of the whole microfluidic chip is enhanced, the chip body has stress bending towards the front surface during packaging, the reinforcing ribs can resist the stress of bending deformation of the chip body, the bending deformation of the chip body during packaging is reduced, the flatness of the chip body after packaging is improved, namely the flatness of the whole microfluidic chip is improved, particularly the microfluidic chip with larger area is provided with a precondition for matching the microfluidic chip with a porous plate matched instrument.
Description
Technical Field
The invention relates to the technical field of microfluidics, in particular to a microfluidic chip, a packaging method thereof and a packaging accessory for packaging the microfluidic chip.
Background
Currently, in the field of high throughput analysis, such as cell culture, immune ELISA reactions, nucleic acid amplification, etc., widely used reaction carriers or consumables are multi-well plates, such as 96-well plates and 384-well plates. Around the well plate are a series of mating instruments such as well plate centrifuges, gun pipettes, PCR instruments, enzyme labeling instruments, etc. The cooperation of these instruments and multi-well plates plays an important role in the field of high throughput analysis, but the current automated operation of liquids in multi-well plates also requires large instruments such as pipetting stations for operation, which are expensive and inflexible in use in many situations.
The microfluidic technology is a technology for controlling fluid to complete various biological and chemical reactions under the assistance of external hardware through a micro pipeline and a micro structure, and has the advantages of high throughput and low reagent consumption analysis. Furthermore, microfluidic technology has advantages in fluid manipulation, and is therefore expected to solve the shortcomings of existing high-throughput assays based on multi-well plates. However, the micro-fluidic chip is utilized to solve the defects of the existing porous plate and simultaneously solve some problems. For example, how microfluidic chips are compatible with existing high-throughput analytical instruments based on multi-well plates. Users are used to sample application by a gun pipette, and biological reaction and detection are performed by a PCR instrument, an enzyme-labeled instrument and the like. If the microfluidic chip cannot adapt to these devices, it may be necessary to purchase devices dedicated to the microfluidic chip and to be familiar with the related techniques of use, resulting in increased costs and inconvenience of use.
Therefore, how to design and process microfluidic chips that are compatible with existing multi-well plate-matched instruments and are used for high-throughput analysis is an important need. However, for good heat conduction and optical detection, the microfluidic chip generally needs to be thinner, and the area of the porous plate is larger, so that the area of the microfluidic chip which is adapted to the porous plate is larger.
When the micro-fluidic chip is packaged, the front surface of the micro-fluidic chip is heated, and as the micro-fluidic chip is usually a high-molecular polymer, the high-molecular polymer is heated and then is cooled again to shrink, so that the micro-fluidic chip has stress bending towards the front surface, the flatness of the micro-fluidic chip after packaging is poor, and the packaging quality is affected. The larger the area of the microfluidic chip, the greater the influence of the package on the flatness of the microfluidic chip. Therefore, how to improve the flatness of a large-area microfluidic chip, so as to provide a premise for adapting the microfluidic chip to a porous plate matched instrument, is an important problem to be solved.
Disclosure of Invention
The invention aims to provide a microfluidic chip for high-throughput analysis, so as to improve the flatness of the microfluidic chip, thereby providing a premise that the microfluidic chip can be matched with a porous plate matched instrument. The invention further aims to provide a packaging accessory for packaging the microfluidic chip and a packaging method of the microfluidic chip.
In order to achieve the above object, the present invention provides the following technical solutions:
a microfluidic chip for high throughput analysis, comprising: the chip comprises a chip body and reinforcing ribs arranged on the back of the chip body.
Preferably, the reinforcing ribs comprise edge reinforcing ribs and/or inner reinforcing ribs, wherein the edge reinforcing ribs are annular and are arranged along the edge of the chip body, and the edge of the chip body surrounds the inner reinforcing ribs.
Preferably, when the reinforcing rib includes an internal reinforcing rib, the reaction hole of the microfluidic chip and the projection of the internal reinforcing rib on the front surface of the chip body have no overlapping portion.
Preferably, when the reinforcing bars include an edge reinforcing bar and an inner reinforcing bar, the height of the edge reinforcing bar is higher than the height of the inner reinforcing bar.
Preferably, the height of the inner reinforcing ribs is 1mm-3mm, and the width of the inner reinforcing ribs is 0.5mm-2mm.
Preferably, the reinforcing ribs include inner reinforcing ribs including first inner reinforcing ribs and second inner reinforcing ribs disposed alternately.
Preferably, the reinforcing ribs include internal reinforcing ribs which are strip-shaped and are arranged along the length direction or the width direction of the chip body.
Preferably, the reinforcing rib comprises an edge reinforcing rib, a bulge is arranged on the back of the chip body, the bulge is flush with the edge reinforcing rib, sample adding holes are formed in the front of the chip body, and the sample adding holes are distributed on the bulge.
Preferably, the front surface of the chip body is provided with a groove and a main flow path, the groove is trapped in the bulge, and one end of the main flow path, which is communicated with the sample adding hole, is positioned in the groove.
