Method for manufacturing sample detection device

Abstract

The present invention relates to a method for manufacturing a sample detection device, comprising the steps of: providing a first plate and a second plate in which a microfluidic channel structure is formed so that a microfluidic channel can be formed between the first plate and the second plate when the first plate and the second plate are sealed together; a reagent application step in which a predetermined amount of a liquid reagent is placed in one or more selected reagent regions in the microfluidic channel structure, and after the reagent is placed, the reagent is freeze-dried; performing surface treatment on the first plate and the second plate so as to enable the treated surfaces to have hydrophilicity; and sealing the first plate and the second plate together to form the sample detection device. According to the manufacturing method provided by the invention, the reagent can be reliably kept at a selected position in the microfluidic channel through a process sequence of first performing reagent application and then performing hydrophilic treatment, so that subsequent detection and analysis of a sample are facilitated.

Description

Technical Field

[0001] The present application relates to the field of medical technology, and more specifically, to a method for manufacturing a sample detection device for a biological fluid sample, wherein the sample detection device is especially a disposable consumable. Background Art

[0002] With the continuous deepening of microfluidic technology research, the manufacturing process of microfluidic chips has also been rapidly developed. Microfluidic chips are an integrated sample detection device that seals the detection reagent inside the chip structure. It can avoid the pre-treatment of samples. That is, in the process of detecting samples, only the sample needs to be added to the microfluidic chip, and the sample and the reagent can be mixed and reacted by itself inside the chip, and finally flow into the detection area for detection. Microfluidic chips can integrate multiple functions, such as liquid self-driving, reagent embedding, sample mixing, reagent reaction, and uniform distribution of samples. Therefore, the manufacturing process of microfluidic chips is also different from traditional consumables for biological sample detection. As the main raw material of disposable microfluidic chips, polymers can be selected, which have the advantages of a wide variety, low manufacturing cost, and batch production. At present, microfluidic chips prepared by polymers have been widely used in many fields such as biochemical analysis, drug screening, and clinical medical testing, and have achieved good results. Summary of the invention

[0003] An object of the present invention is to provide a method for manufacturing a sample detection device, which has simple steps and reasonable cost, and can manufacture a sample detection device that can be used conveniently and reliably.

[0004] To this end, the present invention relates to a method for manufacturing a sample detection device, the manufacturing method comprising the following steps performed in sequence:

[0005] Providing a first plate and a second plate, wherein at least one of the first plate and the second plate has a microfluidic channel structure formed therein, so that when the first plate and the second plate are sealed together, a microfluidic channel can be formed between the first plate and the second plate through the microfluidic channel structure;

[0006] a reagent applying step, in which a predetermined amount of liquid reagent is placed in one or more selected reagent areas in the microfluidic channel structure, and after placing the reagent, the reagent is freeze-dried so that the reagent adheres to the corresponding reagent area of ​​the microfluidic channel structure;

[0007] a surface treatment step, in which the first plate and the second plate are surface treated at least in the region of the microfluidic channel structure so that the treated surfaces of the first plate and the second plate have hydrophilicity;

[0008] A sealing step, in which the first plate and the second plate are sealed together so that a microfluidic channel is formed between the first plate and the second plate, thereby forming the sample detection device.

[0009] The inventor unexpectedly discovered during the research and development process that the manufacturing method of the present invention can be used to manufacture a sample detection device that is convenient for detecting and analyzing samples, wherein the unnecessary diffusion of the reagent on the surface of the cover plate and the main body can be avoided by placing the reagent first and freeze-drying the reagent, and then performing a hydrophilic treatment on the surface of the cover plate and the main body. The inventor unexpectedly discovered during the research and development process that if the surface of the cover plate and the main body is first hydrophilicized and then the reagent is applied, the liquid reagent is easily diffused due to the hydrophilicity of the surface of the cover plate and the main body, and cannot be reliably maintained at the selected position in the microfluidic channel, which is not conducive to the subsequent detection and analysis of the sample.

[0010] In some embodiments, the first plate is provided in the form of a body of a sample detection device, and the second plate is provided in the form of a cover plate of the sample detection device.

[0011] In some embodiments, the main body has a microfluidic channel structure, and the cover plate has or does not have a microfluidic channel structure.

[0012] In some embodiments, in the freeze-drying step, the reagent is freeze-dried at a temperature ranging from -20°C to 0°C, and / or the freeze-drying step is performed in a closed environment.

