阅读说明：本技术 一种微流控pcr反应芯片 (Microfluidic PCR reaction chip ) 是由 周志图 于 2020-12-01 设计创作，主要内容包括：本发明公开了一种微流控PCR反应芯片,包括芯片本体、分布贴覆于芯片本体上、下表面的上封口膜及下封口膜；所述芯片本体为避光材质,所述芯片本体上布设加样孔、透气孔及反应池,所述反应池在芯片本体中间区域集中布设,所述加样孔和透气孔在反应池布设区域周边分散布设,并通过流道与反应池连通,所述上封口膜为避光材质,贴覆在芯片本体上表面,对应反应池布设区域开设供激发光穿过的孔,所述加样孔和透气孔上方开口,通过上封口膜实现闭合,所述反应池和流道下方开口,通过下封口膜实现闭合。芯片本体内部流道采用避光材质隔离,采集荧光信号的时候,降低孔间会互相串光形成的信号干扰,提高检测结果的准确性。(The invention discloses a microfluidic PCR reaction chip, which comprises a chip body, an upper sealing film and a lower sealing film, wherein the upper sealing film and the lower sealing film are distributed and attached to the upper surface and the lower surface of the chip body; the utility model discloses a chip, including chip body, addition hole, bleeder vent and reaction tank, the chip body is the light-resistant material, lay addition hole, bleeder vent and reaction tank on the chip body, the reaction tank is concentrated at chip body middle zone and is laid, addition hole and bleeder vent are laid regional peripheral dispersion in the reaction tank and are laid to communicate through runner and reaction tank, go up the sealing film and be the light-resistant material, paste and cover at chip body upper surface, and the corresponding reaction tank is laid regional seting up the hole that supplies the exciting light to pass, addition hole and bleeder vent top opening realize the closure through last sealing film, reaction tank and runner below opening realize the closure through sealing film down. The internal flow channel of the chip body is isolated by light-resistant materials, so that the signal interference caused by light cross among holes is reduced when fluorescent signals are collected, and the accuracy of detection results is improved.)

1. A microfluidic PCR reaction chip is characterized in that: comprises a chip body (1), an upper sealing film (2) and a lower sealing film (3) which are distributed and adhered on the upper surface and the lower surface of the chip body (1);

chip body (1) is the light-resistant material, lay on chip body (1) and add appearance hole (101), bleeder vent (102) and reaction tank (103), reaction tank (103) are concentrated at chip body (1) middle zone and are laid, add appearance hole (101) and bleeder vent (102) and lay regional peripheral dispersion in the reaction tank and lay to communicate with reaction tank (103) through runner (104), go up sealing film (2) and for the light-resistant material, paste and cover at chip body (1) upper surface, correspond the reaction tank and lay the regional hole of offering the confession exciting light and passing, add appearance hole (101) and bleeder vent (102) top opening, realize the closure through last sealing film (2), reaction tank (103) and runner (104) below opening realize the closure through sealing film (3) down.

2. The microfluidic PCR reaction chip of claim 1, wherein: and a condensing lens is arranged above the reaction tank (103).

3. The microfluidic PCR reaction chip of claim 1, wherein: the chip body (1) and the upper sealing film (2) are both made of black plastic materials.

4. The microfluidic PCR reaction chip of claim 1, wherein: the upper sealing film (2) and the lower sealing film (3) are bonded on the surface of the chip body (1) by adopting one of ultraviolet glue, epoxy resin glue, phenolic resin glue or silicon rubber.

5. The microfluidic PCR reaction chip of claim 1, wherein: each reaction tank (103) corresponds to one sample adding hole (101) and one air vent (102), and the sample adding holes (101) and the air vents (102) are arranged in a staggered and spaced mode.

6. The microfluidic PCR reaction chip of claim 1, wherein: the reaction tank (103) is of a circular counter bore structure.

7. The microfluidic PCR reaction chip of claim 1, wherein: the flow channel (104) is a capillary flow channel, and in the flow channel (104), the width of the sample-adding flow channel is larger than that of the air-permeable flow channel.

8. The microfluidic PCR reaction chip of claim 2, wherein: the condensing lens is a convex lens directly arranged on an opening of the reaction tank (103), or is a convex lens structure formed by wrapping a layer of light-transmitting material shell outside the chip body (1) and cutting the shell at a position corresponding to the reaction tank (103).

Technical Field

The invention relates to the technical field of molecular biology detection, in particular to a microfluidic PCR reaction chip.

Background

In the prior art, the whole PCR reaction chip is made of transparent materials, and when fluorescent signals are collected, holes can be mutually crossed to form signal interference, so that the accuracy of detection results is influenced; secondly, because the sample adding holes are generally arranged on one side or two sides of the reaction chip in parallel, the arrangement mode leads to shorter distance between the sample adding holes, and leads to overflow and hole string of reaction liquid between the sample adding holes. Meanwhile, the reaction tank is also arranged in parallel on the reaction chip, so that the space for receiving the light source is in a straight shape, the reaction tank is suitable for common light sources of the machine, such as a halogen tungsten lamp light source, a hydrogen ion exciter, a Light Emitting Diode (LED) light source and the like, and the problem of low detection repeatability is caused due to the fact that the intensity of the light source received by the reaction hole of the reaction chip is not uniform.