Preferably, the main flow path includes: a first distribution segment communicated with the sample adding hole, a second distribution segment communicated with the reaction hole through a connecting flow path, and a third distribution segment communicated with the first distribution segment and the second distribution segment; wherein the cross-sectional area of the first distribution segment is greater than the cross-sectional area of the third distribution segment.
Preferably, the microfluidic chip is provided with a positioning structure for positioning with a porous plate matched instrument.
Preferably, the distance between the sample adding holes of the microfluidic chip is 4.5+/-0.2 mm, or the distance between the sample adding holes is 9+/-0.2 mm;
the micro-fluidic chip is rectangular, and the reaction holes of the micro-fluidic chip are distributed in a matrix form;
The reaction holes are distributed at equal intervals in a 16 multiplied by 24 mode, the row spacing of the reaction holes is 4.3mm-4.7mm, and the column spacing of the reaction holes is 4.3mm-4.7mm;
or the reaction holes are distributed at equal intervals in an 8 multiplied by 12 mode, the row interval of the reaction holes is 8.8mm-9.2mm, and the column interval of the reaction holes is 8.8mm-9.2mm;
or the reaction holes are distributed at equal intervals in the form of 8 multiplied by 24, the row interval of the reaction holes is 8.8mm-9.2mm, and the column interval of the reaction holes is 4.3mm-4.7mm;
Or the reaction holes are distributed at equal intervals in a 16 multiplied by 12 mode, the row interval of the reaction holes is 4.3mm-4.7mm, and the column interval of the reaction holes is 8.8mm-9.2mm.
Preferably, the microfluidic chip further comprises an exhaust structure disposed on the chip body, the exhaust structure comprising: an exhaust passage, and an exhaust hole communicating with the exhaust passage.
Preferably, the inlet section of the sample adding hole of the microfluidic chip is a divergent section, and the inlet section diverges from the bottom end of the sample adding hole to the inlet end of the sample adding hole.
According to the microfluidic chip provided by the invention, the reinforcing ribs are arranged on the back of the chip body, so that the structural strength of the whole microfluidic chip is enhanced, when the microfluidic chip is packaged, the chip body has stress bending towards the front, the reinforcing ribs can resist the stress of bending deformation of the chip body, the bending deformation of the chip body during packaging is reduced, the flatness of the chip body after packaging is effectively improved, namely the flatness of the whole microfluidic chip is improved, and particularly the microfluidic chip with larger area provides a precondition for the microfluidic chip to be matched with a porous plate matched instrument.
Based on the microfluidic chip provided above, the embodiment of the invention further provides a packaging fitting for packaging the microfluidic chip, the packaging fitting for packaging the microfluidic chip comprises a support body which is an elastic piece, the support body is provided with a support structure which can be completely attached to the back of the chip body, and the support structure comprises: and the matching structure can be matched with the reinforcing rib.
Preferably, the reinforcing ribs comprise edge reinforcing ribs which are annular and are arranged along the edge of the chip body; the back of the chip body is provided with a bulge, the bulge is flush with the edge reinforcing rib, the front of the chip body is provided with sample adding holes, and the sample adding holes are distributed on the bulge;
The mating structure includes: and the abdication groove is matched with the inner reinforcing rib, the first step structure is matched with the edge reinforcing rib, and the second step structure is matched with the bulge.
Preferably, the support body comprises a first support split body and a second support split body which are connected, and the abdication groove and the first step structure are arranged on the first support split body;
the top surface of the second support split body is lower than the top surface of the first support split body, and the second support split body and the first support split body are matched to form the second step structure;
wherein a gap is provided between the first support sub-body and the second support sub-body.
Based on the microfluidic chip and the packaging accessory for packaging the microfluidic chip, the invention also provides a packaging method of the microfluidic chip, and the packaging method of the microfluidic chip comprises the following steps:
Processing a first substrate and a second substrate, wherein the front surface of the first substrate is provided with a reaction hole, and the back of the first substrate is provided with a reinforcing rib;
adding materials required by the reaction into the reaction holes;
placing the first substrate on a packaging accessory, wherein the back of the first substrate is attached to the packaging accessory;
placing a second substrate on the front side of the first substrate, and packaging the first substrate and the second substrate;
the packaging fitting is used for packaging the microfluidic chip, and the first substrate and the second substrate are elastic pieces.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and that other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art.
Fig. 1 is a schematic back view of a microfluidic chip according to an embodiment of the present invention;
FIG. 2 is a schematic front view of the microfluidic chip of FIG. 1;
FIG. 3 is an enlarged view of a portion of FIG. 2;
fig. 4 is a partial enlarged view of a flow path in a microfluidic chip according to an embodiment of the present invention;
fig. 5 is a partial view of a perspective view of a microfluidic chip provided by an embodiment of the present invention;
Fig. 6 is a schematic diagram of a microfluidic chip according to an embodiment of the present invention during sample loading;
fig. 7 is a schematic structural diagram of a packaging fitting for packaging a microfluidic chip according to an embodiment of the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The microfluidic chip provided by the embodiment of the invention is used for high-throughput analysis and comprises a chip body.