[0013] In some embodiments, in the surface treatment step, the treated surfaces of the first plate and the second plate are provided with hydroxyl groups or carboxyl groups.

[0014] In some embodiments, in the surface treatment step, the treated surfaces of the first plate and the second plate are made hydrophilic by plasma treatment, electroplating treatment, or chemical coating treatment.

[0015] In some embodiments, the surface treatment step is performed such that the treated surface of the second plate has a higher degree of hydrophilicity than the treated surface of the first plate.

[0016] In some embodiments, in the sealing step, the first plate and the second plate are sealed together by one or more of bonding, heat compression sealing, ultrasonic welding, laser welding, and adhesive bonding.

[0017] In some embodiments, the first plate and the second plate are sealed together by bonding, wherein bonding between corresponding surfaces of the first plate and the second plate to be sealed is achieved based on the following reaction formula: 2Si-OH→Si-O-Si+2H2O.

[0018] In some embodiments, before bonding, the surfaces to be sealed of the first plate and the second plate are subjected to plasma treatment.

[0019] In some embodiments, the first plate and the second plate are respectively made of a polymer material, and the polymer material is selected from polymethyl methacrylate, polydimethylsiloxane, polypropylene, polycarbonate, and polystyrene.

[0020] In some embodiments, the first plate and the second plate are made of different types of polymer materials.

[0021] In some embodiments, in the step of providing the first plate and the second plate, the first plate and the second plate are manufactured by injection molding a polymer material, and the injection molding process satisfies one or more of the following:

[0022] The melting point of the polymer material is between 200°C and 300°C;

[0023] The temperature of the mold used for injection molding is between 20°C and 100°C;

[0024] The injection pressure is between 60MPa and 180MPa;

[0025] The injection molded material is dried between 40°C and 100°C;

[0026] The drying time of the injection molded material is 3 to 8 hours;

[0027] The molding shrinkage of the material after injection molding is between 0.3% and 1.3%.

[0028] In some embodiments, the manufacturing method further includes a reduction treatment step after the sealing step, in which the sample detection device is placed in a sealed space and an inert gas is introduced into the sealed space.

[0029] In some embodiments, in the reduction step:

[0030] The inert gas is argon and / or helium, and / or

[0031] The duration of the reduction treatment step is more than 48 hours.

[0032] In some embodiments, one of the first plate and the second plate is made of polydimethylsiloxane, and the other is made of polymethyl methacrylate, and bonding is achieved between the corresponding surfaces to be sealed of the first plate and the second plate based on the following reaction formula: 2Si-OH→Si-O-Si+2H2O, wherein, in the surface treatment step, the treated surfaces of the first plate and the second plate are provided with hydroxyl groups.

[0033] The various technical features mentioned above and the various technical features to be mentioned below as well as the technical features that can be derived from the drawings can be combined with each other arbitrarily, as long as the individual technical features combined with each other are not contradictory to each other. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] The present invention is described in more detail below with reference to the accompanying drawings by means of exemplary embodiments, but the present invention is not limited thereto.

[0035] Figure 1 is a schematic flow chart of a method for manufacturing a sample detection device according to some embodiments of the present invention.

[0036] Figure 2 is used Figure 1 Schematic diagram of a sample detection device manufactured by the manufacturing method. DETAILED DESCRIPTION

[0037] Figure 1 A schematic flow chart of a method 1 for manufacturing a sample detection device according to some embodiments of the present invention is shown. Figure 2 It shows the use of Figure 1 Schematic diagram of a sample detection device 2 manufactured by the manufacturing method 1. Figure 2 As shown, the sample detection device 2 can mainly include a cover plate 3 and a main body 4, and at least one of the cover plate 3 and the main body 4 can be formed with a microfluidic channel structure 5, so that when the cover plate 3 and the main body 4 are sealed together, a microfluidic channel can be formed between the cover plate 3 and the main body 4. More advantageously, as Figure 2 As further shown, the microfluidic channel structure 5 can be arranged only in the main body 4, the cover plate 3 may not have a microfluidic channel structure, and thus the surface facing the main body 4 is flat, or structureless, and the cover plate 3 is used to adhere to and close the microfluidic channel structure 5 to form a microfluidic channel. In addition, structures such as a sample addition hole 6 and a discharge outlet 7 may be provided in the cover plate 3. Alternatively, the microfluidic channel structure may also be provided in the cover plate 3. Alternatively, microfluidic channel structures may be provided not only in the main body 4 but also in the cover plate 3, respectively, and they respectively form a part of the microfluidic channel.