Disclosure of Invention

The purpose of the invention is as follows: in order to overcome the defects of the background art, the invention discloses a microfluidic PCR reaction chip capable of effectively reducing the mutual crosstalk interference among holes.

The technical scheme is as follows: the microfluidic PCR reaction chip comprises a chip body, an upper sealing film and a lower sealing film, wherein the upper sealing film and the lower sealing film are distributed and attached to the upper surface and the lower surface of the chip body;

the utility model discloses a chip, including chip body, addition hole, bleeder vent and reaction tank, the chip body is the light-resistant material, lay addition hole, bleeder vent and reaction tank on the chip body, the reaction tank is concentrated at chip body middle zone and is laid, addition hole and bleeder vent are laid regional peripheral dispersion in the reaction tank and are laid to communicate through runner and reaction tank, go up the sealing film and be the light-resistant material, paste and cover at chip body upper surface, and the corresponding reaction tank is laid regional seting up the hole that supplies the exciting light to pass, addition hole and bleeder vent top opening realize the closure through last sealing film, reaction tank and runner below opening realize the closure through sealing film down.

On one hand, the light-shielding material is selected, so that the internal flow channel can be isolated, when the fluorescent signal is collected, the signal interference formed by mutual light crosstalk among holes is reduced, and the accuracy of the detection structure is improved; on the other hand, the sample adding holes and the air holes are distributed in a scattered mode, and pollution of samples among the holes is effectively prevented.

Furthermore, a condensing lens is arranged above the reaction tank.

The chip body and the upper sealing film are both made of black plastic materials.

Furthermore, the upper sealing film and the lower sealing film are bonded on the surface of the chip body by adopting one of ultraviolet glue, epoxy resin glue, phenolic resin glue or silicon rubber, so that the phenomenon that the sealing film is scratched and falls off in the experimental process and loses the sealing property is prevented.

Furthermore, each reaction tank corresponds to one sample adding hole and one air vent, and the sample adding holes and the air vents are arranged in a staggered and spaced mode.

Furthermore, the reaction tank is of a circular counter bore structure, the detected sample is concentrated in the circular reaction tank, the fluorescence signal is most concentrated, and the fluorescence signal can be efficiently collected.

Furthermore, the flow channel is a capillary flow channel, and in the flow channel, the width of the sample adding flow channel is larger than that of the air-permeable flow channel, so that the reaction liquid can quickly flow into the reaction tank for reaction through the capillary action after entering the sample adding hole.

Furthermore, the condensing lens is a convex lens directly arranged on the upper opening of the reaction tank, or a layer of transparent material shell is wrapped outside the chip body, and the convex lens structure is formed on the shell by cutting corresponding to the position of the reaction tank. The device is beneficial to gathering exciting light, and the exciting light is emitted into the reaction tank after being focused, so that the detection sensitivity is improved; meanwhile, the sample reaction pool is arranged in a centralized mode, and light path design is facilitated.

Work as condensing lens is for the convex lens integrated into one piece structure that corresponds the cutting of reaction tank position and form on the shell, only uses a holistic panel to realize setting up of sample liquid storage space promptly and the closure of reaction tank top, overall structure is simple, directly adopt the mould shaping can, easy manufacturing, cost greatly reduced.

Has the advantages that: compared with the prior art, the invention has the advantages that: firstly, the flow channel in the chip body is isolated by adopting a light-shading material, so that when fluorescent signals are collected, the signal interference formed by mutual light crosstalk among holes is reduced, and the accuracy of a detection result is improved; secondly, the sample adding holes and the air holes are arranged in a staggered mode, so that sample pollution among the holes is effectively prevented, a lower sealing film is attached to the bottom surface of the chip body, when the PCR chip is placed on the temperature control module, the lower sealing film is tightly attached to the temperature control module, after sample adding, the PCR reaction liquid in a reaction tank above the lower sealing film and the temperature control module are only separated by one film, the refrigerating capacity or the heating capacity of the temperature control module can quickly penetrate through the lower sealing film to be conducted to the PCR reaction liquid, and compared with a plastic plate, the heat conducting speed is higher, the heat conducting performance is more uniform, and quick; then, the lens design is adopted above the reaction pool area of the chip body, so that the reaction pool is an accommodating space which is shielded above, pollutants are prevented from falling into the reaction pool from the upper side, the exciting light is favorably collected, the exciting light is emitted into the reaction pool after being focused, the detection sensitivity is improved, and meanwhile, the reaction pool area is intensively arranged, and the light path design is favorably realized; and finally, the upper sealing film is made of a light-shading material, and when images are collected, except a fluorescence emission area, other areas are black backgrounds, so that fluorescence signals can be collected conveniently, and subsequent software image processing is facilitated.

Drawings

FIG. 1 is a schematic diagram of a separation configuration of the present invention;

FIG. 2 is a plan view of the top surface of the chip body of the present invention;

FIG. 3 is a plan view of the bottom surface of the chip body of the present invention.