In order to keep good heat conduction characteristics and good optical permeability of the microfluidic chip, the thickness of the chip body is set to be thinner, and the thickness of the chip body is preferably selected to be 0.5mm-3mm. In order to improve the flatness of the micro-fluidic chip with a larger area after packaging, the back of the chip body is provided with the reinforcing ribs, as shown in fig. 1.
The back of the chip body refers to the portion of the chip body on the same side as the sample addition end of the sample addition hole 7. The back of the chip body is the surface of the chip body, which is positioned on the same side as the sample adding end of the sample adding hole 7; the front surface of the chip body is opposite to the back surface of the chip body.
According to the microfluidic chip, the reinforcing ribs are arranged on the back of the chip body, so that the structural strength of the whole microfluidic chip is enhanced. When the packaging modes such as hot-press sealing, laser welding and ultrasonic welding are adopted for packaging, the chip body manufactured by the high polymer is heated and then cooled to shrink, so that the chip body has stress bending towards the front, the reinforcing ribs can resist the stress of bending deformation of the chip body, the bending deformation of the chip body during packaging is reduced, the flatness of the chip body after packaging is effectively improved, namely the flatness of the whole microfluidic chip is improved, and particularly the microfluidic chip with larger area is provided with a precondition for adapting the microfluidic chip to a porous plate matched instrument.
It is understood that the porous plate matching instrument comprises a sample adding device such as a gun, and a reaction detecting device such as a PCR instrument and an enzyme label instrument. The specific type of the instrument matched with the porous plate is selected according to actual needs, and the embodiment of the invention is not limited to the specific type.
The positions, the shapes and the number of the reinforcing ribs are designed according to actual needs. Preferably, the reinforcing ribs include edge reinforcing ribs 4, and the edge reinforcing ribs 4 are annular and are disposed along the edge of the chip body.
In the practical application process, the reinforcing ribs can also be selected to comprise inner reinforcing ribs, and the edges of the chip body surround the inner reinforcing ribs. When the reinforcing bars include the edge reinforcing bars 4, the edge reinforcing bars 4 are located at the periphery of the inner reinforcing bars, i.e., the inner reinforcing bars are located inside the edge reinforcing bars 4.
When the reinforcing ribs comprise the internal reinforcing ribs, the non-overlapping parts of the reaction holes 11 of the microfluidic chip and the projection of the internal reinforcing ribs on the front surface of the chip body are preferentially selected, so that adverse effects on heat transfer and optical detection of the reaction holes 11 caused by the existence of the internal reinforcing ribs are effectively avoided.
Of course, on the premise of ensuring heat transfer of the reaction holes 11 and enabling normal optical detection, it is also possible to select that the projections of the reaction holes 11 and the internal reinforcing ribs on the front surface of the chip body have an overlapping portion that should be as small as possible.
When the flatness of the micro-fluidic chip is ensured to be maintained, the micro-fluidic chip is prevented from shrinking at the inner reinforcing rib as much as possible during injection molding. To achieve this, the internal reinforcement is selected to be of a narrower, taller construction. Preferably, the height of the inner reinforcing ribs is 1mm-3mm, and the width of the inner reinforcing ribs is 0.5mm-2mm.
The specific structure of the inner reinforcing ribs is also designed according to practical needs, for example, in order to effectively improve the strength, the inner reinforcing ribs include a first inner reinforcing rib 5 and a second inner reinforcing rib 6 which are arranged in a crossing manner. The number of the first internal reinforcing ribs 5 may be one or two or more; the number of the second internal reinforcing ribs 6 may be one or two or more.
The internal reinforcing ribs are strip-shaped. Further, the internal reinforcing ribs are provided along a length direction or a width direction of the chip body. At this time, if the internal reinforcing ribs include the first internal reinforcing rib 5 and the second internal reinforcing rib 6 which are disposed alternately, the first internal reinforcing rib 5 is preferably disposed along the length direction of the chip body, and the second internal reinforcing rib 6 is disposed along the width direction of the chip body. It is understood that the chip body is rectangular at this time.
Of course, the inner rib may be formed in other shapes and arranged in other directions, and is not limited to the above embodiment.
When the above-mentioned reinforcing ribs include the edge reinforcing ribs 4, the edge reinforcing ribs 4 can be directly used for clamping the microfluidic chip, so that the height of the edge reinforcing ribs 4 is selected to be high, and preferably the height of the edge reinforcing ribs 4 is 4mm-8mm. Further, when the above-mentioned reinforcing ribs include the edge reinforcing ribs 4 and the inner reinforcing ribs, the height of the edge reinforcing ribs 4 is higher than the height of the inner reinforcing ribs.
When the reinforcing rib comprises the edge reinforcing rib 4, the back of the chip body is provided with the bulge 8, the bulge 8 is flush with the edge reinforcing rib 4, the chip body is provided with the sample adding holes 7, and the sample adding holes 7 are distributed on the bulge 8, as shown in fig. 1. Thus, the supporting surface of the back of the chip body is enlarged, and the microfluidic chip is convenient to place.