[0038] The cover plate 3 and the main body 4 can be made of polymer materials respectively, and the above-mentioned polymer materials can be selected from polymethyl methacrylate (PMMA), polydimethylsiloxane (PDMS), polypropylene, polycarbonate, polystyrene, etc. These polymer materials have the characteristics of strong rigidity, weak adsorption force and good optical properties, and are suitable for use as materials for the cover plate 3 and the main body 4 used to make the sample detection device 1. In some embodiments, the cover plate 3 and the main body 4 can be made of the same material. In other embodiments, the cover plate 3 and the main body 4 can also be made of different materials, so that the complementary advantages between different materials can be utilized, so that the formed sample detection device 2 can have the advantages of multiple materials and improve the adaptability to the detection of multiple biological samples. The materials of the cover plate 3 and the main body 4 can be selected based on the requirements brought about by the characteristics of the fluid used to flow in the microfluidic channel.

[0039] like Figure 1 As shown, the manufacturing method 1 may first include a step 10 of providing a cover plate 3 and a main body 4. In this step, the cover plate 3 and the main body 4 may be manufactured by injection molding a polymer material. During the injection molding process, different temperature and pressure conditions may be used according to the melt index and shrinkage of different materials. In some embodiments, the melting point of the polymer material used for injection molding may be controlled between 200°C and 330°C, the mold temperature used for injection molding may be controlled between 20°C and 100°C, and the injection pressure may be controlled between 60MPa and 180MPa. The injection molded material may be dried at 40°C to 100°C, and the drying time may be 3 to 8 hours. Advantageously, the molding shrinkage of the injection molded material may be between 0.3% and 1.3%.

[0040] like Figure 1 As shown, the manufacturing method 1 may then include a reagent application step 20. The reagent is a reagent to be reacted with the sample to detect the sample. The reagent application step 20 may include a reagent placement step 201 in which a predetermined amount of liquid reagent can be placed in one or more selected reagent areas in the microfluidic channel structure 5. The predetermined amount of reagent is usually in the microliter level. The placement of the reagent can be automatically completed, for example, by an automatic spotting machine. The components of the reagent can be, for example, trypan blue, acridine orange, etc. After the reagent placement step 201, the reagent application step 20 may also include a reagent freeze-drying step 202 so that the reagent can be fixed and attached to the corresponding reagent area of ​​the microfluidic channel structure 5. Advantageously, the freeze-drying step 202 can be performed under a temperature condition between -20°C and 0°C. Advantageously, the freeze-drying step 202 can be performed in a closed environment. In the freeze-drying step 202, the reagent and the main body 4 and / or the cover plate 3 carrying the reagent can be freeze-dried together.

[0041] To facilitate the flow of the sample to be tested in the microfluidic channel, the manufacturing method 1 may include a surface treatment step 30 for the cover plate 3 and the main body 4 after the reagent application step 20, so that the treated surfaces of the cover plate 3 and the main body 4 (and correspondingly, the treated surface of the microfluidic channel) are activated and modified to be hydrophilic. The hydrophilic surface has a relatively stable adsorption capacity for cells and a binding capacity with proteins, is suitable for cell adherence culture, and can effectively improve the adhesion and extension effect of cells. The surface treatment step 30 treats the treated surfaces of the cover plate 3 and the main body 4 so that the treated surface of the main body 4 has a higher degree of hydrophilicity than the cover plate 3. In the surface treatment step 30, the treated surfaces of the cover plate 3 and the main body 4 can be provided with hydrophilic groups, such as hydroxyl groups, carboxyl groups, etc. In the surface treatment step, the treated surfaces of the cover plate 3 and the main body 4 can be made hydrophilic by plasma treatment, electroplating treatment, chemical coating treatment, etc.

[0042] By placing the reagent first to freeze-dry it and then subjecting the surface of the cover plate 3 and the main body 4 to hydrophilic treatment, it is possible to avoid unnecessary diffusion of the reagent on the surface of the cover plate 3 and the main body 4, thereby helping to reliably retain the reagent at the selected position in the microfluidic channel. On the contrary, if the surface of the cover plate 3 and the main body 4 is hydrophilic treated first and then the reagent is applied, the liquid reagent is easy to diffuse due to the hydrophilicity of the surface of the cover plate 3 and the main body 4, and cannot be reliably retained at the selected position in the microfluidic channel, which is not conducive to the subsequent detection and analysis of the sample.