Detailed Description

The technical solution of the present invention is further described below with reference to the accompanying drawings and examples.

The microfluidic PCR reaction chip shown in FIG. 1 comprises a chip body 1, and an upper sealing film 2 and a lower sealing film 3 which are distributed and adhered on the upper surface and the lower surface of the chip body 1, wherein one of ultraviolet glue, epoxy resin glue, phenolic resin glue or silicon rubber is adopted for adhesion during adhering.

As shown in fig. 2 and fig. 3, the chip body 1 is made of a light-shielding material, and may be one or a combination of several materials selected from silicon, glass, polymethyl methacrylate, polycarbonate, polystyrene, polyethylene terephthalate, polyethylene naphthalate, cyclic olefin copolymer, and epoxy resin, and is preferably made of black plastic, which has the advantages of low cost, easy processing, and low requirement for molding process. The chip comprises a chip body 1, and is characterized in that sample adding holes 101, air holes 102 and reaction tanks 103 are distributed on the chip body 1, the reaction tanks 103 are distributed in a concentrated manner in the middle area of the chip body 1, the sample adding holes 101 and the air holes 102 are distributed and distributed at the periphery of the distribution area of the reaction tanks, each reaction tank 103 corresponds to one sample adding hole 101 and one air hole 102, and the sample adding holes 101 and the air holes 102 are distributed at intervals in a staggered manner. The sample adding hole 101 and the air hole 102 are communicated with the reaction cell 103 through a flow channel 104, the flow channel does not penetrate through the chip body 1, and the length of the sample adding flow channel and the length of the air permeable flow channel can be controlled to distinguish the sample adding flow channel and the air permeable flow channel. The flow channel 104 is a capillary flow channel, and in the flow channel 104, the width of the sample adding flow channel is larger than that of the air permeable flow channel, so that the PCR reaction solution can quickly flow into the reaction tank for reaction through capillary action after entering the sample adding hole. The reaction tank 103 is of a circular counter bore structure, the detected sample is concentrated in the circular reaction tank, the fluorescence signal is most concentrated, and the fluorescence signal can be efficiently collected. The sample adding hole 101 is funnel-shaped, so that sample introduction is facilitated.

The upper sealing film 2 is used for preventing reaction liquid in the reaction tank from volatilizing into the external air during heating and reaction, which not only affects the detection result, but also causes the nucleic acid in the external air to pollute the reaction system of the next reaction, which causes laboratory pollution, the upper sealing film 2 is made of light-proof material, preferably black plastic film such as PE (polyethylene)/PP (polypropylene)/PVC (polyvinyl chloride) with the thickness of 0.1-0.5mm, and is pasted on the upper surface of the chip body 1, holes for exciting light to pass are arranged in the corresponding reaction tank area, openings are arranged above the sample adding holes 101 and the vent holes 102, the upper sealing film 2 is closed, the upper sealing film has the main functions of sealing and avoiding light, the sample adding holes 101 and the vent holes 102 can be shielded, the reaction tank 103 area is exposed, the reaction liquid in the reaction tank is prevented from volatilizing into the external air during heating and reaction, which not only affects the detection result, the nucleic acid in the outside air can pollute the reaction system of the next reaction, so that the laboratory is polluted, and the light can be avoided, so that the fluorescence can be collected and the incident light source can be received more effectively. A condenser lens is arranged above the reaction tank 103, openings are formed below the reaction tank 103 and the flow channel 104, and the lower sealing film 3 is used for realizing closing. The lower sealing film 3 is made of heat-conducting material and is selected from one or combination of PVC, PE, PP or PS. The lower sealing film 3 of the PCR reaction chip adopts a light-transmitting material, can be PVC, PE, PP, PS and the like, and has the thickness of 0.02-0.3 mm. The lower sealing film 3 is thin, the heat conduction speed is higher, and the heat conduction is more uniform.

The condenser lens above the reaction cell 103 is a convex lens directly disposed on the reaction cell 103, or a convex lens structure formed by wrapping a layer of transparent material on the chip body 1 and cutting the shell corresponding to the reaction cell 103.

The use process of the microfluidic PCR reaction chip comprises the following steps:

1) sample application:

materials required for the reaction are added to the well 101. Biological materials required for the reaction, such as primers, probes, enzymes, etc. for the PCR reaction are added to the wells 101 through a pipetting station, a pipette gun, various contact spotting or spraying devices, and spotting is completed when the liquid therein flows or is freeze-dried and solidified into the reaction wells 103. In addition, the reaction tank 103 may also be filled with solidified biological materials, such as lyophilized pellets of biological reagents.

2) Sealing the sample addition hole 101 and the vent hole 102:

after the sample injection is completed, the sample injection hole 101 and the air vent 102 may be closed by glue or the like.

3) Reaction and detection:

the chip can complete reaction and detection with instruments such as real-time fluorescence PCR instrument, enzyme labeling instrument, etc.

The specific type of the instrument matched with the multi-well plate is selected according to actual needs, and the embodiment of the invention does not limit the type of the matched instrument.