In order to further optimize the above technical solution, as shown in fig. 2 and 3, the front surface of the chip body is provided with a groove 14 and a main flow path 13, the groove 14 is trapped in the protrusion 8, and one end of the main flow path 13, which is communicated with the sample adding hole 7, is located in the groove 14; the front end face of the sample adding hole 7 is flush with the front face of the chip body.
It will be understood that the front end face of the well 7 refers to the end face of the well 7 on the same side as the front face of the chip body. The front face of the main flow path 13 is also flush with the front face of the chip body. The front surface of the main channel 13 is the surface of the main channel 13 on the same side as the front surface of the chip body.
The structure is beneficial to flatness and encapsulation of the micro-fluidic chip from three aspects:
1) The uneven surface of the chip body at the position during injection molding due to the solid structure of the bulge 8 is avoided, and the packaging of the microfluidic chip is not facilitated;
2) The front area of the chip body at the groove is reduced, and the larger plane is more difficult to realize the sealing without trapping bubbles, so that the sealing is convenient, and the packaging is convenient;
3) The grooves 14 allow only the area near the sample addition hole 7 and the area of the portion where the main flow channel 13 communicates with the sample addition hole 7 to be in the plane of the front surface of the chip body, and the chip body is in contact with the heat source only in the area near the main flow channel 13 and the vicinity of the sample addition hole 7 during heat sealing or welding, which is advantageous in that these areas are more firmly heat sealed or welded. Particularly, the chip body is located at the edge, compared with the central area of the chip, the heat dissipation is faster, the existence of the groove 14 can separate the sample adding hole 7, the area of the part where the main flow path 13 is communicated with the sample adding hole 7 and the periphery of the chip body from each other by air, and the difference of different packaging effects of the chip body caused by peripheral heat dissipation can be effectively reduced.
The number of the grooves 14 may be one or two or more. To simplify production, the recess 14 is one and is strip-shaped.
As shown in fig. 3 and 4, in the microfluidic chip described above, the main flow channel 13 communicates with the sample well 7 and the reaction well 11. The main flow path 13 includes: the first distribution segment 131 communicating with the well 7, the second distribution segment 132 communicating with the reaction well 11 through the connecting channel 12, and the third distribution segment 133 communicating with the first distribution segment 131 and the second distribution segment 132. The second distribution sections 132 are in one-to-one correspondence with the reaction holes 11.
Preferably, the cross-sectional area of the first distributing section 131 is larger than the cross-sectional area of the third distributing section 133. Two advantages brought by this structure:
1) The first distributing section 131 is directly connected with the sample loading hole 7, and the sample loading hole 7 is located in the protrusion 8, so that the first distributing section 131 can cross the edge of the protrusion 8, as shown in fig. 3, when in heat sealing or ultrasonic welding, the edge part of the protrusion 8 is stressed greatly due to the fact that the thicker edge of the protrusion 8 is directly supported, the deformation of the chip body at the position is large when the chip body is deformed by heat, and if the first distributing section 131 is thinner, the direct deformation is possible to be blocked. Thus, the thicker first distribution segment 131 can avoid this problem;
2) The first distributing section 131 is directly connected with the sample adding port 7, the larger cross section of the first distributing section brings about larger containing volume, and the liquid or air contained in the first distributing section does not participate in the subsequent distribution to the reaction hole 11, so that the liquid or air contained in the first distributing section can play a role in buffering, and the requirements on the accuracy of sample adding amount and sample adding method can be reduced.
The connecting flow path 12 may be partially or entirely blocked by heat sealing deformation, and in order to facilitate blocking thereof, the connecting flow path 12 has a small aspect ratio, preferably, the aspect ratio of the connecting flow path 12 is 0.01 to 0.50.
In order to facilitate the matching of the microfluidic chip and the porous plate matching instrument, the microfluidic chip is provided with a positioning structure 3 for positioning the porous plate matching instrument. It will be appreciated that the above-mentioned perforated plate kit or the fittings adapted to the perforated plate kit also have a structure for positioning with the positioning structure 3 to ensure positioning.
It should be noted that the positioning structure 3 may also be used for correcting deformation of the microfluidic chip.
The positioning structure 3 may be a positioning hole, a positioning column, a positioning groove, etc. If the positioning structure 3 is a positioning column, when the chip body adopts film heat sealing, the film for heat sealing needs to be separated from the positioning column, and requirements on the structure and positioning of the film are met. In order to facilitate the packaging of the chip body, the positioning structure 3 is preferably selected as a positioning hole. At this time, the chip body has no protruding part, namely, the protruding part is not used for blocking the film, and the film can be directly stuck with the film for heat sealing.
The number of positioning structures 3 is chosen according to the actual need. In order to enhance the positioning effect, at least two positioning structures 3 are preferably selected. Further, the number of the positioning structures 3 is four, two positioning structures 3 are located at one end of the microfluidic chip, and the other two positioning structures 3 are located at the other end of the microfluidic chip.