[0043] After the above-mentioned surface treatment step 30, the manufacturing method 1 may subsequently include a sealing step 40 to seal the cover plate 3 and the main body 4 together, so as to form a microfluidic channel between the cover plate 3 and the main body 4, and form a sample detection device 2. Generally speaking, it is hoped that the seal between the cover plate 3 and the main body 4 has sufficient mechanical strength and is not easy to crack and leak, and it is hoped that the microfluidic channel will not be deformed or blocked as much as possible. The sealing step 40 can be completed by one or more of bonding, hot pressing sealing, ultrasonic welding, laser welding and bonding.

[0044] As an example, the cover plate 3 and the main body 4 can be sealed together by bonding, wherein the bonding between the corresponding surfaces to be sealed of the cover plate 3 and the main body 4 is based on the following reaction formula: 2Si-OH→Si-O-Si+2H2O. Before bonding, the corresponding surfaces to be sealed of the cover plate 3 and the main body 4 can be plasma treated. Specifically, for example, when one of the cover plate 3 and the main body 4 is made of polydimethylsiloxane and the other is made of polymethyl methacrylate, a coupling agent can be applied to the surface to be sealed of the component made of polymethyl methacrylate so that the corresponding surface to be sealed has a silicon amino group (Si-NH2 ), and then degrade the silanol group into silanol group (Si-OH) by plasma treatment to react with the silanol group on the surface to be sealed of the component made of polydimethylsiloxane after plasma treatment, so as to achieve the bonding between the corresponding surfaces to be sealed through the reaction formula 2Si-OH→Si-O-Si+2H2O. It should be understood that the reaction formula for achieving bonding is not limited to the reaction formula described above, and the specific steps for achieving bonding based on the reaction formula described above are not limited to the specific steps described above. The reaction formula used for bonding and its specific steps can be adaptively adjusted based on the selected materials for forming the cover plate 3 and the main body 4 and the characteristics of the surface of the microfluidic channel to be achieved.

[0045] In a more advantageous embodiment, the cover plate 3 and the main body 4 can be configured such that one of them is made of polydimethylsiloxane and the other is made of polymethyl methacrylate, and they are sealed together by bonding based on the reaction formula 2Si-OH→Si-O-Si+2H2O to achieve stable and reliable bonding between the cover plate 3 and the main body 4. Polydimethylsiloxane has the advantages of acid and alkali resistance, low toxicity, high tensile strength and good air tightness, and is suitable for packaging the sample detection device 2, which can effectively improve the performance and stability of the sample detection device 2. Polymethyl methacrylate has good comprehensive mechanical properties, acid and alkali resistance and atmospheric aging resistance, and is therefore also suitable for making a part of the sample detection device 2. In addition, in the surface treatment step 30, the treated surfaces of the cover plate 3 and the main body 4 can be provided with hydroxyl groups so that the corresponding surfaces have good hydrophilicity.

[0046] like Figure 1 As further shown, after the sealing step 40, the manufacturing method 1 may further include a reduction treatment step 50. In the reduction treatment step 50, the sample detection device 2 may be placed in a closed space, and an inert gas may be introduced into the closed space. The inert gas may be, for example, argon, helium, etc. Advantageously, the duration of the reduction treatment step 50 may be more than 48 hours.

[0047] The manufacturing method 1 of the present invention can be used to manufacture a sample detection device 2 that is convenient for detecting and analyzing samples. By placing the reagent first and freeze-drying the reagent, and then performing a hydrophilic treatment on the surface of the cover plate 3 and the main body 4, unnecessary diffusion of the reagent on the surface of the cover plate 3 and the main body 4 can be avoided, thereby helping to reliably maintain the reagent at a selected position in the microfluidic channel, which is beneficial to the subsequent detection and analysis of the sample.

[0048] It should be noted that the terms used herein are for the purpose of illustrating specific aspects only and are not intended to limit the disclosure. As used herein, the singular forms "a" and "the one" shall include the plural forms unless the context clearly states otherwise. It is understood that the terms "include" and "comprises" and other similar terms, when used in the application documents, specify the existence of the stated operations, elements and / or parts, without excluding the existence or addition of one or more other operations, elements, parts and / or combinations thereof. As used herein, the term "and / or" includes all arbitrary combinations of one or more related enumerated items. In the description of the drawings, similar reference numerals always represent similar elements.

[0049] Finally, it should be pointed out that the above embodiments are only used to understand the present invention, and do not limit the protection scope of the present invention. For those skilled in the art, modifications can be made based on the above embodiments, and these modifications do not depart from the protection scope of the present invention.