Preferably, the pitch of the loading holes 7 is 4.5.+ -. 0.2mm or 9.+ -. 0.2mm. Thus, the micro-fluidic chip is matched with a gun-discharging liquid-transferring device commonly used by a porous plate, and the like, so that the micro-fluidic chip is convenient to operate.
In order to facilitate matching of the microfluidic chip and the porous plate matched instrument, the microfluidic chip is rectangular, and the reaction holes 11 of the microfluidic chip are distributed in a matrix form. It is understood that the reaction holes 11 are distributed on the chip body. The micro-fluidic chip is rectangular, and a round corner or a chamfer structure can be arranged at the vertex angle of the micro-fluidic chip.
The length and width of the microfluidic chip are selected according to the size of the actual porous plate. For example, microfluidic chips are about 127.76mm long and about 85.48mm wide, identical to standard 96 or 384 well plates. The embodiment of the invention is not limited in this way.
The reaction holes 11 of the microfluidic chip are distributed in accordance with the reaction holes on the porous plate so as to ensure matching with the porous plate matching instrument.
For example, the reaction wells 11 of the microfluidic chip are identical to the reaction well distribution on the 96-well plate, or the reaction wells 11 of the microfluidic chip are identical to the reaction well distribution on the 384-well plate. Specifically, the reaction holes 11 are distributed at equal intervals in the form of 16×24, the row interval of the reaction holes 11 is 4.3mm to 4.7mm, and the column interval of the reaction holes 11 is 4.3mm to 4.7mm; or the reaction holes 11 are distributed at equal intervals in the form of 8 multiplied by 12, the row interval of the reaction holes 11 is 8.8mm-9.2mm, and the column interval of the reaction holes 11 is 8.8mm-9.2mm; or the reaction holes 11 are distributed at equal intervals in the form of 8 multiplied by 24, the row interval of the reaction holes 11 is 8.8mm-9.2mm, and the column interval of the reaction holes 11 is 4.3mm-4.7mm; alternatively, the reaction holes 11 may be equally spaced in the form of 16X 12, with the row spacing of the reaction holes 11 being 4.3mm to 4.7mm and the column spacing of the reaction holes 11 being 8.8mm to 9.2mm.
The dimensions of the above reaction wells 11 are adapted to a standard 96-well plate or 384-well plate kit, such as a real-time fluorescence PCR instrument or an enzyme-labeled instrument.
Of course, the reaction holes 11 may be distributed at other distances, so long as the reaction holes 11 of the microfluidic chip are distributed uniformly with the reaction holes on the porous plate.
When the chip body is in the packaging process, bubbles are easier to generate due to the large area of the chip body. In order to exhaust or avoid generating bubbles, the microfluidic chip further includes a venting structure 2, as shown in fig. 2, the venting structure 2 includes: an exhaust passage, and an exhaust hole communicating with the exhaust passage. It is understood that the vent holes penetrate the chip body to ensure venting.
Preferably, the exhaust channel is disposed at an edge of the chip body; the exhaust holes are multiple and uniformly distributed in the exhaust channel.
Further, the chip body is rectangular, two exhaust channels are provided, one exhaust channel is arranged along the width direction of the chip body, and the other exhaust channel is arranged along the length direction of the chip body.
Of course, the exhaust structure 2 may alternatively be designed in other shapes and structures, and is not limited thereto.
As shown in fig. 5 and 6, the inlet section of the sample loading hole 7 of the microfluidic chip is a divergent section, and the inlet section diverges from the bottom end of the sample loading hole 7 to the sample loading end of the sample loading hole 7.
According to the structure of the sample adding hole 7, when the liquid-transferring suction head is inserted into the sample adding hole 7, the bottom of the inlet section of the sample adding hole 7 can clamp the liquid-transferring suction head, and the end head of the liquid-transferring suction head can be prevented from contacting the second substrate, so that the close contact between the liquid-transferring suction head and the sample adding hole 7 is realized, and the liquid can not leak in the sample adding process; meanwhile, the inlet section of the sample adding hole 7 is convenient for inserting and positioning the liquid-transferring suction head, and can also accommodate a small amount of liquid overflowed when the sample is introduced and pulled out of the liquid-transferring suction head, so that the pollution of a sample is avoided.
The chip main body includes: the first substrate 1 and the second substrate connected are encapsulated. Specifically, the second base cover is disposed on the first base 1 and is connected with the first base 1 in a sealing manner. The second substrate is not shown in the drawing. Of course, the chip body may alternatively be composed of at least three layers of substrates, and is not limited to two layers of substrates, namely, the first substrate 1 and the second substrate. The reaction holes 11 and the flow paths communicating with the reaction holes 11 are all provided in the first substrate 1, and the strength of the first substrate 1 is low, and the reinforcing ribs are preferably provided in the first substrate 1. The specific position of the reinforcing rib is selected according to actual needs. In order to avoid that the reaction holes 11 and the flow paths of the microfluidic chip are influenced by the reinforcing ribs, the reinforcing ribs are preferably arranged on the back of the first substrate 1, i.e. the side of the first substrate 1 away from the second substrate.