Claims

1. A method for manufacturing a sample detection device, characterized in that: The manufacturing method comprises the following steps which are performed in sequence: Providing a first plate and a second plate, wherein at least one of the first plate and the second plate has a microfluidic channel structure formed therein, so that when the first plate and the second plate are sealed together, a microfluidic channel can be formed between the first plate and the second plate through the microfluidic channel structure; a reagent applying step, in which a predetermined amount of liquid reagent is placed in one or more selected reagent areas in the microfluidic channel structure, and after placing the reagent, the reagent is freeze-dried so that the reagent adheres to the corresponding reagent area of ​​the microfluidic channel structure; a surface treatment step, in which the first plate and the second plate are surface treated at least in the region of the microfluidic channel structure so that the treated surfaces of the first plate and the second plate have hydrophilicity; A sealing step, in which the first plate and the second plate are sealed together so that a microfluidic channel is formed between the first plate and the second plate, thereby forming the sample detection device.

2. The method for manufacturing a sample detection device according to claim 1, characterized in that: Providing the first plate in the form of a main body of a sample detection device, and providing the second plate in the form of a cover plate of the sample detection device, preferably, the main body has a microfluidic channel structure, and the cover plate has or does not have a microfluidic channel structure; and / or In the freeze-drying step, the reagent is freeze-dried at a temperature ranging from -20°C to 0°C, and / or the freeze-drying step is performed in a closed environment; and / or In the surface treatment step, the treated surfaces of the first plate and the second plate are provided with hydroxyl groups or carboxyl groups; and / or In the surface treatment step, the treated surfaces of the first plate and the second plate are made hydrophilic by plasma treatment, electroplating treatment, or chemical coating treatment.

3. The method for manufacturing a sample detection device according to claim 1 or 2, characterized in that: The surface treatment step is performed such that: the treated surface of the second plate has a higher degree of hydrophilicity than the treated surface of the first plate; and / or In the sealing step, the first plate and the second plate are sealed together by one or more of bonding, heat compression sealing, ultrasonic welding, laser welding and adhesive bonding; and / or The first plate and the second plate are sealed together by bonding, wherein bonding between corresponding surfaces to be sealed of the first plate and the second plate is achieved based on the following reaction formula: 2Si-OH→Si-O-Si+2H2O.

4. The method for manufacturing a sample detection device according to claim 3, characterized in that: Before bonding, the surfaces to be sealed of the first plate and the second plate are subjected to plasma treatment.

5. The method for manufacturing a sample detection device according to claim 1 or 2, characterized in that: The first plate and the second plate are respectively made of a polymer material, and the polymer material is selected from polymethyl methacrylate, polydimethylsiloxane, polypropylene, polycarbonate, and polystyrene.

6. The method for manufacturing a sample detection device according to claim 5, characterized in that: The first plate and the second plate are made of different types of polymer materials respectively.

7. The method for manufacturing a sample detection device according to claim 1 or 2, characterized in that: In the step of providing the first plate and the second plate, the first plate and the second plate are manufactured by injection molding a polymer material, and the injection molding process satisfies one or more of the following: The melting point of the polymer material is between 200°C and 300°C; The temperature of the mold used for injection molding is between 20°C and 100°C; The injection pressure is between 60MPa and 180MPa; The injection molded material is dried between 40°C and 100°C; The drying time of the injection molded material is 3 to 8 hours; The molding shrinkage of the material after injection molding is between 0.3% and 1.3%.

8. The method for manufacturing a sample detection device according to claim 1 or 2, characterized in that: The manufacturing method further includes a reduction treatment step after the sealing step. In the reduction treatment step, the sample detection device is placed in a sealed space, and an inert gas is introduced into the sealed space.

9. The method for manufacturing a sample detection device according to claim 8, characterized in that: In the reduction process step: The inert gas is argon and / or helium, and / or The duration of the reduction treatment step is more than 48 hours.

10. The method for manufacturing a sample detection device according to claim 1 or 2, characterized in that: One of the first plate and the second plate is made of polydimethylsiloxane, and the other is made of polymethyl methacrylate, and the corresponding surfaces to be sealed of the first plate and the second plate are bonded based on the following reaction formula: 2Si-OH→Si-O-Si+2H2O, wherein, in the surface treatment step, the treated surfaces of the first plate and the second plate are provided with hydroxyl groups.

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