The front surface of the first matrix 1 is heated due to the packaging modes such as hot press sealing, laser welding and ultrasonic welding, and the high polymer is cooled after being heated to shrink, so that the first matrix 1 has stress bending towards the front surface, and at the moment, the reinforcing ribs at the back of the first matrix 1 just can resist the stress of bending deformation of the first matrix 1, so that the flatness of the first matrix 1 after packaging is improved, and the flatness of the whole chip body after packaging is further improved, namely the flatness of the microfluidic chip is improved.
The chip body may be the first substrate 1, or alternatively, the chip body may include a first substrate 1 and a second substrate connected to each other.
In the microfluidic chip, the reaction holes 11 and the sample addition holes 7 are plural, the sample addition holes 7 are located at two ends of the main flow path 13, the connecting flow path 12 and the reaction holes 11 are located on the same surface of the first substrate 1, and the sample addition holes 7 penetrate the first substrate 1. The distribution of the main channels 13 is parallel to the distribution of the reaction wells 11, and the connection channels 12 are in one-to-one correspondence with the reaction wells 11. The number of main channels 13 may be one or two or more. The reaction well 11 has a cavity to hold a liquid to be detected. The reaction hole 11 is formed on one side surface of the first substrate 1, and the reaction hole 11 is a blind hole.
To facilitate centrifugal dispensing and to improve dispensing uniformity, the second dispensing section 132 projects toward the connecting flow path 12. Preferably, the second distribution segment 132 is arcuate or V-shaped. Further, the second distribution segment 132 is semi-circular in shape. It will be appreciated that rounded corners may be provided at the V-shaped corners to facilitate centrifugal dispensing, although other shapes for the second dispensing section 132 may be selected and are not limited to the above-described configuration.
Further, to facilitate more uniform centrifugal distribution of the sample, the cross-sectional area of the second distribution segment 132 is greater than the cross-sectional area of the third distribution segment 133.
Based on the microfluidic chip provided in the foregoing embodiment, the embodiment of the present invention further provides a packaging fitting for packaging a microfluidic chip, as shown in fig. 7, where the packaging fitting for packaging a microfluidic chip includes a support body that is an elastic member, and the support body is provided with a support structure that can be completely attached to the back of the first substrate 1, and the support structure includes: and the matching structure can be matched with the reinforcing ribs on the back of the first matrix 1.
The supporting structure of the supporting body can be completely attached to the back of the first base body 1, so that the first base body 1 is supported; because the supporting body is an elastic piece, the packaging fitting is pressed and deformed after being combined with the first base body 1, and the structure and the shape of the first base body 1 can be self-adapted, so that the first base body 1 can be well packaged under the conditions of certain bending and local unevenness.
If above-mentioned strengthening rib includes edge strengthening rib 4, and edge strengthening rib 4 is annular and sets up along the edge of chip body, and the back of chip body is equipped with protruding 8, protruding 8 and edge strengthening rib 4 parallel and level, and sample application hole 7 has been seted up in the front of chip body, and sample application hole 7 distributes on protruding 8, then above-mentioned cooperation structure includes: the relief groove 01 matched with the inner reinforcing rib, the first step structure 02 matched with the edge reinforcing rib 4 and the second step structure 03 matched with the bulge 8.
In order to improve the elasticity of the support body and ensure that the first base body 1 can be well packaged under the conditions of certain bending and local unevenness, the support body comprises a first support split body 05 and a second support split body 06 which are connected, and a yielding groove 01 and a first step structure 02 are arranged on the first support split body 05; the top surface of the second support split body 06 is lower than the top surface of the first support split body 05, and the second support split body 06 and the first support split body 05 are matched to form a second step structure 03; wherein a gap 04 is provided between the first support sub-body 05 and the second support sub-body 06. The gap 04 is a narrow gap having a large depth.
By providing the above-mentioned gap 04, the degree of freedom of movement of the second step structure is increased, and the elasticity of the whole structure is improved, so that the whole structure can be better adapted to the first substrate 1.
The packaging fitting is preferably an elastic plastic member such as a silica gel member or a rubber member, and is manufactured by machining or injection molding.
Because the packaging accessory is the elastic plastic piece, and the heat conductivity of the elastic plastic piece is poor, in the packaging mode that the microfluidic chip can be heated in hot press sealing, laser welding, ultrasonic welding and the like, the packaging accessory reduces the heat dissipation capacity of the microfluidic chip, so that the energy consumption during packaging is reduced, namely the heating degree of the front surface of the microfluidic chip is reduced, the front surface deformation of the microfluidic chip caused by the heating is reduced, and the flatness of the microfluidic chip after packaging is guaranteed.
Based on the packaging fitting for packaging the microfluidic chip provided by the embodiment, the embodiment of the invention also provides a packaging method of the microfluidic chip, and the packaging method of the microfluidic chip comprises the following steps:
1) Processing a substrate:
Specifically, the first substrate 1 and the second substrate are processed, the front surface of the first substrate 1 is provided with the reaction holes 11, and the back of the first substrate 1 is provided with the reinforcing ribs. The first base 1 and the second base are elastic members. The material of the first substrate 1 is a high molecular polymer. The polymer is polymethyl methacrylate, polycarbonate, polypropylene, or the like. The first substrate 1 is formed by injection molding, laser engraving or machining. The second substrate is the same substrate as the first substrate 1, or the second substrate is a heat-seal film that matches the first substrate 1. For the convenience of detection, the first substrate 1 and/or the second substrate are/is a transparent substrate.
2) Spotting:
The materials required for the reaction are added into the reaction well 11. Biological materials required for reaction, such as primers, probes, enzymes, etc. for PCR reaction, are added into the reaction well 11 through a pipetting workstation, a gun pipette, various contact type spotting or spotting devices, and the spotting is completed after the liquid therein volatilizes or freeze-dries and solidifies into the reaction well 11. In addition, a biomaterial, such as a lyophilized pellet of a biological agent, or the like, which has been solidified, may be added to the reaction well 11.
3) And (3) packaging:
Placing the first base body 1 on the packaging fitting, and attaching the back of the first base body 1 to the packaging fitting; the second substrate is placed on the front side of the first substrate 1, encapsulating the first substrate 1 and the second substrate. The package fitting is the package fitting for packaging a microfluidic chip according to the above embodiment.
The first substrate 1 and the second substrate are elastic materials at normal temperature, and are deformable and elastic after being heated to a certain temperature, which is a characteristic of many high molecular polymers. The first substrate 1 and the second substrate can be packaged and attached by the prior art such as hot press sealing, laser welding, ultrasonic welding or glue sealing, so as to realize sealing connection. For example, one of the first substrate 1 and the second substrate is a substrate having a single-sided tape.
Because the first substrate 1 has a rugged complex structure, the packaging accessory is adopted to assist in packaging. The back surface of the first substrate 1 faces the package assembly to be bonded thereto. The packaging fitting enables the first base body 1 to be well packaged under the conditions of certain bending and partial unevenness.
According to the packaging method of the microfluidic chip, the first substrate 1 and the second substrate are packaged through the packaging fittings, so that the first substrate 1 can be well packaged under the conditions of certain bending and local unevenness, high-efficiency non-leakage packaging is convenient to achieve, and the packaging effect is improved.
In addition, because the packaging accessory is an elastic plastic piece, the heat conductivity of the elastic plastic piece is poor, and in the packaging mode that the microfluidic chip can be heated in the hot press sealing, laser welding, ultrasonic welding and the like, the heat dissipation of the microfluidic chip is reduced by the packaging accessory, so that the energy consumption during packaging is reduced, namely the heating degree of the front surface of the microfluidic chip is reduced, the front surface deformation of the microfluidic chip caused by the heating is reduced, and the flatness of the microfluidic chip after packaging is guaranteed.
The use method of the microfluidic chip comprises the following steps:
1) Sample injection:
the sample to be detected is added into the main flow path 13 of the microfluidic chip through the sample adding hole 7.
2) Sealing the sample adding port:
after the sample injection is completed, the sample injection hole 7 can be sealed by adopting a heat sealing mode or by adopting a viscose mode and the like.
3) Sample was distributed by centrifugation:
The microfluidic chip is placed on a centrifuge, the main flow path of the microfluidic chip is centrifuged toward the center of centrifugation and the reaction well 11 is centrifuged away from the center of centrifugation, and a sample is introduced into the reaction well 11 from the main flow path 13 through the connection flow path 12. In the centrifugal distribution process, the direction of the centrifugal force at the connecting flow path 12 is parallel to the axial direction of the connecting flow path 12, or the angle between the direction of the centrifugal force at the connecting flow path 12 and the axial direction of the connecting flow path 12 is not more than 80 degrees.
4) Isolating the reaction wells:
For the microfluidic chip subjected to centrifugal distribution, the microfluidic chip is placed on a microfluidic chip matching device, and the local deformation of the chip is isolated by connecting part or all of the flow paths 12 through hot pressing, so that each reaction hole 11 is physically isolated, the isolation reliability is effectively improved, and cross contamination and pollution of products after reaction to the environment are avoided.
5) Reaction and detection:
The isolated chip is reacted and detected by an instrument matched with a conventional porous plate, such as a real-time fluorescence PCR instrument, an enzyme-labeled instrument and the like.
The specific type of the instrument matched with the porous plate is selected according to actual needs, and the embodiment of the invention does not limit the type of the matched instrument.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (14)
1. A microfluidic chip for high throughput analysis, comprising: the chip comprises a chip body and reinforcing ribs arranged on the back of the chip body;
The reinforcing ribs comprise edge reinforcing ribs (4) and inner reinforcing ribs, wherein the edge reinforcing ribs (4) are annular and are arranged along the edge of the chip body, and the edge of the chip body surrounds the inner reinforcing ribs;
The back of the chip body is provided with a bulge (8), the bulge (8) is flush with the edge reinforcing rib (4), the chip body is provided with a sample adding hole (7), and the sample adding holes (7) are distributed on the bulge (8); a groove (14) and a main flow path (13) are formed in the front surface of the chip body, the groove (14) is trapped in the bulge (8), and one end, communicated with the sample adding hole (7), of the main flow path (13) is positioned in the groove (14); the front end face of the sample adding hole (7) is flush with the front face of the chip body;
the microfluidic chip is provided with a positioning structure (3) for positioning the instrument matched with the porous plate.
2. The microfluidic chip according to claim 1, wherein the reaction holes (11) of the microfluidic chip and the projections of the internal ribs on the front face of the chip body have no overlapping portions.
3. The microfluidic chip according to claim 1, wherein the height of the internal bead is 1mm-3mm and the width of the internal bead is 0.5mm-2mm.
4. The microfluidic chip according to claim 1, wherein the internal ribs comprise first internal ribs (5) and second internal ribs (6) arranged crosswise.
5. The microfluidic chip according to claim 1, wherein the internal ribs are in a strip shape and are disposed along a length direction or a width direction of the chip body.
6. The microfluidic chip according to claim 1, wherein the height of the edge bead (4) is higher than the height of the inner bead.
7. The microfluidic chip according to claim 1, wherein the main flow path (13) comprises: a first distribution section (131) communicating with the sample application well (7), a second distribution section (132) communicating with the reaction well (11) of the microfluidic chip through a connecting flow path (12), and a third distribution section (133) communicating with the first distribution section (131) and the second distribution section (132); wherein the cross-sectional area of the first distribution section (131) is greater than the cross-sectional area of the third distribution section (133).
8. The microfluidic chip according to claim 1, wherein,
The distance between the sample adding holes (7) of the microfluidic chip is 4.5+/-0.2 mm, or the distance between the sample adding holes (7) is 9+/-0.2 mm;
The micro-fluidic chip is rectangular, and the reaction holes (11) of the micro-fluidic chip are distributed in a matrix form;
The reaction holes (11) are distributed at equal intervals in a 16 multiplied by 24 mode, the row interval of the reaction holes (11) is 4.3mm-4.7mm, and the column interval of the reaction holes (11) is 4.3mm-4.7mm;
Or the reaction holes (11) are distributed at equal intervals in the form of 8 multiplied by 12, the row interval of the reaction holes (11) is 8.8mm-9.2mm, and the column interval of the reaction holes (11) is 8.8mm-9.2mm;
Or the reaction holes (11) are distributed at equal intervals in the form of 8 multiplied by 24, the row interval of the reaction holes (11) is 8.8mm-9.2mm, and the column interval of the reaction holes (11) is 4.3mm-4.7mm;
or the reaction holes (11) are distributed at equal intervals in a 16 multiplied by 12 mode, the row interval of the reaction holes (11) is 4.3mm-4.7mm, and the column interval of the reaction holes (11) is 8.8mm-9.2mm.
9. The microfluidic chip according to claim 1, further comprising an exhaust structure (2) provided to the chip body, the exhaust structure (2) comprising: an exhaust passage, and an exhaust hole communicating with the exhaust passage.
10. The microfluidic chip according to claim 1, wherein the inlet section of the sample application well (7) of the microfluidic chip is a diverging section, and the inlet section diverges from the bottom end of the sample application well (7) to the sample application end of the sample application well (7).
11. The microfluidic chip according to any one of claims 1-10, wherein the chip body comprises a first substrate (1) and a second substrate which are connected in a packaging manner, and the reaction holes (11) of the microfluidic chip are arranged on the first substrate (1); the reinforcing ribs are arranged on the back of the first substrate (1).
12. A packaging fitment for packaging a microfluidic chip as claimed in claim 11, comprising a support body which is an elastic member, the support body being provided with a support structure capable of being fully attached to the back of the first substrate (1), the support structure comprising: a fitting structure capable of fitting with the reinforcing rib;
The mating structure includes: and the yielding groove (01) is matched with the inner reinforcing rib, the first step structure (02) is matched with the edge reinforcing rib (4), and the second step structure (03) is matched with the bulge (8).
13. The packaging kit for packaging a microfluidic chip according to claim 12, wherein,
The support body comprises a first support split body (05) and a second support split body (06) which are connected, and the abdication groove (01) and the first step structure (02) are arranged on the first support split body (05);
The top surface of the second support split body (06) is lower than the top surface of the first support split body (05), and the second support split body (06) and the first support split body (05) are matched to form the second step structure (03);
Wherein a gap (04) is provided between the first support body (05) and the second support body (06).
14. The packaging method of the microfluidic chip is characterized by comprising the following steps:
Processing a first substrate (1) and a second substrate, wherein the front surface of the first substrate (1) is provided with a reaction hole (11), and the back of the first substrate (1) is provided with a reinforcing rib;
adding materials required by the reaction into the reaction holes (11);
Placing the first substrate (1) on a packaging fitting, attaching the back of the first substrate (1) to the packaging fitting, placing the second substrate on the front surface of the first substrate (1), and packaging the first substrate (1) and the second substrate;
wherein the packaging fitting is a packaging fitting for packaging a microfluidic chip according to claim 12 or 13, and the first substrate (1) and the second substrate are elastic members.
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